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B. E.

Easton
President of the A m e r i c a n Society
of Sugar Beet Technologists for t h e
b i e n n i u m 1964-65 is M r . B. E. Easton.
M r . Easton is A g r i c u l t u r a l Superint e n d e n t , C a n a d a a n d D o m i n i o n Sugar
Company Limited, Chatham, Ontario,
Canada.

JOURNAL
of the
American Society of Sugar
Beet Technologists
Volume 13
Number 1
April 1964

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado, U. S. A.

Subscription
S4.50
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M a d e in the U n i t e d States of America

JOURNAL
of the
American Society of Sugar
Beet Technologists
Volume 13
Number 1
April 1964

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado, U. S. A.

Subscription
$4.50
$5.00
$1.25
$1.40

per
per
per
per

year,
year,
copy,
copv.

prices:
domestic
foreign
domestic
foreign

Made in the United States of America

TABLE OF CONTENTS
Author

Page

Presidential Address

Guy Rorabaugh

Not a new, but another look at sugar

C. F. MacLeod

T h e challenges of an e x p a n d i n g beet sugar


industry

G. D. Manuel

Agriculture's

responsibility

in

growing

economy

Eric Thor

13

Nitrogen effects on sugar crops

L. D. Baver

21

Isolates of beet mosaic virus with different


degrees of virulence
Marble leaf of sugar beet, caused by a juice
transmissible virus

C. TV. Bennett

27

C. TV. Bennett

33

Jay L. Haddock
Darrel M. Stuart

42

R. F. Olson

59

Beet leafhopper and curly-top disease survey


in Washakie County

Dale Fullerton

62

Redistribution of nitrate in soils


effects on sugar beet n u t r i t i o n

Myron Stout

68

G. A. Wicks
F. N. Anderson

81

Critical levels versus quantity-quality factors


for assessing nutritional status of sugar
beet plants
A

rapid method for the


analysis of lactic acid

routine

factory

and

its

Effectiveness of PEBC, D A T C , a n d E n d o t h a l l
for controlling weeds in sugar beets in
Western Nebraska
Minutes of the T h i r t e e n t h General Meeting
of the American Society of Sugar Beet
Technologists
Meritorious Service Awards

_ 96
101

Presidential Address
GUY RORABAUGH 1

We are gathered here in one of the world's most beautiful


cities for the 13th General Meeting in the 27th year since the
formation of the American Society of Sugar Beet Technologists.
And as we prepare for the sessions which are to follow perhaps
we should look again at the goals and purposes which drew us
together 27 years ago and continue to inspire our efforts. Let
me quote:
''The objective of this Society shall be to foster all phases of
sugar beet and beet sugar research and to act as a clearing house
for the exchange of ideas resulting from such work."
One of the special characteristics of our age is that we must
work in groupsin larger and larger groups as our world grows
more and more complex. Few of the worthwhile things which
we must accomplish can be achieved by independent individual
action. We have found the frequent need to explore our ideas
with others, usually with groups of others. We have learned
that we must have an inter-change of ideas and fuller understanding among ourselves. We depend upon each other for willing and sympathetic help. And as the demand for and cost of
development and research work have climbed to new heights,
there has been expanding need for full exchange and dissemination of scientific information to avoid unnecessary and wasteful
duplication of effort and a slow-down in getting on with what
must be accomplished.
As the research activities of our industry expand, our Society
must be prepared to meet the challenges. T h e past exchange
of technological ideas and developments within our industry has
beenr m the
key to the beet industry's ability to weather many a
st
w
hich might otherwise have sunk us all. Our Society, and
1
all those who have worked with it and for it over the years, can
be proud of the record of accomplishment through the free exchange of information and ideas. I know of no other industry
in which technological information is as widely and freely exchanged as in our own.
Today we are on the threshold of a great period of expansion
of our industry. We face a challenge and a test, both in processing and agriculture, to make this expansion successful. Less
tnan two months ago in our nation's capital, representatives of
25 would-be sugar beet production areas requested reserve acrel

V i c e President, Holly Sugar Corporation. Colorado Springs, Colorado.

JOURNAL OF THE A. S. S. B. T.

age allotments in 18 states from Maine to Texas and virtually


from the Mexican to the Canadian border. T h e great interest
in expansion has been stimulated in large part by the United
States policy to increase its reliance on domestic sugar-producing
areas to assure adequate supplies of this necessary commodity
at all times. History shows clearly that establishment of new
production areas demands as a pre-requisite a considerable
amount of research and development.
In the established industry, there is an intensification of the
search for ways to increase production and reduce costs; and it
falls upon those whose responsibility is primarily in the fields
of research to seek out, by combining their efforts, ways to increase both the quality and yields in field and factory. Along
with gaining better yields and quality, over-all production costs
must be decreased. We haven't been standing still in these areas
of endeavor. Looking back over the period since the formation
of this Society, the gains are clear-cut, the accomplishments many
and great. Looking ahead, much progress remains to be made.
All of us, regardless of the segment of the beet industry we represent, face the challenges and share the motivation for reaching
the goal of producing for an expanding market more sugar per
acre, and at less cost.
Complete field mechanization and the introduction of automation within our factories, as a result of research and development, are leading to another industrial revolution within our
industry. Automation in itself poses numerous problems, not
the least of which has to do with the kind of training and preparation our present and future employees must receive. It is
obvious already that employees of the future will need a higher
degree of technical education and greater training to acquire
skills the majority of our present workers do not possess. Planning ahead demands deep and serious consideration of this fact
of life.
In agriculture, complete mechanization from planting through
harvest has been demonstrated and is practiced in limited areas.
The demand for faster progress is being thrust upon us, however, by the imminent loss of a great proportion of the available
hand labor through expiration of Public Law 78, commonly
referred to as the Bracero Program. After the end of this year,
we are told, there will be no further extension of Public Law 78.
Those who have depended upon availability of Mexican Nationals for thinning and weeding operations must look elsewhere
to get this work accomplished. There seems to be no question
that there will be a painful pinch. Therefore, the problem is
clear: complete control of noxious weeds must be achieved in

VOL.

13, No.

1, APRIL

1964

all areas through economical use of chemical herbicides before


full mechanization is universally practicable. Work to bring
about the desired full mechanization must be pushed with all
speed and in an all-out cooperative effort.
Diseases form still another group of problems. Work to develop beet varieties more resistant to curly top and virus yellows
rates a high priority. New and more virulent strains of the curly
top virus have been isolated, pointing up the need for expanded
research in this field. Virus yellows still is a serious threat to
the beet industry of California. Among the soil pests, nematodes
are increasing in the older areas of beet production and are
spreading into newer areas. Research is needed to stem the
spread and threat of this parasite. Resistance to leaf spot is inadequate in most commercial beet varieties now in use, and
this fact alone is a limiting factor preventing the successful, economical production of the crop, year after year, in some areas.
Further research of a basic nature is needed in the categories
of male sterility, polyploidy, bolting, monogerm seed germination, and the chemical factors in the composition of commercial
beets that affect sugar formation and purity. Sugar production
per acre has been markedly increased, but the extractable sugar
per ton of beets has shown a consistent decline for a number of
years. The effect upon factory operations and costs is serious
in its proportions. Soluble impurities not removed by present
process purification methods have increased. New, economical
purification methods are needed, along with research which will
find us the answers to those unknowns involved in the decline
in beet quality.
As the pressure for increased beet sugar production mounts,
more and more is being demanded of our older processing plants.
Slicing capacities have been increased, in numerous instances,
without comparable expansion of capacities of all stations, particularly on the low raw side. The result very often is higher
molasses losses and lower rates of extraction. Here is presented
the challenge to develop an economical process which will decrease molasses production without necessitating: the replacement
of or additions to present sugar end equipment at the expense
of very heavy capital investment.
As beet acreage is expanded in many areas, storage of beets
in piles is extended over longer periods and losses, generally,
become greater. More knowledge must be developed as to the
factors and mechanisms contributing: to storage losses, and a
way found to cut these losses to a minimum.

JOURNAL OF THE A. S. S. B. T.

There is another big field beckoning to the researcherthe


better and more efficient use of by-products.
How we can attack the new and the old problems of our
industry, how we can meet the challenges will be considered and
discussed in the many Section Meetings to be held during the
next four days. New ideas, results of work thus far accomplished,
and valuable data will be presented. These will stimulate new
thoughts in all our minds, and perhaps help to reshape our perspective. And all of this, fellow members of the American Society
of Sugar Beet Technologists, is why we are here, "to act as a
clearing house for the exchange of ideas." Our membership is
composed of specialists in many fields. If this meeting runs true
to form, approximately 60 per cent of those here today are directly connected with processing companies. Of this group, 56 per
cent are concerned with matters relating to the arrowing of beets
from agronomy to genetics and seed production, from polyploidy to weed control. The balance, or 44 per cent of the
processor-related group are individuals concerned primarily with
the converting of beets to sugarengineers, chemists and technologists. Other members of our Society here, proportionately
speaking, are about as follows: 7.7 per cent are scientists from
the United States Department of Agriculture; 5.2 per cent are
scientists from colleges and universities; 21.9 per cent are engineers and scientists connected with suppliers of our industry; 3
per cent are beet growers or their representatives; and the remaining 3 per cent are unclassified, or miscellaneous.
With such a team, and the priceless privilege of free interchange and expression of ideas and information, we have the
essential wherewithal to rise to any challenge; and I confidently
predict that within the next 10 years more progress will be made
in the beet industry, in both processing and agriculture, than that
which has been achieved in the past forty.

Not a New, but Another Look at Sugar


G. F. MACLEOD 1

Here in our land of plenty three times a day we sit down to


eat. Some of us even have time for coffee breaks in mid-morning
and mid-afternoon. All of us see before us a marvelous storehouse of energy; yet seldom do we give it much thought. We eat
it, use the energy and it goes forth into every facet of endeavor.
This is our sugar.
It seems worthwhile, on occasion, to back off from our intense
focus on our own particular job and look at the total picture.
Most men need to feel a little important, significant and useful.
Let's take not a new look but another look at the sugar bowl,
envelope or cube.
The struggle for energy begins early in the life of man and
early in his history. The club which extended the cave man's
fist-power, the wheel, fire, combustion engines and atomic bombs
all are parts of our struggle for more and more energy. But back
of all these there must be human energy, and where does it
come from? Obviously, from plants and animals which eat the
plants. And where does the plant get its store of energy? From
the endless power of the sun, captured by the green leaf. What
nlant do we use most to capture, store and preserve the sun's
energy most efficiently? The sugar beet whose mysteries we find
most intriguing and at whose shrine we come here to ponder
and debate.
This elementary lesson in biology, so well known to us all,
has other bits of charm which we are prone to forget. Our fossil
fuels, oil, gas, coal and even uranium are finite in amounts. T h e
vast stores of energv which they represent, accumulations of years
almost unnumbered, still have limits. We mine and burn these
stores in ever-increasing amounts. They have an end no matter
how remote. They are depleted with each calorie used.
Not so our sun whose boundless energy the planet receives
in its checkerboard of nights and days. We are profligate with
it. We nurture it, capture it, transform it and store it for our
use each day, each year in the art and science of agriculture.
Thus with each harvest new annual wealth, never before seen
by man comes into use in life as we know it. T h e sugar beet
once lifted from the earth is man's first look at new energy from
1
Consultant Agriculture Department. Union Carbide Chemicals Company, Division of
Carbide Chemicals Company, Division of Union Carbide Corporation, San
California.

Francisco.

JOURNAL OF THE A. S. S. B. T.

the sun. Once started on its way that is the end of it and no
man will ever see it again. Instead he'll have its stored energy
in bowl, in cube and in envelope. This is fuel for life.
In fact, this is a prime reason for saying that agriculture is
basic. T h e production of new, annual wealth from the soil, sun
and air gives man his sustenance and the nation economic
strength from which to draw the military posture necessary to
maintain our way of life. For every dollars worth of agricultural
commodity produced, ten dollars in ancillary jobs are generated.
These are the reasons why everyone has, or should have, an
interest in agriculture. This is the reason we can feel, each of
us, that our job is an important one. This is why we must give
thought to our agriculture in which the production of sugar beets
is a most important, basic business.
Every informed source readily admits a rapidly expanding;
population. Sugar consumption reflects it. We may have reached
a plateau in our per capita consumption, but there are a lot
more mouths to feed. It took from the beginning of recorded
history to 1887 to populate the world with 1.3 billion people.
In my lifetime, plus ten years, another billion were produced.
There will be 4 billion more produced in the rest of the century.
They all have to eat. We'll be fighting for standing room and
sugar if we do not meet this challenge.
Individual farms grow fewer and larger, but the total land
available for food production grows smaller in extent as the
battle for standing room and playing room grows hotter. T h e
youngster born today faces a world with three acres of tillable
land per person. When he is 36 years old there will be less than
half an acre of tillable land per person. Our current surpluses
can vanish in seven months if ever we come to a sudden stop.
This is a real and immediate challenge to every technologist
and it is one of great urgency to you.
These large farms are faced with the need for increased
efficiency. Capital with which to operate is concentrating in a
few great sources. Spurred on by a price-cost squeeze automation
is fast reducing labor's place in agriculture. In 1900 we used
37 percent of our work force in agriculture, but in 1960 less
than 9 percent produced much more food. Even with our present
rate of mechanical replacement there may be less than 100
days of work in a year's time for agricultural workers in the
near future.
You are a part of a rapidly expanding technology. New
approaches to biological problems are engendered by a new
sophistication in research. T h e advent of new techniques in
chemistry, physics, genetics, the whole sub-cellular approach to

VOL.

13, No.

1, APRIL

1964

the secrets of living things will make for rapid changes in biology
and agriculture. The quest for two pounds of sugar where one
is now will lead to rapid changes in hybridization, mechanization
and chemicalization. It is from groups such as this that men
expect to obtain the technical knowledge for survival in a
crowded world. Let us hope that we may find a public policy
which will understand the need and create a climate in which
our studies may flourish rather than be stunted by emotional or
political unrealities.
There can be no question of the need for changing legislation
and regulation in a rapidly expanding, technological agriculture.
The very fact that agriculture has a direct impact upon the health
of the people, their economy and their social problems are proofs
of its basicity in our society. To deal with the dynamic changes
involved there will be many new laws, rules and regulations.
It will require constant watchfulness on the part of those well
informed about agricultural problems lest political expediency
or opportunistic, bureaucratic empire builders delay our progress
unnecessarily. This, too, is our task. The technical fields in
which we spend our days will serve men better if each endeavor
is motivated by an understanding of the socio-economic impact
of our efforts.
If, as some leaders believe, the second half of our century
belongs to biology as the first has to mathematics, physics and
chemistry, then we have yet another task before us. There is
currently a demand for 15,000 new young technologists in agriculture each year. We are producing about half that number.
Each of us in his own circle should try to enthuse these brilliant
youngsters with the desire to prepare themselves for opportunities in the fields of biology and agriculture. If we ourselves can
be enthusiastic about our own work, we can better enlist the
interest of fast on-coming geniuses. If we do not enlist them in
the glamour of living things to study then other groups will and
all mankind may suffer. It is not prudent to leave this job to
counsellors less familiar with the field than we who are working in it.
The basic requirement in research is imagination. While
this characteristic is not the exclusive property of the young,
it is most abundantly evident in youth. True, it may need the
guidance and control of more experienced heads but the fresh,
imaginative attack upon our unsolved problems in agriculture
needs tender loving care and encouragement. We can hardly
hope to stimulate the enthusiasm of a youngster by regaling him
with our own adversities and defeats. Their interest lies in the
challenges and opportunities our studies disclose.

JOURNAL OF THE A. S. S. B. T.

For each of us there must be some event which starts us on


the road of formulating a philosophy for living. There was a
young man to whom many doors were opened in late high school
and early college. The personalities and enthusiasm (or lack
of it) among his teachers confused and annoyed him. His lectures and his labs were motions necessary to conform and stay
with the crowd. His efforts were kept at a minimum needed
to "get by." By process of elimination and attrition along: with
getting nearer to the end of the road and his diploma, he began
to feel that living things were of most interest.
One day, sitting before a fireplace where he had been many
times before, he raised his eyes to read the words engraved in
the mantle. He had seen these often, read them often, but never
until this day did their full import reach him. They were "Ye
forest logs fire back the glow of summer suns of long ago." The
great oak logs glowed silently back at him with a new light. Here
was warmth from the suns of history. Here was nature's storehouse for man's winter needs. Here was mystery, wealth, power
and knowledge. It was all his.
When next you eat, and it will be soon, look lovingly at that
most wonderful of nature's powerhouses, the sugar bowl. And
when you take up the course of your discussions, do so with
greatest self respect life's greatest fuelsugar, new energy from
the sun each year.

The Challenges of an Expanding Beet Sugar Industry


G. D.

MANUEL 1

Mr. President, distinguished guests of the Society, fellow


technologists.
At the outset let me remind you of one point. Although my
membership in the ASSBT dates back to 1942, and I have frequently been involved in various sections of the Society as well
as some of its politics, this is my first opportunity to address the
entire body in open session.
Perhaps after my remarks today, future invitations to speak
to this august body will again stipulate that I confine my remarks
to Section meetings from which one can leave without causing
embarassment to self or speaker.
In any event, I am pleased to participate in this general session
of the 13th biennial meeting of the Society, and it is always a
source of personal pleasure for me to meet with so many old
friends at these gatherings.
Since we are friends, I hope you will bear with me if I exercise a point of personal privilege and range farther afield than
others who have addressed you during this general session.
First of all, I want to offer congratulations. I feel that our
industry's technical gains have been outstanding, and you who
are members of the Society have been the prime movers in making these gains. On the farm we have progressed from high hand
labor requirements to a comparatively high degree of mechanization. Our land is producing more beets and more sugar per
acre. Our farmers now plant monogerm seed which has further
increased our mechanical efficiency. Plant breeding has given
us disease resistance that has helped overcome diseases that can
plague our fields. In our factories we can chart reduced production costs and we can boast of automated processes. And,
while a number of our factories were built many years ago, their
equipment is now, for the most part, the most modern. And,
more importantly we are producing a product of unsurpassed
quality.
The year I was attending my first meeting of the Society,
1942, the beet sugar industry produced 1,725,000 tons of sugar
from 953,000 acres of sugar beets. In 1963 it is estimated that
the industry will produce 3,100,000 tons of sugar from 1,236,000
acres. This is a notable improvement in yield per acre.
1

President, Spreckels Sugar Company, San Francisco, California.

10

JOURNAL OF THE A. S. S. B. T.

The 1942 crop was processed through 82 factories while the


1963 crop is being processed through 60 factories. This marked
decline in the number of factories necessary to process the crop
speaks for bigger and more efficient factory operation.
These farm and factory accomplishments are most noteworthy, and many people have been involved in these changes
and all of you are to be congratulated on what has been accomplished. But, I would caution you that we cannot rest on our
laurels. We need only to analyze where we stand today and
where we must go from here.
While our sugar beet harvest is almost completely mechanized, I do not think we should concede that it is perfect.
Better trash separation, better topping and other harvest aspects
still need to be studied. While yields are high, these high yields
have given rise to new problems in beet quality and in processing. Where we were once only concerned with convincing
growers to use nitrogen, we now must caution them against overusage. All of us who are concerned with the study of disease and
disease resistance know that the job is never done. Where in
1942 curly top and leaf spot were the prime topics for discussion,
virus yellows is now the threat we must struggle against to keep
our industry healthy. I am sure that other problems will arise
through the years to tax the capacity and ingenuity of all of you
because agriculture is a continually changing industry due to
the fact that nature itself is unpredictable.
What all this adds up to, really, is that your jobs are just as
important, just as challenging, and just as rewarding today as
they were twenty years ago. Our industry must continue to improve or fall behind because now, more than ever before, the
domestic industry is influenced by and exerts influence on the
world sugar picture.
I think the circumstances surrounding 1963 brought this
point home to a lot of people. Last year for the first time many
people outside our industry came to recognize the importance
of this domestic beet industry. When world sugar suddenly
turned up scarce, it was our industry that was able to expand its
production by some 20% and in so doing contributed a degree
of stability to an otherwise widely fluctuating market. This sudden expansion of our industry, accomplished without too great
a strain, was possible because of the cumulative technological
advances in agriculture and processing.
Now that we have attained a higher level of production, what
is ahead for us? In our own country, based solely on projected
population trends, our annual consumption of sugar by 1980

VOL. 13, N o .

1, A P R I L 1964

will reach 12.5 million tons. In just 25 years our American consumers will annually require 14 million tons of sugar.
With this increased consumption in this country, we also
look for even more rapid increases in consumption in other
parts of the world. Some of the countries which are now, for
the first time emerging economically are demanding of the world
sugar market needs such as we have never known. Average free
world per capita consumption in 1961 was only 43 pounds per
person as compared to about 100 pounds in the United States.
In some of the underdeveloped countries the consumption is
now less than 10 pounds per person. As these countries develop,
the potential for increased consumption of sugar appears almost
unlimited.
Authorities generally agree that we should expect consumption to increase at the rate of approximately 3 per cent per year
if supplies are available. At this rate, a conservative estimate of
world sugar consumption in only 25 years will be between 100
and 125 million tons. This means, by 1989, the world sugar
industry, if it is to meet minimum world sugar demand must
produce at least 100 per cent more sugar than is being produced
today.
This increase in world sugar needs will keep some of
our foreign suppliers busy, meaning that the beet industry certainly must prepare to meet its share of requirements.
This is, indeed, a real challenge to the entire sugar industry,
and management, reflecting on that challenge, must turn to you
people to point the way.
I should point out that the straight line projections on which
to base consumption estimates may not be valid unless your
technical skills can keep the cost-price ratio to our advantage.
Indeed, the price increases of the past year actually slowed down
the expected increase in sugar usage and, at the same time,
opened the door for other types of sweeteners. In 1963 the
corn sweeteners and other types of sweeteners scored impressive
gains at our expense. Last year, for instance, the expected normal
marketing increase for all sweeteners was 175,000 tons. Of this
expected increase the deliveries of corn sweeteners showed an
overall gain of 90,000 tons during the first three quarters of
1963.
Such growth by competitors cannot and should not be overlooked by our industry. Our prices must be competitive or certainly there will be adverse effects on our long-term growth.
I should also mention that with the higher prices realized
in 1963, many people have become interested in beet sugar pro-

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JOURNAL OF THE A. S. S. B. T.

duction; and, at a recent hearing in Washington, twenty-three


applications were received from various states for creation of
new facilities to further expand our industry. Both the new
and old facilities should be prepared to face lower prices in the
future or we are not expanding on a sound basis.
I feel that the real challenge facing all of us today is to make
this expanding industry as low in production costs as possible
which can be through greater mechanization, higher yields and
more efficient handling of the crop. We must also have higher
quality in our beets to maintain our plant operations at reasonable levels. This may sound like a simple task, but I think all
of us here in this room are involved in this task because it affects
all phases of our industry.
I feel confident that you can do the job and that our industry
will make greater strides in the next twenty years than it has
during the last twenty.

Agriculture's Responsibility in a Growing Economy


ERIC THOR 1

I am glad to have the opportunity to discuss with you agriculture's responsibility in our nation's economy because I know
many of you are thinking seriously about the future of agriculture
and how you, your associations and other agricultural organizations will fit into the changing scene. Today, I would therefore
like to do some thinking with you about the kind of history the
sugar industry and other segments of agriculture are going to
write.
Last spring, at the University of California, we held a symposium on "The Future of California Agriculture." We did
some looking ahead to the 1970's and even to the 1980's and
beyond. First, however, we looked back. We reviewed what had
been done before we discussed the future.
Here, too, I would like to take a quick look at agriculture
today and what is happening to it. We might say that the most
important feature in the agricultural scene is change itself. Our
friends in other fields of endeavor find present-day farming, with
its airplanes, chemicals and great machines, baffling enough
without peering ahead into the vague future of the 1970's and
1980's.
We know that American agriculture today is feeding and
clothing 180 million people in this country alone. In addition,
the production of one out of every six acres of our land is exported to feed and clothe people in foreign lands. In 1963, our
farmers set a new high level of total food and fiber production on
the smallest number of acres harvested at any time in the twentieth
century. We also know that consumers are spending the smallest
share of their income on food in the history of our nation, and
that this share is the smallest of any nation in the world.
Our agriculture has already entered the domain of science,
engineering and business management. The fruit and vegetable
packer, the canner, the freezer, the miller, the creamery operator,
the feed manufacturer, the rice dryer, and the cotton ginner
are all part of agriculture. So are the shippers who send produce
to market and the merchandisers who sell it from the market
shelf, even though most of them do not realize it. Agriculture
also embraces the suppliers who provide feeds, seeds, fertilizers,
farm machinery, petroleum, hardware, lumber, cans and processing equipment, and the bankers who provide credit and
financial guidance to farmers. It includes research scientists and
1
Agricultural Economist. Giannini Foundation of Agricultural Economics, University
of California, Berkeley.

14

JOURNAL OF THE A. S. S. B. T.

the agricultural extension workers of the universities and of the


members of the United States Department of Agriculture.
But most of all, agriculture counts upon those who raise the
livestock, produce the meat, milk, and eggs and till the soil and
grow the crops. They are the people really responsible for the
great contributions agriculture has made to our nation's economy.
Without their willingness to accept and adopt new ideas, the
agriculture of our nation could not have become the envy of
all of the nations of the world.
Agriculture is our largest industry. It employs approximately
7 million workers on the farms and ranches. This is more than
the combined employment in transportation, public utilities, and
the steel and automobile industries. Farm and ranch assets total
about $210 billion, or nearly three-fourths of the value of current
assets of all corporations in the United States and about twothirds of the market value of all corporation stocks on the New
York Stock Exchange.
Our 3.7 million independent farmers and ranchers are perhaps the best customers that our nonagricultural people have,
for they spend about $27 billion a year for goods and services
to produce crops and livestock. Farm and ranch people spend
another $15 billion a year for food, clothing, drugs, furniture,
appliances, and other materials which they use for everyday
living.
Farmers and ranchers are also creators of private employment. We do not have a full count of the people engaged in
each of the associated services and manufacturing industries.
We do know that the figure is huge and that four out of every
10 jobs in private employment relate to agriculture. T e n million people have jobs storing, transporting, processing and merchandising the products of agriculture. Six million have jobs
providing the supplies that farmers and ranchers buy. All in
all, without stretching our figures, we can say that total agriculture employs approximately 26 million people and accounts
for roughly 40 per cent of the national economy.
Agriculture has also contributed greatly to the nonagricultural growth of our nation's economy. First, output per man
hour has been increased to the extent that one hour of farm
labor today produces more than four times as much food and
fiber as it did in 1920. Hundreds of thousands of people have
been released from agriculture to other occupations as plumbers,
carpenters, lawyers, doctors, school-teachers and technicians of
all sorts who produce the products and services associated with
higher standards of living. Second, increased efficiency in agriculture has made it possible for consumers to spend a greater

VOL.

13, No.

1, A P R I L 1964

15

part of their income for products and services other than food.
U. S. consumers are today spending approximately 20 per cent
of their income for food while consumers in England spend 41
per cent, in Norway 44 per cent, in France 38 per cent, and in
the Soviet Union 53 per cent.
Although agriculture is perhaps the nation's most important
single industry and has contributed greatly to the growth of
the nation and to the well-being of our citizens, it is an industry
that has had and is having problems. Its major problem during
the greater portion of the past four decades has been income.
Right now average per capita farm income on a nation wide
basis is barely half that earned by comparable people in other
industries. Moreover, incomes earned in agriculture are far
more variable and unstable than those earned in other industries.
The low per capita income reflects the status of the many underemployed people in agriculture. It also reflects the fact that
farmers and ranchers have not been able to increase their income
or even to keep the savings made possible by the development
and adoption of new technological innovations.
Many people fail to understand how an industry that has so
greatly increased its efficiency has been unable to increase its
per capita income. Farmers have not been able to increase their
income despite greatly improved efficiency because of the competitive structure of agriculture. The agricultural improvements
adopted during the past 40 years have been primarily capitalusing, labor-saving, and output-increasing. When demand does
not increase to offset the increased output, such improvements
initiate a three-stage economic reaction.
Stage 1 is the adoption of new techniques by early innovators.
Once a new technology is developed, it attracts the attention of
a small number of farmers who always keep their eyes and ears
open for anything that will reduce their per unit costs. Once
these farmers find an innovation they think will help them make
a profit, they generally adopt it. Since these early adopters expect
the innovation to increase profit, they also tend to increase their
crop acreage or livestock numbers associated with the new technology. But since few farmers are involved at this stage, their
increase in output is relatively small and has little effect upon
market prices.
Stage 2 is general adoption of the new technique by other
farmers, who have observed the success achieved by the early
adopters. As many more farmers adopt the innovation, output
is increased considerably and prices fall. Sometimes the price
falls to the point where the farmer may actually have less net

16

JOURNAL OF THE A. S. S. B. T.

income than before the idea was developed. T h e savings are


passed on to the consumer, and the farmer is powerless to do
anything about it. From the standpoint of the economy as a
whole, this price drop is very desirable. It enables the consumer
to spend less for food and more for other things associated with
higher standards of living. It also tends to force farmers out of
agriculture into nonagricultural jobs. This was very important
during the 1940's and 1950's, because increased numbers of professional people, skilled workers, and laborers were necessary for
the economic growth of the nonagricultural sector of our economy.
Stage 3 of this economic chain is the movement of people out
of agriculture. The theory is that if farmers move out of agriculture fast enough, the income to those who remain in agriculture will increase. Increasing per capita income is one of
the opportunities for agriculture in the years that lie ahead.
Reducing the number of people in agriculture is the most economical method to increase per capita income.
People have been moving out of agriculture at a rapid rate.
During the 1950's farm population decreased about 6.5 million
from roughly 23 million to about 16.5 million. Getting farmers to leave agriculture will be more difficult in the 1960's and
1970's than it was in the 1950's. Employment opportunities for
farmers in non-agricultural jobs have decreased in the past two
or three years. They will probably continue to decrease in coming years because of the competition for nonagricultural jobs.
The large numbers of young men and women born in the early
1940's are now beginning to enter the labor force. This influx
of young workers is also expected to increase faster than jobs.
Another problem is that the average age of all farm operators
in the U. S. is now over 50 years. Even though farmers are able
to do many things, most of them have no particular skill to
compete in nonagricultural industries with the young people
now entering the labor force. Since this is the case, the greatest
and perhaps the only real opportunity to reduce the number
of people in agriculture lies in training farm young people for
nonfarm employment and encouraging them to leave agriculture.
The present secondary education system in many agricultural
areas is not educating and encouraging enough young people
to leave agriculture. In fact, approximately 45 per cent of the
funds spent for vocational education in the secondary schools
is used exclusively for training in farming skills and home economics. If we are ever to solve the problem of underemployment
in agriculture, then this vocational training program needs to
be reoriented. At the same time that we train youths for job
opportunities available in nonagricultural industries, we must

VOL.

13, N o .

1, APRIL

1964

17

give serious thought to maintaining and building agriculture's


leadership. After all, agriculture in its larger sense represents
about 40 per cent of the nation's economy. The most competent
leadership is needed in research, teaching, and the many business ramifications of agriculture. There are now, and there will
continue to be, excellent and rewarding opportunities for men
who are well trained in professions and businesses serving agriculture. Agricultural leaders of tomorrow will need training in
more than the arts of farming. They need the broadest kind of
basic foundation. They need some understanding of science,
business, political science, and economics. We owe it to ourselves and to those who will come after us to encourage bright
young minds to go beyond the secondary schools to obtain the
best technical and professional training to be had.
Agriculture, represented by organizations such as yours, must
assume the responsibility for leadership in agricultural areas for
reorienting the education and training programs of the farm
youth. The majority have to be trained to leave the farm or
ranch. Some, if they obtain the proper technical and professional
training, can go into industries associated with agriculture.
A second opportunity for agriculture lies in the area of
increasing marketings to both the world and domestic consumers.
The value of U. S. agricultural exports in the fiscal year 196263 advanced to a new record of $5.1 billionup 4 per cent over
the previous 1961-62 peak. The 1963-64 export is expected to
rise to about $6 billion.
Increased competition from other countries is going to put
pressure on world prices. Many of the countries whose economic
development we have aided are just beginning to compete with
us in both the world and our own domestic market. We can
also expect added competition from the regional trading blocs,
such as the European Common Market, "the so-called Outer
Seven on the fringe of the Common Market, and the proposed
Latin American trade groups, as well as other trade groups that
may be formed in the future. In the short run, these countries
are going to make it difficult for us to increase our export volume.
However, as modern asricultural technolosries free some of the
labor resources of the underdeveloped countries from agricultural
work, as has been done in the United States, their industrial
capacity will increase. This will mean higher per capita incomes
and greater demand for food and fiber products.
If the United States is to meet this competition and increase
its share of the food and fiber business in both the world and

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JOURNAL OF THE A. S. S. B. T.

the domestic markets, production costs must be continually reduced. This means continual development and adoption of new
output-increasing technologies.
Two factors are primarily responsible for the great advances
we have made in agriculture during the past few decades. First,
millions of farmers, spurred by the incentive and pride of ownership inherent in the American family-farm economy, have applied
new discoveries and new methods to their own operation. Second,
a tremendous amount of research and development has been
carried on, primarily by the land-grant colleges, the United States
Department of Agriculture, and private industry.
It is estimated that if agriculture were still using the methods
of 20 years ago for producing crops and raising and feeding livestock, it would have cost an extra $13 billion to produce the
nation's food and fiber. To offset this, the average consumer
family in the United States would have had to spend approximately $240 more to obtain the food they purchased last year.
It is agriculture's responsibility, and particularly the responsibility of organizations such as yours, to see that both the familyfarm concept and public research are continued. In fact, if it
were not for public supported research and the dissemination
of its findings to farmers, we probably would not have the familytype agriculture we have today. Without public supported research and the results made available to all farmers, large and
small alike, large corporations would have been able to develop
and capitalize on private research. This would have put the
family farmer at an economic disadvantage and encouraged tremendously huge land holdings.
T h e third opportunity for agriculture is to free itself from
many of the government programs that exist today. This will
not be an easy task, nor can it be done overnight. In fact, if
agriculture frees itself from federal price support and production
control programs by 1970, it will have accomplished a Herculean
task.
T h e major problem in removing government programs is
that we have a much larger production base of agricultural lands
than is needed to meet the demand of domestic and foreign
markets at prices satisfactory to farmers. Many of these lands
were put into production as a result of government programs
starting back as earlv as World War I and reemnhasized during
World War II. During the two wars, food and fiber needs dictated that all effort be made to increase agricultural production.
After World War II. government policy has not encouraged
withdrawal of farm lands from production.

VOL.

13, No.

1, APRIL

1964

19

Competent researchers now estimate that agriculture will be


able to meet food and fiber demands 20 years hence with approximately 50 million fewer crop acres than were available in 1959.
As long as this great production capacity is available for farming, farmers will have a tendency to grow crops and produce
livestock on the land, because there is little other economic
opportunity for its use.
A number of attempts have been made to withdraw rather
large acreages rather permanentlv from agriculture but still to
maintain the land in a manner that would keep it available in
case of emergency or when population growth may need it. However, these attempts have been opposed politically, primarily by
nonagricultural industries. They are concerned about the effect
such reductions might have upon nonagricultural business.
Agriculture must therefore assume the responsibility of putting forth, or at least going along with, some type of program
that will (1) maintain income to agriculture and related business, (2) assume the people of our nation an abundant supply
of food and fiber, (3) enable us to sell products in the world
market at prices that will compete with those of other countries,
and (4) minimize government cost.
A fourth opportunity open to agriculture lies in resource
development for urbanization. Here there are two ways in which
farmers can improve their income. First, they can join with
nonagricultural interests in bringing nonagricultural industries
into agricultural areas. This approach will help provide employment for farm people. Second, they can provide services,
particularly open-space recreation, for urban people.
The development of agricultural area resources for urban
growth is not easy, because farm people and urban people generally do not see eye to eye on what is best for the community. Controversy over land use for urban growth is part of today's daily
life in agricultural areas. Most farmers tend to prefer the complete laissez-faire approach, while urban people tend to favor
an over-all growth plan of some type.
Regardless of what individuals favor, this is truly a critical
time in the history of many communities. In some areas citizens
today may well be facing their last chance to choose their own
pattern of resource development. Once the concrete has been
laid, decisions to put land into subdivisions, shopping areas, freeways, or airports become, for all practical purposes, irreversible.
A bulldozer can scrape away houses; but this is rarely, if ever,
economically feasiblecapital losses are usually too great. Whatever pattern for resource development is chosen by a community,
it will affect the growth of the particular community, the growth

20

JOURNAL OF THE A. S. S. B. T.

of the individual state, and perhaps the long-run status of the


community and the state with respect to the nation and the
world.
Plans for resource development must include a means of
helping those who depend upon farming, ranching and forestry
for their livelihood to assess their needs and work out problems
they can't solve alone. T h e plans must also provide both economic
opportunity and the amenities of life for the farm people who
leave agriculture to become part of the urban sector. There must
be provision, too, for the urban people who move into the agricultural areas to live and raise their families. Although development must be planned and orderly, it must not and need not
infringe unduly upon our system of private enterprise. Moreover, plans must be flexible enough to provide for changing
demands of future generations.
Resource development for urban growth is the responsibility
of all leaders in agricultural areas, city and rural people alike.
Their challenge and responsibility is to point out the alternative
opportunities and to stimulate interest and participation by
citizens of every community in creating and regulating their
economic growth as the majority of the people desire.
Those of you in the sugar industry have contributed greatly
to the growth of American agriculture. You have led the way
in plant breeding, in use of fertilizers, in mechanization of both
production and harvest and in the development of modern processing methods. You have assisted in making American agriculture world renowned. You are aware of the responsibilities
of agriculture to the people of this nation and to the people of
the world.
As we look forward to the decades ahead, each of you realizes
that all of us in agriculture have a job to do this week and all
52 weeks of this year and for many years to come. It is our job
to see that agricultural and urban people alike recognize the
opportunities available. Together with community and industry
leaders, we must assume the responsibilities involved in educating our young people and developing our resources in a manner
that will yield greatest economic opportunity and more of the
amenities of life to all the people of this great nation of ours.
Failure on our part to do this will be paid for by the sweat and
tears of those who will be forced out of agriculture into unemployment and poverty.

Nitrogen Effects On Sugar Crops1


L. D. BAVER2

Nitrogen is an interesting nutritive element. In fertilizer it


occurs generally in two forms: ammonium (including urea) and
nitrate. Ammonia is adsorbed on the soil exchange complex
and does not leach readily; it is not utilized to any great extent
by most crops except, perhaps, in the early stages of growth.
Ammonia is converted to the nitrate form through the action
of soil microorganisms. Nitrates are not adsorbed by the soil
but move freely within the soil water. They can be leached
rather rapidly in the absence of an active root system.
Soil microorganisms, in decomposing organic residues, compete with growing plants for available soil nitrogen. This nitrogen is then tied up within their bodies. Most of this nitrogen
goes through the mineralization process. If the organic residues
undergoing decomposition are highly carbonaceous, nitrogen
deficiency symptoms may appear in the leaves of the growing
plant. If mineralizable nitrogen is released during the latter
stages in the development of the plant, it may be difficult to
ripen the crop.
Thus, the problem of providing the plant with the correct
amount of nitrogen must take into consideration not only the
amount of nitrogen in the applied fertilizer but also the amount
that is being released in the soil through the mineralization
process.
The major impact of applied nitrogen on plant growth is to
increase the rate of top growth more rapidly than the root growth.
There is an increase in the top:root ratio. In the case of small
grains, excessive nitrogen generally causes lodging of the crop.
With potatoes, there is a decrease in tuber formation and the
production of large amounts of tops. Extra nitrogen on tobacco
reduces the sugar content and increases the nicotine. Sugar crops
respond to high amounts of nitrogen bv giving increased tonnages of the plant but a decrease in the percentage of sugar
present. It is the function of this brief discussion to show the
similarity between sugar beets and sugarcane in their behavior
to nitrogen fertilization.
The Effect of Nitrogen on Sugar Beets
The Institute for Sugar Beet Investigations at Gottingen,
Germany has made extensive studies of the effect of the amount
1
Published with the approval of the Dirertor as Paper No. 137 in the Journal Series
of the Experimental Station, Hawaiian Sugar Planters' Association.
2
Director Emeritus and Consulting Scientist, Experiment Station, Hawaiian Sugar
Planters' Association.

22

JOURNAL OF THE A. S. S. B. T.

and timing of nitrogen on the yield and quality of sugar beets3.


Their studies established two rather significant points. In the
first place, there was an increasing concentration of unassimilated
nitrogenous compounds in the beet juice as the amount of nitrogen applied to the beets as fertilizer was increased. This nitrogen,
given the name "harmful nitrogen", consists primarily of amino
acids and related compounds. In the second place, increased
nitrogen fertilization raised the amount of ash in the juice.
Both of these factors had a deleterious effect upon the pol and
the extraction of the sugar.

Figure 1.The effects of increasing a m o u n t s of fertilizer upon sugar


beets (Ludecke).

The data in Figure 1 illustrate the impact of increasing


amounts of fertilizer, containing the same ratio of N-P-K, upon
beet-root and beet-sugar production and the composition of the
juice. The basic fertilizer treatment was 80 lbs N, 70 lbs P2O5,
and 100 lbs K2O per acre. Nitrogen was increased by 40-1 b
increments, with the other nutrients being raised proportionately to keep the same ratio. For the first increment of nitrogen,
the tonnage of beets and tops increased about the same. However, more nitrogen beyond this point resulted in a tremendous
increase in top growth in comparison with the added tonnages
of the roots. In other words, the top:root ratio was increased
significantly. T h e pol of the juice decreased rather significantly
H. Ludecke, Zuckerfabrik Northeim, 1953.

VOL.

13, No.

1, APRIL 1964

23

with all three increments of nitrogen. This decrease, when


coupled with a rather modest increase in. beet yields, resulted
in no increase in total sugar for the second and third increments
of nitrogen. The curves also point out rather clearly the rather
high amounts of ash and the increasing amounts of harmful
nitrogen with the extra nitrogen additions.

Figure 2.The effects of increasing amounts of nitrogen u p o n sugar


beets (Ludecke).

Figure 2 shows the relative effects of increasing the amount


of nitrogen from 80 lbs to 200 lbs of nitrogen per acre. T h e
yields of tops, beets, and sugar for the 80-1 b N application is
equal to 100% in these comparisons. These data corroborate
those in Figure 1. There is a very large increase in the top: root
ratio. The effect of the decrease in pol on sugar yields is more
dramatic, however, than in Figure 1. In this case, there was an
over-all decrease in the amount of sugar produced when nitrogen
applications exceeded 120 lbs per acre. The usual effects on increasing amounts of ash and harmful nitrogen are clearly shown.
Increasing amounts of nitrogen increase the toprroot ratio,
amounts of ash and harmful nitrogen and decrease the percentage
of pol. Late applications of nitrogen also have similar effects;
this is clearly shown in Figure 3. Here, 140 lbs of nitrogen were
applied before seeding. This was compared with the same
amount of nitrogen applied as split applications before seeding,
after thinning, and two weeks after thinning. The delayed

JOURNAL

I COMPOSITION

OF

THE

A.

S.

S.

B.

T.

Before seeding | 4 Q # N 9 5 # N 7 0
After thinning
45HN 35
14 days after
35
thinnina

#N
*N
#N

Figure 3.The effects of the time of application of nitrogen u p o n


sugar beets (Ludecke).

nitrogen increased the top:root ratio as well as the amount of


ash and harmful nitrogen. Pol was decreased, with a slight decrease in sugar produced from the latest application.
This impact of nitrogen on sugar beet quality was considered
as one of the major problems of the industry in 1957. It will
be interesting to find out in a visit to the Institute in 1964 how
they have resolved it.
The Effect of Nitrogen on Sugarcane
Field observations and experiments have indicated that excessive nitrogen increases the amount of cane but may not necessarily increase the amount of sugar produced due to a decrease
in pol. When the cane plant puts on new growth as a result of
nitrogen additions, it could be either through an increase in
the amount of green tops or through the production of new
suckers. In both cases, this green material is high in both unassimilated nitrogen and reducing sugars. The deleterious effects
of nitrogen on sugar yields depend considerably upon the cane
variety and the climatic conditions. Some varieties are relatively
insensitive to nitrogen additions as far as the effect upon juice
quality is concerned. Others are relatively sensitive. With certain
varieties, an increase in nitrogen may even result in a decrease
in the amount of cane produced. Cloudy, cool weather during
the boom
andquality
ripening
the crop's
tends to give
poorer
juice
thanstages
does insunny,
warmgrowth
weather.

VOL. 13, No. 1, A P R I L 1964

Figure 4.The effect of the a m o u n t a n d timing of nitrogen applications upon the stalk population of sugarcane (Stanford).

The effects of nitrogen on sugarcane; therefore, are quite


similar to those discussed for sugar beets. Both the amount and
timing of the nitrogen are important in assessing the impact of
nitrogen on cane4. The data in Figure 4 illustrate the effects on
the increase in sucker growth of increasing the amount of nitrogen from 200 lbs to 600 lbs per acre and delaying application
of nitrogen from 3 months to 8 months of age. With increased
and delayed amounts of nitrogen, the amount of primaries in
the total stalk population decreases and the number of suckers
increases.
The impact of nitrogen upon pol % cane is shown in Figure
5. Effects are shown for four different harvesting dates, from
15 to 24 months. At each harvest date, there is a decrease in
pol % cane as the nitrogen is increased from 200 lbs to 600 lbs
per acre. Similar decreases are observed when the date of application is delayed from 3 months to 12 months.
* George Stanford, Hawaiian Planters' Record, 56:289, 1963.

JOURNAL OF T H E A. S. S. B. X.

Figure 5.Xhe effect of the a m o u n t a n d timing of nitrogen applications u p o n the juice quality of sugarcane (Stanford).

T h e s e d a t a become q u i t e i m p o r t a n t from a practical p o i n t


of view w h e n o n e analyzes the sugar p r o d u c t i o n trends of t h e
i n d u s t r y in relation to fertilization. F r o m 1950 t h r o u g h 1955
the H a w a i i a n i n d u s t r y as a whole showed an increase of a b o u t
18 tons of cane a n d nearly 2 tons of sugar p e r acre. New, higheryielding varieties were c o m i n g i n t o t h e p i c t u r e . T h e r e was a n
increasingly favorable w e a t h e r p a t t e r n . Fertilizer c o n s u m p t i o n
(N +- P 2 O s + K 2 0 ) increased from a b o u t 24,000 tons to 37,000
tons. H o w e v e r , t h e r e was a decrease in b o t h cane a n d sugar
p r o d u c e d for b o t h t h e 1956 a n d 1957 crops. T h i s took place in
spite of t h e fact t h e fertilizer tonnages h a d increased to a b o u t
50,000 for t h e 1957 crop. C o m p a r e d with 1951, in 1957 t h e
irrigated p l a n t a t i o n s , for e x a m p l e , were p r o d u c i n g a b o u t 5 0 %
as m u c h sugar p e r u n i t of fertilizer applied. T h i s a p p a r e n t l y
was t h e c o m b i n a t i o n of over-fertilization w i t h n i t r o g e n a n d
p o o r e r climatic conditions.
A p p a r e n t l y , n i t r o g e n is t h e m a j o r factor affecting t h e q u a l i t y
of sugar crops over which m a n s h o u l d h a v e some c o n t r o l . Exp e r i e n c e b o t h in t h e beet a n d cane fields, as well as in t h e exp e r i m e n t a l plots, has shown t h a t m a n has n o t b e e n d o i n g too
good a j o b exercising this c o n t r o l .

Isolates of Beet Mosaic Virus With Different D e g r e e s


of Virulence
C.
Received

W.

BENNETT1

for publication

June

24,

1963

Introduction
Beet mosaic has b e e n r e p o r t e d from all c o u n t r i e s w h e r e sugar
beets are g r o w n commercially.
It is t h e most c o m m o n a n d
widespread of t h e virus diseases of this crop. It occurs in all the
c o m m e r c i a l b e e t a r e a s o f C a l i f o r n i a a n d m a n y fields h a v e h i g h
percentages of infection at harvest.
Symptoms usually are mild
a n d t e n d to be m a s k e d d u r i n g the w a r m e r part of the season.
In general, the disease has n o t b e e n considered a highly i m p o r t a n t

factor i n b e e t p r o d u c t i o n . I n r e c e n t years, h o w e v e r , m o r e evi_ d e n c e t h a t b e e t m o s a i c is c a p a b l e of c a u s i n g m e a s u r a b l e r e d u c t i o n


n yield u n d e r s o m e c o n d i t i o n s i n b o t h t h e seed a n d t h e r o o t
c r o p has a c c u m u l a t e d .
It is p o s s i b l e t h a t a p a r t of this a p p a r e n t i n c r e a s e in i m p o r t a n c e of m o s a i c as a disease of s u g a r b e e t is d u e to i n c r e a s e d p r e v a lence o f m o r e v i r u l e n t s t r a i n s o f t h e causal v i r u s .
Observations a n d studies have b e e n m a d e d u r i n g the past
few years t o d e t e r m i n e w h e t h e r m o r e h i g h l y v i r u l e n t s t r a i n s o f
t h e v i r u s m a y b e i n v o l v e d i n losses c a u s e d b y t h i s disease. B e e t s
in the seed-producing area n e a r Xehachapi, California, have
s h o w n u n i f o r m l y m o r e s e v e r e m o t t l i n g a n d leaf d i s t o r t i o n t h a n
i n o t h e r areas. I n 1959 a n d 1960 p l a n t s s h o w i n g m a r k e d s y m p t o m s o f m o s a i c w e r e s e l e c t e d a t X e h a c h a p i a n d tested o n p o t t e d
plants in the greenhouse. Symptoms produced by virus from
s o m e o f t h e s e s e l e c t i o n s p r o v e d t o b e m o r e severe t h a n t h o s e
| p r o d u c e d b y isolates f r o m b e e t s f r o m t h e S a l i n a s Valley.
In other parts of California beet plants with symptoms m o r e
I severe t h a n a v e r a g e h a v e a l s o b e e n f o u n d . O n e o f t h e s e p l a n t s ,
which showed u n u s u a l l y severe symptoms, was selected from a
field n e a r S t o c k t o n , C a l i f o r n i a , i n 1961 for m o r e e x t e n s i v e s t u d y .
| X h e v i r u s was t r a n s f e r r e d f r o m t h i s p l a n t t o g r e e n h o u s e p l a n t s
for f u r t h e r c o m p a r i s o n w i t h a stock v i r u s c u l t u r e f r o m t h e
Salinas Valley. X h e r e s u l t s o f t h e s e c o m p a r i s o n s a r e p r e s e n t e d
i n t h i s r e p o r t . X h e S a l i n a s V a l l e y isolate i s d e s i g n a t e d " c o m m o n
m o s a i c v i r u s isolate ( C ) " a n d t h e isolate f r o m S t o c k t o n , " s e v e r e
m o s a i c v i r u s isolate (S)."
1

Plant Pathologist, Crops Research Division, Agricultural Research Service, U S


Department of Agriculture .

28

JOURNAL OF THE A. S. S. B. T

It is of interest in this c o n n e c t i o n t h a t in 1962 M a r x a n d


Beiss ( l ) 2 r e p o r t e d t h e occurrence of a m o r e v i r u l e n t strain of
beet mosaic virus in G e r m a n y w h e r e beet mosaic has b e e n con
sidered a factor of i m p o r t a n c e in beet p r o d u c t i o n for several
years.
Descriptions of Symptoms
Common Mosaic. In the field, symptoms are e v i d e n t on sugar
beet p l a n t s t h r o u g h o u t t h e year. T h e y t e n d t o b e m o r e m a r k e d ,
however, in the s p r i n g a n d fall or d u r i n g periods of r a p i d g r o w t h .
T y p i c a l symptoms consist of different p a t t e r n s of m o t t l i n g varying from small m o r e or less c i r c u l a r chlorotic spots to larger
chlorotic areas t h a t collectively cover most of the leaf surface.
T h e r e is little leaf d i s t o r t i o n a n d t h e disease usually does n o t
noticeably dwarf t h e p l a n t in the field (Figure 1A).
In t h e greenhouse, first s y m p t o m s of disease a p p e a r on t h e
y o u n g leaves a n d may consist of translucency of veins similar
to that p r o d u c e d by curly top virus except t h a t t h e t r a n s l u c e n t
areas p r o d u c e d by mosaic are b r o a d e r . Soon t h e y o u n g leaves
show chlorotic areas of various sizes, shapes, a n d degrees of yellowing. F r e q u e n t l y no vein translucency is evident a n d chlorotic
spots on t h e y o u n g leaves are the first evidence of disease.
Severe Mosaic. As w i t h c o m m o n mosaic, t h e first s y m p t o m of
disease may consist of vein translucency or chlorosis. In some
plants, veins may show necrosis. M o t t l i n g of y o u n g leaves, sometimes in the absence of vein chlorosis, soon appears. Leaves may
be deformed a n d show g r e e n blisters (Figure IB). As infectedplants c o n t i n u e to grow, leaves show various stages a n d degrees
of deformity a n d m o t t l i n g , or they may show m o t t l i n g w i t h o u t
deformity. M o t t l i n g is m o r e intense t h a n in plants w i t h comm o n mosaic. Some p l a n t s show degrees of leaf deformity a n d
chlorosis c o m p a r a b l e to those p r o d u c e d on beets by c u c u m b e r
mosaic virus. C h l o r o t i c areas are d e e p e r yellow t h a n similar
areas p r o d u c e d by the C isolate.
Comparison of T w o Mosaic Virus Isolates
Transmission. B o t h isolates a r e readily juice transmissible
b u t a h i g h e r percentage of infection was o b t a i n e d with isolate
S t h a n w i t h isolate C. Both isolates p r o d u c e local lesions on
sugar beet a n d Chenopodtum amaranticolor.
T e s t s w i t h sugar
beet in w h i c h half of t h e leaf was i n o c u l a t e d w i t h isolate C a n d
t h e o t h e r half was i n o c u l a t e d with isolate S, gave an average of
16 local lesions p e r half-leaf with isolate C a n d 31 local lesions
with isolate S in 30 single-leaf tests. Similar tests with C. amaranticolor gave averages of 22 a n d 41 for t h e C a n d S isolates, respectively.
2

Number in parentheses refers to literature cited.

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30

JOURNAL OF THE A. S. S. B. X

Incubation Period in the Plant. Several tests were m a d e in


which beet plants of the same size a n d age were inoculated by
r u b b i n g o n e leaf of each p l a n t with juice from plants infected
with isolates C or S. T h e results of one such test in which the
plants were inoculated at 3 different ages, are shown in T a b l e
1. T h e average t i m e for a p p e a r a n c e of symptoms on t h e inoculated plants was longer in plants inoculated with isolate C
t h a n with isolate S in each age class. T h i s may reflect m o r e
r a p i d m u l t i p l i c a t i o n of the S-isolate or it may indicate a greater
p l a n t sensitivity to this isolate.
Table I.Incubation periods of severe (S) and common (C) mosaic virus isolates
in sugar beet plants inoculated in different stages of growth.
Stage of growth
of beet plants
at time of
inoculation

2 leaf
6-leaf
12-leaf

Plants infected
of 40 inoculated with
indicated isolate
(Number)

38
39
40

22
29
36

Average incubation period


(Days)
S

7.3
13.6
13.3

9.3
16.3
17.1

Effect of Isolates C and S on Grozvth of Sugar beet in the


Greenhouse. Seven greenhouse tests were m a d e in which plants
of the variety US 75 in 6-inch pots were inoculated in a b o u t the
4-leaf stage with the respective virus isolates. T h e plants were
held for 49 to 82 days after i n o c u l a t i o n a n d t h e n harvested a n d
weighed.
T h e S-isolate p r o d u c e d m o r e severe symptoms in all tests
a n d t h e plants were somewhat m o r e yellow t h a n those inoculated
with the C-isolate. M a n y plants h a d deformed leaves a n d the
plants were m o r e obviously s t u n t e d t h a n those inoculated with
the C-isolate. R e d u c t i o n in weight varied considerably in the
different tests b u t the S-isolate caused m o r e injury t h a n the Cisolate in all tests. Each isolate caused appreciable r e d u c t i o n
in yield ( T a b l e 2).

T h e percentage r e d u c t i o n in weight of p l a n t s in these tests


is u n d o u b t e d l y h i g h e r t h a n that expected in t h e field. However,
t h e r e is some evidence t h a t very early infection w i t h mosaic virus
in t h e field m i g h t lead to a p p r e c i a b l e r e d u c t i o n in weight of
roots at harvest. Infection of field plants after they have attained
considerable size p r o b a b l y causes m u c h less d a m a g e . R a t e of
g r o w t h of plants may also influence t h e percentage reduction
in yield. T h i s may account for the smaller a m o u n t of damage J

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Table 2.Reduction in weight of sugar beet plants caused by severe (S) and common
i (C) mosaic virus isolates under greenhouse conditions.

Xest No.

T i m e from
inoculation
to harvest
(Days)

Plants
with each
treatment 1
(Number)

Average weight at harvest


o f plants inoculated with
indicated virus isolate
(Grams)
S~~

2
3
4
5
6
7

I
f

73
62
70
49
64
82

io
20
20
20
20
20
48

V7A~
24.1
62.4
65.3
137.2
76.0
259.9

_ _
18^6
26.4
68.5
68.4
149.2
90.5
269.8

Check
26.6
37.2
79.2
81.5
164.7
103.3
277.0

* All plants were in 6-inch pots: 1 p l a n t per pot in test 7, 2 plants per pot in test 5,
and 4 plants per pot in all other tests.

to inoculated plant in Xest 7, in which there was only 1 plant


p e r p o t , w h i c h p r o v i d e d space for g o o d g r o w t h f r o m t i m e o f inoculation to harvest.
Properties of Virus Isolates. T h e t w o isolates, C a n d S, p r o d u c e such m a r k e d l y different s y m p t o m s u n d e r s o m e c o n d i t i o n s
t h a t i t s e e m e d d e s i r a b l e t o m a k e p r o p e r t y tests t o o b t a i n f u r t h e r
information as to w h e t h e r they actually are strains of the same
v i r u s o r d i s t i n c t a n d u n r e l a t e d viruses.
T h e r m a l i n a c t i v a t i o n tests w e r e m a d e b y p l a c i n g j u i c e f r o m
b e e t p l a n t s i n f e c t e d w i t h t h e r e s p e c t i v e isolates i n s m a l l t h i n | walled test t u b e s a n d i m m e r s i n g these i n a w a t e r b a t h h e l d a t t h e
d e s i r e d t e m p e r a t u r e for 1 0 m i n u t e s . T h e j u i c e was t h e n u s e d
t o i n o c u l a t e b e e t p l a n t s i n t h e 4 - t o 6-leaf stage. T h e r e was less
infection from j u i c e h e l d 10 m i n u t e s at 56 a n d 5 8 C a n d no inf e c t i o n was o b t a i n e d f r o m j u i c e h e l d a t 6 0 C . T h e r e w a s n o
significant difference b e t w e e n t h e t h e r m a l i n a c t i v a t i o n p o i n t s
of t h e t w o isolates.
I n tests i n w h i c h b e e t j u i c e f r o m i n f e c t e d p l a n t s w a s d i l u t e d
with water to various degrees, a slight a m o u n t of infection was
o b t a i n e d w i t h e a c h isolate a t a d i l u t i o n o f 1-4,000, b u t n o i n fection was o b t a i n e d f r o m d i l u t i o n s of 1-10,000.
a n d h e l d a t r o o m t e m p e r a t u r e ( 2 1 - 2 3 C ) a n d u s e d for i n o c u l a t i n g b e e t p l a n t s at d a i l y i n t e r v a l s , i n d i c a t e d a g r a d u a l d e c l i n e in
infectivity o v e r a p e r i o d of 6 days a n d loss of i n f e c t i v i t y a f t e r
7 days. H i g h e r p e r c e n t a g e s o f i n f e c t i o n w e r e o b t a i n e d , h o w e v e r ,
w i t h t h e S-isolate t h a n w i t h t h e C-isolate o v e r t h e 6-day p e r i o d
of activity.

$2

J O U R N A L OF T H E A.

S. S.

B. T.

T h e results of property studies show that t h e t w o isolates are


s i m i l a r in the properties tested, i n d i c a t i n g that both are strains
of beet mosaic virus.
Summary and Conclusions
Results of studies of beet mosaic virus over the past 2 years
at t h e U. S. A g r i c u l t u r a l Research Station at Salinas, California,
indicate t h a t this virus p r o b a b l y occurs u n d e r n a t u r a l conditions
in the form of a n u m b e r of variants or strains that differ in
v i r u l e n c e on sugar beet. H i g h l y v i r u l e n t strains of this virus
may be capable of causing measurable r e d u c t i o n s in yield of
sugar beets, particularly if infection occurs in the early stages
of growth of the beet plant.
Literature Cited
1)

MARX, R U T H , and U. BEISS. 1962. Nachweis von Stammen beim Mosaikvirus der Beta-Ruben. Phytopath. Z. 4 4 ( 1 ) : 94-100.

M a r b l e L e a f of Sugar Beet, C a u s e d by a
Juice Transmissible Virus
C.

W.

BENNETT1

Received for publication July 19,

196}

Introduction
In S e p t e m b e r 1959, s u g a r b e e t p l a n t s {Beta vulgaris L.) w e r e
received f r o m a field in e a s t e r n O r e g o n for d e t e r m i n a t i o n of
t h e cause o f y e l l o w i n g o f o l d e r leaves. T h e y e l l o w i n g p r o d u c e d
o n leaves o f t h e s e p l a n t s r e s e m b l e d t h a t p r o d u c e d b y b e e t
y e l l o w s a n d b e e t w e s t e r n yellows i n c e r t a i n r e s p e c t s , b u t t h e r e
was a type of m o t t l i n g n o t characteristic of symptoms of e i t h e r
o f these diseases. T e s t s s h o w e d t h a t t h e p l a n t s w e r e i n f e c t e d
with a juice-transmissible virus, a p p a r e n t l y n o t previously des c r i b e d o n s u g a r b e e t . T h e disease a n d its causal v i r u s w e r e
studied further a n d the results are presented herein. Because of
the type of d i s c o l o r a t i o n on m a t u r e leaves of affected p l a n t s t h e
name b e e t m a r b l e leaf is suggested as a c o m m o n n a m e for thedisease.
Symptoms and Host R a n g e
G r e e n h o u s e s y m p t o m s o f m a r b l e leaf h a v e b e e n s t u d i e d o n
s e v e r a 1 species o f p l a n t s i n o c u l a t e d w i t h j u i c e f r o m diseased b e e t
P l a n t s . T h e v i r u s m a y p r o d u c e b o t h local lesions a n d systemic
infection.
Local
Symptoms
local lesions a r e p r o d u c e d on j u i c e - i n o c u l a t e d leaves of Beta
vulgaris
L. (sugar b e e t ) , B. macrocarpa Guss., Chenopodium
amaranticolor C o s t e & R e y n . , a n d C. murale L. ( s o w b a n e ) . L e s i o n s
a r e s i m i l a r o n all o f t h e s e hosts. T h e y b e g i n t o a p p e a r o n s u g a r
b e e t leaves a s c h l o r o t i c s p o t s a b o u t 1 m m i n d i a m e t e r a b o u t 9
d a y s a f t e r i n o c u l a t i o n . L e s i o n s i n c r e a s e i n size slowly a n d m a y
attain a d i a m e t e r of 2-3 mm ( F i g u r e 1A a n d B ) . A s m a l l n e c r o t i c
s p o t m a y d e v e l o p i n t h e c e n t e r o f t h e lesion, s o m e t i m e s surrounded by o n e or m o r e rings r a n g i n g in color from yellow to
green. If lesions a r e n u m e r o u s , i n o c u l a t e d leaves of C. amaranticolor and C. murale often y e l l o w a n d d r o p . In s u c h leaves t h e
l e s i o n s m a y b e s u r r o u n d e d b y a r i n g o f g r e e n tissue.
Systemic
Effects
T h e m a r b l e leaf disease h a s b e e n f o u n d o n l y o n s u g a r b e e t .
Its effects have been studied on sugar beet a n d other host plants
under greenhouse conditions.
1

Pathologist, Crops Research Division, Agricultural Research Service, U. S.


department of Agriculture.

32

JOURNAL OF THE A. S. S. B. T.

T h e results of property studies show that the two isolates are


similar in the properties tested, indicating that both are strains
of beet mosaic virus.
Summary and Conclusions
Results of studies of beet mosaic virus over the past 2 years
at the U. S. Agricultural Research Station at Salinas, California,
indicate that this virus probably occurs u n d e r n a t u r a l conditions
in the form of a n u m b e r of variants or strains that differ in
virulence on sugar beet. Highly virulent strains of this virus
may be capable of causing measurable reductions in yield of
sugar beets, particularly if infection occurs in the early stages
of growth of the beet plant.
Literature Cited
(1) MARX, RUTH, and U. BEISS. 1962. Nachweis von Stammen beim Mosaikvirus der Beta-Ruben. Phytopath. Z. 4 4 ( 1 ) : 94-100.

M a r b l e Leaf of Sugar Beer, Caused by a


Juice Transmissible Virus
C.

W.

BENNETT1

Received for publication July 19,

1963

Introduction
In S e p t e m b e r 1959, sugar beet plants (Beta vulgaris? JL.) w e r e
received from a field in eastern O r e g o n for d e t e r m i n a t i o n of
t h e cause of yellowing of o l d e r leaves. T h e yellowing p r o d u c e d
on leaves of these plants r e s e m b l e d t h a t p r o d u c e d by beet
yellows a n d b e e t western yellows in c e r t a i n respects, b u t t h e r e
was a type of m o t t l i n g n o t characteristic of s y m p t o m s of e i t h e r
of these diseases. T e s t s showed t h a t t h e p l a n t s w e r e infected
with a juice-transmissible virus, a p p a r e n t l y n o t previously described o n sugar beet. T h e disease a n d its causal v i r u s w e r e
studied further a n d t h e results a r e p r e s e n t e d h e r e i n . Because of
the type of discoloration on m a t u r e leaves of affected plants t h e
n a m e beet m a r b l e leaf is suggested as a c o m m o n n a m e for t h e
disease.
Symptoms a n d Host R a n g e
G r e e n h o u s e symptoms of m a r b l e leaf have b e e n studied on
several species of p l a n t s i n o c u l a t e d w i t h j u i c e from diseased b e e t
plants. T h e virus may p r o d u c e b o t h local lesions a n d systemic
infection.
Local
Symptoms
Local lesions are p r o d u c e d on juice-inoculated leaves of Beta
vulgaris L.
(sugar beet), B. macrocarpa Guss., Chenopodium
amaranticolor Coste 8c Reyn., a n d C. murale L. (sowbane). Lesions
are similar on all of these hosts. T h e y b e g i n to a p p e a r on sugar
beet leaves as chlorotic spots a b o u t 1 mm in d i a m e t e r a b o u t 9
days after i n o c u l a t i o n . Lesions increase in size slowly a n d m a y
a t t a i n a d i a m e t e r of 2-3 mm ( F i g u r e 1A a n d B). A small n e c r o t i c
spot m a y d e v e l o p in t h e c e n t e r of t h e lesion, sometimes surr o u n d e d b y o n e o r m o r e r i n g s r a n g i n g i n color from yellow t o
green. If lesions are n u m e r o u s , i n o c u l a t e d leaves of C. amaranticolor a n d C. murale often yellow a n d d r o p . In such leaves t h e
lesions m a y be s u r r o u n d e d by a r i n g of green tissue.
Systemic
Effects
T h e m a r b l e leaf disease has b e e n f o u n d only o n sugar b e e t .
Its effects h a v e b e e n studied on sugar b e e t a n d o t h e r host p l a n t s
under greenhouse conditions.
1
Plant Pathologist, Crops Research Division, Agricultural Research Service, U. S.
Department of Agriculture.

34

JOURNAL OF THE A. S. S. B. T.

Figure 1.Sugar beet leaves showing local lesions following juice


inoculation. A, leaf with lesions in early stage of development; B, leaf
with older lesions showing necrotic centers surrounded by chlorotic rings.

Beta vulgaris L. Sugar beet. Systemic effects appear on sugar


beet plants 10-30 days after inoculation. First systemic symptoms
consist of vein chloiosis or m o t t l i n g on young leaves a n d various
patterns of chlorosis on half-grown leaves. Often the chlorotic
areas are broader than the veins and they may be continuous
or broken a n d accompanied by an indefinite mottle in some
cases (Figure 2A). As the leaf matures, vein chlorosis becomes
less conspicuous and the leaf becomes mottled. T h e chlorotic
areas, however, are not conspicuous or well defined (Figure
2B). As the leaf ages the m o t t l i n g becomes indefinite a n d the
tissue between the m a i n veins turns yellow prematurely (Figure
3). T h e leaves are not thickened as with beet yellows, b u t they
appear dry and papery. Affected plants show no recovery. T h e
range of symptoms described on leaves of different ages continues
to be produced as long as affected plants r e m a i n reasonably vigorous. Except for the effects on young leaves, m a r b l e leaf could
easily be mistaken for beet yellows or beet western yellows.
Beta atriplicifolia R o u y . Symptoms are similar to those described on sugar beet.
Beta macrocarpa Guss. A b o u t 20 days after inoculation,
young leaves begin to curl d o w n w a r d at the tips a n d vein clear
ing is marked. N e w leaves are curled a n d stunted a n d growth
of axillary buds is stimulated, often resulting in the production

VOL.

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35

1964

of a type of rosette. Plants are s t u n t e d a n d t h e o l d e r leaves


yellow p r e m a t u r e l y . Seed yield is r e d u c e d drastically, a n d seeds
from diseased p l a n t s are m u c h smaller t h a n those from h e a l t h y
plants.

Figure 2.A, young sugar beet leaf showing vein chlorosis and mottle;
|B, leaf approaching maturity, showing marbled type of mottling.

Figure 3Sugar beet leaves showing yellowing


on older leaves by the marble leaf disease.

and

necrosis

produced

JOURNAL OF THE A. S. S. B. T.

Beta patellar is Moq. Leaves t u r n yellow or brownish, new


growth yellows prematurely, and small dark spots appear on
young leaves. No mottling is produced and there is little stunting of affected plants.
Beta patula Ait. Leaves begin to yellow as they approach
maturity a n d may show a type of mottling or splotching. Leaves
yellow a n d die prematurely.
Beta procumbens Chr. Sm. Yellowing, which is rather uniform without mottling, begins to appear when the leaves are
a b o u t half-grown. Older leaves die prematurely a n d the plants
are stunted.
Atriplex coronata S. Wats. (Crown saltbush). First symptoms
begin to appear a b o u t 20 days after inoculation as broad chlorotic
areas along the m a i n veins. Leaves produced later show m o t t l i n g
a n d yellowing. Plants are not appreciably stunted.
Atriplex pacifica Nels. (Dot scale). Plants begin to produce
leaves with reddish splotches on the blades and petioles and on
the stems about 30 days after infection. R e d d e n i n g is most intense at the leaf tips and a r o u n d the edges of the leaves. Plants
are not markedly dwarfed.
Chenopodium amaranticolor Coste & Reyn. Systemic infection begins to appear a b o u t 20 days after infection. First
leaves that show symptoms may be only partially affected. Invaded portions show marked vein clearing: of continuous or
broken lines. Leaves produced later show distinct vein clearing
a n d a type of mottle or splotchino. Leaves are dwarfed a n d tend
to b e n d downward a n d to roll slightly. G r o w t h is reduced b u t
infected plants produce seeds. T h e inflorescences of diseased
plants have considerably m o r e red p i g m e n t than healthy plants.
Seeds of diseased plants are small and germination is reduced.
Chenopodium ambrosioides L. (Mexican-tea).
Plants show
marked m o t t l i n g of a diffuse type. T i p s of branches are distorted
a n d leaves yellow a n d die. Seed production is m u c h reduced.
Chenopodium murale L. (Sowbane). Systemic infection soon
follows the production of local lesions on inoculated leaves.
Y o u n g leaves are mottled a n d mav show necrosis. Usually the
blossoms become necrotic a n d the flowering parts die. Few seeds
are produced on plants inoculated in t h e 5 to 10-leaf stage. In
plants that are larger when inoculated, systemic infection usually
does n o t become evident before the plants m a t u r e , or symptoms
are produced on only the axillary shoots in t h e inflorescence.
Plants are dwarfed a n d stems t e n d to b e n d a n d become distorted.
Stems of parts of the plant showing leaf symptoms are brittle.

V O L . 13, N o . 1, A P R I L 1964

37

Chenopodium urbicum L. (City goosefoot).


L a r g e chlorotic
areas are p r o d u c e d on leaves t h a t d e v e l o p after infection. P l a n t s
are s t u n t e d a n d leaves b e c o m e necrotic.
Plants on which no Injection was Observed
In host-range studies t h e following species in t h e i n d i c a t e d
families, i n o c u l a t e d by m e a n s of r u b b i n g j u i c e from infected
beet plants over leaves t h a t h a d b e e n lightly s p r i n k l e d w i t h an
abrasive, showed no evidence of infection.
A I Z O A C E A E . T e t r a g o n i a tetragonoides (Pall.) Ktze. ( N e w
Zealand spinach).
A M A R A N T H A C E A E . A m a r a n t h u s californicus (Moq.)
S.
Wats., A. caudatus L. (Love-lies-bleeding), A. cruentus L. ( P u r p l e
a m a r a n t h ) , A. retroflexus L. ( R e d r o o t a m a r a n t h ) .
C H E N O P O D I A C E A E . A triplex coulteri (Moq.) D. Dietr.,
A. expansa (D. & H.) S. W a t s . (Fogweed), A. polvcarpa ( T o r r . )
S. Wats. (Cattle saltbush), Beta lomatogona Fisch. & Mey., B.
maritima L., B. procumbens C h r . Sm., B. trigyna W a l d s t . & Kit.,
B. webbiana Moq., Chenopodium bonus-henricus L.
(Good K i n g
H e n r y ) , C. capitatum (L.) Aschers. ( S t r a w b e r r y - b l i t e ) , a n d
Monolepis nuttaliana
(Schult.)
Greene.
C O M P O S I T A E . S o n c h u s oleraceus L. a n d Zinnia elegans
Jacq. (Zinnia).
CRUCIFERAE.Capsella
bursa-pastoris
L.,
(Shepherd's
purse) a n d Armoracia rusticana G a e r t n . , B. Mey., & Scherb.
(Horseradish).
C O N V O L V U L A C E A E . i p o m o e a purpurea
(L.)
Roth
( M o r n i n g glory).
G E R A N I A C E A E . E r o d i u m cicutarium
(L.)
L'Her.
(Filaree).
L E G U M I N O S A E . P h a s e l o u s vulgaris L. (Bean) a n d Vigna
sinensis ( T o r n e r ) Savi (Cowpea).
M A L V A C E A E . M a l v a parviflora L. (Little mallow) a n d M.
sylvestris L. ( H i g h mallow).
P R I M U L A C E A E . S a m o l u s parviflorus Raf.
( W a t e r pimpernel).
P O L Y G O N A C E A E . R u m e x crispus L. (Yellow dock).
S O L A N A C E A E . C a p s i c u m frutescens L.
( P e p p e r ) , Datura
meteloides D C .
( T o l g u a c h a ) , D. stramonium L.
(Jimsonweed),
Lycopersicon esculentum
Mill.
(Tomato),
Nicandra physalodes
(L.) Pers., (Apple-of-Peru), Nicotiana bigelovii S. W a t s . (Indian
tobacco), N. glutinosa L., a n d Solanum dulcamara L.
(Bitter
c l i m b i n g nightshade).
U R T I C A C E A E . U r t i c a dioica L. (Stinging nettle).

38

J O U R N A L OF T H E A. S. S. B. T.

Transmission of Causal Virus


Several m e t h o d s of i n o c u l a t i o n h a v e b e e n used w i t h t h e
m a b l e leaf v i r u s over a p e r i o d of 3 years.
Juice Transmission.
M a r b l e leaf virus is r e a d i l y t r a n s m i s s i b l e
by juice inoculation by the r u b b i n g method of inoculation.
Usually, h i g h p e r c e n t a g e s of i n o c u l a t e d p l a n t s of susceptible
species b e c o m e infected.
Insect Transmission.
T h e species of insects used in t r a n s mission tests w e r e Circulifer tenellus
(Baker), Myzus persicae
(Sulz.), Aphis fabae Scop., Hyalopterus atriplicis L., Pemphigus
betae, D o a n e , a n d Macrosiphum euphorbiae
(Thomas).
T r a n s m i s s i o n was o b t a i n e d w i t h M. persicae, A. fabae, a n d
M. euphorbiae. T h e t r a n s m i s s i o n level was low w i t h each of
these species. In tests w i t h M. persicae no infection was o b t a i n e d
w i t h single a p h i d s . W i t h 2, 5, a n d 10 a p h i d s p e r p l a n t infection
increased w i t h n u m b e r o f a p h i d s , b u t less t h a n 2 0 % infection
was o b t a i n e d w i t h 10 a p h i d s p e r p l a n t . Fifty a p h i d s p e r p l a n t
gave a b o u t 5 0 % infection. E v e n l o w e r p e r c e n t a g e s o f infection
were o b t a i n e d w i t h A. fabae a n d Al. euphorbiae.
No infection
was o b t a i n e d w i t h t h e o t h e r species of insects e v e n w h e n used
in very large n u m b e r s . It seems u n l i k e l y t h a t a n y of t h e species
shown t o b e vectors o f t h e v i r u s w o u l d b e a b l e t o p r o d u c e w i d e s p r e a d transmission from b e e t to b e e t unless very large p o p u l a tions were present.
Tests for Seed Transmission.
P l a n t s of Beta vulgaris
(an
a n n u a l type), B.
macrocarpa,
Chenopodium amaranticolor,
and
C. urbicum w e r e i n o c u l a t e d w i t h m a r b l e leaf v i r u s in early stages
o f d e v e l o p m e n t a n d r e t a i n e d u n t i l seeds w e r e harvested. Seeds
from these p l a n t s w e r e p l a n t e d in flats in t h e g r e e n h o u s e a n d
seedlings w e r e w a t c h e d t o d e t e r m i n e w h e t h e r thev showed s y m p t o m s of disease. O b s e r v a t i o n s w e r e m a d e on 692 B. vulgaris seedlings, 439 B. macrocarpa seedlings, 742 C. amaranticolor seedlings a n d 740 C. urbicum seedlings. No e v i d e n c e of infection
was f o u n d on p l a n t s of a n y of these species. Since p l a n t s of each
of these 4 species show m a r k e d s y m p t o m s w h e n infected, it was
c o n c l u d e d t h a t t h e m a r b l e leaf virus p r o b a b l y is n o t seedtransmitted.
Tests for Transmission by Dodder. Cuscuta californica H. &
A. was established on b e e t p l a n t s w i t h m a r b l e leaf a n d stems
were t r a i n e d from diseased t o h e a l t h y s e e d l i n g s u g a r b e e t p l a n t s .
I n o t h e r tests, stem t i p s w e r e b r o k e n from t h e d o d d e r p l a n t s
c r o w i n g o n diseased beets a n d p l a c e d o n h e a l t h y s e e d l i n g beet
plants w h e r e t h e y soon b e c a m e established. F o r t y p l a n t s w e r e
inoculated by each of these m e t h o d s , b u t no e v i d e n c e of v i r u s

VOL.

13, N o .

1, A P R I L 1964

39

transmission was o b t a i n e d . J u i c e i n o c u l a t i o n s from d o d d e r g r o w ing on diseased beet p l a n t s w e r e m a d e to Beta macrocarpa. No


infection was o b t a i n e d . It seems p r o b a b l e , therefore, that C.
californica is n o t a host of m a r b l e leaf virus a n d t h a t colonies
growing on diseased beet p l a n t s c o n t a i n little or no virus.
P r o p e r t i e s of C a u s a l V i r u s
In studies of p r o p e r t i e s of m a r b l e leaf virus, juice was pressed
f r o m diseased leaves of sugar beet plants a n d i n o c u l a t e d i n t o
leaves of h e a l t h y plants of Beta macrocarpa after t h e i n d i c a t e d
t r e a t m e n t . Results w e r e recorded on t h e basis of n u m b e r of
local lesions p r o d u c e d .
Thermal inactivation. In t r e a t m e n t s of beet j u i c e for 10 m i n .
at 5C intervals, t h e r e was a progressive decrease in active v i r u s
from 50 t h r o u g h 6 0 . T h e r m a l i n a c t i v a t i o n p o i n t of t h e virus
appears to lie b e t w e e n 60 a n d 65.
Tolerance of dilution. In tests in which beet juice was d i l u t e d
with water, progressively fewer lesions w e r e p r o d u c e d as d i l u t i o n
was increased, b e g i n n i n g w i t h a 1-10 d i l u t i o n . T h e d i l u t i o n
endpoint of t h e virus in beet j u i c e a p p e a r s to lie b e t w e e n 1-500
and 1-1000, i n d i c a t i n g a low c o n c e n t r a t i o n of active v i r u s in t h e
beet plant.
Resistance to aging. Persistence of active v i r u s in expressed
juice of sugar beet varied widely in different tests at r o o m
t e m p e r a t u r e . In some tests no v i r u s was recovered after 4 h o u r s ,
whereas in others a small a m o u n t of infection was o b t a i n e d
after 24.
D a m a g e Produced By Marble Leaf
Since the d i s t r i b u t i o n a n d incidence of m a r b l e leaf a r e u n known, it is n o t possible to evaluate accurately t h e a c t u a l or
potential economic i m p o r t a n c e of t h e disease in t h e field. It has
been d e t e r m i n e d , however, that m a r b l e leaf is c a p a b l e of producing a p p r e c i a b l e stunting- of plants in a g r e e n h o u s e .
In 4 g r e e n h o u s e tests, p l a n t s grrowing in 6-inch pots, 4 p l a n t s
per pot, were i n o c u l a t e d in a b o u t t h e 4-leaf stage a n d h a r v e s t e d
and weighed 60 to 78 days after i n o c u l a t i o n . In a second t y p e
of test, plants g r o w i n g singly in 3-gallon crocks a n d w a t e r e d w i t h
Hoagland solution were i n o c u l a t e d in t h e 10 to 14-leaf stage.
They were harvested a n d weighed in lots of 10 at m o n t h l y intervals 2 to 5 m o n t h s after i n o c u l a t i o n . In these tests ( T a b l e
1 ) m a r b l e leaf p r o d u c e d a p p r e c i a b l e r e d u c t i o n s i n total p l a n t
weight, particularly in p l a n t s i n o c u l a t e d in t h e s e e d l i n g stage,
These losses a r e a b o u t t h e same as those p r o d u c e d by b e e t mosaic
under similar conditions. Such losses i n d i c a t e t h a t m a r b l e leaf

40

J O U R N A L OF T H E A. S. S. B. T.

Table 1.Effect of marble leaf


greenhouse conditions.

Test number

2
3
4
5

on weight

Time from
inoculation
to harvest

of

sugar

beet

plants

(US

75)

under

Average weight of plants 1


Healthy

Diseased

Days

Grams

Grams

60
60
60
78
60
90
120
150

28
33
30
54
994
1331
1690
1861

24
24
25
48
848
1230
1401
1595

1
In tests 1 to 4, inclusive, 40 plants in 6-inch pots (4 plants per p o t ) were inoculated
in the 4 to 6-leaf stage; in test 5, plants (singles) in 3-gallon crocks were inoculated in
the 10 to 14-leaf stage and 10 plants were harvested after each indicated time interval.
Inoculated plants were compared with equal numbers of check (non-inoculated) plants in
each test.

w o u l d b e c a p a b l e o f c a u s i n g m e a s u r a b l e yield r e d u c t i o n s i n t h e
field if v e c t o r s w e r e a v a i l a b l e to p r o d u c e w i d e s p r e a d d i s s e m i n a t i o n of t h e causal v i r u s in early stage of d e v e l o p m e n t of b e e t
plants.
T e s t s w e r e m a d e also t o d e t e r m i n e t h e effects o f m a r b l e leaf
on seed p r o d u c t i o n in Beta macrocarpa. P l a n t s w e r e i n o c u l a t e d
i n a b o u t t h e 6-leaf stage a n d r e t a i n e d for seed p r o d u c t i o n a l o n g
w i t h a p p r o p r i a t e h e a l t h y c h e c k p l a n t s . I n 2 tests ( T a b l e 2 ) t h e r e
w e r e r e d u c t i o n s i n seed yield a n d w e i g h t o f m o r e t h a n 5 0 p e r c e n t . T h e effect of t h e disease on yield of seed of s u g a r b e e t has
not been determined.
Table 2.Effect of marble leaf on seed production and seed size in Beta macrocarpa.
Test
number

Weight of seeds from


10 plants

Weight of 100 seeds

Healthy
plants

Diseased
plants

Healthy
plants

Diseased
plants

Grams

Grams

Grams

Grams

68.5

31.2

3.0

1.3

82.4

36.2

3.5

1.2

VOL.

13, N o .

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41

1964

Summary
A juice-transmissible virus which causes vein-yellowing a n d
m o t t l i n g of i m m a t u r e leaves a n d distinct yellowing of m a t u r e
leaves of sugar beet p l a n t s was isolated from beets from O r e g o n .
T h e virus p r o d u c e s local lesions on i n o c u l a t e d leaves of Beta
vulgaris, B.
macrocarpa^
Chenopodiurn
amaranticolor, a n d
C.
murale. Systemic infection was o b t a i n e d in 6 species of Beta, 4
species of Chenopodiurn a n d 2 species of Atriplex.
T h e virus
appears to have a l i m i t e d host r a n g e .
Myzus persicae, Aphis
fabae, a n d Macrosiphum euphorbiae a r e inefficient vectors of t h e
virus. No evidence of seed transmission was found. T h e virus
has a t h e r m a l inactivation p o i n t b e t w e e n 60 a n d 65 C, a d i l u t i o n
e n d p o i n t of a b o u t 1-1,000, a n d it r e m a i n s active in e x t r a c t e d
juice at r o o m t e m p e r a t u r e for 24 h o u r s , or less. U n d e r greenhouse conditions t h e disease caused a b o u t a 10 p e r c e n t r e d u c t i o n
in p l a n t g r o w t h . Yield a n d size of seeds of Beta macrocarpa w e r e
greatly reduced. T h e disease a p p a r e n t l y has a very l i m i t e d dist r i b u t i o n a n d is n o t k n o w n to be c a u s i n g m e a s u r a b l e loss to
the sugar beet c r o p in t h e areas w h e r e it was f o u n d .

Critical Levels Versus Q u a n t i t y - Quality Factors for


Assessing Nutritional Status of Sugar Beet Plants 1
JAY L . H A D D O C K AND D A R R E L M . S T U A R T 2
Received for publication September 30, 1963

T h e ideal p l a n t n u t r i e n t c o n c e n t r a t i o n a n d b a l a n c e for sugar


beet p l a n t tissue has n o t b e e n p r o p o s e d . M a n y p l a n t physiologists
a n d agronomists, d o u b t t h e possibility of establishing stable reference values for p l a n t n u t r i e n t c o n c e n t r a t i o n i n specific p l a n t
tissue t h a t will h o l d even in a restricted c l i m a t e .
M u c h effort has b e e n d i r e c t e d t o f i n d t h e k i n d a n d q u a n t i t y
of p l a n t n u t r i e n t s t h a t m u s t be a d d e d to soils to o b t a i n h i g h
yields of h i g h q u a l i t y sugar beets. D u r i n g t h e past forty years,
sugar beet yields in most areas h a v e s h o w n a g r a d u a l increase.
U n f o r t u n a t e l y , d u r i n g this same p e r i o d t h e r e has b e e n a p e r sistent d e c l i n e i n quality. E v e n w i t h t h e m a r k e d i m p r o v e m e n t
in yield of roots p e r acre, m a n y s t u d e n t s of p r o d u c t i o n p r o b l e m s
feel t h a t yield can b e d o u b l e d a n d q u a l i t y i m p r o v e d .
Before i m p o r t a n t a d d i t i o n a l progress can b e m a d e t o w a r d s
increasing yield a n d q u a l i t y of c o m m e r c i a l sugar beets by fertilization, it will be necessary to establish s t a n d a r d s of reference t h a t
will sharply distinguish b e t w e e n well-nourished a n d i n a d e q u a t e l y
n o u r i s h e d beet plants.
G o o d a l l a n d Gregory (3) 3 discussed t h e r e l a t i o n of yield to
n u t r i e n t supply a n d u p t a k e a n d p r o p o s e d t h e use o f t h e t e r m
intensity level. They stated " F o r each factor in t u r n t h e r e is an
optimum level. G r o w t h is increased if i n t e n s i t y is b r o u g h t to
this level a n d decreased if raised f u r t h e r . " Macy (8) p r o p o s e d
t h e c o n c e p t of t h e " p o v e r t y a d j u s t m e n t z o n e " as a m e c h a n i s m
w h i c h m a y e x p l a i n w h y n u t r i e n t c o m p o s i t i o n o f p l a n t tissue m a y
vary a n d g r o w t h be c o m p e n s a t e d for, by t h e relatively favorable
effects of o t h e r factors. He f u r t h e r suggested t h e use of critical
a n d minimum percentages as precise values of n u t r i e n t c o m p o s i t i o n w h i c h set t h e u p p e r a n d lower l i m i t s respectively of " p o v e r t y
a d j u s t m e n t z o n e . " H e d i d n o t h o l d t h a t t h e critical a n d m i n i m u m
percentages of n u t r i e n t s in p l a n t tissue are i n v a r i a b l e . Shear a n d
C r a n e (11) observed t h a t "optimum nutritional status as reflected
by leaf analysis is different for each c r o p a n d s t a n d a r d s of c o m p a r i s o n m u s t b e set u p for different tissues a n d for each c r o p . "
They
(1943-47) f u r t h e r stated " T h e first a n d only infallible
s y m p t o m of t h e deficiency of any e l e m e n t is e v i d e n c e d by a red u c e d r a t e of g r o w t h . "
1
Contribution from the Southwest Branch, Soil and Water Conservation Research
Division, Agricultural Research Service, USDA, in cooperation with the Utah Agricultural
Experiment
Station, Logan, Utah. Utah Experiment Station Journal Paper No. 231.
2
Research Soil Scientist and Soil Scientist, respectively, USDA, Logan, Utah.
3
Numbers in parentheses refer to literature cited.

VOL.

13, N o .

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1964

43

T h o m a s (13) s u m m a r i z e d his conclusions on the application


of foliar diagnosis as a tool for diagnosing m i n e r a l r e q u i r e m e n t
of plants by t h e s t a t e m e n t "Soil a n d p l a n t testing m e t h o d s are
necessarily valid only w h e n each is used in a c o m p a r a t i v e m a n ner a n d are usable only w i t h a key or reference to some s t a n d a r d . "
T h o m a s (14) c o n c l u d e d t h a t the use of l i m i t i n g values of each
n u t r i e n t is i n a d e q u a t e as a basis of ascertaining relationships of
composition to yield. T h e most effective p r o c e d u r e is o n e t h a t
will give n o t only a q u a n t i t a t i v e m e a s u r e of these n u t r i e n t s
present in t h e leaf at t h e m o m e n t of sampling, b u t in a d d i t i o n ,
an i n d e x of t h e q u a l i t a t i v e relations r e s u l t i n g from t h e i r i n t e r actions. T h o m a s was of t h e o p i n i o n that there was no fundamental physiological e x p e r i m e n t a l basis to s u p p o r t the use of
tissues o t h e r t h a n w h o l e active leaves.
U l r i c h (17) c o m b i n e d Macy's m i n i m u m percentage a n d
critical percentage i n t o o n e value which he identified as critical
level. He identified this c o n c e n t r a t i o n of p l a n t n u t r i e n t s by saying " W h e n the n u t r i e n t c o n c e n t r a t i o n s of t h e plants are above
the critical level a n d r e m a i n there t h r o u g h o u t t h e e n t i r e growth
period, t h e r e is very little chance of a response in g r o w t h from
addition of m o r e n u t r i e n t s . " U l r i c h , et al. (18) proposed the
following critical levels for sugar beet petioles from recently
m a t u r e d leaves: n i t r a t e - n i t r o g e n 1,000 p p m , p h o s p h o r u s 750
p p m a n d potassium 10,000 p p m . T h e s e values were proposed
for a 2 p e r c e n t acetic acid extract of oven-dry, finely-ground leaf
petioles.
M a n y s t u d e n t s of sugar beet p r o d u c t i o n p r o b l e m s believe that
the most i m m e d i a t e a n d significant advance in yield a n d quality
of sugar beets can be o b t a i n e d by i m p r o v i n g t h e n u t r i t i o n a l
conditions in c o m m e r c i a l fields. T h e a u t h o r s a p p r o a c h e d a study
of this p r o b l e m w i t h the following assumptions w h i c h they obtained from a review of available l i t e r a t u r e on m e t h o d s of appraising n u t r i t i o n a l status of p l a n t :
1. T h e r e is an established casual r e l a t i o n s h i p b e t w e e n internal n u t r i e n t c o n c e n t r a t i o n a n d p l a n t growth.
2 . T h e pot c u l t u r e m e t h o d i n which r o o t t e m p e r a t u r e s are
m a i n t a i n e d in a m a n n e r similar to that f o u n d in commercial
sugar beet f i e l d s , provides t h e most r a p i d a n d c o n v e n i e n t
a p p r o a c h t o t h e identification o f t h e o p t i m u m m i n e r a l n u t r i tion of sugar beets.
3. T h o s e sugar beet plants which m a k e t h e best growth m u s t
b e t h e best n o u r i s h e d . A n a d e q u a t e n u t r i t i o n m u s t b e the
best balanced n u t r i t i o n .

44

JOURNAL OF THE A. S. S. B. T.

4. If t h e r e is a well-defined m i n i m u m p e r c e n t a g e of m i n e r a l
e l e m e n t i n p l a n t tissue t h e r e m u s t also b e a n o p t i m u m p e r centage.
A study of t h e available l i t e r a t u r e led t h e a u t h o r s to select
t h e U l r i c h (17) t e c h n i q u e a n d a modification of t h e T h o m a s
(13) p r o c e d u r e as t h e most fruitful m e t h o d s for d i a g n o s i n g n u t r i tional status of sugar beet plants.
T h e d a t a p r e s e n t e d i n this p a p e r are t h e r e s u l t o f a n
a t t e m p t t o d e t e r m i n e w h i c h o f t h e t w o m e t h o d s u n d e r observat i o n was m o r e s u i t a b l e as a basis for d i a g n o s i n g significant n u t r i tional d i s t u r b a n c e s in sugar beets. A second closely r e l a t e d objective was t o observe t h e o p t i m u m n u t r i e n t c o n c e n t r a t i o n a n d
balance o f t h e t h r e e p r i m a r y p l a n t n u t r i e n t s , n i t r o g e n , p h o s p h o r u s , a n d potassium, i n sugar beet petioles c o n d u c i v e t o h i g h
yields of roots a n d sugar.
Methods and Procedure
W h i l e ten n u t r i e n t solutions w e r e used i n this study, o n l y
six of these are referred to in this p a p e r in o r d e r to simplify prese n t a t i o n of data ( T a b l e 1). All of these w e r e modifications of H o a g land's (6) n u t r i e n t s o l u t i o n N o . 1 . T h e n u t r i e n t s o l u t i o n s w e r e
m a d e w i t h t a p water which c o n t a i n e d 0, 2, 16, 44 p p m of K, N a ,
M g a n d Ca, respectively, a n d f u r t h e r modified b y t h e v e r m i c u l i t e
s u b s t r a t u m w h i c h at e q u i l i b r u m w i t h t h e w a t e r p r o v i d e d 18, 5,
19, a n d 46 p p m of K, N a , Mg a n d Ca, respectively. T h e s e c o n c e n t r a t i o n s w e r e constantly b e i n g modified b y n u t r i e n t s o l u t i o n
additions, unequal water and plant nutrient withdrawal, a n d
chemical p r e c i p i t a t i o n . T e n - g a l l o n cans filled w i t h N o . 2 v e r m i c u l i t e were b u r i e d in soil to w i t h i n 1 inch of t h e t o p r i m in
o r d e r t o m a i n t a i n r o o t t e m p e r a t u r e s c o m p a r a b l e t o n o r m a l soil
t e m p e r a t u r e s . Five holes w e r e p u n c h e d i n t h e b o t t o m o f each
c a n t o p r o v i d e a d e q u a t e d r a i n a g e . T w e n t y sugar b e e t seeds o f
a c o m m e r c i a l m o n o g e r m variety, S L C 126, w e r e p l a n t e d A p r i l
15, 1960. T h e s e w e r e t h i n n e d J u n e 29, to leave a final s t a n d of
t h r e e p l a n t s p e r can. T h e r e w e r e t e n cans i n each t r e a t m e n t
w h i c h w e r e r a n d o m i z e d i n each row. T h e cans w e r e spaced o n
40-inch centers so t h a t each p o t h a d 11 sq ft of surface. T h e
n u t r i e n t s o l u t i o n s w e r e p r e p a r e d similarly all season e x c e p t for
t r e a t m e n t Check-N w h i c h was r e d u c e d i n n i t r o g e n t o one-half
n o r m a l c o n c e n t r a t i o n o n S e p t e m b e r 1 a n d t o zero n i t r o g e n
O c t o b e r I . O n e gallon o f each n u t r i e n t s o l u t i o n was a p p l i e d t o
its respective can daily, except d u r i n g h o t w e a t h e r i n m i d - J u l y
a n d m i d - A u g u s t w h e n o n e a n d one-half gallons w e r e used.
T h o m a s (14) b e l i e v e d i t i n c o n g r u o u s t o u s e p l a n t tissue
o t h e r t h a n w h o l e leaf tissue as a basis for c h a r a c t e r i z i n g n u t r i -

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tional status of plants. Nevertheless, in o r d e r to e x t e n d t h e comparison of the U l r i c h a n d T h o m a s t e c h n i q u e s t h e a u t h o r s m o d i fied t h e T h o m a s p r o c e d u r e to i n c l u d e soluble extract of beet
leaf petioles as well as total composition of leaf blades.
Q u a n t i t y a n d q u a l i t y factors are expressed in this p a p e r in
terms of m i l l i e q u i v a l e n t s p e r 100 grams of oven-dry p l a n t tissue.
To express the intensity of n u t r i t i o n on a q u a n t i t a t i v e basis, the
milliequivalents p e r 100 grams of N + P . + K are s u m m e d . T h e
quality n i t r o g e n value is expressed as a percentage of the

blades were used, these p l a n t n u t r i e n t values were expressed on


the basis of total composition. W h e n sugar beet petioles were
used, the q u a n t i t y a n d q u a l i t y factors were calculated from the
concentration of soluble n i t r o g e n ( N 0 3 N + organic N
+ N H 4 N) p h o s p h o r u s a n d potassuim in acid extract (1 gram
petiole tissue oven d r i e d at 70 C, g r o u n d to pass 40 mesh sieve
and extracted w i t h 100 ml solution.) T h e general o u t l i n e of the
Ulrich (17) t e c h n i q u e was used.
Leaf blades a n d petiole samples from the most recently matured leaves were o b t a i n e d from each p l a n t every two weeks beginning July 1. T h e s e tissues were d r i e d rapidly at 70 C, g r o u n d to
pass a 40-mesh screen, a n d e x a m i n e d chemically to d e t e r m i n e
the c o n c e n t r a t i o n of acetic acid soluble n u t r i e n t s in petioles.
T h e 2 p e r c e n t acetic acid extract (1 g r a m of p l a n t tissue per
100 ml of solution) from petioles was analyzed for nitrate-nitrogen
using the d e p h e n y l a m i n e spot plate m e t h o d . Soluble a m m o n i a
and organic-nitrogen were o b t a i n e d by t h e H i l l e b r a n d , et al. (6)
procedure. T o t a l n i t r o g e n was d e t e r m i n e d in leaf blades by
Perrin's (9) m e t h o d . P h o s p h o r u s was d e t e r m i n e d in both leaf
blades a n d petioles by Barton's (2) p r o c e d u r e . T h e flame p h o t o meter, direct intensity m e t h o d , was used for potassium d e t e r minations. Sucrose a n d p u r i t y were d e t e r m i n e d on t h e sugar
beet p u l p , o b t a i n e d w i t h the Keil rasp, by t h e cold water digestion
Sachs-Le Docte m e t h o d .
Experimental Results
Quality of Sugar Beets as Affected by
Nutritional
Environment
T h e d a t a on t h e yield of sugar beet roots are presented in
4
Figure l . T r e a t m e n t s N H 4 , 1/4 N a n d L o w K, gave root yields
significantly lower t h a n Check-N t r e a t m e n t , a n d t h e l/2 N a n d
1/2 K t r e a t m e n t s showed a s t r o n g tendency for r e d u c e d growth.
Yield

and

4
Duncan's multiple range test for a comparison of six means at the .05 level of significance is used throughout this paper.

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JOURNAL OF THE A. S. S. B. T.

T h e n u t r i t i o n a l e n v i r o n m e n t s used i n this study e x e r t e d a s


m u c h or greater influence on t h e yield of t o p s as on r o o t s as
shown in F i g u r e 2. W h e n yield of roots a n d tops a r e g r a p h e d
in ascending a r r a n g e m e n t s as in Figures 1 a n d 2, it will be n o t e d
t h a t t h e relative g r o w t h rates of roots a n d tops a r e affected differently by different n u t r i e n t solutions. H o w e v e r , f o u r t r e a t m e n t s giving t h e lowest r o o t g r o w t h also gave t h e lowest t o p
g r o w t h . T h e t o p g r o w t h from t r e a t m e n t 1 / 2 K was significantly
g r e a t e r t h a n from 1 / 2 N , N H 4 , L o w K , a n d 1 / 4 N .

Figure 1.Yield of sugar beet roots as affected by nutritional environment 1960.

Figure 2.Yield of sugar beet tops as affected by nutritional environment 1960. (Fresh weight.)

T h e yield of gross sugar m a y be of g r e a t e r significance t h a n


e i t h e r yield of r o o t s or tops as an i n d i c a t i o n of t h e influence of
nutritional environment on total growth rate. T h e s e data are
shown in Figure 3. Again, four of the five nutritional treatments
adversely i n f l u e n c i n g yields of r o o t s w e r e r e s p o n s i b l e for depressed
yields o f sucrose w h e n c o m p a r e d t o C h e c k - N t r e a t m e n t . T h e
1/2
N t r e a t m e n t d i d n o t depress yield of gross s u g a r significantly
b e l o w C h e c k - N , h o w e v e r , t h e t r e n d for this t r e a t m e n t was s t r o n g ly in t h e d i r e c t i o n of d e p r e s s i o n .

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Figure 3-Yield of sugar as affected by n u t r i t i o n a l environment 1960.

Yield and Quality Related to Critical Nutrient Level


T h e concentrations of the three primary plant nutrients, in
recently m a t u r e d leaf petioles, are shown in Figures 4, 5, a n d
6. It is obvious from t h e graphical data in F i g u r e 4, that the
nitrate-nitrogen c o n c e n t r a t i o n in sugar beet petioles was above
the critical level for all t r e a t m e n t s . T h e 14 N t r e a t m e n t reached
the critical level at two s a m p l i n g periods b u t was n o t below it
at any t i m e .

Figure 4.Seasonal concentration of nitrate nitrogen in sugar beet


petioles as influenced by n u t r i e n t environment 1960.

T h e graphical r e p r e s e n t a t i o n of t h e p h o s p h o r u s concentration in sugar beet petioles as given in F i g u r e 5 shows a d e q u a t e


n u t r i t i o n w i t h respect to this n u t r i e n t for the six n u t r i e n t c u l t u r e d plants.
T h e seasonal potassium c o n c e n t r a t i o n in sugar beet petioles
shown in F i g u r e 6 suggests t h a t all plants were well supplied
with this n u t r i e n t .

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J O U R N A L OF T H E A.

S. S.

B.T.

Figure 5.Seasoal soluble p h o s p h o r u s concentration


petioles as influenced by n u t r i t i o n a l e n v i r o n m e n t 1960.

in

sugar

beet

F i g u r e 6.Seasonal soluble potassium c o n c e n t r a t i o n


petioles as influenced by n u t r i t i o n a l e n v i r o n m e n t 1960.

in

sugar

beet

Quantity factor of nutrition in


beet petiolesThe d a t a in
F i g u r e 7 for q u a n t i t y factor for s o l u b l e c o n s t i t u e n t s in sugar
b e e t petioles a r e p l o t t e d b y l i n e g r a p h s a n d cover t h e p e r i o d
from J u l y 1 to harvest. T h e solid h o r i z o n t a l lines r e p r e s e n t t h e
e x t r e m e s in seasonal v a r i a t i o n for i n t e n s i t y of n u t r i t i o n for welln o u r i s h e d sugar b e e t plants. T h e s e a r b i t r a r y b o u n d a r i e s a p p e a r
to serve t h e useful p u r p o s e of s e p a r a t i n g w e l l - n o u r i s h e d from
i n a d e q u a t e l y - n o u r i s h e d plants. T h e seasonal r a n g e i s established
on t h e basis of t h e c o m p o s i t i o n of s u g a r b e e t p l a n t s g r o w i n g in
t h e C h e c k - N t r e a t m e n t ( H o a g l a n d ' s 1 / 2 s t r e n g t h s o l u t i o n dep l e t e d i n n i t r o g e n t o 5 0 p p m o f n i t r a t e - n i t r o g e n from S e p t e m b e r
1 to O c t o b e r 1 a n d d e v o i d of n i t r o g e n after O c t o b e r 1). T h r e e
treatments (1/2 N, N H 4 , and 1/4 N) produced plants with quantity
b e l o w d e s i r a b l e limits.
Quality factor for nitrogen
in
beet petiolesThe g r a p h i c a l
d a t a in F i g u r e 8 r e p r e s e n t t h e q u a l i t y factor for n i t r o g e n . It is

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Figure 7-Seasonal q u a n t i t y factor of sugar beet petioles as affected


by nutritional environment 1960.

Figure 8.Seasonal quality factor for nitrogen in sugar beet petioles


as influenced by nutritional environment 1960.

noted that two t r e a t m e n t s (1/4 N a n d 1/2 N) are well below t h e


b o u n d a r y limits. O n l y t h e L o w K t r e a t m e n t p r o d u c e d plants
frequently above desirable limits. T h e ideal for n i t r o g e n q u a l i t y
appears to be a b o u t 50. H o w e v e r , a seasonal r a n g e b e t w e e n 45
and 55 is characteristic of well-nourished plants.
Quality factor for phosphorus nutrition in
beet petiolesThe
data in F i g u r e 9 show the p h o s p h o r u s q u a l i t y factor in petioles
from t h e Check-N t r e a t m e n t to vary from 2.5 to 6 t h r o u g h o u t
the season. T h e c o m p o s i t i o n of petioles from t h e 1 / 2 K t r e a t m e n t
is close to these same values. If these are ideal values, it is evident

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JOURNAL OF T H E A. S. S. B. T.

Figure 9.Seasonal quality factor for p h o s p h o r u s in sugar beet petioles


as influenced by n u t r i t i o n a l e n v i r o n m e n t 1960.

F i g u r e 10.Seasonal quality factor for potassium in sugar beet petioles


as influenced by n u t r i t i o n a l e n v i r o n m e n t 1960.

t h a t leaf petioles from t r e a t m e n t 1 / 2 N, 1 / 4 N a n d N H 4 a n d to


s o m e e x t e n t L o w K h a d excessive c o n c e n t r a t i o n s of p h o s p h o r u s .
Quality factor for potassium
nutrition
in
beet petiolesThe
d a t a in F i g u r e 10 show t h e ideal seasonal r a n g e in q u a l i t y factor
for s o l u b l e p o t a s s i u m in b e e t petioles of w e l l - n o u r i s h e d p l a n t s
t o b e b e t w e e n 4 0 a n d 50.
Quantity factor of nutrition in beet leaf bladesData showi n g t h e seasonal q u a n t i t y factor for s u g a r b e e t leaf b l a d e s a r e
given i n F i g u r e 11. W h i l e t h e r e w e r e differences b e t w e e n t r e a t m e n t s , n o n e varied widely from t h e C h e c k - N t r e a t m e n t . T h e
ideal q u a n t i t y factor for leaf blades a p p e a r s to h a v e a seasonal
v a r i a t i o n b e t w e e n 400 a n d 500 m e q p e r 100 g r a m s in d r y leaf
b l a d e tissue.

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1964

Figure 11.Seasonal quantity factor


affected by nutritional environment 1960.

of

sugar beet

leaf-blades

as

Figure 12.Seasonal quality factor for nitrogen in sugar beet leafblades as affected by nutritional environment 1960.

Quality factor for nitrogen in sugar beet leaf bladesThe data


for n i t r o g e n q u a l i t y in F i g u r e 12 clearly show t h a t t r e a t m e n t s
1/4
N, a n d 1 / 2 N w e r e low in n i t r o g e n . N o n e of the o t h e r treatments can be identified as b e i n g o u t of n i t r o g e n balance.
Quality factor for phosphorus in sugar beet leaf bladesThe
graphical d a t a i n F i g u r e 1 3 indicate t h a t two treatments, N H 4
a n d L o w K, p r o d u c e d plants which c o n t a i n e d relatively high concentrations of leaf p h o s p h o r u s .
Quality factor for potassium in sugar beet leaf bladesThe
graphical d a t a in F i g u r e 14 clearly segregate plants from treatments N H 4 , L o w K, 1/4 N a n d 1 / 2 N as b e i n g t o o high or too low
in potassium for ideal growth, relative to t r e a t m e n t Check-N.

52

J O U R N A L OF T H E A. S. S. B. T.

Figure 13.Seasonal quality factor for p h o s p h o r u s in sugar beet leafblades as affected by n u t r i t i o n a l e n v i r o n m e n t 1960.

Figure 14.Seasonal quality factor for potassium in sugar beet leafblades as affected by n u t r i t i o n a l e n v i r o n m e n t 1960.

Nitrogen : Potassium ratio in beet leaf bladesThe d a t a in


F i g u r e 15 s h o w i n g t h e seasonal n i t r o g e n : p o t a s s i u m r a t i o in
sugar beet leaf blades i n d i c a t e t h a t this factor is r e m a r k a b l y sensitive to n u t r i t i o n a l d i s t u r b a n c e s in t h e sugar b e e t p l a n t . It is a
s i m p l e m a t t e r t o identify p l a n t s w i t h n u t r i t i o n a l d i s t u r b a n c e s b y
this m e t h o d , if o n e can safely assume t h a t t h e t r e a t m e n t C h e c k - N
is an ideal n u t r i e n t c u l t u r e for t h e sugar beet p l a n t .
Nutrient Quantity and Quality
ValuesThe d a t a in T a b l e
2 r e p r e s e n t a s u m m a r y of t e n t a t i v e q u a n t i t y a n d q u a l i t y values
w h i c h a p p e a r best s u i t e d for m a x i m u m g r o w t h of sugar b e e t r o o t s
a n d yield o f sugar. F u r t h e r s t u d y m a y r e s u l t i n c o n f i r m a t i o n o r

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Figure 15.Seasonal nitrogenrpotassium ratio in sugar beet leaf-blades


as affected by n u t r i t i o n a l environment 1960.

modification of these t e n t a t i v e references. It is possible that a


study of o t h e r sugar beet varieties or significant climatic changes
from those involved in this study m i g h t dictate small modifications of these suggested values.
Discussion
T h e a u t h o r s have used t h e Ulrich (17) p r o c e d u r e for a
n u m b e r of years as a m e a n s of identifying the n u t r i e n t status of
sugar beet plants grown u n d e r field e x p e r i m e n t a l conditions.
It has served a very good p u r p o s e u n d e r c o n d i t i o n s w h e r e n u t r i e n t
compositions a m o n g plants varied widely.
Yield data in Figures 1, 2 a n d 3 show that relatively m i n o r
variations in available n u t r i e n t s , n i t r o g e n a n d potassium, may
result in significant yield differences. W h i l e p h o s p h o r u s was
applied to t h e six t r e a t m e n t s in e q u a l concentrations, the presence of variable c o n c e n t r a t i o n s of n u t r i e n t s o t h e r t h a n phosphorus, physiological changes in the substrate modifying the p H ,
and variable root a n d top p r o d u c t i o n s resulted in a significant
range in p h o s p h o r u s c o n c e n t r a t i o n in p l a n t petioles (see Figure
5).
C o m m e r c i a l sugar beets generally will show a w i d e r seasonal
fluctuation in n u t r i e n t composition t h a n d i d these six n u t r i e n t cultured beets. Nevertheless, since factors other t h a n available
n u t r i e n t s were k e p t u n i f o r m a n d since significant yield differences
were o b t a i n e d a m o n g t r e a t m e n t s , p l a n t material from this studv
appeared to be suitable for a comparison of critical level a n d
quantity-quality
factor m e t h o d s .
It is obvious that t h e critical level for nitrate-nitrogen shown
in F i g u r e 4 w o u l d have to be raised above 1,000 a n d p r o b a b l y
u p t o 5,000 p p m i n o r d e r t o p r o p e r l y identify p l a n t s with t h e
m i n i m u m of n i t r o g e n n u t r i t i o n a l d i s t u r b a n c e which could be

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J O U R N A L OF T H E A. S. S. B. X.
Table 1.Nutrient concentration in various nutrient solutions, 1960.
Parts per million of various nutrients 1

No.

Description

Check-N
1/2 N S e p t . 1 to O c t . 1
N o N O c t . 1 t o O c t . 15
1/2 K
LowK
1/2N
1/4 N
NH4

2
3
4
5
6

NO3-N

NH4-N

Ca

Mg

Na

pH

75

15
15
15
15
15
15

110
55
15
110
110
110

145
165
205
145
145
145

50
50
50
50
50
50

18
18
18
18
18
18

7.6-8.0
7.7-8.0
7.6-8.1
7.8-8.1
7.7-8.0
7.4-6.0

105
105
105
70
30

Minor elements added to all nutrient solutions:


Cu = .01, Mo = .004, and Fe= 4.5 ppm.

B = 0.25, Mn

0.25, Zn =

.028,

Table 2.Nutrient quantity and quality values for sugar beet leaf blades a n d leafpetioles which resulted in maximum sugar yields.
Factors
Quantity Factor
med/100g
Quality Factors
Nitrogen
Phosphorus
Potassium
N:K ratio

Leaf blades
Range
400 500
62-68
4.8-8.0
28-32
1.9-2.4

Ideal
450
65
6.4
30
2.15

Leaf petioles
Range
Ideal
300-500
45-55
3-7
40-56
0.9-1.4

410
50
5
45
1.1

identified as affecting r o o t yield. T h e low yield p e r f o r m a n c e


of plants g r o w n in t r e a t m e n t 1/4 N as s h o w n in F i g u r e s 1, 2, a n d
3 is closely associated w i t h , if n o t a result of, low n i t r o g e n c o n t e n t of leaf petioles. P r e s e n t s t a n d a r d s of critical levels (18) do
n o t clearly identify these p l a n t s as n i t r o g e n deficient.
Critical levels are n o t helpful in i d e n t i f y i n g n u t r i t i o n a l dist u r b a n c e s related to p h o s p h o r u s in this study.
It does n o t a p p e a r possible to identify a n y of t h e p e t i o l e
tissue in F i g u r e 6 as m a n i f e s t i n g c h e m i c a l c o m p o s i t i o n s y m p t o m s
of pota ssium n u t r i t i o n a l d i s t u r b a n c e . A r e v i e w of t h e d a t a in
T a b l e 1 a n d F i g u r e s 1, 2 a n d 3 w o u l d i m p r e s s o n e t h a t t h e r e a s o n
for p o o r p e r f o r m a n c e of p l a n t s in L o w K t r e a t m e n t m u s t be related to i n a d e q u a t e p o t a s s i u m , b u t deficiency of p o t a s s i u m is n o t
e v i d e n t in t h e d a t a in F i g u r e 6.
T h e d a t a o n q u a n t i t v factor, shown i n F i g u r e 7 , t h r o w s o m e
d o u b t o n t h e n u t r i t i o n a l a d e q u a c y o f p e t i o l e tissue from t h r e e
t r e a t m e n t s , viz, 1/4 N. N H , a n d 1/2 N if t h e a r b i t r a r y b r a c k e t
lines a r e accented a s i n c l u d i n g o p t i m u m i n t e n s i t y o f n u t r i t i o n .
T h e graphical data in Figure 8 lend support to the statement
by T h o m a s , et al., (16) " F o l i a r diagnosis can be a m o r e a c c u r a t e
i n d e x of t h e effect of a fertilizer t h a n a r e yields." W h i l e t h e

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yield of sugar from t r e a t m e n t 1/2 N was n o t significantly different


from that of Check-N, t h e q u a l i t y factor for n i t r o g e n suggests
the possibility of slight i n a d e q u a c y in n i t r o g e n all season.
T h e a u t h o r s do n o t have clear-cut evidence that a relatively
high c o n c e n t r a t i o n of p h o s p h o r u s is d e t r i m e n t a l to growth of
sugar beets. It is evident, however, that high q u a l i t y factor for
phosphorus is associated w i t h low p r o d u c t i o n performance in
the case of N H 4 a n d i/4 N t r e a t m e n t s in F i g u r e 9. Goodall a n d
Gregory (3) in discussing t h e r e l a t i o n of yield to n u t r i e n t uptake state, " F o r each factor in t u r n there is an o p t i m u m level.
G r o w t h is increased if intensity is b r o u g h t to this level a n d decreased if raised f u r t h e r . "
T h e data in F i g u r e 10 direct one's a t t e n t i o n to the high
rather t h a n low relative c o n c e n t r a t i o n of potassium with particular reference to t r e a t m e n t s 1/4 N a n d 1/2 N. T h o m a s (15) observed in his studies ". . . w h e n o u t of physiological balance
luxury c o n s u m p t i o n will occur. L u x u r y c o n s u m p t i o n results in
disturbances of n o r m a l m e t a b o l i s m sufficient to affect growth
and r e p r o d u c t i o n . "
In a discussion of t h e functional aspects of m i n e r a l n u t r i t i o n
of green p l a n t s Pirson (10) states " I t has n o t yet been clarified
w h e t h e r excess of an e l e m e n t causes disturbances which represent
direct a n d specific consequences of this p a r t i c u l a r excess w i t h i n
the cell or w h e t h e r they are d e p e n d e n t u p o n the exclusion of
a n o t h e r e l e m e n t in t h e simplest case according to the p a t t e r n
of competitive i n h i b i t i o n . "
T h e a u t h o r s d o n o t k n o w w h e t h e r the relativelv high n i t r o gen found in petioles from L o w K t r e a t m e n t (Figure 8), arid
high p h o s p h o r u s in petioles from t r e a t m e n t s N H 4 . 1/4 N a n d
1/4 N (Figure 9) were t h e result or cause of vield disturbances.
Some students of p l a n t n u t r i t i o n (1,4.5.7) believe an excess of
specific n u t r i e n t s m a y be as h a r m f u l to growth processes as is a
deficiency. If this be t r u e , the use of t h e duality factor concept
may be a v a l u a b l e tool in diagnosing n u t r i t i o n a l disturbances.
T h e graphical d a t a shown in Figures 11 to 14 were o b t a i n e d
according to T h o m a s ' (13) proposals w i t h t h e following modifications: (A) p h o s p h o r u s a n d potassium were considered on the
elemental r a t h e r t h a n o x i d e basis t h r o u g h o u t a n d (B) t h e q u a n tity factor was calculated on the basis of milliequivalents of each
n u t r i e n t p e r 100 grams of d r y p l a n t tissue r a t h e r t h a n percentage
of N, P 2 O 5 , a n d K 2 0 in d r y p l a n t tissue.
T h e d a t a in F i g u r e 11 do n o t clearly indicate intensitv of
n u t r i t i o n as a serious p r o b l e m in any of t h e n u t r i e n t solutions
u n d e r observation in this study. T h i s criterion of n u t r i t i o n a l
intensity d i s t u r b a n c e m a y be m o r e useful in identifying diffi-

56

J O U R N A L OF T H E A. S. S. B. T.

culties a m o n g c o m m e r c i a l l y g r o w n beets t h a n a m o n g e x p e r i m e n t a l l y g r o w n p l a n t s w i t h a l i m i t e d r a n g e in n u t r i t i o n a l intensity.


T h e q u a l i t y factors for n i t r o g e n i n leaf b l a d e s s h o w n i n
F i g u r e 12 emphasize t h a t p l a n t tissues from t r e a t m e n t s 1/4 N
a n d 1/2 N a r e i n a d e q u a t e l y s u p p l i e d w i t h n i t r o g e n m o s t of t h e
season. C o n c l u s i o n s r e a c h e d from a s t u d y of these d a t a a r e s i m i l a r
to those d r a w n from a study of t h e p e t i o l e d a t a in F i g u r e 8.
Q u a l i t y factor d a t a for p h o s p h o r u s p r e s e n t e d in F i g u r e 13
i n d i c a t e t h a t tissues from t h e two t r e a t m e n t s N H 4 a n d L o w K
m a y h a v e l u x u r y - c o n s u m e d p h o s p h o r u s . T h i s i s c o n c l u d e d from
t h e fact t h a t these t r e a t m e n t s gave low p r o d u c t i o n p e r f o r m a n c e
a n d therefore t h a t p h o s p h o r u s at levels f o u n d in tissue from
these t r e a t m e n t s was n o t r e q u i r e d for r a p i d p l a n t g r o w t h .
G r a p h i c a l data o n q u a l i t y for p o t a s s i u m a r e s h o w n i n F i g u r e
14. It is o b v i o u s t h a t t r e a t m e n t s N H 4 a n d L o w K p r o d u c e d leaf
blades low in potassium a n d t r e a t m e n t s 1/4 N a n d 1/2 N p r o d u c e d
tissue high in potassium r e l a t i v e to t h a t f o u n d in leaf b l a d e s
from Check-N t r e a t m e n t . It m a y be of passing i n t e r e s t to n o t e
t h a t h i g h seasonal potassium values for t r e a t m e n t s 1/4 N a n d
1/2 N w e r e clearly segregated in p e t i o l e tissue ( F i g u r e 10) w h i l e
low potassium values for N H 4 a n d L o w K t r e a t m e n t s w e r e distinctly separated in leaf blades ( F i g u r e 14). T h e r e a r e s o m e
discrepancies b e t w e e n t h e c o n c l u s i o n a n d i n t e r p r e t a t i o n s o n e
w o u l d m a k e from t h e chemical c o m p o s i t i o n s of p e t i o l e a n d
b l a d e tissues. In most cases these differences in i n t e r p r e t a t i o n
a p p e a r to be in t h e r e a l m of d e g r e e of deficiency or excess r a t h e r
than adequate or inadequate balance. However, the graphical
d a t a s h o w i n e q u a l i t y factor for p h o s p h o r u s i n d i c a t e t h a t it is t o o
h i g h in leaf blades ( F i g u r e 13) a n d a b o u t r i g h t in leaf petioles
('Figure 9). T h e a u t h o r s h a v e n o satisfactory e x p l a n a t i o n for
this p a r t i c u l a r discrepancy.
W h i l e T h o m a s (13) d i d n o t p r o p o s e t h e use o f n i t r o gen : potassium ratios as an essential f e a t u r e of his foliar diagnosis, t h e a u t h o r s have i n c l u d e d F i g u r e 15 as a s u p p l e m e n t to
t h e T h o m a s m e t h o d . T h e g r a p h i c a l d a t a i n F i g u r e 1 5 a r e calc u l a t e d from d a t a already available in t h e d e v e l o p m e n t of this
m e t h o d . A r b i t a r a r y b o u n d a r y lines a r e u s e d i n F i g u r e 1 5 t o
i n c l u d e t h e eight seasonal values for n i t r o g e n : p o t a s s i u m r a t i o
of leaf b l a d e s from t r e a t m e n t C h e c k - N . T h i s g r a p h i c a l analysis
indicates t h a t leaf b l a d e tissue from t r e a t m e n t s N H 4 a n d L o w
K is t o o h i g h a n d from t r e a t m e n t s 1/2 N a n d 1/4 N t o o l o w in
N : K ratio. It throws some d o u b t on treatments 1/2 K. T h u s it
m a y be possible to identify a w i d e r a n g e of v a r i a t i o n in t h e

VOL.

13, N o .

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57

n u t r i t i o n a l d i s t u r b a n c e of sugar beets in o n e graph, w h e n it is


certain that only t h e t w o n u t r i e n t s , n i t r o g e n a n d potassium, are
involved.
Conclusions
T h e critical level p r o c e d u r e as proposed by U l r i c h (17) for
diagnosing n u t r i t i o n a l disturbances in the sugar beet has been
d e m o n s t r a t e d to be a useful tool in c o m m e r c i a l fields. In its
present form a n d for n u t r i e n t c u l t u r e studies this t e c h n i q u e
appears to lack sensitivity. P o t c u l t u r e studies revealed that concentrations of p l a n t n u t r i e n t s in p l a n t tissue m a y be considerably above proposed critical levels a n d yet n o t high e n o u g h to
support m a x i m u m yield of roots a n d sugar.
A modification of the T h o m a s (13) m e t h o d appears to be
well suited to t h e diagnosis of n u t r i t i o n a l disturbances which
adversely influence yield of sugar beet roots a n d sugar. N o t only
is the q u a n t i t y - q u a l i t y t e c h n i q u e sensitive as a m e a s u r e of m i n i m u m c o n c e n t r a t i o n s which l i m i t p l a n t performance, b u t i t
appears to be suitable as a device for t h e recognition of l u x u r y
c o n s u m p t i o n of p l a n t n u t r i e n t s .
It appears from this l i m i t e d study t h a t t h e essential n u t r i e n t
concentrations in p l a n t tissue responsible for o p t i m u m p r o d u c t i o n
of p l a n t m a t e r i a l may be relatively n a r r o w .
A d d i t i o n a l study is n e e d e d to d e t e r m i n e m o r e precisely the
influence of season a n d a wide r a n g e in n u t r i e n t availability on
plant p e r f o r m a n c e a n d composition.
Acknowledgements
T h e a u t h o r s are i n d e b t e d t o California Chemical C o m p a n y
for financial assistance in c o n d u c t i n g this study.
L i t e r a t u r e Cited
(1)

(2)
(3)

(4)
(5)

ALEXANDER, J.

X.,

C.

C.

SCHMER,

L.

P.

ORLEANS,

and R.

H.

COTTON.

1954. T h e effect of fertilizer application on leaf analysis and yield


of sugar beets. Proc. Am. Soc. Sugar Technol. 8: 370-385.
BARTON, CHARLES J. 1948. Photometric analysis of phosphate rock.
Analyt. Chem. 20: 1068-1073.
GOODALL, D. W., and F. G. GREGORY. 1947. Chemical composition of
plants as an index of their nutritional status. Imperial Bureau of
Horticultural a n d Plantation crops. Tech-Communication No. 17.
GREGORY, F. G. 1937. Mineral nutrition of plants. Ann. Rev. Biochem. 6: 557-578.
HADDOCK, J A Y L. 1962. T h e influence of irrigation regime on yield
and quality of potato tubers and nutritional status of plants. Am.
Potato J. 38: 423-434.

J O U R N A L OF T H E A. S. S. B. T.
(6)

(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)
(15)

(16)

HILLEBRAND,

W.

F.,

H.

A.

BRIGHT,

J.

K.

HOFFMAN,

and

G.

E.

F.

L U N D E L L . 1944. A p p l i e d Inorganic Analysis. J o h n Wiley a n d Sons.


N e w York. Kjeldahl m e t h o d p p . 636-641.
LUNDEGARDH, H. 1951. Leaf Analysis. Hilger a n d Watts, L t d . L o n d o n ,
England.
M A C Y , P A U L . 1936. T h e quantitative mineral n u t r i e n t requirements
of plants. Plant Phys. 11: 741-764.
PERRIN, C. H. 1953. R a p i d modified procedure for d e t e r m i n i n g
Kjeldahl nitrogen. Analyt. Chem. 25: 968-971.
PIRSON, A. 1955. Functional aspects in mineral n u t r i t i o n of green
plants. A n n . Review of P l a n t Phys. 6: 71-114.
SHEAR, C. B. and H. L. CRANE. 1946. Nutrient-element balance: A
fundamental concept in p l a n t n u t r i t i o n . Am. Soc. H o r t . Sci. Proc.
5 1 : 319-326.
SHEAR, C. B. and H. L. CRANE. 1943-47. Nutrient-element balance, p p .
592-601. Yearbook of Agriculture.
T H O M A S , W A L T E R . 1947. Principles and application of the m e t h o d
of foliar diagnosis. I n t e r n a t i o n a l Congress of P u r e a n d A p p l i e d
Chemistry. 3: 233-334.
T H O M A S , W A L T E R . 1945. Present status of diagnosis of mineral requirements of plants by means of leaf analysis. Soil Sci. 59: 353-374.
T H O M A S , W A L T E R . 1932. T h e reciprocal effects of nitrogen, phosphorus
and potassium as related to the absorption of these elements by
plants. Soil Sci. 33: 1-20.
T H O M A S , W A L T E R , W . B. M A C K a n d R. H . C O T T O N .

1942.

Foliar diag-

nosis in relation to irrigation. Am. Soc. H o r t . Sci. Proc. 40: 531-535.


(17) U R I C H , ALBERT. 1948. Plant analysis methods and interpretations of
results. Chapter IV. Diagnostic T e c h n i q u e s for Soils a n d Crops.
American Potash Institute, Washington, D. C.
(18)

U L R I C H , A., D.

RIRIE,

F. J.

HILLS,

A.

G.

GEORGE,

and

M.

D.

1959. P l a n t Analysis: A Guide to Sugar Beet Fertilization.


Calif. Agr. E x p t . Sta. Bull. 766 (1) : 1-25.

MORSE.

Univ.

A Rapid Method for the Routine Factory Analysis


of Lactic Acid
R. F. OLSON1
Received for publication June 14, 1963

Lactic acid d e t e r m i n a t i o n s in sugar beet processing l i q u o r s


have b e e n utilized to m e a s u r e t h e e x t e n t of sucrose d e g r a d a t i o n
(2, 4, 5) 2 . T h e majority of investigators use t h e colorimetric
method, described b y B a r k e r a n d S u m m e r s o n (1). This m e t h o d
is accurate a n d relatively simple, however it is l i m i t e d in its
ability to h a n d l e large n u m b e r s of samples. Stark, G o o d b a n a n d
Owens (3) p o i n t e d o u t t h a t p a p e r c h r o m a t o g r a p h y is a rapid,
relatively accurate m e t h o d by w h i c h organic acids can be i d e n t i fied. The p u r p o s e of this p a p e r is to present a n e w p a p e r c h r o m atographic m e t h o d for d e t e r m i n i n g lactic acid. This m e t h o d is
being used at several factories w i t h confidence t h a t an accurate
picture of lactic acid c o n d i t i o n s is b e i n g recorded.
Materials and Methods
A c h r o m a t o g r a m is o b t a i n e d in t h e following m a n n e r :
a. W h a t m a n #4 paper181/4" X 221/2"
b . descending i r r i g a t i o n
c. r o o m t e m p e r a t u r e d e v e l o p m e n t
d. calcium lactate p e n t a h y d r a t e as s t a n d a r d
e. solvent
A parallel line is d r a w n 14 cm from o n e e n d of paper, each
spot b e i n g separated by 3 cm on this line. The standards are
located in t h e six m i d d l e positions a n d the u n k n o w n s are spotted
on either side. The v o l u m e of sample spotted is c o n t i n g e n t u p o n
the lactic acid c o n c e n t r a t i o n . For h i g h concentrations of lactic
acid, 15 l a m b d a are spotted a n d for low c o n c e n t r a t i o n s of lactic
acid i n c r e m e n t s of 15 l a m b d a are spotted.
Extensive w o r k was d o n e in selecting a suitable solvent. The
solvent finally selected has the following m a k e - u p :
1. Stock S o l u t i o n
70 p a r t s isopropyl alcohol
25 parts b e n z e n e
10 parts n-butyl alcohol
2. To 800 ml of stock solution a d d
200 ml distilled w a t e r
50 ml glacial acetic acid
.3 g bromcresol g r e e n
.1 g b r o m p h e n o l b l u e
1
2

Research Chemist, American Crystal Sugar Company, Rocky Ford, Colorado.


Numbers in parentheses refer to literature cited.

60

J O U R N A L OF T H E A. S. S. B. T.

T h i s yellow-orange, non-corrosive solvent has a p l e a s a n t o d o r


a n d i s easily p r e p a r e d . I t does n o t leave i n t e r f e r i n g r e s i d u e s u p o n
e v a p o r a t i o n . L a c t i c acid i s r e a d i l y s e p a r a t e d from o t h e r c o n s t i t u e n t s i n b e e t sugar processing l i q u o r s .
After spotting, t h e p a p e r s a r e placed i n t h e solvent t r o u g h s
a n d allowed to r e m a i n for 1 to 1 1 / 2 h o u r s ; they a r e t h e n r e m o v e d
from t h e c a b i n e t s a n d allowed t o dry. T h e solvent r e m o v a l i s
facilitated w h e n t h e c h r o m a t o g r a m is exposed to w a t e r v a p o r .
T h i s can b e a c c o m p l i s h e d b y h a n g i n g t h e c h r o m a t o g r a m b e t w e e n
sheets of wet canvas in t h e d r y i n g oven or by c o n s t r u c t i n g a canvas l i n e d frame in w h i c h t h e c h r o m a t o g r a m is placed. T h e
c h r o m a t o g r a m develops q u i t e r e a d i l y if t h e canvas frame is
placed in a sink c o n t a i n i n g a small a m o u n t of h o t w a t e r . It is
n o t e d t h a t spraying o r d i p p i n g o f t h e c h r o m a t o g r a m has b e e n
e l i m i n a t e d by t h e a d d i t i o n of t h e acid-base dyes to t h e solvent.
The c h r o m a t o g r a m will have a b l u i s h - g r e e n b a c k g r o u n d w i t h
lactic acid as a yellow spot. T h e sensitivity of t h e m e t h o d is 1
g a m m a of lactic acid p e r 15 l a m b d a of v o l u m e .
Results and Discussion
A c o m p a r a t i v e analysis was m a d e b e t w e e n t h e c o l o r i m e t r i c
a n d c h r o m a t o g r a p h i c m e t h o d s for t h e d e t e r m i n a t i o n of lactic
acid. T h e samples w e r e o b t a i n e d from T h e G r e a t W e s t e r n Sugar
C o m p a n y ' s L o v e l a n d factory. T h e results w e r e :
Sample
Cossette Prses Juice
Diffusion Juice
Cell 3
Cell 5
Cell 7
Cell 9
Cell 11
Cell 13
Cell 15
Cell 17
P u l p Press W a t e r

Lactic Acid, p p m
G.W. Colorimetric A.C.S. C h r o m a t o g r a p h i c
27
33
64
58
72
66
85
83
75
100
89
108
93
95
77
116
79
71
88
71
63
47

T h e t i m e r e q u i r e d for t h e d e v e l o p m e n t o f c h r o m a t o g r a m s
enables the worker to make evaluations t h r o u g h o u t the entire
day a n d t h e d a t a will reflect a large n u m b e r of samples. T h e
simplicity o f t h e p a p e r c h r o m a t o g r a p h i c m e t h o d h a s l e d t o t h e
i n t r o d u c t i o n o f t h e a p p a r a t u s i n t o r o u t i n e factory analysis. T e c h n i c i a n s a r e easily t r a i n e d t o spot a n d e v a l u a t e t h e c h r o m a t o g r a m s .
Certain precautions have to be taken in order to provide
adequate chromatograms. T h e room in which the chromatog r a m s a r e s p o t t e d a n d d e v e l o p e d s h o u l d be free of e x t r a n e o u s

VOL.

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61

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odors, d u s t a n d traffic. A m m o n i a fumes a n d lime d u s t a r e extremely deleterious to t h e d e v e l o p m e n t of a c h r o m a t o g r a m . An


e x p e n d i t u r e of $100 or less will p r o v i d e t h e c o m p l e t e a p p a r a t u s
necessary for the c h r o m a t o g r a p h y of lactic acid.
Summary
A c h r o m a t o g r a p h i c m e t h o d has b e e n described for t h e det e r m i n a t i o n of lactic acid in sugar beet processing liquors. T h e
m e t h o d is r a p i d a n d a d a p t a b l e to r o u t i n e factory analysis.
L i t e r a t u r e Cited
(1) BARKER, S. B. and W. H. SUMMERSON. 1941. T h e colorimetric determination of lactic acid in biological material. J. Biol. Chem. 138: 535554.
(2)

N O R M A N , LLOYD W . a n d G U Y O. RORABAUGH.

acid in the Silver Diffuser.


V I I I : 242-247.
(3)

STARK, J. B., A. E. GOODBAN, a n d H . S. O W E N S .

graphy of organic acids. Anl. Chem.


(4)

1954.

Studies of lactic

Proc. Am. Soc. Sugar Beet Technol.


1951.

Paper chromato-

2 3 ( 3 ) : 413-415.

STARK, J . B., A. E. GOODBAN a n d H . S. O W E N S .

1952.

Composition of

certain beet diffusion juices from the 1950 campaign. Proc. Am. Soc.
Sugar Beet T e c h n o l . V I I : 688-691.
(5)

VERHART, M. L. A. a n d N . H . M . DE VISSER.

Zucherindustries, p 446.

1959.

Zeitschrift fur die

Beet Leafhopper and Curly-Top Disease Survey


in W a s h a k i e County 1
DALE

FULLERTON2

Received for publication May J, 1963

Sugar beets i n W a s h a k i e C o u n t y o f N o r t h e r n W y o m i n g h a v e
b e e n seriously d a m a g e d by curly t o p of sugar beets each y e a r
since 1960. Yield losses h a v e b e e n h i g h to growers in an a r e a
t h a t has b e e n a n d still is e c o n o m i c a l l y d e p e n d e n t on sugar beets.
Losses of 2 tons p e r acre in 1960, 4 to 5 tons in 1961, a n d 2 to 3
tons in 1962 h a v e b e e n e s t i m a t e d by c o u n t y a g r i c u l t u r a l officials.
W a s h a k i e C o u n t y is w i t h i n t h e t e r r i t o r y of H o l l y Sugar C o r p o r a t i o n , o n e o f t h e i r factories b e i n g located a t W o r l a n d , W y o m i n g . Resistant sugar beet seed h a s b e e n u s e d i n this c o u n t y
for m a n y years a n d u n t i l 1960 a p p e a r e d to be a d e q u a t e in w i t h s t a n d i n g curly t o p disease.
T h e c u l t i v a t e d area of t h e c o u n t y is s i t u a t e d in an i r r i g a t e d
valley f o r m e d b y t h e Big H o r n R i v e r , t r a v e r s i n g t h e c o u n t y s o u t h
to n o r t h . C u l t i v a t e d lands on b o t h t h e west a n d east sides of
t h e r i v e r m e r g e w i t h a r i d r a n g e l a n d o n t h e west, east, a n d s o u t h
sides of t h e c o u n t y . T h e a r i d r a n g e l a n d is c h a r a c t e r i z e d by sparse
vegetation.
C u r l y t o p is a virus disease t h a t d a m a g e s sugar beets. T h e
only k n o w n m e t h o d of transmission is by t h e b e e t l e a f h o p p e r ,
Circulifer tenellus (Baker)
(l)3.
In o t h e r s u g a r - b e e t - g r o w i n g
regions t h e beet l e a f h o p p e r o v e r w i n t e r s in desert areas a n d migrates i n t o c u l t i v a t e d valleys in t h e s p r i n g (2). H o w e v e r , its
h a b i t s a n d b e h a v i o r a r e i n c o m p l e t e l y k n o w n , a n d this lack o f
i n f o r m a t i o n has c o n t r i b u t e d t o i n a d e q u a t e c o n t r o l a n d p r e v e n t i v e
measures. I n f o r m a t i o n a s t o w h e r e a n d h o w t h e b e e t l e a f h o p p e r
o v e r w i n t e r s a n d on t h e t i m e of m i g r a t i o n is necessary to i n i t i a t e
controls. L i m i t s of t h e insect's host r a n g e w o u l d also be of v a l u e .
T h i s survey was c o n d u c t e d t o d e t e r m i n e t h e e x t e n t o f c u r l y
t o p infection a n d t o answer some o f t h e q u e s t i o n s p e r t a i n i n g t o
t h e h a b i t s of t h e b e e t leafhopper. It was h o p e d t h a t t h e survey
w o u l d p r o v i d e a c l u e or s t a r t i n g p o i n t in t h e search for a b e t t e r
u n d e r s t a n d i n g o f t h e b e e t leaf h o p p e r - c u r l y t o p r e l a t i o n s h i p i n
Washakie County.
1
Published with approval of the Director, Wyoming Agricultural Experiment Station
as Journal
Paper No. 207.
2
3 Instructor of Entomology, University of Wyoming, Laramie, Wyoming.
Numbers in parentheses refer to literature cited.

VOL. 13, No.

63

1, APRIL 1964

Methods
Sugar beet fields in W a s h a k i e C o u n t y were surveyed for t h e
beet leafhopper on five occasionsApril 17, May 8, J u n e 16,
J u n e 21, a n d J u l y 11, 1962. T h i r t y - e i g h t fields were designated
as check fields a n d each of these fields was e x a m i n e d d u r i n g
every survey (Figure 1). Fifteen a d d i t i o n a l fields were e x a m i n e d
on each of t h e five surveys, using different fields each time. T h e
38 p e r m a n e n t fields were picked largely at r a n d o m ; however, a
few were picked on the basis of curly t o p infection in 1960 a n d
1961.
T w o m e t h o d s were used to survey each field: (a) a standard
insect n e t a n d (b) a square-foot sampler. T h e sampler is a m o d i fication of the Hills S q u a r e Foot Sampler (3). It is a cage con-

SUGAR

BEET

FIELDS

SUGAR

BEET

FIELDS

USED

Figure 1.Location of sugar beet fields used as checks in the survey


tor beet leafhoppers.

J O U R N A L OF T H E A. S. S. B. T.

64

sisting of a c i r c u l a r f r a m e w r a p p e d w i t h cheesecloth a n d is o p e n
at b o t h e n d s . W h e n a desired p l a n t is found, t h e o p e r a t o r places
t h e s a m p l e r over t h e p l a n t . T h e o p e r a t o r t h e n d i s t u r b s t h e p l a n t ,
c a u s i n g t h e leaf h o p p e r s to j u m p to t h e cheesecloth. The insects
c a n b e c o u n t e d o r p i c k e d off t h e sides w i t h a n a s p i r a t o r , w h i c h
c a n be a n y form of suction or v a c u u m p u m p . The insects a r e
t h e n r e m o v e d from t h e aspirator, c o u n t e d , a n d p r e s e r v e d .
Fifty square-foot samples w e r e t a k e n at r a n d o m locations in
each field w i t h t h e sampler, a n d 200 sweeps w e r e t a k e n in each
field w i t h t h e insect net. W e e d hosts of t h e b e e t l e a f h o p p e r t h a t
w e r e n e a r t o b e e t f i e l d s , o r b o r d e r e d t h e m , w e r e also e x a m i n e d ,
u s i n g b o t h m e t h o d s of survey.
X h e curly t o p survey was c o n d u c t e d i n W a s h a k i e C o u n t y
d u r i n g t h e week of A u g u s t 6 to 10, 1962, a n d was d e s i g n e d to
i n c l u d e t h e e n t i r e sugar b e e t acreage in t h e c o u n t y . A total of
187 fields was e x a m i n e d , a few fields m a y h a v e b e e n missed because of t h e i r h i d d e n or u n s e e n location.
A total of 1,000 plants was visually i n s p e c t e d by e x a m i n i n g
100 plants at each of 10 locations selected at r a n d o m in each
field. The infected p l a n t s from each of t h e l o c a t i o n s w e r e t h e n
c o u n t e d t o give t h e total n u m b e r o f infected p l a n t s , t h u s d e t e r m i n i n g t h e p e r c e n t a g e of infected plants. E a c h field was g i v e n
a degree of infection r a t i n g based on a scale of five r a t i n g s as
follows:
Percent infection
0
1-10
11-20
21-50
51-100

Degree of infection
none
light
moderate
severe
very severe

Plants showing questionable curly t o p symptoms were n o t


c o u n t e d . S y m p t o m s w e r e d e t e r m i n e d e n t i r e l y b y visual observations i n t h e f i e l d . A m a p was c o n s t r u c t e d t o show t h e l o c a t i o n
of t h e sugar b e e t field a n d t h e d e g r e e of infection in each field.
N o a t t e m p t was m a d e t o show t h e location o f host p l a n t s o t h e r
t h a n sugar beets. I t i s p r o b a b l e t h a t s o m e infection o c c u r r e d
after t h e survey was c o m p l e t e d , h o w e v e r , it is u n l i k e l y t h a t these
new infections w o u l d a p p r e c i a b l y c h a n g e t h e g e n e r a l conclusions.
Results a n d Discussion
The first b e e t leaf h o p p e r s w e r e collected d u r i n g a survey on
J u l y 11. X h e f i r s t s y m p t o m s o f c u r l y t o p , f o u n d d u r i n g t h e survey
c o n d u c t e d on J u n e 2 1 , o c c u r r e d 21 days before discovery of t h e
f i r s t b e e t leafhoppers. N e i t h e r t h e b e e t leafhoppers n o r s y m p t o m s
of t h e disease w e r e n o t e d in surveys m a d e A p r i l 17, M a y 8, or
J u n e 16. O n J u l y 11, w h e n t h e b e e t l e a f h o p p e r was f i r s t dis-

VOL. 13, N o .

1, APRIL 1964

65

covered, an average of t h r e e i n d i v i d u a l s p e r 200 sweeps were


taken. M o r e beet leafhoppers were f o u n d at t h e e n d of t h e fields
nearest to t h e desert area t h a n at t h e opposite end. Symptoms of
curly t o p o c c u r r e d first in t h e e n d of the fields b o r d e r i n g the
desert area i n d i c a t i n g that m i g r a t i o n from t h e desert area to
cultivated areas h a d just b e g u n .
T h e first symptoms of the disease a n d t h e first beet leafhoppers were n o t e d in fields b o r d e r i n g t h e desert area on the east
side of the cultivated area. C u r l y t o p infection was n o t f o u n d on
the west side of t h e Big H o r n R i v e r u n t i l 14 days after t h e first
symptoms a p p e a r e d on t h e east side.

Figure 2.Distribution of sugar beet fields and degree of curly top


infection in Washakie County.

66

JOURNAL OF T H E A.

S.

S.

B.

T.

All of t h e sugar b e e t fields h a d some infected p l a n t s ; in no


case was a field r a t e d absolutely free. T h e p e r c e n t a g e of p l a n t s
infected in each field r a n g e d from 1.2 to 73.5 p e r c e n t . E i g h t
fields or 4.3 p e r c e n t were very severely infected, 17 fields or 9.1
p e r c e n t w e r e severely infected, 31 fields or 16.6 p e r c e n t w e r e
m o d e r a t e l y infected, a n d 131 fields or 70 p e r c e n t w e r e considered to have a light infection.
T h e m a p ( F i g u r e 2) shows the d i s t r i b u t i o n of sugar b e e t
fields a n d t h e degree of infection for each field. T h e most heavily
infected f i e l d s w e r e f o u n d o n t h e e x t r e m e b o u n d a r i e s , w h e r e
t h e cultivated areas m e e t t h e a r i d r a n g e l a n d . Most of t h e fields
in t h e c e n t e r of t h e area h a d only light infections. Fields considered to be very severely infected were f o u n d in t h e s o u t h eastern c o r n e r of the beet-growing area. T h e r e a p p e a r e d to be
a g r a d u a l decrease in t h e d e g r e e of infection from the o u t e r
b o r d e r s toward t h e c e n t e r of t h e c u l t i v a t e d area.
Fields on t h e east side of t h e r i v e r h a d m o r e infection t h a n
those on t h e west side. Five fields on t h e west side of t h e r i v e r
were considered to be m o d e r a t e l y infected, o n e was severely infected, n o n e was very severely infected, a n d t h e rest h a d only
light infections.
Conclusions
T h e fact that beet leafhoppers w e r e n o t f o u n d u n t i l 21 days
after detection of curly t o p infection indicates t h a t t h e m e t h o d
of survey a n d detection m a y n o t have b e e n as effective as desired.
It is a p p a r e n t that small p o p u l a t i o n s of b e e t leafhoppers d i d
exist in t h e fields before d e t e c t i o n of t h e disease s y m p t o m s . M o r e
intensive s a m p l i n g m a y have p i c k e d u p t h e insects m u c h s o o n e r
t h a n was r e p o r t e d . A s h o r t e r t i m e lapse in days b e t w e e n surveys
m i g h t also have detected t h e b e e t l e a f h o p p e r before t h e July
11 date.
D e t e c t i o n of s y m p t o m s a n d beet leafhoppers on t h e east side
of the r i v e r before d e t e c t i o n on t h e west side indicates t h a t t h e
beet leafhopper p o p u l a t i o n or at least t h e m a j o r i t y of t h e p o p u l a t i o n exists in t h e desert area on t h e east side of t h e river.
L a r g e n u m b e r s o f beet leafhoppers w e r e f o u n d o n t h e e n d
of t h e fields b o r d e r i n g t h e desert area, w h i c h indicates t h a t b e e t
leafhoppers m i g r a t e d from t h e desert area i n t o t h e c u l t i v a t e d
area. T h e m i g r a t i o n w o u l d also a c c o u n t for t h e most severe
infection in fields on t h e o u t e r edges of t h e c u l t i v a t e d area.
A decreasing degree of infection o c c u r r e d from t h e o u t e r m o s t fields i n w a r d , a n d fields in t h e m i d d l e of t h e valley h a d
o n l y light infection. A p p a r e n t l y t h e b e e t leafhoppers a r e e i t h e r
o v e r w i n t e r i n g i n t h e desert area o r are m i g r a t i n g from o t h e r
o v e r w i n t e r i n g g r o u n d s i n t o t h e desert area. M i g r a t i o n t o t h e
c u l t i v a t e d area occurs as suitable host p l a n t s b e c o m e available.

VOL.

13, N o .

1, A P R I L 1964

67

A larger p o p u l a t i o n of leafhoppers occurred on the eastern


side or the river t h a n on the western side. T h e most severely
infected fields were f o u n d southeast of W o r l a n d a n d may i n d i cate that a m o r e dense p o p u l a t i o n of weed hosts are on the desert
near these fields t h a n occurs along the b o r d e r s of the r e m a i n i n g
fields. Visual observations before the survey showed t h a t the
earliest signs of curly t o p were also f o u n d in this area.
Summary
A survey was c o n d u c t e d in W a s h a k i e C o u n t y on A p r i l 17,
May 8, J u n e 16, J u n e 2 1 , a n d July 11, 1962, to detect t h e earliest
possible presence of t h e beet leafhopper. T h e first beet leafhoppers were f o u n d on July 11, or 21 days after t h e first symptoms
of curly t o p a p p e a r e d .
T h e first indication of curly t o p a n d t h e presence of beet
leafhoppers occurred on the east side of t h e Big H o r n R i v e r in
fields b o r d e r i n g the desert area. A p p a r e n t l y the b u l k of the
population exists on t h e desert on the east side of the river.
A survey of sugar beet fields in W a s h a k i e C o u n t y to d e t e r m i n e
the degree of curly t o p infection was c o n d u c t e d on A u g u s t 6-10,
1962. A total of 187 sugar beet fields were surveyed, a n d 1,000
plants in each field were e x a m i n e d visually.
T h e most severely infected fields were found along the o u t e r
edges of the cultivated areas adjacent to the arid rangeland. T h e
eastern edge was m o r e severely infected t h a n was the western
edge. An area southeast of W o r l a n d h a d t h e most severely infected fields.
Seventy p e r c e n t of the fields h a d light infections, 16.6 percent
were moderately infected, 9.1 p e r c e n t were severely infected, a n d
4.3 percent were very severely infected.
L i t e r a t u r e Cited
(1) DOUGLASS, J. R. and W. C. COOK. 1954. T h e Beet Leafhopper. USDA
Circ. 942: 1-21.
(2) Fox, D. E. 1938. Occurrence of the Beet Leafhopper and Associated
Insects on Secondary Plant Successions in Southern Idaho. USDA
Tech. Bui. 607: 1-14.
(3) H I L L S , O. A. 1933. A new method for collecting samples of insect p o p ulations. J. Econ. Ent. 26: 906-910.

Redistribution of N i t r a t e in Soils and Its Effects


on Sugar Beet Nutrition 1
MYRON

STOUT2

Received for publication October 31, 1963

Introduction
N i t r o g e n n u t r i t i o n has b e e n s h o w n t o b e a n i m p o r t a n t factor
in sugar beet yield a n d q u a l i t y (2,3,5,6,7,8) 3 . If n i t r o g t n is t o o
low, u n p r o f i t a b l e yields result, b u t if it is t o o h i g h at harvest,
beets are low i n sucrose a n d p u r i t y . T h u s t i m i n g o f n i t r o g e n
u p t a k e , as well as t h e total a m o u n t , is essential in successful
sugar beet p r o d u c t i o n .
N i t r a t e n u t r i t i o n is o n e of t h e m o r e difficult p r o d u c t i o n
variables to c o n t r o l because it is influenced by so m a n y e n v i r o n m e n t a l conditions. T h e i n c o r p o r a t i o n i n t o soil o f p r e v i o u s c r o p
residues, t i m e of a p p l i c a t i o n of o r g a n i c or i n o r g a n i c forms of
nitrogen, temperature, and the method and a m o u n t of water
application affect t h e availability of n i t r a t e to g r o w i n g c r o p
plants.
Table 1.Precipitation and Evaporation, Salt Lake Valley, 1959-1960 and Cache
Valley, 1961.
Evaporation (inches)
Precipitation (inches)
19613
19613
I960 4
19594
Month
19591
I9602
April
May

June
July
August
September
October
Total
1
2
3
4

1.61
2.05
1.38
0.19
1.76
1.66
0.22
8.87

0.40
1.09
0.30
0.10
0.66
0.70
1.23
4.48

0.69
0.66
0.59
0.53
0.72
1.89
1.64
6.72

7.34
8.71
12.95
14.12
12.94
8.09
4.94
69.09

7.42
10.31
13.65
16.16
13.11
9.76
5.37
75.81

3.96
5.65
6.59
7.53
6.65
3.87
no data
34.25

Salt Lake Airport W. B. (Salt Lake Valley)


Midvale Station (Salt Lake Valley)
Greenville Farm, U.S.U. (Cache Valley)
Morton Salt Company (Salt Lake Valley)

T h e w i d e discrepancy b e t w e e n a m o u n t s o f rainfall a n d e v a p o r a t i o n ( T a b l e 1) causes t h e r e d i s t r i b u t i o n of soluble salts in


soils. T h i s results i n t h e b u i l d - u p o f h i g h surface c o n c e n t r a t i o n s
a n d is t h e p r i m a r y cause of salinity p r o b l e m s in a r i d climates.
E x t e n s i v e research has r e s u l t e d i n s o m e i n g e n i o u s practical
1
The field studies involving deep vs. shallow furrowing were done in cooperation with
members
of the Research Staff of the Utah-Idaho Sugar Co.
2
Physiologist, Crops Research Division, Agricultural Research Service, United States
Department
of Agriculture.
3
Numbers in parentheses refer to literature cited.

VOL.

13, N o .

1, A P R I L

1964

69

measures for m a n i p u l a t i n g t h e localization of high salt concentrations (1). Soluble p l a n t n u t r i e n t s , especially n i t r a t e , follow the
same general p a t t e r n s of m o v e m e n t a n d c o n c e n t r a t i o n in soils.
T h e m o v e m e n t a n d localized p a t t e r n s of r e d i s t r i b u t i o n of n i t r a t e
strongly affect n u t r i t i o n of t h e sugar beet c r o p .
T h e p u r p o s e of this r e p o r t is to provide a d d i t i o n a l information on the e x t e n t of n i t r a t e m o v e m e n t in soils a n d to suggest
some c u l t u r a l practices that may alleviate some u n d e s i r a b l e effects
from excess n i t r a t e supply at t h e w r o n g t i m e .
Chemical and Soil-Sampling Methods
Chemical. T h e colorimetric p h e n o l d i s u l p h o n i c acid m e t h o d
described by M. L. Jackson (1958 edition) was used for soil
nitrate analyses. E i g h t grams of screened (20 Mesh), well-mixed
soil were leached for 20 m i n u t e s by end-over-end r o t a t i o n in
50-ml test tubes w i t h 40 ml of d i l u t e C u S O 4 solution a n d a small
a m o u n t of C a ( O H ) 2 . T h e soil suspension was t h e n filtered
through close-textured filter p a p e r a n d an a l i q u o t of filtrate used
for n i t r a t e estimation.
Soil sampling and preparation.
E x t r e m e gradients in concentration of n i t r a t e usually e n c o u n t e r e d in cultivated, a r i d soils
make s a m p l i n g a n d sample h a n d l i n g u n u s u a l l y subject to errors
(4,5). To r e d u c e errors, it was f o u n d c o n v e n i e n t to lightly press
irregular soil surfaces a n d t h e n to use a flat-bottom scoop with
sides 1/4 inch h i g h to take the 0 to 1/2-inch surface samples. T h e
soil t u b e was t h e n inserted to the 6-inch d e p t h a n d any dry surface soil scraped away w i t h t h e scoop. T h e soil t u b e was t h e n
rotated a n d pressed against t h e t o p sides of t h e hole to insure
a r a t h e r conical-shaped, firm t o p before w i t h d r a w i n g the t u b e .
If dry soil fell i n t o t h e hole, a n o t h e r 1/2-inch of core was removed a n d discarded before t a k i n g the n e x t sample.
T h e soil was m i x e d in pans (a separate p a n for each d e p t h
increment) a n d a sample of a b o u t 60 to 100 grams was sealed
in small, w i d e - m o u t h bottles w i t h 2 to 3 ml of toluene. Soil
samples were d r i e d in m e t a l dishes at 65 C overnight, t h e n
passed b e t w e e n steel surfaced rolls a n d lightly r u b b e d t h r o u g h
a 20-mesh screen by m e a n s of a large r u b b e r stopper. T h e
screened soil was t h e n rolled t h o r o u g h l y a n d stored in p a p e r
bags for analysis. T h e r a t h e r e x t r e m e p r e c a u t i o n s in sampling
and sample p r e p a r a t i o n were taken because m o r e t h a n t h o u s a n d fold differences in n i t r a t e c o n c e n t r a t i o n s may be e n c o u n t e r e d
between surface 1/2-inch a n d lower layers of soil. F o r this reason,
similar d e p t h - i n c r e m e n t s of different soil samples were usually
grouped a n d processed. T h e e q u i p m e n t was t h o r o u g h l y cleaned
before p r e p a r a t i o n of a n o t h e r d e p t h - i n c r e m e n t g r o u p of samples.
Depth of samples was u n i f o r m in all tests. T h e r e f o r e , to simplify
presentation a d e p t h code is used in p r e s e n t i n g data. D e p t h in-

JOURNAL OF THE A.

70

S. S.

B.

T.

c r e m e n t s w e r e as follows: A, 0" to 1/2"; B, 1/2" to 6"; C, 6" to 12";


D , 12" t o 2 4 " a n d E, 2 4 " to 36".
Experimental Results
Holden
Plot
Studies, 1959-1960
B o t h vertical a n d lateral m o v e m e n t o f n i t r a t e w e r e s t u d i e d
o n 40-inch d o u b l e - r o w beds u n d e r f r e q u e n t i r r i g a t i o n s d u r i n g
late s u m m e r in 1959 a n d 1960. Studies w e r e m a d e on a fine
sandy l o a m p r e p a r e d a n d p l a n t e d t o sugar b e e t seed d u r i n g
A u g u s t . I r r i g a t i o n furrows were 40 inches a p a r t a n d a b o u t 2 to
3 inches d e e p . T h e p l o t was usually i r r i g a t e d at intervals of
a b o u t five days w i t h a small stream of water. T h e r e was no
f l o o d i n g o f b e d s w h e r e n i t r a t e studies w e r e m a d e . After p r e l i m i n a r y observations showed a m a r k e d lateral m o v e m e n t of nit r a t e in t h e surface, samples w e r e t a k e n to a d e p t h of 36 inches
across t h e beds at f r e q u e n t spacings as s h o w n in tables 2 a n d 3.
P r e l i m i n a r y samples i n d i c a t e d m o r e t h a n a fourfold increase
in n i t r a t e c o n c e n t r a t i o n in t h e c e n t e r of t h e b e d w h e r e m o i s t
surface zones barely coalesced in c o n t r a s t to a s i m i l a r l o c a t i o n
w h e r e m o i s t u r e d i d n o t traverse so far. D a t a ( T a b l e 2) also
Table 2.Vertical and lateral movement of nitrate in relation to irrigation, rainfall
and evaporation. Fine sandy loam; 40-inch double beds. Holden plot, 1959.
Distance from Furrow ( inches)
Rainfall*
inches

Date
S e p t . 10
13
14
15

0,03
0.60
0.19

19
20
21
23

0.19
0.05
0.18
0.03

25
26
27
28

0.27
0.01
0.29
0.05

30

Depth
code

14

20~

Nitrate N
ppm

ppm

ppm

ppm

525

1570

1800

A
B
C
D
E

3
10
9
7
4

2
1
2
2
4

8
88
25
11
7

6
57
29
12
8

A
B
C
D
E

22
22
11
5
4

188
19
1
2
4

460
66
15
10
6

1900
118
18
10
4

A
B
C
D
E

2
3
3
2
2

5
5
3
4
4

550
186
25
7
7

170
69
16
7
5

Rainfall 7200 South 3rd East 1.4 miles from plots


Irrigations September 1, 5, 9, and 14

VOL. 13, N o . 1, A P R I L 1964

71

indicate a p r o n o u n c e d lateral m o v e m e n t of n i t r a t e toward the


centers of beds below t h e surface. Rainfall a n d e v a p o r a t i o n p r o duced strong effects on vertical m o v e m e n t of n i t r a t e . Surface
concentrations of n i t r a t e in furrows were p r o b a b l y m o r e dep e n d e n t on l e n g t h of t i m e of s a m p l i n g after irrigation t h a n on
other variables.
Table 3.Vertical and lateral movement of nitrate in relation to irrigation, rainfall and
evaporation. Fine sandy loan; 40-inch double beds. Holden plot, 1960.
Distance from Furrow (inches)
Rainfall 1
inches

Date

Aug. 12

Aug. 19

Aug. 22
A u g . 23

Depth
code

10

20
Nitrate N

10

ppm

ppm

ppm

ppm

ppm

A
B
C
D
E

1
1
4
11
16

370
9
6
10
11

230
12
10
13
11

210
6
4
18
17

1
1
3
15
13

A
B
C
D
E

3
1
3
11
15

340
7
5
13
13

700
8
18
18

510
2
3
18
21

3
1
2
11
10

A
B
C
D
E

1
2
2
11
4

43
38
14
18
19

108
90
15
26
20

50
12
4
16
16

1
1
1
7
19

0.66

A u g . 28 F e r t i l i z e d i n b o t t o m o f f u r r o w s w i t h a m m o n i u m n i t r a t e ;; t h e n irr i g a t e d
Aug. 30

Sept. 1
3
6
12

0.13
0.39
0.01
0.04

A
B
C
D
E

5
32
27
19
13

368
7
10
17
14

464
37
11
20
16

504
4
5
18
15

3
11
5
6
10

A
B
C
D
E

21
12
32
20
16

350
9
19
24
12

1210
28
22
18
17

470
2
4
14
17

6
4
10
12
14

Rainfall, Midvale, Utah

Data ( T a b l e 3) indicate essentially the same p a t t e r n s of


m o v e m e n t in 1960 as those in table 2 except t h a t surface n i t r a t e
concentrations were h i g h e r in 1959. Field laborers a p p l i e d amm o n i u m n i t r a t e to t h e p l o t on A u g u s t 28. It is evident t h a t
the u n a u t h o r i z e d a d d i t i o n of n i t r a t e to o n e furrow was twice

72

JOURNAL OF THE A. S. S. B. T.

as m u c h as in t h e o t h e r . T h e rainfall of 0.66 i n c h , A u g u s t 22
o c c u r r e d d u r i n g a very short p e r i o d . T h i s heavy s h o w e r d i d
n o t leach n i t r a t e from t h e surface n e a r l y so c o m p l e t e l y as t h e
less r a p i d rainfall of S e p t e m b e r 13-15, 1959 ( T a b l e 2) or t h a t
of O c t o b e r 8-12, 1960 ( T a b l e 6). Studies r e p o r t e d in T a b l e 3
were t e r m i n a t e d by t h e accidental c u l t i v a t i o n of surface soil
after samples were t a k e n S e p t e m b e r 12.
Effect

of deep

vs.

shallow furrows on
yield and quality

nitrate

redistribution;,

D e e p vs. shallow-cultivated strips w e r e c o m p a r e d on two fields


in 1959. U n t i l a b o u t J u l y 1, t h e fields w e r e u n i f o r m l y fertilized,
c u l t i v a t e d a n d irrigated. D e e p - f u r r o w e d strips w e r e m a d e a b o u t
J u l y 1 at t h e t i m e of final c u l t i v a t i o n . B o t h fields w e r e i r r i g a t e d
in t h e same a l t e r n a t e furrows all s u m m e r . T h e soil was h a r d
a n d relatively dry after A u g u s t 11. N o f l o o d i n g o c c u r r e d n e a r
a n y s a m p l e d areas b u t t h e r e was little difference in d e p t h of
irrigation furrows at harvest d u e to w a s h i n g a n d silting of soil.
Soil a n d beet s a m p l i n g sites were chosen a b o u t 1/4 t h e distance
from the t o p a n d 1/4 the distance from t h e b o t t o m of each field.
Because of usual differences in yield a n d q u a l i t y b e t w e e n t o p
a n d b o t t o m of i r r i g a t e d fields, c o m p a r i s o n s a r e possible only between adjacent positions.
Soil below t h e surface was consistently m o r e moist a n d
s a m p l i n g was easier in deeply-furrowed strips w h e r e w a t e r p e n e t r a t i o n was better. T h e s e differences a r e b e l i e v e d to be significant at least with respect to differences in yield at harvest.
Presence of an average of n e a r l y 400 p p m of n i t r a t e n i t r o g e n
in relatively d r y surface soil after a series of r a i n s t h a t o c c u r r e d
d u r i n g t h e s a m p l i n g p e r i o d seems very significant ( T a b l e 4).
Foliar p r o t e c t i o n of this excess n i t r a t e in t h e surface d u r i n g
precipitations t o t a l i n g 3.25 inches may be an i m p o r t a n t factor
in r e d u c i n g hazards of late rainfall in some relatively d r y climates
p r o v i d e d t h a t soils a r e n o t flooded by p r e c i p i t a t i o n .
Soil samples w e r e t a k e n in r i d g e d soil b e t w e e n furrows a n d
b e t w e e n a p p r o x i m a t e l y n o r m a l l y spaced beets. D a t a ( T a b l e 5)
i n d i c a t e t h a t t h e r e was a fairly consistent difference in b o t h
yield a n d q u a l i t y o f beets b e t w e e n d e e p a n d shallow-furrowed
strips of each field. T h e r e w e r e also consistent differences in
yield, sugar p e r c e n t a g e , a n d p u r i t y in favor of deeply-furrowed
strips. Differences i n a m i n o n i t r o g e n , s o d i u m , a n d p o t a s s i u m
w e r e n o t sufficiently consistent to justify a n y definite conclusions.
T w o tests o n t h e effect o f d e e p vs. n o r m a l c u l t i v a t i o n w e r e
r u n in C a c h e Valley in 1961. Surface c o n c e n t r a t i o n s of n i t r a t e
w e r e n o t so g r e a t as t h e y w e r e in Salt L a k e Valley in 1959 because

73

VOL. 13, N o . 1, A P R I L 1964

the e v a p o r a t i o n rates a n d t e m p e r a t u r e s were lower. Surfacenitrate c o n c e n t r a t i o n was greater in deeply-furrowed strips. Average yield of sugar p e r acre, tons of beets p e r acre, sugar p e r c e n t age, a n d p u r i t y values were h i g h e r in deeply cultivated strips.
Table 4.Nitrate nitrogen content of soil on two farms, deep vs. shallow furrows.
All samples taken between beets in the beds between furrows, 1959.
LESLIE JONES FARM

MELVIN JONES FARM

*4

VOL. 13, N o . 1, A P R I L 1964


Table 6.Nitrate nitrogen
Toppenish, Washington, 1960.

75
distribution

in

furrow-irrigated

fields

of

sugar beets,

MACK HOUSTON FARM


Location

Position

Depth
code

Nitrate N at dates indicated


5/14

6/30

ppm

ppm

8/5

9/8

10/17

ppm

7/19

ppm

ppm

ppm

Beet row

A
B
C
D
E

340
81
82
17
6

840
36
133
51
39

3400
130
32
28
10

2700
312
71
30
12

4200
103
71
4
2

1720
137
38
4
4

Furrow

A
B
C
D
E

370
119
90
19
8

260
11
15
11
7

320
6
17
5
4

99
7
2
2
2

78
4
1
1
1

170
13
3
3
2

Beet row

A
B
C
D
E

260
88
68
16
7

290
34
76
46
16

1650
112
56
33
53

1900
24
107
6
7

1110
17
11
1
1

710
52
102
34
15

Furrow

A
B
C
D
E

620
107
65
17
14

160
10
10
5
4

33
5
3
4
2

81
8
3
2
25

94
14
1
1
1

32
13
2
2
1

BILL PARRISH FARM


4/29
6/22

ppm

ppm

8/5

9/8

ppm

7/19

ppm

ppm

Beet row

A
B
C
D
E

94
56
31
18
12

210
25
24
32
16

1250
25
19
10
4

1400
93
21
37
18

3650
55
6
9
6

Furrow

A
B
C
D
E

99
42
35
15
6

128
13
9
8
6

240
5
3
20
4

90
7
3
3
7

36
4
2
3
5

Beet R o w

A
B
C
D
E

55
52
28
15
5

84
69
36
32
32

1900
200
121
59
59

1700
43
88
23
28

1920
126
13
23
9

Furrow

A
B
C
D
E

188
48
26
14
12

91
20
12
14
11

57
7
4
4
6

118
10
4
4
7

290
13
S
6
2

JOURNAL OF T H E A. S. S. B. T.

76

Table 7.Nitrate nitrogen distribution in sprinklerirrigated sugar beet fields, Walla


Walla, Washington, 1960.
FRANK RIZZUTE FARM
Nitrate N at dates indicated
Location
I

Position

Depth
code

5/31

7/1

7/29

8/11

10/16

ppm

ppm

ppm

ppm

ppm
1
1
1

Beet row

A
B
C
D
E

150
54
14
4

67
22
25
14
2

72
3
1
1

35
2
2
1
1

Furrow

A
B
C
D
E

130
94
21
4
2

41
69
47
3
1

11
2
1
1
1

10
1
11
1
1

1
3
1
1
1

7/27

8/11

W. F. SCHIFFMAN FARM
5/31

N . S . no sample

7/1

10/16

VOL. 13, N o . 1, A P R I L 1964

Soil

Nitrate

Studies on
Sugar

77

Sprinkler vs.
beet Fields

Furrow-irrigated

Soil samples w e r e t a k e n d u r i n g s u m m e r a n d early fall of 1960


o n t w o f u r r o w - i r r i g a t e d a n d t w o sprinkler-irrigated sugar beet
f i e l d s . T h e M a c k H o u s t o n a n d Bill P a r r i s h farms i n T o p p e n i s h ,
W a s h i n g t o n , w e r e furrow-irrigated, w h i l e those of W. F. Schiffm a n a n d F r a n k R i z z u t i i n W a l l a W a l l a , W a s h i n g t o n , were irrigated by s p r i n k l i n g . Samples w e r e t a k e n in beet rows a n d in
the furrows.
Data ( T a b l e s 6 a n d 7) i n d i c a t e large differences in surface
nitrate c o n c e n t r a t i o n r e s u l t i n g from t h e t w o m e t h o d s of irrigation. C o n c e n t r a t i o n s of n i t r a t e in surface soils w e r e low w h e r e
beets were i r r i g a t e d by s p r i n k l i n g a n d nearly all n i t r a t e in t h e
soil was periodically l e a c h e d to active r o o t zones of plants.
U n d e r f u r r o w i r r i g a t i o n , h i g h c o n c e n t r a t i o n s of n i t r a t e were
found in d r y surface soil w h e r e beets could n o t use it. Concentrations of m o r e t h a n 4,000 p p m of n i t r a t e n i t r o g e n w e r e observed
o n the Mack H o u s t o n F a r m . A l t h o u g h surface c o n c e n t r a t i o n s
were m u c h g r e a t e r w h e r e samples w e r e t a k e n t h a n i n furrows
some idea of t h e a m o u n t s involved m a y be realized by the fact
that only a little m o r e t h a n 600 p p m of n i t r o g e n in a 1/2-inch
layer of soil is e q u i v a l e n t to 100 p o u n d s of n i t r o g e n p e r acre.
If we estimate s a m p l e d areas to r e p r e s e n t 1/3 of the surface, m o r e
than 200 p o u n d s of n i t r o g e n p e r acre m i g h t be c o n c e n t r a t e d in
this area w h e r e it is n o t available to beets unless a r a i n occurs
to leach it i n t o t h e r o o t zone.
An e x t r e m e case of e v a p o r a t i v e r e d i s t r i b u t i o n of n i t r a t e in
part of a 10-acre c o m m e r c i a l field of beets was observed in 1960.
T h e field h a d b e e n p l a n t e d to g r a i n in 1959 a n d a fairly heavy
cover of straw a n d s t u b b l e was p l o w e d u n d e r . N i t r o g e n was
applied as a n h y d r o u s a m m o n i a at t h e r a t e of 135 p o u n d s of N
per acre. P h o s p h a t e was also a p p l i e d in the fall w i t h t h e anhydrous a m m o n i a . An early p l a n t i n g of sugar beets was frozen
and the field was r e p l a n t e d A p r i l 22. A l t h o u g h t h e r e was a good
stand of sugar beets, they g r e w very slowly a n d were obviously
very deficient in n i t r o g e n as s h o w n by tests w i t h d i p h e n y l a m i n e .
T h e field was furrow-irrigated in t h e same a l t e r n a t e furrows all
season. A l t h o u g h furrows w e r e r a t h e r shallow, t h e r e was little
or no flooding. A heavy surface a p p l i c a t i o n of a m m o n i u m n i t r a t e
was m a d e to t h r e e strips, crosswise to t h e furrows, a b o u t J u l y 2.
T h e s e cross-strips w e r e a b o u t 10-feet w i d e a n d 200-feet long.
Response of beets to a m m o n i u m n i t r a t e was very striking a n d
sharply d e l i n e a t e d ( F i g u r e 1). Foliage of sugar beets in t h e fertilized cross-strips was m o r e t h a n k n e e h i g h a n d very d a r k green,

78

JOURNAL OF THE A. S. S. B. T.

Figure 1.Sugar beets on the Joseph J. Schmidt farm, West J o r d a n ,


U t a h , October 17, 1960. Beets in foreground have been harvested. The
cross strip received a heavy application of a m m o n i u m n i t r a t e in July. At
harvest there was m o r e n i t r a t e in the surface of the area where the beets
were deficient b u t it was in the dry soil a n d u n a v a i l a b l e to the plants.

w h i l e foliage of adjacent beets d i d n o t cover m o r e t h a n half


t h e 22-inch rows a n d were only a b o u t ankle-high. S y m p t o m s
i n d i c a t e d a n e x t r e m e n i t r o g e n deficiency.
On O c t o b e r 8, soil samples w e r e t a k e n to a d e p t h of t w o feet
in i r r i g a t e d furrows a n d b e t w e e n furrows in fertilized cross-strips
a n d i n t h e nitrogen-deficient sugar b e e t area. T h e soil was
m o d e r a t e l y d r y w h e n s a m p l e d b u t a light, persistent rainfall,
a m o u n t i n g to 1.23 inches, o c c u r r e d d u r i n g t h e p e r i o d of O c t o b e r
8 to 12. T h e same areas w e r e r e s a m p l e d on O c t o b e r 12. In addit i o n to soil-profile samples, four surface samples w e r e t a k e n bet w e e n furrows w h e r e nitrogen-deficient beets were located a n d
five surface samples were t a k e n in fertilized cross strips w h e r e
heavy foliage h a d p r o t e c t e d surface soil from rainfall. T h e s e
Table 8.Nitrate nitrogen in soil profile before and after a prolonged light rainfall
of 1.23 inches, J. J. Schmidt Farm, West Jordan, Utah, 1960.

VOL. 13, N o . 1, A P R I L 1964

79

latter samples w e r e n o t t a k e n m i d w a y b e t w e e n rows, b u t closer


to beet rows w h e r e foliar cover was b e t t e r .
Data ( T a b l e 8) show t h a t befort t h e rainfall t h e r e was m o r e
nitrate in surface soil w h e r e t h e deficient beets w e r e located t h a n
in heavily fertilized cross strips. T h e 1260 p p m of n i t r a t e n i t r o g e n
observed in surface soil, w h e r e t h e deficient beets w e r e located,
is e q u i v a l e n t to a b o u t 200 p o u n d s of n i t r o g e n p e r a c r e i n t h e
top l/2-inch of soil. Rainfall leached all b u t 4 p p m i n t o lower
layers of u n p r o t e c t e d soil, b u t , w h e r e t h e r e was good foliar cover,
5 separate samples h a d an average of 372 p p m t h a t r e m a i n e d in
surface soil after p r e c i p i t a t i o n . T h e s e observations, c o n c e r n i n g
protection by g o o d foliar cover in p r e v e n t i n g l e a c h i n g surface
nitrate to l o w e r levels, w e r e similar to observations in 1959
(Table 4).
Discussion
A p p l i c a t i o n of t h e p r i n c i p l e s g o v e r n i n g salt m o v e m e n t a n d
zones of c o n c e n t r a t i o n in soils to p l a n t n u t r i t i o n has received
little a t t e n t i o n . Most c o n c e r n has b e e n d i r e c t e d t o w a r d solving
adverse effects t h a t t h r e a t e n to cause a b a n d o n m e n t of soils. H o w ever, some of this basic k n o w l e d g e can be used in u n d e r s t a n d i n g
seasonal v a r i a t i o n s in n u t r i e n t a v a i l a b i l i t y a n d p r o b a b l y in
developing t e c h n i q u e s t o modify n u t r i e n t u p t a k e a n d t o m o r e
nearly a p p r o a c h t h e o p t i m u m needs of sugar beets.
T h e e x t e n t o f r e d i s t r i b u t i o n p a t t e r n s observed i n a r i d soils
u n d e r furrow-irrigation or b e d - p l a n t e d beets offers a lucid explanation of t h e e r r a t i c s u g a r c o n c e n t r a t i o n a n d q u a l i t y of sugar
beets. F r e q u e n t heavy r a i n s before harvest can depress q u a l i t y
of beets by c a u s i n g an u n f a v o r a b l e a b u n d a n c e of n i t r o g e n to
become available a t t h e w r o n g t i m e . Dry preharvest c o n d i t i o n s
may result i n s u g a r c o n c e n t r a t i o n s t h r e e o r m o r e p e r c e n t h i g h e r
than w h e n wet w e a t h e r occurs before harvest. T h e s e f l u c t u a t i o n s
in availability of n i t r a t e m a k e it difficult to estimate o p t i m u m
a m o u n t s of n i t r o g e n fertilizers to a d d to soil even t h o u g h total
a m o u n t s p r e s e n t m a y b e d e t e r m i n e d before p l a n t i n g . However,
knowledge o f e v a p o r a t i v e r e d i s t r i b u t i o n patterns, relative
amounts, a n d e x t e n t of foliar p r o t e c t i o n from r a i n s h o u l d be
useful in d e v e l o p i n g c u l t u r a l m e t h o d s to utilize these p h e n o m e n a
to i m p r o v e sugar b e e t c u l t u r e . Sprinkler-irrigated beets a n d
those in areas w h e r e f r e q u e n t l a t e - s u m m e r a n d fall rains occur
should be h a n d l e d so as to utilize nearly all n i t r o g e n before
harvest. In some of o u r a r i d climates excess n i t r o g e n may be
relatively i m m o b i l i z e d i n d r y soil u n d e r a n a d e q u a t e protective
foliar cover.
New products are being introduced to reduce mobility of
nitrogen n u t r i e n t s by e i t h e r r e d u c i n g t h e r a t e of nitrification of

80

JOURNAL OF THE A.

S.

S.

B.

T.

a m m o n i u m salts or by r e d u c i n g solubility of n i t r a t e by incorp o r a t i n g i t i n t o m o r e slowly s o l u b l e forms. T h e a u t h o r believes


that the data presented on patterns of nitrate movement in the
soil i n d i c a t e t h a t a n y s u p p l e m e n t a l n i t r a t e a d d e d t o r o w crops
after t h e first i r r i g a t i o n s h o u l d be placed b e l o w t h e b o t t o m of
i r r i g a t i o n furrows in o r d e r to l e n g t h e n t h e p e r i o d of availability
to p l a n t s before it reaches d r y surface layers of soil.
Summary
X h e n i t r a t e ion moves very freely w i t h m o i s t u r e i n soils. I t
m a y b e easily l e a c h e d b u t i n a r i d o r semi-arid climates t h e movem e n t is p r e d o m i n a n t l y to t h e surface as sub-surface m o i s t u r e
moves u p w a r d a n d evaporates. Surface c o n c e n t r a t i o n s of n i t r a t e
m a y be as m u c h as a t h o u s a n d t i m e s t h a t of subsurface concentrations u n d e r furrow i r r i g a t i o n . T h i s r e d i s t r i b u t i o n has a p r o f o u n d
effect on sugar beet n u t r i t i o n . By decreasing n i t r a t e availability
late in t h e season, sugar c o n c e n t r a t i o n a n d q u a l i t y of t h e r o o t s
is i m p r o v e d . Late-season rainfall or s p r i n k l e r i r r i g a t i o n usually
results i n lower sugar c o n c e n t r a t i o n s a n d q u a l i t y .
Literature Cited
(1) BERNSTEIN, L E O N . 1959. Salt tolerance of vegetable crops in the west.
U. S. D e p t . Agr. Inf. Bull. 205. 5 p p .
(2) HADDOCK, J. L., et al. 1956. Nitrogen constituents associated with reduction of sucrose percentage a n d purity of sugar beets. J. Am. Soc.
Sugar Beet T e c h n o l . 9: 110-117.
(3) HADDOCK, J. L., et al. 1959. X h e influence of cultural practices on the
quality of sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 10 (4) : 290301.
(4) NIELSON, R. F. a n d L. A. BANKS. 1960. A new look at nitrate movem e n t in soils. U t a h Agr. E x p . Sta. Farm a n d H o m e Science 21 (1) :
2-3, 19.
(5) STOUT, M Y R O N . 1961. A new look at some nitrogen relationships affecting the quality of sugar beets. J. Am. Soc. Sugar Beet T e c h n o l .
11: 288-298.
(6) U L R I C H , ALBERT. 1942. T h e relationship of nitrogen to the formation
of sugar in sugar beets. Proc. Am. Soc. Sugar Beet T e c h n o l . 3: 66-80.
(7) U L R I C H , ALBERT and F. J. H I L L S . 1960. Mineral nutrition a n d sugar
beet quality. 1960 California Sugar Beet, p p . 36-37.
(8) W E N T , F. W. 1957. Climate a n d agriculture. Sci. Am. 1 9 6 ( 6 ) : 82-94.

Effectiveness of PEBC, D A T C , ond Endothall


for Controlling W e e d s in Sugar Beets
in W e s t e r n Nebraska 1
G.

A.

WICKS

AND F .

N.

ANDERSON2

Received for publication October 4, 1963

Introduction
O n e of t h e first selective h e r b i c i d e s used on sugar beets was a
concentrated salt s o l u t i o n by B a k k e (3) 3 in 1947; since t h e n
many chemicals h a v e b e e n t r i e d to selectively r e m o v e weeds from
sugar beets (4,9,11). D a l a p o n (2,2-dichloropropronic acid) has
been consistently effective in t h e c o n t r o l of a n n u a l grasses in
sugar beets w h i l e 3 , 6 - e n d o x o h e x a h y d r o p h t h a l i c acid (endothall)
and trichloroacetic acid ( T C A ) h a v e given v a r i a b l e c o n t r o l o n
a n n u a l grasses a n d broadleaf weeds (1,4). P r o p y l ethyl-n-butylthiolcarbamate ( P E B C ) has given acceptable weed control in
California, C o l o r a d o , W y o m i n g a n d M o n t a n a (2,5,8). A n o t h e r
chemical 2,3-dichloroallyl d i i s o p r o p y l t h i o l c a r b a m a t e ( D A T C ) has
proven to be an excellent h e r b i c i d e for wild oat control (9).
E x p e r i m e n t s w e r e i n i t i a t e d in western N e b r a s k a at Hershey,
Mitchell, a n d N o r t h P l a t t e , N e b r a s k a , t o d e t e r m i n e suitable
herbicides for c o n t r o l l i n g weeds in sugar beets.
Material and Methods
In 1961, e n d o t h a l at 2, 4, 6, a n d 8 l b / A , P E B C at 2, 4, a n d
8 l b / A tert-butyl d i - n - p r o p y l t h i o l c a r b a m a t e (R-1856) at 2.5, 5.0,
and 10.0 l b / A , a n d D A X C at 1, 2, a n d 4 l b / A were a p p l i e d preplant as broadcast t r e a t m e n t s a n d i m m e d i a t e l y i n c o r p o r a t e d on
April 12 a n d 13, A p r i l 14 a n d 15, a n d A p r i l 25 at Mitchell,
Hershey, a n d N o r t h P l a t t e , N e b r a s k a , respectively. Sugar beets
were p l a n t e d A p r i l 13, 15, a n d 26 at these p a r t i c u l a r locations.
In 1962, e n d o t h a l l at 2, 4, a n d 8 l b / A , P E B C at 2, 4, a n d 8 l b / A
and D A T C at 1, 2, a n d 4 l b / A w e r e a p p l i e d p r e p l a n t a n d soil
incorporated as b r o a d c a s t t r e a t m e n t s A p r i l 12 a n d 13 at Mitchell,
and A p r i l 31 a n d M a y 1 at N o r t h Platte. Sugar beets were
planted A p r i l 13 a n d M a y 4 at t h e respective locations. Incorporation to a d e p t h of 2 to 3 inches was accomplished w i t h i n
one m i n u t e after s p r a y i n g in 1961, a n d w i t h i n five m i n u t e s after
spraying in 1962, by a p o w e r - d r i v e n r o t a r y tiller m o u n t e d on a
Gravely g a r d e n tractor.
1

Published with the approval of the directors as paper No. 1423, Journal Series
Nebraska
Agricultural Experiment Station.
2
Assistant
Professors of Agronomy, University of Nebraska Experiment Station at
North
Platte Mitchell, Nebraska, respectively.
3
Numbers in parentheses refer to literature cited.

82

J O U R N A L OF T H E A.

S. S. B. T.

E n d o t h a l l was a p p l i e d p r e e m e r g e n c e in 1961 at rates of 2,


4, a n d 8 l b / A A p r i l 13, 19, a n d 27, respectively, at M i t c h e l l ,
H e r s h e y , a n d N o r t h Platte. I n a d d i t i o n e n d o t h a l l was a p p l i e d
p o s t e m e r g e n c e 26, 44, a n d 33 days after p l a n t i n g at t h e respective
locations. Sugar beets w e r e in t h e 4- to 6-leaf stage at t h e t i m e
of t h e p o s t e m e r g e n c e spraying.
H a n d w e e d i n g at weekly intervals a n d check ( n o w e e d control) w e r e used as t r e a t m e n t s in 1961 a n d 1962. W e e d s w e r e
r e m o v e d from t h e n o weed c o n t r o l plots following w e e d harvest.
In 1961 t h e c u l t u r a l practice of t h e c o o p e r a t i n g f a r m e r was inc l u d e d as a t r e a t m e n t . T h e farmer's o p e r a t i o n s at M i t c h e l l inc l u d e d t w o c u l t i v a t i o n s ; a t H e r s h e y e n d o t h a l l was a p p l i e d a n d
soil i n c o r p o r a t e d in a b a n d a h e a d of t h e p l a n t e r a n d t h e sugar
beets w e r e c u l t i v a t e d f i v e times. T h e N o r t h P l a t t e location
i n c l u d e d p r e p l a n t a n d soil i n c o r p o r a t e d a p p l i c a t i o n of e n d o t h a l l
a n d two cultivations. T h e farmer's practices a t t h e t h r e e locations were h a n d w e e d e d a n d t h i n n e d b y M e x i c a n laborers.
A r a n d o m i z e d block design with four r e p l i c a t i o n s was used
at each location in 1961 a n d a split-plot design w i t h five replications was used at each location in 1962. In t h e split-plot design,
one-half of each m a i n plot received t h e first c u l t i v a t i o n at t h e
t i m e t h e s u r r o u n d i n g f i e l d was c u l t i v a t e d a n d t h e o t h e r half
received all c u l t i v a t i o n s except t h e f i r s t . T h e plots t h a t received
t h e first c u l t i v a t i o n a r e referred to as n o r m a l , t h e others a r e called
delayed. T h e farmer's t r e a t m e n t s in 1961 w e r e c o m p o s e d of
t h r e e strips six rows w i d e , o n e o n each side a n d o n e d o w n t h e
m i d d l e of t h e e x p e r i m e n t a l area. Plots to be i n c l u d e d in t h e
e x p e r i m e n t w e r e selected at r a n d o m from t h e strips.
In 1961, visual notes w e r e t a k e n 4 3 , 47, a n d 38 davs after
p l a n t i n g a t M i t c h e l l , H e r s h e y , a n d N o r t h P l a t t e , respectively.
In 1962, c o u n t s w e r e t a k e n from t w o p e r m a n e n t l y m a r k e d areas.
1 ft by 9 ft, directly over t h e sugar b e e t row, 36 a n d 53 davs after
p l a n t i n g at M i t c h e l l a n d 36 days after at N o r t h P l a t t e . Also an
area .5 ft by 18 ft (9 sq ft) was c o u n t e d over t h e r o w at M i t c h e l l
68 days after p l a n t i n g . T h e l a t t e r c o u n t o c c u r r e d 5 days after
beets w e r e t h i n n e d o n c e by m e c h a n i c a l t h i n n e r . A dry seedbed
p r e v e n t e d susrar b e e t seed g e r m i n a t i o n so t h e y w e r e i r r i g a t e d t h e
first week of M a y at M i t c h e l l . R a i n on M a y 13 was sufficient for
sugar b e e t seed g e r m i n a t i o n a t N o r t h P l a t t e .
W e e d yields w e r e o b t a i n e d in 1961 a n d 1962 bv h a r v e s t i n g
above g r o u n d p o r t i o n s from 6.5, 9, a n d 18 scmare feet. W e e d
harvest o c c u r r e d 69, 65, a n d 58 davs after plantino- r^s*>ectivelv.
a t M i t c h e l l , H e r s h e y a n d N o r t h P l a t t e i n 1961. T h e 1962 harvest o c c u r r e d 83 a n d 59 to 60 davs after p l a n t i n g , respectively,
a t M i t c h e l l a n d N o r t h P l a t t e . W e e d s w e r e s e p a r a t e d a s t o grasses

VOL.

13,

No.

1,

APRIL

1964

83

and broadleaf w e e d s in 19.61, a n d in 1962 they were seperated


by species at w e e d harvest. W e e d s w e r e oven-dried a n d r e p o r t e d
as p o u n d s p e r acre. A t r a n s f o r m a t i o n of log (x + 1) was used
o n weed c o u n t s a n d w e e d weights. T h e d a t a were analyzed a n d
expressed as g e o m e t r i c m e a n s . In 1962 weeds were harvested
only from t h e delayed c u l t i v a t e d plots. All plots were weeded
following w e e d harvest a n d k e p t weed-free t h e rest of t h e season.
T h e n o r m a l c u l t i v a t e d plots w e r e w e e d e d at t h e t i m e of the
first cultivation.
In 1961 plots w e r e located on t h e following soil types:
Bridgeport l o a m a t N o r t h P l a t t e a n d Mitchell, a n d B r i d g e p o r t
sandy clay l o a m at H e r s h e y . T h e 1962 plots were located on the
following soil types: B r i d g e p o r t loam at Mitchell, a n d H a l l
loam at N o r t h P l a t t e .
Results
Climatology
T h e soil t e m p e r a t u r e ( d e p t h t h r e e inches) at t h e t i m e of
application a n d i n c o r p o r a t i o n of t h e p r e p l a n t t r e a t m e n t s in 1961
was 4 5 F at M i t c h e l l , 49-61F at Hershey, a n d 54-58F at N o r t h
Platte. T h e soil t e m p e r a t u r e in 1962 at t h e t i m e of application
and i n c o r p o r a t i o n of t h e p r e p l a n t t r e a t m e n t s was 44-56 F at
Mitchell a n d 46-58F a t N o r t h Platte.
T h e soil m o i s t u r e in t h e t o p t h r e e inches of t h e soil at t h e
time p r e p l a n t t r e a t m e n t s w e r e m a d e in 1961 was 1 7 . 4 % at
Mitchell, 1 7 . 9 % a t H e r s h e y , a n d 1 9 . 7 % a t N o r t h Platte. T h e soil
moisture in 1962 at t h e t i m e of t h e p r e p l a n t t r e a t m e n t s was 1 2 . 2 %
a t Mitchell a n d 9 . 8 % a t N o r t h P l a t t e .
Table 1.Accumulative precipitation in inches for 42 days following planting of
sugar beets at experimental locations in 1961 and 1962.

84

JOURNAL OF THE A. S. S. B. T.

T h e average m i n i m u m a n d m a x i m u m a i r t e m p e r a t u r e s for
o n e a n d t w o weeks after p l a n t i n g , respectively, in 1961 w e r e
28-58F a n d 31-61F at M i t c h e l l , 33-65F a n d 34-63F at H e r s h e y ,
a n d 34-58F a n d 40-56F a t N o r t h P l a t t e . T h e average m i n i m u m
a n d m a x i m u m a i r t e m p e r a t u r e s i n 1962 for o n e a n d two weeks
after p l a n t i n g , respectively, w e r e 37-75F a n d 41-74F at M i t c h e l l
a n d 54-88F a n d 53-78F a t N o r t h P l a t t e .
The p r e c i p i t a t i o n data, from p l a n t i n g to 42 days later, a r e
p r e s e n t e d in T a b l e 1 for 1961 a n d 1962.
Effect on prethinning sugar beet stands
In 1961 P E B C at 4 a n d 8 l b / A r e d u c e d sugar beet stands
b e l o w t h e check b y 2 0 a n d 3 9 % . D A T C a t 2 a n d 4 l b / A r e d u c e d
stands by 17 a n d 3 3 % . S t a n d losses on plots t r e a t e d with R-1856
at 5 a n d 10 l b / A w e r e 5 a n d 1 1 % . E n d o t h a l l a p p l i e d p r e p l a n t
at 4 a n d 8 l b / A r e d u c e d stands by 5 a n d 1 7 % ; all m e t h o d s of
a p p l i c a t i o n w e r e a b o u t t h e same. In 1962 plots t r e a t e d w i t h
P E B C at 4 a n d 8 l b / A lost 19 a n d 5 2 % of t h e p r e t h i n n i n g
stand, D A X C at 2 a n d 4 l b / A lost 11 a n d 2 7 % a n d stand loss
on t h e e n d o t h a l l at 8 l b / A plots was 1 0 % .
Annual
grass
control
X h e a n n u a l grasses i n 1961 a n d 1962 w e r e p r e d o m i n a t e l y
Setaria spp. Visual ratings t a k e n in 1961 a r e s h o w n in Table 2.
Visual c o n t r o l w i t h P E B C a t t h e t h r e e locations r a n g e d from
75 to 9 4 % w i t h 2 l b / A , 95 to 1 0 0 % w i t h 4 l b / A , a n d 1 0 0 % w i t h
8 l b / A . P e r c e n t control w i t h D A T C r a n g e d from 3 2 t o 8 2 %
w i t h 1 l b / A , 65 to 9 5 % for 2 l b / A , a n d 98 to 1 0 0 % for 4 l b / A .
C o n t r o l o b t a i n e d w i t h R-1856 was as follows: 35 to 6 8 % w i t h
2.5 l b / A , 7 2 t o 9 9 % w i t h 5.0 l b / A , a n d 9 2 t o 9 8 % for 10.0 l b / A .
Visual c o n t r o l w i t h e n d o t h a l l a p p l i e d p r e p l a n t was as follows: 0 to 9 2 % for 2 l b / A , 15 to 9 8 % for 4 l b / A , 28 to 9 8 %
for 6 l b / A , a n d 22 to 1 0 0 % for 8 l b / A . R e s u l t s at M i t c h e l l w e r e
excellent, at H e r s h e y c o n t r o l was fair; at N o r t h P l a t t e , it was
poor.
C o n t r o l w i t h e n d o t h a l l a p p l i e d p r e e m e r g e n c e r a n g e d from
0 t o 5 8 % for 2 l b / A , 15 to 7 4 % for 4 l b / A , a n d 70 to 8 8 % for
8 l b / A . X h e r e was little difference b e t w e e n c o n t r o l at M i t c h e l l
a n d H e r s h e y , at N o r t h P l a t t e it was p o o r , e x c l u d i n g t h e 8 l b / A
rate.
The p o s t e m e r g c n c e t r e a t m e n t of e n d o t h a l l gave c o n t r o l of
5 t o 5 5 % at 2 l b / A , 48 t o 8 0 % for 4 l b / A , a n d 70 t o 9 8 % for
8 l b / A . E x c e p t for t h e low rates at N o r t h P l a t t e , c o n t r o l was
a b o u t t h e s a m e for t h e t h r e e locations.
The grass yields at t h e t h r e e locations ( T a b l e 2) showed t h a t
several t r e a t m e n t s c o m p a r e d favorably to h a n d w e e d i n g at week-

Table 2.The effect of various herbicides on annual grasses as measured by visual estimations and oven dry weights at Mitchell, Hershey,
and North Platte, Nebraska in 1961.

1
2

Number of days after planting.


Numbers followed by the same letter do not differ significantly at the 5 percent level using Duncan's multiple range tests.

86

J O U R N A L OF T H E A.

S. S.

B.

T.

ly intervals. T h e s e w e r e : 2, 4, a n d 8 l b / A of P E B C , 5.0 a n d
10.0 l b / A of R-1856, 2 a n d 4 l b / A of D A T C , a n d p o s t e m e r g e n c e
t r e a t m e n t of e n d o t h a l l at 8 l b / A .
Grass c o u n t s a n d yields for 1962 a r e p r e s e n t e d in T a b l e 3.
C o u n t s 36 days after p l a n t i n g at M i t c h e l l show t h a t t h e r e was
no significant r e d u c t i o n in t h e n u m b e r of a n n u a l grasses by
P E B C a n d D A T C t r e a t m e n t s . C o u n t s t a k e n 5 3 days after planti n g s h o w e d significantly fewer p l a n t s on t h e D A T C at 2 a n d 4
l b / A a n d 2, 4, a n d 8 l b / A of P E B C t h a n on t h e check. Grass
yields o n t h e D A T C a n d P E B C t r e a t m e n t s were e q u a l t o o r
lower t h a n t h e h a n d w e e d t r e a t m e n t .
A t N o r t h P l a t t e w h e n c o u n t s w e r e t a k e n 3 6 days after planti n g P E B C a n d D A T C h a d e l i m i n a t e d significantly m o r e a n n u a l
grasses t h a n t h e check. C o n t r o l was still effective 60 days after
planting.
Results w i t h e n d o t h a l l were o u t s t a n d i n g from t h e f i r s t readi n g a t Mitchell, b u t b y t h e second r e a d i n g t h e r e was a n increase
i n p l a n t n u m b e r s o t h a t t h e r e was n o significant difference bet w e e n e n d o t h a l l t r e a t m e n t s a n d t h e check. T h e N o r t h P l a t t e
c o u n t s show n o difference b e t w e e n check a n d a n y e n d o t h a l l
t r e a t m e n t . T h e r e was n o significant difference b e t w e e n check
a n d e n d o t h a l l t r e a t m e n t s in t h e grass yields harvested 83 to 60
days after p l a n t i n g a t M i t c h e l l a n d N o r t h P l a t t e , respectively.
Grass yields for locations show t h a t all rates of D A T C a n d
P E B C w e r e s i m i l a r t o t h e h a n d w e e d t r e a t m e n t . T h e r e was a
highly significant location X t r e a t m e n t i n t e r a c t i o n caused by
b e t t e r p e r f o r m a n c e of e n d o t h a l l at M i t c h e l l .
Broadleaf
weed
control
P r i n c i p l e broadleaf species in 1961 w e r e : kochia (Kochia
scoparia L.)
a n d r o u g h pigweed
(Amaranthus retroflexus L.).
Kochia was p r e d o m i n a t e a t M i t c h e l l a n d N o r t h P l a t t e , a n d r o u g h
p i g w e e d a t Hershey. I n 1962, kochia a n d r o u g h pigweed w e r e
t h e p r e d o m i n a t e broadleaf weeds a t M i t c h e l l a n d N o r t h P l a t t e .
S e e d l i n g alfalfa was p r e s e n t in sufficient q u a n t i t y to c o u n t . Black
nightshade
(Solanum
nigraum L.) was p r e s e n t at t h e N o r t h
P l a t t e site.
Visual r a t i n g s w e r e t a k e n in 1961 a n d results a r e s h o w n in
T a b l e 4. P E B C at 2 l b / A c o n t r o l l e d 30 to 9 5 % , 4 l b / A controlled 8 8 t o 9 8 % , a n d 8 l b / A c o n t r o l l e d 9 2 t o 1 0 0 % . C o n t r o l
w i t h R-1856 r a n g e d from 0 to 1 8 % for 2.5 l b / A , 0 to 6 2 % for
5.0, a n d 2 5 a n d 8 1 % for 1 0 l b / A . C o n t r o l for D A T C r a n g e d
from 0 to 4 5 % for 1 l b / A , 20 to 6 5 % for 2 l b / A , a n d 52 to 9 4 %
for 4 l b / A . O n l y P E B C at 8 l b / A was e q u a l to h a n d - w e e d i n g
at weekly intervals for broadleaf w e e d c o n t r o l .

Table 3.The effect of various herbicides on annual grasses as measured by plant counts and oven-dry weights at Mitchell, and North Platte,
Nebraska in 1962.

1
A nine square foot area was harvested, counted and over-dried 83 days after planting at Mitchell and 60 days after at North Platte for the delayed
cultivation. Normal was counted 68 days after planting.
2
Numbers followed by the same letter do not differ significantly at the 5 percent level using Duncan's multiple range tests.

Table 4.The effect of various herbicides on broadleaf weeds as measured by visual estimations and oven-dry weights at Mitchell, Hershey, and
North Platte, Nebraska, in 1961.

1
2

Number of days after planting.


Numbers followed by the same letter do not differ significantly at the 5 percent level using Duncan's multiple range tests.

VOL. 13, N o . 1, A P R I L 1964

89

E n d o t h a l l a p p l i e d p r e p l a n t a n d soil i n c o r p o r a t e d , c o n t r o l l e d
0 to 9 1 % for 2 l b / A , 12 to 9 8 % for 4 l b / A , 18 t o 1 0 0 % for 6
l b / A , a n d 38 to 9 8 % for 8 l b / A . P r e e m e r g e n c e applications of
e n d o t h a l l c o n t r o l l e d 10 to 4 9 % for 2 l b / A , 8 to 7 2 % for 4 l b / A ,
and 40 to 9 0 % for 8 l b / A . P o s t e m e r g e n c e applications of endothall c o n t r o l l e d 0 to 4 5 % for 2 l b / A , 0 to 6 0 % for 4 l b / A ,
a n d 1 2 t o 9 2 % for 8 l b / A . T h e p r e p l a n t soil i n c o r p o r a t e d a n d
the p r e e m e r g e n c e m e t h o d of a p p l y i n g e n d o t h a l l p e r f o r m e d best
a t M i t c h e l l b u t t h e p o s t e m e r g e n c e t r e a t m e n t was best a t Hershey.
T h e weed yields show t h a t t h e only s t r i k i n g results w i t h endothall w e r e w i t h t h e p o s t e m e r g e n c e t r e a t m e n t s a t Hershey.
T a b l e 5 shows t h e d a t a t a k e n on r o u g h pigweed at Mitchell
a n d N o r t h P l a t t e in 1962. P E B C was m o r e effective by the second
c o u n t i n g a n d all rates w e r e significantly b e t t e r t h a n any o t h e r
t r e a t m e n t . T h e t w o areas c o u n t e d over t h e row a t Mitchell 5 3
a n d 6 8 days after p l a n t i n g r e s p o n d e d similarly. T h e N o r t h
Platte c o u n t s s h o w e d P E B C t o b e t h e o u t s t a n d i n g herbicide
for c o n t r o l of r o u g h pigweed. W e e d yields were c o m p a r a b l e to
the h a n d w e e d t r e a t m e n t at b o t h locations. P E B C at 4 a n d 8
l b / A w e r e s u p e r i o r t o h a n d w e e d i n g a t weekly intervals measured
by D u n c a n ' s m u l t i p l e r a n g e tests on location means.
C o u n t s a n d w e e d weights t a k e n a t b o t h locations indicate
that D A T C was ineffective i n c o n t r o l l i n g r o u g h pigweed.
E n d o t h a l l at 8 l b / A was very effective in c o n t r o l l i n g r o u g h
pigweed at t h e first o b s e r v a t i o n at Mitchell. T h e r e was no significant difference by t h e second r e a d i n g b e t w e e n t h e check a n d
the e n d o t h a l l t r e a t m e n t s on t h e 12-inch w i d e area over t h e
row. T h e r e was a significant difference b e t w e e n t h e check a n d
e n d o t h a l l t r e a t m e n t s o n t h e 6-inch w i d e area. W e e d yields o n
the 4 l b / A plots w e r e significantly less t h a n t h e check. T h e
e n d o t h a l l t r e a t m e n t s at N o r t h P l a t t e w e r e n o t significantly different t h a n t h e check at any t i m e . Most of t h e highly significant
location X t r e a t m e n t i n t e r a c t i o n was caused by b e t t e r e n d o t h a l l
performance at Mitchell.
T a b l e 6 shows t h e d a t a t a k e n on kochia at Mitchell a n d
N o r t h P l a t t e i n 1962. P E B C a n d D A T C d i d n o t control kochia
at e i t h e r location. E n d o t h a l l gave g o o d c o n t r o l of kochia at
Mitchell as i n d i c a t e d by t h e first w e e d counts, a n d by t h e counts
a n d weed weights 8 3 days after p l a n t i n g . T h e second r e a d i n g
did n o t a p p e a r as g o o d as t h e first or t h a t t a k e n 15 days later
on the 6-inch area. C o n t r o l of kochia at N o r t h P l a t t e with
e n d o t h a l l was p o o r , a l t h o u g h t h e r e was a significant r e d u c t i o n
in weed weights b u t n o n e for c o u n t s . A g a i n t h e highly significant l o c a t i o n X treatment interaction was due to better endothall performance at Mitchell.

Table 5.Effect of various herbicides on rough pigweed as measured by plant counts and oven-dry weights at Mitchell and North Platte,
Nebraska in 1962.

1
A nine square foot area was harvested, counted and oven-dried 83 days after planting at Mitchell and 60 days after at North Platte for the delayed
cultivation. Normal was counted 68 days after planting.
2
Numbers followed by the same letter do not differ significantly at the 5 percent level using Duncan's multiple range tests.

Table 6.Effect of various herbicides on kochia as measured by plant counts and oven-dry weights at Mitchell and North Platte, Nebraska
in 1962.

1
A nine square foot area was harvested, counted and oven-dried 83 days after planting at Mitchell and 60 days after at North Platte for the delayed
cultivation. Normal was counted 68 days after planting.
2
Numbers followed by the same letter do not differ significantly at the 5 percent level using Duncan's multiple range tests.

92

JOURNAL OF THE A. S. S. B. T.

Table 7.Effect of various herbicides on seedling alfalfa and black nightshade as


measured by plant counts at Mitchell and North Platte, Nebraska in 1962.

1
Numbers followed by the same letter do not differ significantly at the 5 percent level
using Duncan's multiple range tests.

Seedling alfalfa c o u n t s are p r e s e n t e d in T a b l e 7 for b o t h


locations. T h e e n d o t h a l l t r e a t m e n t s a t M i t c h e l l e l i m i n a t e d t h e
alfalfa. T h e alfalfa was n o t affected b y D A T C a n d P E B C .
E n d o t h a l l t r e a t m e n t s a t N o r t h P l a t t e killed m a n y seedling alfalfa.
N u m b e r of alfalfa on t h e 4 a n d 8 l b / A rates was significantly
less t h a n t h e check. D A T C a t 4 l b / A a n d t h e P E B C t r e a t m e n t s
r e d u c e d alfalfa stands, b u t 8 l b / A of P E B C was t h e only treatm e n t t h a t was significantly less t h a n t h e check.
C o u n t s t a k e n o n black n i g h t s h a d e a t N o r t h P l a t t e a r e g i v e n
i n T a b l e 7 . N o significant difference was n o t e d b e t w e e n treatments.
Discussion
E n d o t h a l l ' s p o o r p e r f o r m a n c e is r e l a t e d to t h e rainfall patt e r n , a s a m o u n t o f rainfall increases results b e c o m e p o o r e r . I n
1961 it was 30 days after p l a n t i n g before an i n c h of r a i n was received at M i t c h e l l . W e e d notes w e r e t a k e n 43 days after p l a n t i n g .
At H e r s h e y it was 17 days after p l a n t i n g before an inch of r a i n
was received; visual notes w e r e t a k e n 47 days after p l a n t i n g .
At N o r t h P l a t t e it was 5 days after p l a n t i n g before an inch of
r a i n fell; visual n o t e s w e r e t a k e n 38 days after p l a n t i n g . It is
p r e s u m e d t h a t e n d o t h a l l was leached o r h a d dissipated from t h e
u p p e r surface of t h e soil after an i n c h of r a i n . C o m e s et al. (6)
have f o u n d t h a t e n d o t h a l l was leached to a d e p t h of t h r e e inches
in sandy l o a m , sandy clay l o a m a n d clay l o a m soils w i t h t w o
i n c hes of water. E n d o t h a l l dissipated q u i c k e r in t h e sandy l o a m

VOL. 13, N o . 1, A P R I L 1964

93

and sandy clay l o a m t h a n t h e clay l o a m soil. T h e y also f o u n d


that t e m p e r a t u r e influenced t h e b r e a k d o w n of e n d o t h a l l . Extended t e m p e r a t u r e s of 6 8 F a n d above hastened dissipation.
O t h e r workers (4,10) h a v e r e p o r t e d p o o r c o n t r o l with e n d o t h a l l .
T h e r e was little difference b e t w e e n t h e p r e p l a n t a n d t h e
preemergence a p p l i c a t i o n s of e n d o t h a l l in weed yields, a l t h o u g h
there was s o m e a d v a n t a g e in t h e p r e p l a n t t r e a t m e n t by visual
observations. Possibly w e e d yields w e r e t a k e n too late to show
this difference. T h e p o s t e m e r g e n c e t r e a t m e n t response was variable. A t H e r s h e y c o n t r o l was m u c h b e t t e r t h a n a t Mitchell a n d
N o r t h Platte. T h i s is p r o b a b l y d u e to differences in weed species
present a n d stage of d e v e l o p m e n t at t i m e of spraying.
In 1962 it was 34 a n d 9 days after p l a n t i n g before an inch
of r a i n was received at M i t c h e l l a n d N o r t h Platte. A p p a r e n t l y ,
the r a i n at N o r t h P l a t t e leached e n d o t h a l l below t h e zone of
g e r m i n a t i n g weeds so few weeds were killed. F u r r o w irrigation
a t Mitchell was early e n o u g h t o g e r m i n a t e most weeds. T h e
weeds w e r e killed before t h e r a i n leached the e n d o t h a l l too
deep. T h i s w o u l d a c c o u n t for t h e significant location X treatment interaction.
T h e furrows t h a t w e r e m a d e for i r r i g a t i o n caused differences
in control w h i c h w e r e n o t i c e a b l e in t h e e n d o t h a l l treatments.
T h i s is i n d i c a t e d by a significant difference in broadleaf weed
counts b e t w e e n t h e e n d o t h a l l t r e a t m e n t s a n d the check o n the
6-inch strip b u t n o t on t h e 12-inch strip. T h e 12-inch strip included a p o r t i o n of t h e soil m o v e d from the furrow. It is
assumed t h a t t h e e n d o t h a l l was e i t h e r m o v e d closer to the row or
had dissipated from t h e furrow edges.
P E B C was effective in c o n t r o l l i n g grasses a n d r o u g h pigweed
in b o t h years at all locations u n d e r extremely variable climatic
conditions. I t d i d n o t c o n t r o l kochia o r nightshade. Kochia has
been r e p o r t e d to h a v e b e e n c o n t r o l l e d by P E B C (7). E i t h e r the
kochia in w e s t e r n N e b r a s k a is resistant to P E B C or conditions
existed t h a t allowed t h e kochia to g e r m i n a t e a n d d e v e l o p fast
enough so t h a t P E B C was ineffective. F o r example, t h e P E B C
could have dissipated from t h e u p p e r q u a r t e r inch of soil a n d
the kochia g e r m i n a t e d i n this area. T h e n t h e small r o o t grew
t h r o u g h t h e t r e a t e d a r e a of P E B C . P E B C was m u c h m o r e effective on broadleaf weeds t h a n R-1856.
D A T C gave excellent c o n t r o l of a n n u a l grasses b o t h years.
T w o year's d a t a i n d i c a t e t h a t broadleaf weeds controlled with
D A T C i s n o t a d e q u a t e . T h i s agrees with o t h e r research work
(9,10).

94

J O U R N A L OF T H E A. S. S.

B.

T.

Summary
1. In western Nebraska endothall, D A T C , and P E B C were
a p p l i e d p r e p l a n t a n d soil i n c o r p o r a t e d to a d e p t h of 2 to
3 inches in 1961 a n d 1962; R-1856 was a p p l i e d in a similar
m a n n e r i n 1961. E n d o t h a l l was also a p p l i e d p r e e m e r g e n c e
a n d p o s t e m e r g e n c e i n 1961. R e s u l t s from t h r e e locations
in 1961 a n d t w o locations in 1962 a r e r e p o r t e d .
2. P E B C was t h e o u t s t a n d i n g h e r b i c i d e t r i e d in this study,
a l t h o u g h i t d i d n o t c o n t r o l kochia o r black n i g h t s h a d e .
T h e c o n t r o l o f a n n u a l grasses a n d r o u g h p i g w e e d b y P E B C
was c o m p a r a b l e t o h a n d w e e d i n g a t weekly intervals.
3. D A T C gave acceptable c o n t r o l of grass b u t n o t broadleaf
weeds.
4 . E n d o t h a l l d i d n o t give consistant w e e d c o n t r o l u n d e r t h e
c o n d i t i o n s of this study.
5. P E B C , R-1856 a n d D A T C were less affected by precipitat i o n t h a n e n d o t h a l l . Soil m o i s t u r e s b e t w e e n 9.8 a n d 1 9 . 7 %
at i n c o r p o r a t i o n t i m e h a d little affect on t h e c o n t r o l of
a n n u a l grasses a n d r o u g h pigweed o b t a i n e d b y P E B C i n
this study.
Acknowledgements
T h i s research was partially s u p p o r t e d by a g r a n t from the
G r e a t W e s t e r n Sugar C o m p a n y . T h e a u t h o r s wish t o t h a n k the
fieldmen a n d t h e m a n a g e r s of t h e factories of t h e G r e a t W e s t e r n
Sugar C o m p a n y a t M i t c h e l l , N e b r a s k a , a n d O v i d , C o l o r a d o , for
t h e i r h e l p i n t h e e x p e r i m e n t . T h a n k s are also e x t e n d e d t o the
c o o p e r a t i n g farmers: Kei M a t s u t a n i a t H e r s h e y ; a n d D e l b e r t
Nicholas a n d A m o s M e h l a t N o r t h P l a t t e , N e b r a s k a .
Literature Cited
(1) ANDERSON, R. N. 1962. Progress a n d problems in sugar beet weed
control. Crystalized Facts A b o u t Sugar Beets. 1 6 ( 1 ) : 13-20.
(2) ANTOGNINI, J. 1961. T i l l a m ( T M ) for weed control in sugar beets
in California. Holly Agricultural News. 9 (1) : 6-7.
(3) BAKKE, A. L. 1947. Spraying beets with salt solutions. N C W C C Rep o r t 4: 26.
(4)

BANDEEN, J . D., G. E. J O N E S , a n d C. M. SWITZER.

1960.

F u r t h e r studies

on the control o weeds in sugar beets with herbicides. J. Am. Soc.


Sugar Beet T e c h n o l . 11 (2) ; 160-163.
(5) B I S C H O F F , K. 1963. T h e T i l l a m story. Holly Agricultural News.
1 1 ( 1 ) : 32-34.

VOL. 13, N o . 1, A P R I L 1964


(6)

95

COMES, R . D., D . W . B O H M O N T , a n d H . P . A L L E Y .

1961.

Movement

and persistence of e n d o t h a l (3,6-endoxohexahydrophthalic acid) as


influenced by soil texture, temperature, and moisture levels. J. Am.
Soc. Sugar Beet T e c h n o l . 11 (4) : 287-293.
(7) D A Y , H. M. 1962. T i l l a m ( T M ) use in the Midwest and Rocky Mountain areas. Crystalized Facts A b o u t Sugar Beets. 1 6 ( 1 ) : 20-22.
(8) NELSON, R. T. 1958. Chemical weed control in sugar beets1958.
T h r o u g h the Leaves. X L V I (5) : 19.
(9) SEXSMITH, J. J. 1960. Chemical control of wild oats (Avena fatua L.)
in sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 11 (3) : 268-278.
(10)

SULLIVAN, E. F.,

R . L. ABRAMS a n d R . R. W O O D .

1963.

W e e d control

in sugar beets by combinations of thiolcarbamate herbicides. Weeds.


11 (4) : 258-260.
(11)

SWANSON, C. R., E. A. HELGESON, a n d L. M. STAHLER.

1948.

Summary

of weed investigations in sugar beets at Fargo, North Dakota, and


East G r a n d Forks, Minnesota, conducted d u r i n g the summer, 1948.
Preemergence treatments. N C W C C Res. R e p t . 4: 42.

Minutes of the T h i r t e e n t h General M e e t i n g


of the
American Society of Sugar Beet Technologists
T h e business m e e t i n g o f t h e T h i r t e e n t h G e n e r a l M e e t i n g
of t h e A m e r i c a n Society of S u g a r Beet T e c h n o l o g i s t s was called
to o r d e r by M r . G u y R o r a b a u g h , P r e s i d e n t of t h e Society, at
11:10 a.m. on W e d n e s d a y , F e b r u a r y 5, 1964, in t h e G o l d Ballr o o m of t h e Sheraton-Palace H o t e l , San Francisco, California.
M r . R o r a b a u g h , serving as c h a i r m a n of t h e m e e t i n g , ann o u n c e d t h a t t h e t h i r t e e n t h b i e n n i a l business m e e t i n g o f t h e
Society was called for t h e p u r p o s e of h e a r i n g c o m m i t t e e r e p o r t s
a n d t o transact such business a s m a y b e a p p r o p r i a t e .
T h e c h a i r m a n called for t h e r e a d i n g of t h e m i n u t e s of t h e
T w e l f t h G e n e r a l Business Session h e l d i n D e n v e r , C o l o r a d o , o n
F e b r u a r y 7 , 1962. U p o n m o t i o n m a d e , seconded a n d u n a n i m o u s ly carried, r e a d i n g of t h e m i n u t e s of such m e e t i n g was dispensed
with.
T h e c h a i r m a n t h e n asked for t h e r e p o r t o f t h e SecretaryT r e a s u r e r . T h e S e c r e t a r y - T r e a s u r e r briefly r e p o r t e d o n t h e business activities of t h e Society for t h e b i e n n i a l p e r i o d e n d i n g
D e c e m b e r 3 1 , 1963. U p o n m o t i o n m a d e , seconded a n d u n a n i m o u s l y carried, t h e r e p o r t of t h e S e c r e t a r y - T r e a s u r e r was a d o p t e d ,
o r d e r e d placed on file a n d m a d e a p a r t of these m i n u t e s . T h e
r e p o r t of t h e Secretary a n d t h e r e p o r t of t h e T r e a s u r e r consisting
of a b a l a n c e sheet a r e h e r e w i t h m a d e a p a r t of these m i n u t e s .
R e p o r t of the Secretary
D e c e m b e r 3 1 , 1963
T h e Society m e m b e r s h i p a t t h e close o f t h e 1962-63 b i e n n i u m
was 737 i n d i v i d u a l s a n d c o m p a n i e s . T h e m e m b e r s h i p i s r e p r e s e n t e d in 38 states, t h e District of C o l u m b i a , o n e t e r r i t o r i a l
possession, 5 p r o v i n c e s of C a n a d a a n d 22 foreign c o u n t r i e s outside t h e c o n t i n e n t a l l i m i t s o f N o r t h A m e r i c a . T h e n u m b e r o f
m e m b e r s has increased by 105 or 16 1/2% over t h e p r e v i o u s bie n n i u m . T h i s i s t h e largest increase i n m e m b e r s h i p ever exp e r i e n c e d by t h e Society. A list of t h e m e m b e r s h i o by geographical division is m a d e a p a r t of this r e p o r t (page 97).
N e a r l y 1100 copies of t h e J o u r n a l of t h e A m e r i c a n Society
o f Sugar Beet T e c h n o l o g i s t s a r e m a i l e d t o t h e m e m b e r s h i p a n d
subscribers each p r i n t i n g to 41 states a n d 43 foreign c o u n t r i e s .
In o r d e r to m a i n t a i n a reserve s u p p l y of n u m b e r s , 1400 copies
of each issue a r e c u r r e n t l y b e i n g p r i n t e d . E a c h year a s u r p r i s i n g
n u m b e r o f o r d e r s a r e received for back v o l u m e s . T h e supply

of Proceedings p r i o r to 1946 is e x h a u s t e d a n d a l i m i t e d n u m b e r
of other Proceedings a n d e a r l i e r J o u r n a l s is available.
T h e increased n a t i o n a l interest in sugar beets a n d
creased Society m e m b e r s h i p has t r a n s f o r m e d t h e Office
Secretary i n t o a b e e h i v e of activity. A p p r o x i m a t e l y o n e
the business h o u r s of t h e Secretary is spent on Society
in a d d i t i o n to o n e full-time office assistant.

t h e inof t h e
half of
matters

It is p r o p e r for t h e Society to express its thanks to t h e Beet


Sug^ar D e v e l o p m e n t F o u n d a t i o n for p e r m i t t i n g t h e M a n a g e r of
the F o u n d a t i o n to s p e n d this m u c h of his t i m e a n d t h e t i m e of
his office assistants in c o n d u c t i n g t h e business affairs of t h e
Society. In a d d i t i o n t h e r e t o , t h e F o u n d a t i o n has provided an
a n n u a l g r a n t of $1,000 to t h e Society to h e l p with the cost of
publishing the J o u r n a l .
Respectfully
JAMES

H.

submitted,

FISCHER

Secretary-Treasurer

JOURNAL OF THE A. S. S. B. T.

98

R e p o r t of the Treasurer
Balance Sheet
D e c e m b e r 3 1 , 1963
Cash Balance, J a n u a r y 1, 1962
Savings A c c o u n t Balance, J a n u a r y 1, 1962
1962 I n t e r e s t E a r n e d on Savings A c c o u n t
1962 Cash R e c e i p t s
1963 I n t e r e s t E a r n e d on Savings A c c o u n t
1963 Cash R e c e i p t s

895.21
5,328.09
242.46
12,855.83
162.99
9,175.41

Less: T r a n s f e r from Savings to C h e c k i n g A c c o u n t


I n t e r e s t on S h o r t - T e r m L o a n

$28,659.99
4.500.00

2.00

1962 Cash D i s b u r s e m e n t s
1963 Cash D i s b u r s e m e n t s
Savings A c c o u n t Balance, D e c e m b e r 3 1 , 1963
Cash Balance, D e c e m b e r 3 1 , 1963

$24,157.99
11,620.44
10,244.77
1,231.54
1,061.24
$24,157.99

NOTE:

N e t decrease i n cash a n d savings J a n u a r y


1, 1962 t h r o u g h D e c e m b e r 3 1 , 1963

$3,930.52

T h e c h a i r m a n t h e n called for a r e p o r t from t h e Steering


C o m m i t t e e a p p o i n t e d t o m a k e p r e l i m i n a r v a r r a n g e m e n t s for t h e
Second Toint M e e t i n g of t h e A S S B T - I I R B to be h e l d in the
U n i t e d States a n d C a n a d a d u r i n g t h e m o n t h s o f M a y a n d J u n e
1965. M r . B . E . Easton, co-chairman w i t h M r .Bion T o l m a n .
p r o c e e d e d w i t h t h e r e p o r t of plans to d a t e . At t h e conclusion
of t h e c o m m i t t e e ' s r e p o r t t h e following r e s o l u t i o n was p r o p o s e d :
Resolution
Whereas
t h e A m e r i c a n Society of Sugar Beet T e c h n o l o g i s t s at
its E l e v e n t h G e n e r a l M e e t i n g in Salt L a k e City on F e b r u a r v 4,
1960, u n a n i m o u s l y accepted a r e p o r t p r o p o s i n g j o i n t m e e t i n g s
between the ASSBT and the I I R B , and
Whereas t h e I I R B s u b s e q u e n t l v hosted t h e A S S B T in Engl a n d for a w e e k ' s t o u r e n d i n g w i t h a j o i n t m e e t i n g on Friday,
M a y 19, 1961, in L o n d o n .
Be It Therefore Resolved
that the ASSBT plan and conduct a
t o u r a n d m e e t i n g for r e p r e s e n t a t i v e s o f t h e I I R B c o m m e n c i n g
in t h e eastern b e e t - g r o w i n g areas of t h e U n i t e d States a n d term-

VOL.

13, N o .

1, A P R I L

1964

99

inating in the western beet-growing areas beginning approximately Monday, May 24, 1965 and ending approximately June
8, 1965.
Be It Further Resolved
that the Executive C o m m i t t e e and
Board of Directors be authorized to proceed immediately with
final plans, such plans to include a tour time schedule, sites and
manner of conducting demonstrations, meals and lodging, modes
of transportation and other details pertinent to an informative
and successful tour and meeting.
Be It Further Resolved
that the m a n a g e m e n t of all North
American Beet Sugar Companies be informed of the entire plan
and solicit their united cooperation in planning and hosting this
fine group of visitors.

Upon motion made, seconded and unanimously carried, the


above resolution was unanimously adopted by the Society.
The chairman then asked for a report from the Site Selection
Committee appointed to select the city and the hotel in which
to hold the Fourteenth General Meeting of the Society in 1966.
Mr. P. B. Smith, speaking on behalf of the other committee
members, Mr. Aldrich Bloomquist and Mr. M. G. Frakes, reported that the Hotel Leamington in Minneapolis, Minnesota,
was herewith being proposed as the site for the 1966 biennial
meeting of the Society. Upon motion made, seconded and unanimously carried, Minneapolis and the Hotel Leamington were
selected as the meeting hotel and the hosting city for our 1966
Fourteenth General Meeting of the Society.
The President of the Society, on behalf of the Executive
Committee and Board of Directors, placed the names of Dr. A.
Carruthers, British Sugar Corporation, Nottingham, England
and J. Earl Coke, Vice President, Bank of America, San Francisco,
California, in nomination as recipients of the Honorary Membership award. Upon motion made, seconded and unanimously
carried, the above two individuals, Dr. A. Carruthers and J.
Earl Coke, were unanimously elected as honorary members of
the American Society of Sugar Beet Technologists for life.
The chairman then reported that the Tally Committee, consisting of Robert S. Gaddie, Dewey Stewart, E. F. Blackwelder
and R. M. McCready, was in the process of counting ballots cast
in favor of nominees for each of the Society offices, such elected
officers to hold office for the biennium 1964-65. Although the

100

JOURNAL OF THE A. S. S. B. T.

results o f t h e tally w e r e n o t m a d e k n o w n u n t i l t h e b a n q u e t t h e
e v e n i n g of t h e day of this business m e e t i n g , t h e results a r e herewith duly recorded:
Executive Committee
President
B.
I m m e d i a t e Past P r e s i d e n t
Guy
Vice P r e s i d e n t
Lloyd
Secretary-Treasurer
James

E. Easton
Rorabaugh
X. Jensen
H . Fischer

B o a r d of D i r e c t o r s
Pacific Coast R e g i o n
E. M. D a n i e l s
Intermountain Region
Ronald C. Johnson
Eastern Rocky Mountain Region
J . L . Porterfield
North Central and Great
Lakes Region
Max C. Henderson
Canada
J. W. Hall
Processing a t L a r g e
R. A. McGinnis
Agricultural at Large
J. C. T a n n e r
On behalf of t h e R e s o l u t i o n s C o m m i t t e e , t h e Society Presid e n t a c k n o w l e d g e d w i t h t h a n k s t h e m a n y Society m e m b e r s a n d
c o n t r i b u t o r s w h o d e v o t e d t i m e a n d effort to t h e success of t h e
T h i r t e e n t h G e n e r a l M e e t i n g . H e specifically a c k n o w l e d g e d t h e
P r o g r a m C h a i r m a n , Russell T . J o h n s o n , his Section C h a i r m e n ,
J o h n T . A l e x a n d e r , R a l p h F i n k n e r , C h a r l e s Schneider, A u s t i n
A r m e r , R o b e r t S . G a d d i e , V a r o n J e n s e n a n d W h i t n e y Newton. I I .
H e a c k n o w l e d g e d t h e Local A r r a n g e m e n t s C o m m i t t e e , A l d e n
Stock, H u g h M e l v i n , C u r z o n Kay a n d R . J . T i n g l e y ; t h e N o m i n a t i n g C o m m i t t e e , R . S . G a d d i e , H . P . H . J o h n s o n , Dewey
Stewart, R . M . M c C r e a d y a n d E . F . Blackwelder; t h e A w a r d s
C o m m i t t e e ,J. S . M c F a r l a n e , L e R o y Powers, L l o y d T . J e n s e n ,
P. B. Smith and A. R. Downie; Publications Committee, James
H . Fischer, M . M . Afanasiev, J . L . H a d d o c k , J u l i a n J o h n s o n a n d
L e R o y Powers. H e t h a n k e d t h e speakers a t o u r g e n e r a l sessions
a n d a t all t h e sectional m e e t i n g s , t h e San Francisco C o n v e n t i o n
a n d Visitors B u r e a u , t h e staff of t h e Sheraton-Palace H o t e l , t h e
Silver E n g i n e e r i n g W o r k s , I n c o r p o r a t e d for p r o v i d i n g refreshm e n t s d u r i n g session breaks, t h e H o l l y Sugar C o r p o r a t i o n , U n i o n
Sugar Division of C o n s o l i d a t e d Foods C o r p o r a t i o n , Spreckels
Sugar C o m p a n y a n d t h e A m e r i c a n Crystal Sugar C o m p a n y for
s u p p l y i n g special e n t e r t a i n m e n t a n d e n t e r t a i n m e n t for t h e ladies
attending the T h i r t e e n t h General Meeting.
T h e r e b e i n g n o f u r t h e r business t h e m e e t i n g was d e c l a r e d
a d j o u r n e d a t 11:40 a.m.
JAMES

H.

FISCHER

Secretary-Treasurer

VOL. 13, N o . 1, A P R I L 1964

101

Meritorious Service
A w a r d Presented
to

GUY

RORABAUGH

G u y R o r a b a u g h was b o r n at Salida, C o l o r a d o , M a y 20, 1911


and a t t e n d e d h i g h school a t C r i p p l e C r e e k , C o l o r a d o . H e attended C o l o r a d o State U n i v e r s i t y a n d was g r a d u a t e d from t h e
University of C o l o r a d o in 1935. U p o n g r a d u a t i o n he was employed by t h e H o l l y S u g a r C o r p o r a t i o n a n d h a s served as special
chemist at Sidney, M o n t a n a , assistant c h e m i s t at H a m i l t o n City,
California, assistant c h e m i s t a n d chief c h e m i s t at H a r d i n , M o n tana, chief c h e m i s t at W o r l a n d , W y o m i n g , m a n a g e r of t h e research l a b o r a t o r y a t C o l o r a d o Springs, b e c a m e g e n e r a l c h e m i s t
and d i r e c t o r of research at C o l o r a d o Springs, was p r o m o t e d to
general s u p e r i n t e n d e n t a n d recently elected vice p r e s i d e n t of
operations. He is a m e m b e r of t h e A m e r i c a n C h e m i c a l Society,
Institute of F o o d T e c h n o l o g i s t s , A m e r i c a n Society of Sugar Beet
Technologists, I n d u s t r y Advisory C o m m i t t e e S u g a r R e s e a r c h
F o u n d a t i o n , Sugar Beet Process Advisory C o m m i t t e e of t h e Beet
Sugar D e v e l o p m e n t F o u n d a t i o n a n d t h e Secretary of A g r i c u l t u r e ' s
Oilseed, P e a n u t a n d S u g a r C r o p s A d v i s o r y C o m m i t t e e . H e i s a
director of t h e Beet Sugar D e v e l o p m e n t F o u n d a t i o n a n d is listed
i n both t h e A m e r i c a n M e n o f Science a n d W h o ' s W h o i n C h e m istry. M r . R o r a b a u g h has b e e n a m e m b e r of t h e A m e r i c a n Society
of sugar beet T e c h n o l o g i s t s since 1954, served as c h a i r m a n of
the Chemistry a n d Factory O p e r a t i o n Section in 1956-57; served
o n t h e n o m i n a t i n g c o m m i t t e e i n 1956-57; served o n t h e A d v i s o r y
c o u n c i l i n 1960-61; a n d was g e n e r a l p r o g r a m c h a i r m a n for t h e
12th G e n e r a l M e e t i n g a n d p r e s i d e n t of t h e Society for t h e bie n n i u m 1962-63.

102

J O U R N A L OF T H E A. S. S. B. T.

Meritorious Service
Award Presented
to

E. JACKSON M A Y N A R D

M r . E . Jackson M a y n a r d was b o r n i n East W e y m o u t h , Massachusetts i n 1892. F o l l o w i n g g r a d u a t i o n from N e w M e x i c o


A g r i c u l t u r a l College in 1915, he served t w o years as assistant
professor at t h e same college. F o l l o w i n g a brief p e r i o d in W o r l d
W a r I , h e e a r n e d a n M.S. i n a n i m a l h u s b a n d r y a t Iowa State
U n i v e r s i t y in 1919. T h e n e x t 12 years w e r e s p e n t at F o r t Collins,
C o l o r a d o , as assocaite professor in a n i m a l investigations, C o l o r a d o
State University, d e v o t i n g m o s t of his t i m e to s t u d y i n g t h e feedi n g v a l u e of v a r i o u s sugar b e t t by-products. In 1931, he b e c a m e
h e a d o f t h e a n i m a l h u s b a n d r y d e p a r t m e n t a t U t a h State Agric u l t u r a l College a n d s u b s e q u e n t l y b e c a m e d e a n of t h e schools
of a g r i c u l t u r e a n d forestry, w h e r e he c o n t i n u e d h i s studies on
b e e t by-products. I n 1936 h e j o i n e d t h e G r e a t W e s t e r n Sugar
C o m p a n y a t Billings, M o n t a n a , a n d h a n d l e d t h e sale a n d distrib u t i o n o f t h e c o m p a n y ' s b y - p r o d u c t feeds i n M o n t a n a a n d W y o ming through the Maynard Brokerage Company. He wrote the
b o o k "Beets a n d M e a t " i n 1945 a n d revised t h e e d i t i o n w i t h
K n a u s in 1959. Since 1945, he has b e e n g e n e r a l livestock consultant. He is d i r e c t o r of t h e N a t i o n a l W e s t e r n Stock Show a n d
a m e m b e r of t h e a g r i c u l t u r a l h o n o r societies A l p h a Zeta, G a m m a
Sigma D e l t a a n d P h i K a p p a P h i .
M r . M a y n a r d h a s b e e n a m e m b e r of t h e Society since its
i n c e p t i o n , served as its general p r o g r a m c h a i r m a n for its fifth
g e n e r a l m e e t i n g , was a m e m b e r of t h e advisory c o u n c i l in 19505 1 a n d served o n t h e n o m i n a t i n g c o m m i t t e e i n 1952-53 a n d o t h e r
n u m e r o u s society duties.

VOL. 13, N o .

1, A P R I L

103

1964

Meritorious Service
Award Presented
to

AUSTIN A. ARMER

Austin A. A r m e r received his B.S. degree in electric engineering in 1925 a n d his M.S. d e g r e e in physics in 1926, b o t h from
the University of California at Berkeley. F r o m 1926 he served
as design a n d sales e n g i n e e r for t h e M a g n a v o x C o m p a n y in
several m a j o r U. S. cities. In 1939 he s t a r t e d a c a r e e r in agriculture w i t h t h e a g r i c u l t u r a l e n g i n e e r i n g d e p a r t m e n t o f t h e
University of California at Davis as a research associate. In t h i s
capacity he designed sugar beet h a r v e s t i n g m a c h i n e r y a n d contributed to t h e studies by t h e u n i v e r s i t y on b e e t seed processing
and treating. M a n y of his d e s i g n e d p r i n c i p l e s in b e e t h a r v e s t i n g
e q u i p m e n t are n o w i n general use. I n 1943 h e j o i n e d Spreckels
Sugar C o m p a n y as a g r i c u l t u r a l e n g i n e e r a n d was i n s t r u m e n t a l
in establishing m e c h a n i c a l h a r v e s t i n g in California. He has contributed substantial e n g i n e e r i n g a n d design assistance t o m a n u facturers of e q u i p m e n t to i m p r o v e t h e m e c h a n i z e d p r o d u c t i o n
of sugar beets. In 1951 he served as a t e c h n i c a l c o n s u l t a n t in
Ireland w h e r e he d e v e l o p e d a sugar b e e t harvester to m e e t t h e
special needs of t h a t c o u n t r y . He h a s m o r e r e c e n t l y d e v o t e d his
efforts to i m p r o v e t h e design a n d c o n s t r u c t i o n of s u g a r b e e t
receiving e q u i p m e n t . H e has b e e n e d i t o r o f t h e Spreckels S u g a r
Beet B u l l e t i n since 1947, serves as a p h o t o g r a p h e r a n d a m o t i o n
picture p r o d u c e r , h a v i n g p r o d u c e d o v e r a dozen s o u n d films
relating to a g r i c u l t u r a l subjects. M r . A r m e r is c u r r e n t l y agricultural e n g i n e e r , Spreckels S u g a r C o m p a n y , is past p r e s i d e n t
of the A m e r i c a n Society of S u g a r Beet T e c h n o l o g i s t s a n d past
chairman of t h e Pacific Coast Section, A m e r i c a n Society of Agricultural E n g i n e e r s .

JOURNAL OF T H E A. S. S. B. T.

104

Forty

Year

Veteran

Azvards

GEORGE H. COONS, U n i t e d States D e p a r t m e n t of Agriculture


CHARLES E. CORMANY, Holly Sugar Corporation
C. ELIASON, Stearns-Roger Corporation
LOUIS R. GARCIA, Holly Sugar Corporation
DOUGLAS W. LOVE, Utah-Idaho Sugar Company
H U G H F. MELVIN, Spreckels Sugar Company
J. F. PRICE, American Crystal Sugar Company
J O H N A. R A T E K I N , Holly Sugar Corporation
K U R T SCHREIBER, Manitoba Sugar Company, Limited
P H I L L I P B. S M I T H , T h e Great Western Sugar Company

JOURNAL
of the
American Society of Sugar
Beet Xechnologists
Volume 13
Number 2
July 1964

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P . O . Box 538
F o r t Collins, C o l o r a d o , U. S. A.

Subscription
$4.50
S5.00
$1.25
$1.40

per
per
per
per

year,
year,
copy,
copy,

prices:
domestic
foreign
domestic
foreign

M a d e in the U n i t e d States of America

TABLE OF CONTENTS
Author
T h e fate
juices

of

formaldehyde

in

sugar

Page

beet
A. Carrtithers
J. F. T. Old

Influence of size of fruit a n d seed on germination of a monogerni sugar beet variety

field

105

F. W. Snyder
G. J. Hogaboam

116

Merle G. Payne
L.eRoy Poivers
Grace W. Maag

127

LeRoy
Powers
Merle G. Payne

138

Variation in emergence of sugar beet plants


a t t r i b u t a b l e to different drill units of the
same p l a n t e r

H. L. Bush

151

Evaporator heat transfer coefficients for beet


sugar solution

Allison S. Chang

153

Influence of prolonged association of sugarbeet n e m a t o d e a n d t o m a t o on intensity of


parasitism

Arnold E. Steele

170

D. IV. Robertson
Mildred L. Thornton

177

Levels o f total nitrogen, p o t a s s i u m a n d


sodium in petioles a n d in thin juice of
sugar beets

Associations of levels of
sium, a n d sodium in
juice with weight of
centage sucrose a n d
purity in sugar beets

total nitrogen, potaspetioles a n d in thin


roots per plot, perpercentage a p p a r e n t

Longevity of sugar beet seed

Sucrose d e t e r m i n a t i o n in sugar beets using


p a p e r chromatography a n d spectrophotometry
.

Beet p u l p in all-barley rations

Spiros
M.
Constantinides
C. L. Bedford
185
W. A. Harris

192

The Fate of Formaldehyde in Sugar Beet Juices


A . C A R R U T H E R S AND J . F . T . O I . D F I E L D 1
Received

for publication

February

6, 1964

F o r m a l d e h y d e s o l u t i o n is o n e of t h e m o s t effective a n d convenient r e a g e n t s for c o n t r o l l i n g m i c r o b i o l o g i c a l activity in t h e


beet factory process. T h e total f e r m e n t a t i o n in c o n t i n u o u s diffusion is f r e q u e n t l y less t h a n in b a t t e r y o p e r a t i o n b u t t h e corrosive effects of acid p r o d u c t i o n a r e m o r e a p p a r e n t in c o n t i n u o u s
diffusers as t h e f e r m e n t a t i o n t e n d s to be localized t o w a r d s t h e
juice e n d of the dilTuser. W h e r e a s heavy f o r m a l d e h y d e dosing
was c o m m o n in t h e r e t u r n w a t e r systems of b a t t e r y diffusers,
with t h e a d o p t i o n of c o n t i n u o u s diffusers t h e use of formaldehyde at t h e juice e n d has greatly increased a n d m a y exceed I lb
of formalin ( 4 0 % f o r m a l d e h y d e ) p e r ton of beet. In t e r m s of
a n h y d r o u s f o r m a l d e h y d e , this is e q u i v a l e n t to a b o u t 0.1 p e r 100
sugar, or a b o u t 1% of t h e non-sugars in juice a n d . since m u c h
of the f o r m a l d e h y d e is c a r r i e d forward in t h e r a w j u i c e to process,
the fate of f o r m a l d e h y d e in t h e purification stages is of considerable i m p o r t a n c e .
Determination of formaldehyde
F o r m a l d e h y d e was d e t e r m i n e d c o l o r i m e t r i c a l l v b y t h e m e t h o d
of Xash (1)- w h i c h d e p e n d s on t h e synthesis of d i a c e t v l d i h y r o l u tidine by t h e H a n t z s c h r e a c t i o n b e t w e e n f o r m a l d e h y d e a n d acetvlacetone in t h e presence of excess a m m o n i u m salt. Each test
solution was d i l u t e d to a f o r m a l d e h y d e c o n c e n t r a t i o n in t h e
range 1 to 8 p p m a n d t h e a b s o r p t i o n was m e a s u r e d at 410 m in a
1 cm cell.
Free formaldehyde in raw juice
A l t h o u g h f o r m a l d e h y d e is a very reactive s u b s t a n c e , t h e bacterial effect is n o t sensibly d i m i n s h e d even after several h o u r s
contact w i t h r a w juice. C o n s e q u e n t l y t h e i n h i b i t i v e effect of
formaldehyde can b e t r a n s m i t t e d b y t h e j u i c e f l o w t o relatively
inaccessible p a r t s of t h e c o u n t e r - c u r r e n t diffusion svstetn to an
extent n o t possible w i t h , for e x a m p l e , c h l o r i n e w h i c h is an extremely efficient b a c t e r i c i d e w h e n a d d e d t o r a w juice b u t reacts
so rapidly w i t h juice c o n s t i t u e n t s t h a t after a few m i n u t e s contact no i n h i h i t i v e effect r e m a i n s .
It is t h e r e f o r e a p p a r e n t t h a t t h e m a j o r i t y of t h e f o r m a l d e hyde in t h e j u i c e stream is n o t d e c o m p o s e d by or irreversibly
with juice c o n s t i t u e n t s .
1 Research Laboratories. British Sugar Corporation Limited, T h e Grange, Brameote,
Nottingham. England.
2 Numbers in parentheses refer to literature cited.

106

J O U R N A L OF T H E A. S.

S. B. T.

T h e validity of this d e d u c t i o n was investigated by a d d i t i o n


of known amounts of formaldehyde to raw juice; the samples
w e r e h e a t e d a t 8 0 o C for 3 0 m i n u t e s a n d t h e f o r m a l d e h y d e content was then d e t e r m i n e d by the Nash procedure. A formalin
usage of 1 lb p e r ton of beet is e q u i v a l e n t to an average formaldehyde a d d i t i o n of a b o u t - 0 2 % on juice but, since formalin is
generally applied by i n t e r m i t t e n t shock dosing, the e x p e r i m e n t
was c o n d u c t e d w i t h f o r m a l d e h y d e a d d i t i o n s r a n g i n g from . 0 0 1 %
to 0 . 1 5 % on juice. T h e recovery of formaldehyde as a percentage
of the applied dosage is recorded in F i g u r e 1.

F i g u r e I.Recover)' of formaldehyde from r a w juice.

E v e n a t t h e lowest dosage, 5 0 % o f t h e a p p l i e d f o r m a l d e h y d e
was recovered a n d t h e recovery rose r a p i d l y t o a l m o s t 9 0 % with
a f o r m a l d e h y d e a d d i t i o n o f . 0 2 % o n juice, a n d t o a b o u t 9 8 %
at h i g h e r dose rates. F o r m a l d e h y d e a p p l i e d d u r i n g diffusion is
in c o n t a c t w i t h a m i x t u r e of juice a n d partially e x h a u s t e d cossettes so t h a t t h e possibility arises of c o m b i n a t i o n w i t h p r o t e i n
or other p u l p constituents. On treating mixtures of juice and
p u l p with f o r m a l d e h y d e at a level of 0 . 0 1 % on total l i q u i d s however, t h e recoveries r a n g e d from 7 8 t o 8 3 % w h i c h a r e very similar t o those o b t a i n e d w i t h j u i c e a l o n e .
It is t h e r e b y c o n f i r m e d t h a t t h e m a j o r i t y of t h e formaldeh y d e i n t r o d u c e d i n t o t h e diffuser r e m a i n s in a free or very
lightly b o u n d form.
T h i s c o n c l u s i o n is p e r h a p s r a t h e r u n e x p e c t e d in view of the
c o m m o n s u p p o s i t i o n that t h e bactericidal effect of f o r m a l d e h y d e
may b e d u e t o c o m b i n a t i o n w i t h b a c t e r i a l p r o t e i n . D e s p i t e the
large excess of p r o t e i n n i t r o g e n o v e r a d d e d f o r m a l d e h y d e , very
little if a n y of t h e f o r m a l d e h y d e r e a c t e d w i t h p u l p c o n s t i t u e n t s .

VOL.

13, No. 2, J I I Y

107

1964

T h e slight m e a s u r e d loss of- f o r m a l d e h y d e in j u i c e a p p e a r s


to he real a n d was n o t due to i n t e r f e r e n c e by t h e juice a m i n o
acids in the H a n t z s c h r e a c t i o n since 0 . 0 2 % f o r m a l d e h y d e c o u l d
be recovered q u a n t i t a t i v e l y from s y n t h e t i c s o l u t i o n s c o n t a i n i n g
20 moles of e i t h e r g l u t a m i n e or glycine p e r m o l e of f o r m a l d e h y d e .
Probably a small a m o u n t of t h e f o r m a l d e h y d e c o m b i n e s w i t h
some j u i c e c o n s t i t u e n t s since t h e form of t h e . c u r v e a p p r o x i m a t e s
to that of a second o r d e r r e a c t i o n a n d t h e u n a c c o u n t e d formaldehyde at t h e h i g h dose levels increased with i n c r e a s i n g r a w j u i c e
brix. T h i s loss w o u l d h a v e only a slight effect in r e d u c i n g t h e
amount of formaldehyde carried through with the juice to
process.
Since a m i n o acids can b e d e t e r m i n e d b y t h e S o r e n s e n formol
titration, based o n t h e reaction b e t w e e n excess f o r m a l d e h y d e a n d
amino acid to form an N - m e t h y l e n e d e r i v a t i v e , t h e e x t e n t of t h i s
reaction in juice was investigated. As t h e r e a c t i o n w i t h a m i n o
acids is reversible, t h e H a n t z s c h r e a c t i o n c a n n o t be used to estimate free f o r m a l d e h y d e , since t h e progressive r e m o v a l of formaldehyde leads t o c o m p l e t e recovery. T h e r e a c t i o n was t h e r e f o r e
investigated by p o t e n t i o m e t r i c t i t r a t i o n to pH 8.05 of s o l u t i o n s
c o n t a i n i n g 33 m - m o l e s of glycine p e r l i t r e w i t h f o r m a l d e h y d e
addition i n t h e r a n g e 0.06 t o 2 . 5 % . T h e c o n c e n t r a t i o n o f m e t h ylene d e r i v a t i v e was estimated from t h e t i t r a t i o n a n d t h e concentrations of free a m i n o acid a n d f o r m a l d e h y d e w e r e c a l c u l a t e d
by difference. T h e s e values t o g e t h e r w i t h t h e calculated concentration e q u i l i b r i u m c o n s t a n t s a r e r e c o r d e d i n T a b l e 1 .
Table I.Interaction of formaldehyde and glycine.
per litre).
Formaldehyde
added
20.7
41.2
78.8
158.6
241
320
485
6S0
777

N-methylene
derivative
1.8
5.1
H.2
17.9
21.4
23.7
26.5
28.1
29.0

Residual
formaldehyde
18.9
36.1
67.6
140.7
220
296
459
602
748

(Concentrations in m-moles

Residual
glycine
32.6
29.1
21.4
18.1
11.8
9.3
6.9
4.5
3.1

K
X 103
m-moles- 1
.litres
2.9
4.8
7.7
7.1
8.2
8.6
8.4
10.4
12.5

T o o b t a i n m e a s u r a b l e yields o f t h e m e t h y l e n e d e r i v a t i v e , t h e
e x p e r i m e n t a l c o n c e n t r a t i o n s o f a m i n o acid a n d f o r m a l d e h y d e
were h i g h e r t h a n o c c u r i n factory p r a c t i c e a n d even so. t h e
a m o u n t of m e t h y l e n e d e r i v a t i v e f o r m e d was very low at t h e
lowest f o r m a l d e h y d e a d d i t i o n w h e r e a s very little free a m i n o
acid r e m a i n e d a t t h e highest f o r m a l d e h y d e level. C o n s e q u e n t l y

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JOURNAL OF THE A. S. S. B. T.

t h e e q u i l i b r i u m c o n s t a n t s c a n n o t b e c a l c u l a t e d very precisely
at these two f o r m a l d e h y d e c o n c e n t r a t i o n s b u t nevertheless it
a p p e a r s that t h e e q u i l i b r i u m c o n s t a n t becomes s m a l l e r as t h e form a l d e h y d e c o n t e n t i s r e d u c e d . F o r a n average a m i n o acid conc e n t r a t i o n in j u i c e of 15 m m o l e s p e r litre a n d f o r m a l d e h y d e
c o n c e n t r a t i o n s of e i t h e r . 0 3 % or . 0 0 3 % a K v a l u e of 0.0029
w o u l d c o r r e s p o n d t o c o m b i n a t i o n o f only a b o u t 3 % o f t h e
i o r m a k l e h y d e a n d even if t h e e q u i l i b r i u m c o n s t a n t was as high
as 0.008, o n l y 1 0 % of t h e f o r m a l d e h y d e w o u l d be c o m b i n e d
with a m i n o acids.
It is therefore c o n c l u d e d t h a t t h e g r e a t e r p a r t of t h e formaldehyde i n t r o d u c e d i n t o the juice e n d of the diffuser will be carried
t h r o u g h as free f o r m a l d e h y d e i n t o t h e c a r b o n a t a t i o n system.
Degradation of formaldehyde in carbonatation
T h e effect of l i m i n g a n d c a r b o n a t a t i o n on f o r m a l d e h y d e in
r a w juice was e x a m i n e d by clarification of juice c o n t a i n i n g formaldehyde-C14.
P a r a f o r m a l d e h y d e - C 1 4 (1.16 mg = 100 .C) p l u s inactive
p a r a f o r m a l d e h y d e (4.8 mg) was s u s p e n d e d in 1.1 ml w a t e r a n d
c o n v e r t e d to f o r m a l d e h y d e - C 1 4 by h e a t i n g at 1 0 0 C in a sealed
t u b e for 5 h o u r s . A test e x p e r i m e n t w i t h i n a c t i v e p a r a f o r m a l d e hyde showed t h a t m o r e t h a n 90%, c o n v e r s i o n was o b t a i n e d in
30 minutes u n d e r the experimental conditions.
1 ml (91 c) of the r a d i o a c t i v e f o r m e l d e h y d e s o l u t i o n was
a d d e d to 30 ml of raw juice from t h e l a b o r a t o r y micro-battery
to give a total f o r m a l d e h y d e c o n c e n t r a t i o n of a b o u t 0 . 0 2 % a n d
samples of t h e u n t r e a t e d m i x t u r e w e r e r e m o v e d for c o u n t i n g ,
Since f o r m a l d e h y d e is volatile, a p p r e c i a b l e losses o c c u r r e d if the
radioactive j u i c e was d r i e d directly for c o u n t i n g b u t this loss
was p r e v e n t e d b y c o n v e r t i n g t h e f o r m a l d e h y d e i n t o t h e nonvolatile d i m e d o n e d e r i v a t i v e o n t h e s a m p l e c o u n t i n g p a n . 2 5 l
a l i q u o t s of t h e r a d i o a c t i v e j u i c e w e r e t r a n s f e r r e d to a tared
s a m p l e p a n , 40 g of inactive f o r m a l d e h y d e was a d d e d together
with 100 l of a 1% s o l u t i o n of d i m e d o n e in m e t h a n o l . W a t e r
was a d d e d to give a total v o l u m e of 600 l to fill t h e source
area of t h e s a m p l e p a n a n d t h e s o l u t i o n was e v a p o r a t e d to dryness u n d e r a n infra r e d l a m p . T h e s a m p l e was r e w e i g h e d and
c o u n t e d u s i n g a P h i l i p s e n d w i n d o w G . M . t u b e type 18505 under c o n d i t i o n s of s t a n d a r d g e o m e t r y as r e p o r t e d previously (2)
T h e c o u n t r a t e was c o r r e c t e d for self a b s o r p t i o n to give a calc u l a t e d zero a b s o r p t i o n r a t e of 7580 c o u n t s m i n p e r 25 l of

juice.

Vol. 13, No. 2, July 1964

109

A s a m p l e of t h e r a d i o a c t i v e j u i c e (15 m l ) was h e a t e d to 8 0 C
in a c e n t r i f u g e t u b e in a w a t e r b a t h a n d s t i r r e d w i t h a fine jet
of air. T h e juice was defecated u s i n g d r y l i m e ( 1 . 6 % ) for 5 m i n utes a n d gassed w i t h c a r b o n d i o x i d e from a low p r e s s u r e reservoir
to the first a n d t h e n to the second c a r b o n a t a t i o i i end p o i n t s . T h e
carbonatatioii p r e c i p i t a t e s w e r e r e m o v e d bv c e n t r i f u g a t i o n . A
sample of t h e second c a r b o n a t a t i o i i juice was d r i e d a n d c o u n t e d
as before a n d it was f o u n d t h a t 7 5 % of the initial r a d i o a c t i v i t y
had passed t h r o u g h i n t o t h e second c a r b o n a t a t i o i i juice.
A l i q u o t s o f t h e second c a r b o n a t a t i o n juice w e r e a p p l i e d
to a s t r i p of W h a t m a n X o . 1 filter p a p e r , 50 cm in l e n g t h , a n d
subjected to h i g h voltage e l e c t r o p h o r e s i s at 100 v / c m for 15 m i n utes i n 1 % a m m o n i u m c a r b o n a t e . T h e filter p a p e r was t h e n exposed for 24 h o u r s for a u t o r a d i o g r a p h y with Ilford X-ray I n d u s trial G film. T h e positions of t h e b a n d s of r a d i o a c t i v e p r o d u c t s
revealed on t h e film were r e m a r k e d l y s i m i l a r to those r e p o r t e d
previously (2) for t h e acidic p r o d u c t s p r o d u c e d by a l k a l i n e degradation of fructose or glucose: t h e e x p e r i m e n t was t h e r e f o r e
repeated w i t h o u t f o r m a l d e h y d e b u t s u b s t i t u t i n g u n i f o r m l y labelled fructose-C14 (4.9 mg == 100 c) p l u s i n a c t i v e fructose
(30 mg) . T h e two second c a r b o n a t a t i o i i juices w e r e a p p l i e d side
by side to t h e s a m e p a p e r s t r i p for electrophoresis, a n d a u t o r a d i o graphs o f t h e p r o d u c t s a r e r e p r o d u c e d i n F i g u r e 2 a n d 3 . T o
obtain clear s e p a r a t i o n of t h e p r o d u c t b a n d s , t h e a m o u n t of:
radioactive m a t e r i a l was r e d u c e d to 15 m c for t h e s e p a r a t i o n
shown in F i g u r e 2 w i t h an a u t o r a d i o g r a p h i c e x p o s u r e of 7 days,
while t h e a p p l i c a t i o n was d o u b l e d for F i g u r e 3 to show t h e less
intense p r o d u c t s at t h e e x p e n s e of s o m e o v e r l o a d i n g of t h e m o r e
active b a n d s .
During liming and carbonatatioii both the formaldehyde and
the fructose w e r e c o n v e r t e d a l m o s t exclusively i n t o a n i o n i c products. X o c a t o n i c c o m p o n e n t s c o u l d b e d e t e c t e d i n e i t h e r system
and t h e r e was no d e t e c t a b l e loss of activity from e i t h e r second
carbonatatioii juice on t r e a t m e n t with a s t r o n g c a t i o n e x c h a n g e
resin ( Z e o K a r b 225) w h e r e a s 9 2 % o f t h e activity a r i s i n g from
f o r m a l d e h y d e d e g r a d a t i o n a n d 9 5 % o f t h e activity a r i s i n g from
fructose d e g r a d a t i o n c o u l d be a b s o r b e d on a s t r o n g anion exchanger (De-Acidite F F ) . ^
T h e p r i n c i p a l p r o d u c t b a n d s h a v e b e e n l a b e l l e d 1 to 8, t h e
higher n u m b e r s r e p r e s e n t i n g t h e m o s t m o b i l e o f t h e e l e c t r o p h o r e tic b a n d s . T h e e l e c t r o p h c r o g r a m s h o w n in Figure 3 was c u t i n t o
strips t o isolate t h e p r o d u c t b a n d s , i n c l u d i n g s o m e faint i n t e r m e diate b a n d s which were visible o n t h e o r i g i n a l a u t o r a d i o g r a p h .
The strips w e r e e l u t e d and counted to determine the r e l a t i v e
yields a n d these v a l u e s a r e also r e c o r d e d o n F i g u r e 3 .

110

J O U R N A L OF T H E A. S. S. B. T.

Figures 2 a n d 3.Autoradiographs following electrophoresis of deg r a d a t i o n products of formaldehyde C14 a n d fructose C14.

At t h e p r e s e n t stage of t h e i n v e s t i g a t i o n , t h e fructose degradation products have been examined in m o r e detail than the
p r o d u c t s from f o r m a l d e h y d e a n d only a few of t h e l a t t e r have
b e e n positively identified, b u t from e l e c t r o p h o r e t i c a n d c h r o m a t o g r a p h i c s e p a r a t i o n s it a p p e a r s t h a t t h e m a j o r i t y of t h e b a n d s
consist of t h e same acids, or closely r e l a t e d isomeric acids, regardless of w h e t h e r t h e source was f o r m a l d e h y d e or fructose. T h e
relative p r o p o r t i o n s of t h e p r o d u c t s a r e clearly different w i t h the
different sources a n d it is p r o b a b l e that, as w i t h fructose, the
p r o p o r t i o n s of t h e f o r m a l d e h y d e p r o d u c t s will vary a c c o r d i n g
t o t h e initial c o n c e n t r a t i o n a n d t h e d e f e c a t i o n c o n d i t i o n s .
B a n d 8 consists of formic acid a n d was o b t a i n e d in h i g h e r
yield from f o r m a l d e h y d e t h a n from fructose. Most of t h e rem a i n i n g b a n d s do n o t consist of a single s u b s t a n c e a n d after elu-

VOL. 13, No. 2, JULY

1964

111

tion from t h e e l e c t r o p h e r o g r a m t h e b a n d s can b e f u r t h e r re


solved b y c h r o m a t o g r a p h y . T h e c a r b o n c h a i n l e n g t h increases
with d e c r e a s i n g e l e c t r o p h o r e t i c m o b i l i t y a n d , a l t h o u g h s i m p l e
carboxylic acidvS c o n t r i b u t e t o t h e m o r e m o b i l e b a n d s , t h e princi
pal c o n s t i t u e n t s a r e t h e hydroxy-acids f o r m i n g t h e l o w e r h o m o
logues of t h e s a c c h a r i n i c a c i d series.
T h e acids f o r m i n g b a n d 7 c o n t a i n two c a r b o n a t o m s a n d can
he resolved i n t o s e p a r a t e b a n d s of glycollic a n d acetic a c i d by
c h r o m a t o g r a p h y i n n - p r o p a n o l : .880 a m m o n i a 70:30.
B a n d G consists p r e d o m i n a n t l y of lactic acid w h i c h r e p r e s e n t s
the m a j o r single p r o d u c t of a l k a l i n e d e g r a d a t i o n of fructose a n d
is also o n e of t h e p r i n c i p a l p r o d u c t s of t h e d e g r a d a t i o n of formaldehyde.
T h e faint b a n d 5 in t h e fructose s e p a r a t i o n is p r o b a b l y gly
ceric acid. T h i s b a n d was less clearly resolved from lactic acid
in the f o r m a l d e h y d e s e p a r a t i o n .
B a n d 4 c o n t a i n s 2:4 d i h v d r o x v b u t y r i c acid a n d this b a n d rep
resents a h i g h e r p r o p o r t i o n of t h e p r o d u c t s from f o r m a l d e h y d e
d e g r a d a t i o n t h a n from fructose d e g r a d a t i o n . T h e b a n d i s m o r e
diffuse from t h e f o r m a l d e h y d e t r e a t m e n t a n d m a y c o n t a i n o t h e r
4 c a r b o n a t o m acids.
B a n d 3 is again t h e m o r e p r o m i n e n t in t h e f o r m a l d e h y d e
d e g r a d a t i o n . T h i s b a n d p r o b a b l v c o n t a i n s p e n t o s a c c h a r i n i c acids.
B a n d 2 is a p r o m i n e n t p r o d u c t of t h e fructose d e g r a d a t i o n
and in t h e b a n d f r o m this system several of t h e i s o m e r i c glucosaccharinic acids have b e e n i d e n t i f i e d . T h i s b a n d is n o t a m a j o r
p r o d u c t of t h e f o r m a l d e h y d e d e g r a d a t i o n b u t a series of b a n d s
of low i n t e n s i t y o c c u r in this r e g i o n .
B a n d 1 is a l m o s t i m m o b i l e a n d may be d u e e i t h e r to l a c t o n e s
or unreacted carbohydrates. T h i s b a n d does not contain any unreacted f o r m a l d e h y d e a n d n o n e o f t h e o t h e r b a n d s c o u l d b e de
fected by e l e c t r o p h o r e s i s of t h e r a d i o a c t i v e r a w j u i c e before
liming.
C a r b o h y d r a t e s c a n be p r o d u c e d by a l d o l c o n d e n s a t i o n s from
f o r m a l d e h y d e , a n d even i n t h e n i n e t e e n t h c e n t u r y s y r u p s con
t a i n i n g hexoses h a d b e e n p r e p a r e d from p a r a f o r m a l d e h y d e b v
Butleroff. a n d from f o r m a l d e h y d e b y L o e w ; a l t h o u g h t h e overall
yields w e r e low, these syrups c o n t a i n e d -acrose w h i c h was
shown by F i s c h e r to be DL-fructose.
I t was t h e r e f o r e t o b e e x p e c t e d t h a t s o m e h v d r o x v acids w o u l d
be formed by a l k a l i n e d e g r a d a t i o n of f o r m a l d e h y d e , p a r t i c u l a r l v
as it is n o t essential for t h e c o n d e n s a t i o n to p r o c e e d as far as hexoses before d e g r a d a t i o n is i n i t i a t e d . It is h o w e v e r r a t h e r sur
prising t h a t t h e v a r i o u s acids s h o u l d b e o b t a i n e d s o q u i c k l y a n d

112

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in such good yield, a n d that t h e p r o d u c t s s h o u l d be so s i m i l a r


to those formed by a l k a l i n e d e g r a d a t i o n of fructose.
R a t e of decomposition of formaldehyde d u r i n g l i m i n g
The r a t e of d e c o m p o s i t i o n of f o r m a l d e h y d e was e x a m i n e d
initially by l i m i n g an a q u e o u s s o l u t i o n of f o r m a l d e h y d e at 8 0 C
a n d residual f o r m a l d e h y d e was m e a s u r e d b y t h e N a s h p r o c e d u r e .
Xo facilitate analysis t h e c o n c e n t r a t i o n of f o r m a l d e h y d e was
h i g h e r t h a n in factory practice a n d , as t h e t i m e r e q u i r e d for carb o n a t a t i o n was e x p e c t e d to be l o n g relative to t h e r a t e of decomposition, t h e reaction was s t o p p e d a t t h e r e q u i r e d t i m e intervals
by n e u t r a l i z a t i o n with h y d r o c h l o r i c acid. Because of t h e analogy
with fructose d e g r a d a t i o n , r e d u c i n g m a t e r i a l s w e r e also m e a s u r e d
with t r i p h e n y l t e t r a z o l i u m (3) d u r i n g t h e d e g r a d a t i o n .
O n e m l o f f o r m a l i n ( a p p r o x i m a t e l y 3 3 % f o r m a l d e h y d e ) was
d i l u t e d to 25 ml with w a t e r a n d t h e s o l u t i o n was h e a t e d in a
water b a t h to 8 0 C . Dry l i m e (0.4 g) was a d d e d to t h e s o l u t i o n
at 8 0 C a n d at 1 m i n u t e intervals a 1 ml a l i q u o t was w i t h d r a w n ,
n e u t r a l i z e d i m m e d i a t e l y with 0.7 ml of N h y d r o c h l o r i c acid,
a n d d i l u t e d to 25 ml for d e t e r m i n a t i o n of t h e c o n c e n t r a t i o n s
of f o r m a l d e h y d e a n d r e d u c i n g m a t e r i a l . X h e c o n c e n t r a t i o n s of
t h e latter w e r e expressed relative to fructose s t a n d a r d s .
The e x p e r i m e n t a l results are r e c o r d e d in F i g u r e 4. It was
observed t h a t t h e s o l u t i o n s u d d e n l y d e v e l o p e d a yellow coloration
5 m i n u t e s 10 seconds after l i m i n g .
A slow d e c o m p o s i t i o n of f o r m a l d e h y d e o c c u r r e d d u r i n g the
first 4 m i n u t e s after l i m i n g b u t t h e r e a f t e r t h e f o r m a l d e h y d e was
rapidly d e c o m p o s e d , t h e c o n c e n t r a t i o n falling t o z e r o w i t h a n

TIME (MINUTES)

Figure 4.Degradation

of

formaldehyde during liming

(no

additives).

VOL.

13,

No.

2, JULY

196-1

113

almost e q u i v a l e n t p r o d u c t i o n o f r e d u c i n g m a t e r i a l . T h e r e d u c i n g
material was n o t stable a n d color was p r o d u c e d soon after t h e
concentration o f r e d u c i n g m a t e r i a l b e c a m e significant. T h e s u b sequent d e c o m p o s i t i o n of t h e r e d u c i n g m a t e r i a l followed a c u r v e
which was a p r o l o n g a t i o n of t h e i n i t i a l f o r m a l d e h y d e d e c o m p o s i tion.
T h e slope o f t h e f o r m a l d e h y d e d e c o m p o s i t i o n c u r v e does n o t
follow any s i m p l e 1st or 2 n d o r d e r r e a c t i o n a n d some o t h e r species
must be involved to i n i t i a t e t h e r a p i d stage of d e c o m p o s i t i o n .
Possibilities for such a " s t a r t e r " c o u l d i n c l u d e m a n y of t h e acidic
products d e t e c t e d in the r a d i o c h e m i c a l e x p e r i m e n t s b u t s o m e of
the m o r e reactive non-acidic p r o d u c t s of r e d u c i n g sugar degradation s e e m e d m o r e p r o b a b l e . T h e e x p e r i m e n t was t h e r e f o r e
repeated in t h e presence of a d d e d g l y c e r a l d e h y d e at an i n i t i a l
concentration of 0 . 0 0 S o . G l y c e r a l d e h y d e is n o t stable u n d e r alkaline c o n d i t i o n s a n d is in e q u i l i b r i u m w i t h d i h y r o x y a c e t o n e w h i c h
has been t e n t a t i v e l y i d e n t i f i e d in low yield as a p r o d u c t of hexose
degradation. T h e d e c o m p o s i t i o n o f t h e f o r m a l d e h y d e , t h e production a n d s u b s e q u e n t d e g r a d a t i o n o f r e d u c i n g m a t e r i a l a n d
the color p r o d u c t i o n followed a s i m i l a r p a t t e r n w i t h glyceraldehyde a d d i t i o n , as shown in F i g u r e 5, b u t t h e r e a c t i o n rates w e r e
greatly increased; t h e f o r m a l d e h y d e c o n c e n t r a t i o n fell to zero
in a b o u t \l/2 m i n u t e s a n d t h e r e d u c i n g m a t e r i a l was d e c o m p o s e d
more rapidly.
T h e c o n c e n t r a t i o n o f r e d u c i n g m a t e r i a l does n o t fall e x p o n entially to zero s h o w i n g that e i t h e r t h e initial r e d u c i n g m a t e r i a l
is a m i x t u r e of different substances, or t h a t s o m e of t h e p r o d u c t s

Figure 5.Degradation of
yccraldchyde starter 0,008%.

formaldehyde

during

liming

(effect of


114

JOURNAL OF THE A. S. S. B. T

of t h e intial d e g r a d a t i o n of r e d u c i n g m a t e r i a l a r e also reducing.


A s i m i l a r t y p e of c u r v e is o b t a i n e d on m e a s u r i n g t h e r a t e of
c h a n g e of r e d u c i n g m a t e r i a l d u r i n g a l k a l i n e d e g r a d a t i o n of fructose or glucose.
P r o b a b l y m a n y o t h e r u n s t a b l e a l d e h y d e s a n d k e t o n e s will
accelerate t h e a l k a l i n e d e g r a d a t i o n of f o r m a l d e h y d e a n d it was
f o u n d t h a t a d d i t i o n of e v e n a m i n u t e a m o u n t of fructose
( 0 . 0 0 4 % ) was sufficient to r e d u c e t h e t i m e of c o m p l e t e formaldeh y d e d e c o m p o s i t i o n to a b o u t 2 1/2 m i n u t e s .
T h e d e g r a d a t i o n o f f o r m a l d e h y d e a t t h e l o w e r concentrations
realized in factory p r a c t i c e was also e x a m i n e d in s y n t h e t i c solutions. All t h e s o l u t i o n s w e r e t r e a t e d w i t h 1.6% l i m e at 80C
a n d t h e d e g r a d a t i o n rates for 0 . 0 2 1 % f o r m a l d e h y d e w i t h o u t additives, w i t h 1 0 % sucrose o r w i t h 0 . 0 0 7 % fructose a n d w i t h 0.12%
fructose a r e r e c o r d e d in F i g u r e 6. At this low c o n c e n t r a t i o n it
was n o t possible to d e t e c t color p r o d u c t i o n w h i c h c o u l d be attributed to the formaldehyde degradation.

Figure 6.Degradation of formaldehyde at low concentration (effect of


fructose and sucrose).
At c o n c e n t r a t i o n s s i m i l a r to those in r a w j u i c e , t h e rate of
a l k a l i n e d e g r a d a t i o n of f o r m a l d e h y d e in a q u e o u s s o l u t i o n without
a d d i t i v e s was very slow; less t h a n 5% b e i n g d e c o m p o s e d in 10
m i n u t e s . In 1 0 % sucrose s o l u t i o n h o w e v e r t h e r e a c t i o n rate was
greatly i n c r e a s e d : this increase may be d u e to t h e g r e a t e r solubility of l i m e in sucrose s o l u t i o n , or to t h e p r e s e n c e of trace imp u r i t i e s i n t h e sucrose w h i c h i n i t i a t e t h e c h a i n r e a c t i o n o r perhaps
to a l k a l i n e d e g r a d a t i o n of t h e sucrose itself to yield s u c h starters.
T h e increased r e a c t i o n r a t e was n o t d u e to traces of fructose in
t h e s u g a r since t h e c o n c e n t r a t i o n of r e d u c i n g sugars in solution
was- less t h a n 0 . 0 0 0 3 % a n d w i t h o u t t h e sucrose t h i s a m o u n t of

VOL. 13, No, 2, JULY 1964

115

fructose d i d n o t m e a s u r a b l y increase t h e r e a c t i o n r a t e . E v e n without sucrose h o w e v e r . 0 . 0 0 7 % fructose b r o u g h t a b o u t a very r a p i d


d e g r a d a t i o n o f t h e f o r m a l d e h y d e a n d w h e n t h e fructose concentration was increased to a b o u t that of total r e d u c i n g sugars in
raw j u i c e , a l m o s t c o m p l e t e d e s t r u c t i o n of t h e f o r m a l d e h y d e occ u r r e d w i t h i n 1 m i n u t e of l i m i n g .
A l t h o u g h t h e r e m a y well be m a n y c o m p o u n d s in raw juice
capable of i n i t i a t i n g t h e c h a i n r e a c t i o n , it is not nccessarv to invoke substances o t h e r t h a n sucrose a n d r e d u c i n g sugars in juice
in o r d e r to e x p l a i n t h e c o m p l e t e d e g r a d a t i o n of f o r m a l d e h y d e
during liming and carbonatation.
Conclusions
1. W h e n f o r m a l d e h y d e is i n t r o d u c e d t o w a r d s t h e h e a d e n d
of the diffuser. very little if any of t h e f o r m a l d e h y d e reacts with
p u l p c o n s t i t u e n t s . O n l y small a m o u n t s c o m b i n e irreversibly with
juice c o n s t i t u e n t s o r c o m b i n e i n t h e r e v e r s i b l e c o n d e n s a t i o n with
a m i n o acids a n d t h e g r e a t e r p a r t is c a r r i e d f o r w a r d as free form a l d e h y d e i n t o t h e r a w j u i c e t o process.
2. T h e formaldehyde is rapidly and completely decomposed
d u r i n g l i m i n g . A b o u t 7 0 % o f t h e f o r m a l d e h y d e a d d e d t o raw
juice c o u l d b e a c c o u n t e d for a s a c i d i c p r o d u c t s i n second c a r b o n atation j u i c e .
3 . T h e acidic p r o d u c t s can b e s e p a r a t e d b y e l e c t r o p h o r e s i s
into a series of p r o d u c t s i n c r e a s i n g in c a r b o n c h a i n l e n g t h w i t h
decreasing m o b i l i t y . T h e p r o d u c t s a r e very s i m i l a r t o t h e p r o d u c t s
of a l k a l i n e d e g r a d a t i o n of fructose, c o n s i s t i n g of s i m p l e c a r b o x ylic acids a n d t h e l o w e r h o m o l o g u e s of t h e s a c c h a i i n i c acid
series.
4. F o r m a l d e h y d e a l o n e is c o m p a r a t i v e l y stable in a l k a l i n e
solution. In t h e p r e s e n c e of d e g r a d a t i o n p r o d u c t s of f o r m a l d e h y d e
itself, or of fructose or of s i m p l e u n s t a b l e a l d e h y d e s , t h e formaldehyde is r a p i d l y d e c o m p o s e d to g i v e an a l m o s t e q u i v a l e n t yield
of r e d u c i n g m a t e r i a l . T h i s r e d u c i n g m a t e r i a l is u n s t a b l e in alkaline s o l u t i o n a n d d e c o m p o s e s t o yield t h e acidic m a t e r i a l t o g e t h e r
with small a m o u n t of y e l l o w p r o d u c t s .

Influence of Size of Fruit and Seed on Germination


of A Monogerm Sugar Beet Variety 1
G. J. HOGABOAM- AND F. W. S.NYDER
Received

for

publication

November

29, 1963

T h e a d v e n t of t h e m o n o g e r m sugar beet has accelerated the


t r e n d toward completely m e c h a n i z e d p r o d u c t i o n . H o w e v e r , mechanized c u l t u r a l o p e r a t i o n s in the s p r i n g d e p e n d on the successful
solution of such p r o b l e m s as precision space-planting, adequate
e m e r g e n c e , weed control, a n d disease c o n t r o l . A sugar beet variety
that p r o d u c e d u n i f o r m l y sized a n d shaped fruits 3 , each containing
a single seed that g e r m i n a t e s rapidly, w o u l d seem to be t h e most
desirable for c o m p l e t e m e c h a n i z a t i o n .

In a study of b u l k e d fruits (seedballs) of m u i t i g e r m varieties


Price a n d Carsner (4)' observed that seeds g e r m i n a t e d m o r e rapidly a n d completely a n d grew m o r e vigorously from seedballs rem a i n i n g on 4- a n d 3.5-mm screens t h a n those from seedballs passing t h r o u g h a 3 m m . b u t r e m a i n i n g on a 2.5 mm screen. In contrast. U s t i m e n k o (9) , in studying b u l k e d fruits of monogerm
strains, observed that seeds in fruits with a high weight per 1,000
fruits g e r m i n a t e d slower a n d less completely t h a n those in fruits
having less weight. In o n e m u i t i g e r m a n d two m o n o g e r m varieties he found that t h e large fruits c o n t a i n e d , b o t h absolutely
a n d relatively, a larger mass of p e r i c a r p than the smaller fruits.
He ascribed t h e slower g e r m i n a t i o n to t h e relatively greater
a m o u n t of pericarp. Decorticating the fruits hastened emergence.
W a t e r a b s o r p t i o n by decorticated fruits was c o m p l e t e after 40
to 50 hours, while non-decorticated fruits r e q u i r e d 80 to 90 hours
O n l y one-half to o n e - t h i r d as m u c h water was i m b i b e d by decorticated fruits.
Savitsky (5) observed that t h e weight of seeds increased
in p r o p o r t i o n to t h e size of t h e m o n o g e r m fruits. Savitsky et al
(6) r e p o r t e d that seeds in large fruits of i n b r e d m o n o g e r m lines
g e r m i n a t e d m o r e slowly t h a n those in small fruits.
Sedlmayr (7) , u s i n g seedballs of a u n i f o r m size-class, demonstiated that speed of g e r m i n a t i o n of the m u i t i g e r m sugar beet
variety US 401 is h e r i t a b l e . Differences in speed of germination

1 Cooperative investigations of the Crops Research Division. Agricultural Research


Service. U. S. Department of Agriculture, and the Michigan Agricultural Experiment Station. Approved for publication as Journal article 3264 .Michigan Agricultural Experiment
Station.
2 Research Agonomist and Plant Physiologist, respectively. Crops Research Division,
Agricultural Research Service, U. S. Department of Agriculture. East Lansing. Michigan
3 A fruit is defined as an individual structure which contains one or more true
seeds. In this paper, the term seed refers only to the true seed contained within the fruit.
4 Numbers in parentheses refer to literature cited.

VoL.

13, N o . 2, J U L Y 1964

117

were d u e m a i n l y to t h e physical a n d chemical n a t u r e of t h e maternal tissues of t h e fruit which s u r r o u n d the t r u e seed (7, 8).
P o x t a t o r a n d H e l m e r i c k (1) observed that large-fruited p a r e n t s
produce p r o g e n i e s w i t h large fruits.
Before b r e e d i n g for u n i f o r m fruits t h e b r e e d e r s h o u l d k n o w
whether t h e r e is a most d e s i r a b l e size a n d w h a t o t h e r characteristics of fruit a n d seed may i n f l u e n c e his choice of size. Since previous work suggested that t h e size of fruit influences g e r m i n a t i o n
performance, this research was u n d e r t a k e n to s e p a r a t e t h e effect
of fruit size from t h e effect of seed size on g e r m i n a t i o n w h e n
the seed is g e r m i n a t e d w i t h i n t h e intact fruit.
Methods and Materials
T h e d a t a w e r e collected on an i n d i v i d u a l p l a n t basis as a
control on v a r i a b i l i t y that exists b e t w e e n p l a n t s for q u a l i t y a n d
concentration of i n h i b i t o r s in t h e i r f r u t s as well as t h e m e c h a n i cal p r o p e r t i e s of t h e fruit affecting g e r m i n a t i o n . S i m p l e a n d
partial c o r r e l a t i o n s were t h e n used to e v a l u a t e the d a t a for w i t h i n
plants, for b e t w e e n p l a n t s , a n d for t h e variety.
In the e x p e r i m e n t a l p r o c e d u r e s (3) e m p l o y e d to i m p r o v e
the m o n o g e r m variety SP 5832-0. each root of 141 selections was
planted in a soil b e d in t h e g r e e n h o u s e at East L a n s i n g . Michigan, in D e c e m b e r 1958. M a t u r e fruits w e r e h a r v e s t e d separately
from each of 131 p l a n t s . All fruits w e r e t r e a t e d w i t h fungicide b u t
received no o t h e r t r e a t m e n t . Eleven to 22 g r a m s of fruits from
each of 19 p l a n t s w e r e sized for d i a m e t e r a n d t h e n for thickness.
T h e p e r c e n t a g e s by w e i g h t of fruits w i t h a single o v a r i a n cavity
(multiple o v a r i a n cavities d i s c a r d e d ) in each of t h e size classes
were listed by p l a n t source. In this study, the 12/64-inch diameter class refers to fruits that passed t h r o u g h a 13 (54-inch r o u n d hole screen a n d r e m a i n e d o n a n l l / 6 4 - i n c h screen. T h e 9 / 6 4 - i n c h
thickness class refers to fruits t h a t passed t h r o u g h a 1 0 / 6 4 - i n c h
slotted screen a n d r e m a i n e d on an 8 64-inch screen.
An i n d e x of u n i f o r m i t y was c a l c u l a t e d b o t h for d i a m e t e r a n d
thickness of fruit. T h e i n d e x e s were c a l c u l a t e d from the fruitsize d i s t r i b u t i o n d a t a as follows: T h e m o d a l class was assigned a
value of zero, t h e class on e i t h e r side of t h e m o d e was assigned
a value of 0 . 1 , the second classes from t h e m o d e w e r e assigned
a value of 0.2, t h e t h i r d a' v a l u e of 0.4, a n d t h e f o u r t h a v a l u e
of 0.8. T h e assigned class v a l u e was m u l t i p l i e d by t h e p e r c e n t a g e
by weight of fruits in t h a t class a n d t h e s u m of these p r o d u c t s
is the i n d e x of u n i f o r m i t y . T h u s , t h e s m a l l e r t h e values of t h e
index, t h e m o r e u n i f o r m t h e size of fruits.
V o l u m e of fruit was c a l c u l a t e d since it offered t h e possibility
of b e t t e r r e l a t i n g fruit size to g e r m i n a t i o n response t h a n e i t h e r

118

JOURNAL OF THE A. S. S. B. T.

d i a m e t e r or thickness. T h e f o r m u l a for the v o l u m e of an oblate


spheroid (V 4/3 a 2 b) a p p e a r e d to be the best o n e for an initial
estimation of the v o l u m e p e r fruit w h e r e " a " is the d i a m e t e r and
"b" is the thickness. T h e actual v o l u m e s were d e t e r m i n e d for
each size class by v o l u m e t r i c d i s p l a c e m e n t in S t o d d a r d ' s solvent 5 .
T h e fruits used in this d e t e r m i n a t i o n were from the same variety
b u t n o t necessarily from t h e same 19 p l a n t s as used for radiographing.
Data on t h e r e l a t i o n of fruit d i a m e t e r a n d fruit thickness
to g e r m i n a t i o n t i m e were o b t a i n e d for each p l a n t .
An X-ray t e c h n i q u e (2) was used to d e t e r m i n e the diameter
of the seed while intact w i t h i n its fruit. Samples of fruits from
each of 19 plants, sized for d i a m e t e r a n d thickness, were placed
on sticky c e l l o p h a n e tape by p l a n t source a n d by size class for
X-ray. T h u s it was possible to o b t a i n data on an i n d i v i d u a l basis.
A total of 6,782 single-cavity fruits, r a n g i n g from a m i n i m u m
of 228 to a m a x i m u m of 480 fruits per p l a n t , was radiographed.
In a d d i t i o n to the d i a m e t e r of t h e seed, the n u m b e r of fruits
c o n t a i n i n g no d e v e l o p e d seed a n d those c o n t a i n i n g 1, 2, 3, or 4
seeds in the single o v a r i a n cavity were recorded. T h e identity
of each fruit was m a i n t a i n e d on the g e r m i n a t i o n b l o t t e r a n d the
t i m e r e q u i r e d to g e r m i n a t e was r e c o r d e d for each seed. Weighted
averages for seed size w e r e used to calculate the average seed
size for a plant, since fruit size classes that w e r e radiographed
were n o t always taken in p r o p o r t i o n a l a m o u n t s .
T h e data o b t a i n e d by i n d i v i d u a l fruits were e v a l u a t e d for
inter-relationships by simple a n d partial correlations. T h e data
consisted of: (A) g e r m i n a t i o n t i m e ; ( B ) fruit d i a m e t e r ; (C)
fruit thickness, (D) seed d i a m e t e r ; (E) n u m b e r of seeds in ovarian cavity; a n d (F) v o l u m e of the fruit. C o r r e l a t i o n coefficients
were calculated for w i t h i n - p l a n t samples, for b e t w e e n - p l a n t samples, a n d tor all samples c o m b i n e d to r e p r e s e n t the variety.
Results
Fruit

size

T h e fruit d i a m e t e r a n d thickness d a t a ( T a b l e 1) for this monogerm variety revealed d i a m e t e r s r a n g i n g b e t w e e n 6/64 a n d 17/64


a n d thicknesses from 4 / 6 4 to 11/'64 of an inch. Each p l a n t had at
least a few fruits in t h e smallest d i a m e t e r a n d thickness class
while only a b o u t half of the plants had fruits in t h e largest diam e t e r a n d thickness class. Plants differed in u n i f o r m i t y of fruit
size.
5
Trade names are mentioned for identification only and do not constitute recommen-'
dation by United States Department of Agriculture.

VoL

13, N o . 2, J U L Y 1964

121

T h e calculated v o l u m e exceeded the actual v o l u m e p e r fruit,


however, w h e n the calculated v o l u m e was s u b s t i t u t e d in the
formula Y = 0.598x + 0.0013. this corrected v o l u m e closely approximated the actual v o l u m e p e r fruit as m e a s u r e d by volumetric d i s p l a c e m e n t .
T h e shape of fruits varied by p l a n t s from o n e in w h i c h t h e
fruit, thickness was 56% of t h e d i a m e t e r to o n e in w h i c h it was
72% ( T a b l e 2).
Seed size ayid numbers of seeds per cavity
T h e d i a m e t e r of the seed generally varied directly w i t h t h e
diameter of t h e fruit a n d also fruit thickness. T h i s t r e n d a n d its
exceptions may be n o t e d in the s u m m a r y of fruit a n d seed characteristics in T a b l e 2. T h e s m a l l e r fruits t e n d to c o n t a i n relativelv
larger seeds as s h o w n by the "seed d i a m e t e r as % fruit d i a m e t e r "
column.

]22

JOURNAL

OF THE A. S. S. B. T.

T h e data on t h e c o n t e n t of the o v a r i a n cavities, o b t a i n e d by


m e a n s of the X-ray t e c h n i q u e , are s u m m a r i z e d in Table 3. Individual p l a n t s of this m o n o g e r m variety varied greatly in t h e percentage of fruits c o n t a i n i n g a single seed. Six of the p l a n t s had
m o r e t h a n 10 p e r c e n t seedless fruits (aborted seeds) . F o u r of the
p l a n t s had 10 p e r c e n t or m o r e of t h e i r fruits c o n t a i n i n g two or
m e r e seeds. M o r e than 23 p e r c e n t of the fruits of p l a n t 41 contained m u l t i p l e seeds. Of t h e fruits in t h e 16.. 24 of an inch diam e t e r si/e-class, m o r e t h a n 8 % w e r e seedless.
Speed
of
germination
T h e g e r m i n a t i o n data were used t o calculate the a m o u n t o f
delay in g e r m i n a t i o n that can be expected from an increase in
fruit d i a m e t e r or an increase in fruit thickness. W h e n fruit thickncss was m a i n t a i n e d b e t w e e n (S\A to 8 64th of an inch, t h e time
to a t t a i n 8 0 % g e r m i n a t i o n was a p p r o x i m a t e l y a day l o n g e r for
each increase of 2 64th of an inch in fruit d i a m e t e r . W h e n the
fruit d i a m e t e r was m a i n t a i n e d at e i t h e r 9 to 11 64th or 11 to
13 64th of an inch, an extra clav was r e q u i r e d to a t t a i n 80%
t e r m i n a t i o n by increasing the thickness class from 5 to 6 1/2 to
6i <, to 8 G4th. Except for t h e smallest seeds a n d fruits, g e r m i n a tion t i m e correlated positively with fruit v o l u m e ( T a b l e 4).
Of the 6,782 single-cavity fruits t h a t were r a d i o g r a p h e d , 2.5%
n e v ? sacrificed to era in e x p e r i e n c e in r e a d i n g the r a d i o g r a p h s and
1 0 . 2 % c o n t a i n e d no d e v e l o p e d seeds. Of t h e r e m a i n i n g fruits,
9 7 . 6 % g e r m i n a t e d w i t h i r the 25-day l i m i t of t h e experiment,
1.9% h a d seeds that were fully d e v e l o p e d a n d a p p e a r e d as though

Expressed as cubic centimeters x 10,000.

VoL

13,

No.

2,

JULY

1964

123

they should have g e r m i n a t e d , a n d 0 . 5 % h a d seeds that a p p e a r e d


incompletely d e v e l o p e d a n d were non-viable.
Correlation
studies
T h e w i t h i n - p l a n t c o r r e l a t i o n s w e r e calculated for each p l a n t ,
but only t h e e x t r e m e s a n d the m e a n s of these d a t a are given in.
Table 5. The b e t w e e n - p l a n t a n d within-variety c o r r e l a t i o n s a r e
also listed.
Table 5.Correlation coefficients between various fruit dimensions, seed diameter, and germination
time for samples from 19 plants of monogerm s S .ir beet variety SP 5832-0.

* Significant at 5% level and * * at 1% level.

W i t h i n - p l a n t c o r r e l a t i o n s for g e r m i n a t i o n t i m e versus fruit


size [ d i a m e t e r ( A B ) , thickness ( A C ) , o r v o l u m e (AF)] a n d
also seed d i a m e t e r (AD) revealed 10 p l a n t s w i t h significant
positive c o r r e l a t i o n s , two w i t h no c o r r e l a t i o n ( n u m b e r s 2 a n d
60), a n d o n e w i t h a n e g a t i v e c o r r e l a t i o n . T h e e x t r e m e n e g a t i v e
within-plant c o r r e l a t i o n s of c h a r a c t e r s with g e r m i n a t i o n t i m e
( F a b l e 5) w e r e o b t a i n e d from p l a n t 48 w h i l e the e x t r e m e positive ones w e r e from p l a n t 110.
In all cases, t h e r e was a significant positive c o r r e l a t i o n between fruit d i a m e t e r a n d fruit thickness ( B C ) , fruit v o l u m e
(BF), a n d seed d i a m e t e r ( B D ) . F r u i t thickness was positively
correlated, w i t h seed d i a m e t e r (CD) a n d fruit v o l u m e was positively c o r r e l a t e d w i t h seed d i a m e t e r ( F D ) . S i m p l e c o r r e l a t i o n s
between seed d i a m e t e r a n d g e r m i n a t i o n t i m e (AD) a r e mis-

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JOURNAL OF THE A. S. S. B. T.

l e n d i n g d u e t o t h e h i g h c o r r e l a t i o n o f s e e d d i a m e t e r ( D ) with
f r u i t size c h a r a c t e r i s t i c s ( B , C , & F ) . P a r t i a l c o r r e l a t i o n s o f seed
d i a m e t e r w i t h g e r m i n a t i o n t i m e , b y r e m o v i n g t h e e f f e c t o f fruit
v o l u m e ( A D . F ) , r e v e a l e d f o u r s i g n i f i c a n t c o r r e l a t i o n s within
p l a n t s ( t h r e e w e r e n e g a t i v e a n d o n e p o s i t i v e ) . I n a l l c a s e s their
v a l u e s w e r e less t h a n + 0 . 2 9 . T h e b e t w e e n - p l a n t a n d w i t h i n - v a r i e t y
p a r t i a l c o r r e l a t i o n s f o r t h e s e t h r e e c h a r a c t e r s w e r e n o t significant.
T h e w i t h i n - p l a n t c o r r e l a t i o n s f o r f r u i t d i a m e t e r v e r s u s seedsper-cavity (BE) were n o t o b t a i n e d from t h e electronic computer,
b u t f o r f r u i t t h i c k n e s s v e r s u s s e e d s - p e r - c a v i t y ( C E ) t h e values
r a n g e d f r o m 0 . 0 0 t o + O . 3 6 a n d f o r f r u i t v o l u m e v e r s u s seedsper-cavity ( F E ) f r o m +0.01 to +0.49. N o n e of t h e betweenp l a n t c o r r e l a t i o n s o f s e e d s - p e r - c a v i t y w i t h f r u i t d i a m e t e r (BE),
fruit thickness (CE), or fruit v o l u m e
(FE)
w e r e significant.
T h e w i t h i n - v a r i e t v c o r r e l a t i o n o f s e e d s - p e r - c a v i t y w i t h f r u i t volu m e ( F E ) was h i g h l y s i g n i f i c a n t a t + 0 . 2 2 .
Discussion
T h e d a t a c l e a r l v r e v e a l a s i g n i f i c a n t p o s i t i v e r e l a t i o n s h i p bet w e e n f r u i t size ( d i a m e t e r , t h i c k n e s s , o r v o l u m e ) a n d t h e time
r e q u i r e d f o r g e r m i n a t i o n . O f t h e f r u i t size c h a r a c t e r s , volume
a p p e a r s r o b e t h e b e s t f o r c o r r e l a t i o n s t u d i e s b e c a u s e i t involves
b o t h d i a m e t e r a n d t h i c k n e s s . I n g e n e r a l , fruit, v o l u m e g a v e the
h i g h e s t c o r r e l a t i o n s w i t h t i m e r e q u i r e d f o r g e r m i n a t i o n . The
s i m p l e c o r r e l a t i o n s b e t w e e n g e r m i n a t i o n t i m e a n d seed diameter
g a v e v a l u e s v e r v s i m i l a r t o t h o s e b e t w e e n g e r m i n a t i o n t i m e and
f r u i t size d u e t o t h e h i g h p o s i t i v e c o r r e l a t i o n s b e t w e e n s e e d size
a n d t h e v a r i o u s f r u i t size a t t r i b u t e s .
W h e n t h e e f f e c t s o f f r u i t v o l u m e w e r e r e m o v e d b v a partial
c o r r e l a t i o n t h e n p l a n t s 4 8 . 59, 60. a n d 1 10 h a d rAD.F v a l u e s of
0 . 2 3 , 0 . 1 8 , 0 . 1 9 a n d + 0 . 2 8 r e s p e c t i v e l y . A l l o t h e r p l a n t had
n o n s i g n i f i c a n t v a l u e s . W h e n c o e f f i c i e n t s o f d e t e r m i n a t i o n were
c a l c u l a t e d , t h e m i n o r r o l e o f s e e d d i a m e t e r per s e i n g e r m i n a t i o n
t i m e was a p p a r e n t .
F r o m t h i s s t u d y i t i s e v i d e n t t h a t f r u i t size, w h e t h e r i t b e
d i a m e t e r , t h i c k n e s s , o r v o l u m e , i s a p o o r i n d i c a t o r o f t h e content
o f t h e o v a r i a n c a v i r y . M a n y f r u i t s 9 6 4 i n c h i n d i a m e t e r o r larger
w e r e f o u n d t o c o n t a i n a b o r t e d s e e d s . M u l t i p l e s e e d s w e r e found
i n b o t h s m a l l a n d l a r g e f r u i t s . C o r r e l a t i o n s b e t w e e n f r u i t volume
a n d s e e d s - p e r - c a v i t y w e r e p o s s i b l e w i t h t h e d a t a f r o m a few o f
t h e p l a n t s . T h e r v a l u e s r a n g e d f r o m + 0 . 0 1 t o + 0 . 4 9 which
w o u l d m e a n t h a t for s o m e p l a n t s t h e r e i s a t e n d e n c y for t h e
l a r g e r f r u i t s t o h a v e m o r e s e e d s p e r c a v i t y b u t f o r o t h e r plants
this t e n d e n c y d o e s n o t exist. T h u s , if a b r e e d e r is g o i n g to se-

Vol. 13, N o . 2, J U L Y 1961

125

lect against m u l t i p l e seeds p e r cavity, he m u s t e x a m i n e a d e q u a t e ly all fruit sizes on a p l a n t r a t h e r t h a n just the larger fruits.
T h e shape of the w h o l e m o n o g e r m fruit was e x a m i n e d by
dividing the fruit thickness by the fruit d i a m e t e r ( T a b l e 2) a n d
by the c o r r e l a t i o n s b e t w e e n fruit d i a m e t e r a n d fruit thickness
( T a b l e 5). Since the e x t r e m e s in the thickness by d i a m e t e r r a t i o
only differed from the m e a n by 12.5% a n d since t h e b e t w e e n plant c o r r e l a t i o n for fruit d i a m e t e r versus fruit thickness was
+0.77, t h e r e a p p e a r s to be only a small c h a n c e to c h a n g e the
shape of fruit of this variety by b r e e d i n g .
If the c o m p o s i t e d a t a a r e considered as r e p r e s e n t i n g t h e variety, t h e n general r e l a t i o n s b e t w e e n fruit size a n d seed d i a m e t e r
may be i n d i c a t e d . T h e larger t h e fruit, t h e slower t h e seed w i t h i n
it g e r m i n a t e d . Seed d i a m e t e r had no significant effect on speed
of g e r m i n a t i o n . F o r any given plant in which viable seeds of
different d i a m e t e r s w e r e c o n t a i n e d in fruits of fixed v o l u m e ,
differences in speed of g e r m i n a t i o n varied by as m u c h as 24
hours, b u t g e n e r a l l y t h e v a r i a t i o n averaged 12 h o u r s or less. In
c o m p a r i s o n , seeds of t h e s a m e d i a m e t e r in fruits of different
sizes, from the same p l a n t a n d p a r t i c u l a r l y from different p l a n t s
( p r e s u m a b l y differing in k i n d s a n d a m o u n t s of i n h i b i t o r s ) , differed in speed of g e r m i n a t i o n by as m u c h as 10 days. E x c e p t for
the m a r k e d l y slower g e r m i n a t i o n of t h e seeds in t h e t w o smallest
fruit v o l u m e classes ( T a b l e 4) which a p p e a r s to be c o n t r o l l e d internally by t h e seeds, speed of g e r m i n a t i o n of seeds in l a r g e r fruits
is r e g u l a t e d m a i n l y by t h e m a t e r n a l tissues of the fruit. G e n e r a l l y
because of c h e m i c a l i n h i b i t o r s in t h e fruit, seeds in situ in fruits
g e r m i n a t e m o r e slowly t h a n seeds r e m o v e d from t h e m . H o w e v e r ,
a s t i m u l a t o r y effect of t h e fruit on speed of g e r m i n a t i o n has b e e n
observed (7, 8 ) . If s a m p l e s of fruits h a v i n g only a s t i m u l a t i n g
action c o u l d b e isolated, t h e n larger fruits p r o b a b l y w o u l d b e
desired.
Summary
Samples of w h o l e fruits from 19 p l a n t s of t h e m o n o g e r m sugar
beet variety SP 5832-0 w e r e sized for d i a m e t e r a n d thickness i n t o
a n u m b e r of size classes. An X-ray t e c h n i q u e was used to e x a m i n e
the c o n t e n t s a n d to d e t e r m i n e the d i a m e t e r of t h e seed w i t h i n
each of 6,782 fruits. Seedless o v a r i a n cavities were f o u n d in 10.2%
of t h e fruits. T w o or m o r e seeds p e r cavity were f o u n d w i t h i n
some of t h e fruits from 1.5 of t h e plants. T h e fruits w e r e placed
on blotters a n d t h e t i m e r e q u i r e d for g e r m i n a t i o n was r e c o r d e d
(in 1/2-day i n c r e m e n t s ) for each seed t h a t g e r m i n a t e d w i t h i n 25
days.

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JOURNAL OF T H E A. S. S. B. T.

F r u i t size, as m e a s u r e d by d i a m e t e r , thickness, or v o l u m e , significantly influenced t h e t i m e r e q u i r e d for g e r m i n a t i o n . Generally seeds in the larger fruits g e r m i n a t e d m o r e slowly. F r u i t
size was a poor i n d i c a t o r of e i t h e r seedless fruits or m u l t i p l e seeded fruits.
Since fruit v o l u m e was highly c o r r e l a t e d w i t h seed diameter,
a partial correlation which r e m o v e d the effects of fruit volume
was used to d e t e r m i n e the r e l a t i o n s h i p b e t w e e n seed d i am et er
a n d g e r m i n a t i o n t i m e . T h e r A D . F v a l u e for t h e variety of 0.03
indicated t h a t seed d i a m e t e r per se h a d little effect on t h e germ i n a t i o n rate. W i t h i n i n d i v i d u a l plants, however, t h e r A D . F
values r a n g e d from 0.23 to +0.28 w h i c h i n d i c a t e d t h a t certain
p o p u l a t i o n s c o u l d be isolated in w h i c h seed d i a m e t e r per se could
influence the g e r m i n a t i o n r a t e .
If g e r m i n a t i o n t i m e (speed of g e r m i n a t i o n ) is to be t h e criterion for selection of m o n o g e r m plants, then, in most cases, large
seeds w o u l d n o t be desirable because of t h e h i g h c o r r e l a t i o n between large seeds a n d large fruits.
Literature Cited
(1) . DOXTATOR, C. W. and R. H. H E L M E R I C K . 1962. Selection for seed size
in monogerm varieties. Am. Soc. Sugar Beet T e c h n o l . J. 12 (3):
268-272.
(2) HOGABOAM, G. J. 1961. R a d i o g r a p h i n g as a m e t h o d of observing some
seed characters in monogerm sugar beet fruits. Am. Soc. Sugar
Beet T e c h n o l . J. 11: 605-609.
(3)

HOGABOAM,

G.

J.,

F.

W.

SNYDER

and

H.

W.

BOCKSTAHLER.

1959.

Se-

lecting for yield a n d sucrose i m p r o v e m e n t of a m o n o g e r m sugar


beet variety. Am. Soc. Sugar Beet T e c h n o l . Eastern Reg. Proc.
10: 21-28.
(4) PRICE. C. a n d E. CARSNER. 1916. Seed size in relation to development and
yield of sugar beets. Am. Soc. Sugar Beet T e c h n o l . Proc. 4: 263-269.
(5) SAVITSKY, V. F. 1954. Relation between the weight of fruit and weight
of germ in mono- a n d multigerm beets. Am. Soc. Sugar Beet Technol. Proc. 8 (2) : 16-22.
(6)

SAVITSKY.

V.

F.,

G.

K.

RYSER.

G.

E.

RUSH

and

C.

P.

PARRISH.

1954.

Inter-relation between weight of seed a n d fruit and utilitarian characters in inbred lines and hybrids of monogerm sugar beets. Am. Soc.
Sugar Beet T e c h n o l . Proc. 8 (2) : 399-403.
(7) SEDLMAYR, T. E. 1960. Inheritance of speed of germination in sugar
beets (Beta vulgaris L.). Doctoral dissertation. Michigan State University.
(8) SNYDER. F. W. 1959. Influence of the seedball on speed of germination
of sugar beet seeds. Am. Soc. Sugar Beet T e c h n o l . J. 10: 513-520.
(9) U S T I M E N K O . S. P. 1957. Effect of pericarp on the sprouting energy of
seeds of monogerm sugar beet. (In Russian) Sakh. Svekla 2 (12) :
24-27.

Levels of Total Nitrogen, Potassium and Sodium in


Petioles and in Thin Juice of Sugar Beets 1,4
MERLE O.

PAYNE2,
Received

LEROY
for

POWERS3

publication

AND G R A C E

January

W.

MAAG2

16. 1964

In 1961 studies w e r e c o n d u c t e d to d e t e r m i n e levels of total


nitrogen, p o t a s s i u m a n d s o d i u m in the petioles as c o m p a r e d with
levels of those chemicals in the t h i n juice of sugar beets (Beta
vulgaris L.). T h e p e t i o l e s a r e a s t r u c t u r e of t h e tops of t h e sugar
beet a n d t h e t h i n juice is p r e p a r e d from t h e r o o t s of t h e same
plant. T h e p r i m a r y p u r p o s e of the study was to d e t e r m i n e w h e t h e r
genotypes differ as to levels of these chemical c o n s t i t u e n t s in the
petioles a n d in t h e t h i n juice a n d , if so, w h e t h e r s o m e g e n o t y p e s
tend to have h i g h e r levels in t h e p e t i o l e s a n d lower levels in t h e
thin j u i c e , w h e r e a s for o t h e r g e n o t y p e s t h e reverse is t r u e . In
other Avoids is t h e r e an i n t e r a c t i o n of g e n o t y p e s a n d m a t e r i a l
analysed (petioles or thin j u i c e ) as regards levels of total n i t r o g e n ,
potassium a n d s o d i u m ? Such i n f o r m a t i o n is of great f u n d a m e n t a l
and practical i m p o r t a n c e to t h e beet s u g a r i n d u s t r y as these
chemicals h a v e b e e n f o u n d to be associated w i t h yield a n d q u a l i t y .
Literature R e v i e w
E m m e r t (3, 4, 5,) 5 w o r k i n g with t o m a t o e s , l e t t u c e a n d cucumbers d e v e l o p e d r a p i d m e t h o d s for e s t i m a t i n g n i t r a t e n i t r o g e n ,
phosphate, a n d p o t a s s i u m i n p l a n t s . H e believed t h a t i n a s m u c h
as the n u t r i e n t s d e r i v e d by a g r o w i n g p l a n t from t h e soil m u s t
enter in s o l u t i o n t h r o u g h t h e stem t h a t t h e c o n c e n t r a t i o n of a
given n u t r i e n t i n this m a t u r e c o n d u c t i v e tissue s h o u l d b e directly
proportional to t h e available s u p p l y of t h e n u t r i e n t in the soil.
Hence, a m e a s u r e of t h e c o n c e n t r a t i o n of n u t r i e n t s in this conductive tissue may be a b e t t e r m e a s u r e of t h e a b i l i t y of t h e soil
to s u p p l y n u t r i e n t s to g l o w i n g p l a n t s t h a n c h e m i c a l tests of t h e
soil itself. Also, he felt that an o p t i m u m c o n t e n t of n u t r i e n t s
1
Cooperative investigations of the Colorado Agricultural Experiment Station, the
Crops Research Division, Agricultural Research Service. U. S. Department of Agriculture, and the Beet Sugar Development Foundation. T h e Colorado State University
gratefully acknowledges financial support from the fames G. Boswell Foundation administered by the Agricultural Research Center of Stanford Research Institute, the National
Institutes of Health, the National Plant Food Institute and contract-research funds [1214-100-4549 ( 3 4 ) ] from the Agricultural Research Service of the U. S. Department
of
Agriculture. Approved bv the Colorado Agricultural Experiment Station for publication
as Scientific Series Article No. 884.
2
Professor of Chemistry and Research Assistant Colorado State University, respectively,
Colorado
State University.
3
Geneticist, Crops Research Division, Agricultural Research Service. U. S. Department4 of Agriculture.
T h e writers arc indebted to R. Ralph Wood of the C.reat Western Sugar Company
for obtaining thin juice samples bv an oxalate method standard with his company and
to the Western Data Processing Center at the University of California at Los Angeles
or 5use of the computing facilities for analysing data. J o b No. 398.
Numbers in parentheses refer to literature cited.

128

JOURNAL OF THE A. S. S. B. T.

in this k i n d of tissue exists for the various stages of g r o w t h of


each k i n d of crop, regardless of t h e k i n d of soil in which the
c r o p is growing. H i s researches s u p p o r t e d these d e d u c t i o n s and
hence seemed to justify the practice of analysing t h e m a t u r e cond u c t i n g tissues of the p l a n t to d e t e r m i n e the ability of the soil
t o p r o v i d e the n u t r i e n t r e q u i r e m e n t s . G a r d n e r a n d Robertson
(6) anaylsed petioles of sugar beets a n d f o u n d it useful in determ i n i n g fertilizer needs as regards n i t r a t e , p h o s p h a t e a n d potassium.
Ulrich (16, 17, 18) a n d T o l m a n a n d J o h n s o n (15) conducted
extensive e x p e r i m e n t s with sugar beets, s t u d y i n g yield a n d quality
as regards fertilizer practices. He f o u n d a negative r e l a t i o n between levels of n i t r a t e n i t r o g e n in t h e petioles a n d percentage
sucrose of the roots. He also established t h e o p t i m u m level of
n i t r a t e n i t r o g e n in t h e petioles as regards p e r c e n t a g e sucrose to
be a p p r o x i m a t e l y 1000 p p m . He f o u n d t h a t once t h e critical
n u t r i e n t level for an e l e m e n t has b e e n established for a crop
t h r o u g h m a n y field e x p e r i m e n t s , p l a n t analysis has the following
a p p l i c a t i o n s : (A) d e t e r m i n a t i o n of t h e k i n d of n u t r i e n t that
m i g h t be deficient in the field; (TV) e s t i m a t i o n of the time of
a p p l i c a t i o n a n d the a m o u n t of fertilizer to a p p l y : (C) aid in
selecting the location of fertilizer e x p e r i m e n t s : a n d (D) aid in
m a i n t e n a n c e of t h e p r o p e r level of soil fertilitv. Also, some beet
sugar c o m p a n i e s have used the t e c h n i q u e s he d e v e l o p e d to determ i n e fertilizer practices and established dates of harvest. Probably o n e of the m o r e i m p o r t a n t c o n t r i b u t i o n s of t h e researches
by Ulrich is the s t i m u l a t i o n of r a t h e r extensive researches on the
effects of fertilizer practices on the yield, q u a l i t y a n d processing
of sugar beets.
R o r a b a u g h a n d N o r m a n (12) f o u n d t h e o r d e r in which
some of t h e c o m m o n beet-sirup i m p u r i t i e s adversely affect crystallization of sugar, d u r i n g factorv processing, to be as follows: (A)
c a r b o n a t e a n d c h l o r i d e salts, (B) a m i n o acids, (C) b e t a i n e and
n o n - n i t r o g e n o u s organic acids, a n d (D) sulfate salts. T h e y found
the c a r b o n a t e s a n d chlorides to be strongly mtlassigenic and to
be p r e s e n t in relatively large c o n c e n t r a t i o n s in t h e beet syrups.
H e n c e , t h e c a r b o n a t e s a n d c h l o r i d e s a c c o u n t e d for a large fraction of the total sugar lost in molasses. T h e v c o n c l u d e that the
most fertile g r o u n d for i m p r o v e m e n t of the crystallization characteristic lies in the e l i m i n a t i o n of c a r b o n a t e a n d c h l o r i d e salts
with l o w e r i n g of p y r o l i d o n e c a r b o x y l i c acid a n d that glutamic
acid is t h e second most likely p o i n t of attack. T h e v f u r t h e r point
o u t that the case against p y r o l i d o n e c a r b o x y l i c acid a n d glutamic
acid is a two-edged o n e , since n o t only a r e they major contribu-

Vol.

13,

No.

2,

JULY

1961

129

tors to sugar loss in molasses, but also d e c o m p o s i t i o n of g l u t a m i n e .


the a m i d e from which they originate, causes processing difficulties
through l o w e r i n g of: buffering capacity of j u i c e a n d l o w e r i n g
alkalinity. T h e r a t h e r extensive studies of C a r r u t h e r s a n d O l d field (2) in general agree with those of R o r a b a u g h a n d N o r m a n
(12). In a d d i t i o n C a r r u t h e r s a n d Oldfield present m e t h o d s of
assessing q u a l i t y that a p p e a r to have considerable m e r i t a n d h e n c e
extensive a p p l i c a t i o n .
H a d d o c k , L i n t o n a n d H u r s t (7) found that n i t r o g e n fertilization a n d n i t r o g e n plant c o m p o s i t i o n are closely associated
with sucrose storage in beet roots a n d sugar recoveries from extract j u i c e . Also, they found t h a t the soluble n i t r o g e n c o n s t i t u e n t s
of the sugar beet roots are highly associated w i t h , if n o t responsible for, variations in p u r i t y a n d sucrose p e r c e n t a g e as well as in
dry m a t t e r percentage. F r o m t h e i r studies t h e p a r t i c u l a r components which a p p e a r to be most highly associated with changes
in quality are t h e g l u t a m i n e a n d a m m o n i a fractions. T h e y believe
the g l u t a m i n e n i t r o g e n to be of greatest signiiicance in q u a l i t y
variation, because of its high association, w i t h q u a l i t y factors, a n d
because the c o n c e n t r a t i o n of this form of n i t r o g e n is ten times
that of a m o m n i a n i t o g e n .
R o u n d s et al. (13) found t h a t t h e n i t r o g e n levels caused
greater variations in the a m o u n t s of nonsugars present in the
beet roots t h a n did the varieties tested. Significant interactions of
varieties X n i t r o g e n fertility levels were found for s o d i u m cont e n t . T h e data p r e s e n t e d indicate that b o t h varieties a n d n i t r o g e n
fertility levels can a p p r e c i a b l y influence t h e a m o u n t of nonsugars in beets. T h e association of total n i t r o g e n a n d n i t r o g e n
c o m p o u n d s with r e d u c e d p u r i t y a n d extraction in the roots was
pronounced. Ryser et al. (14) in a c o m p a r i s o n of harvest dates
found that the sugars from the late harvest were h i g h e r t h a n
from the early harvest while t h e p u r i t i e s were lower. T h e unexpected decrease in p u r i t y was n o t a c c o u n t e d for by an increase
in the level of a m i n o n i t r o g e n . Levels of n i t r a t e n i t r o g e n in the
petioles, a m i n o N in t h e roots, a n d Xa c o n t e n t in t h e roots were
greatly influenced by h i g h n i t r o g e n fertilization, b u t varietal differences were j u s t as striking. In Xa c o n t e n t , a four or fivefold
difference b e t w e e n low Xa types a n d h i g h Xa types was n o t
uncommon.
O w e n et al. (8) found t h a t highest p u r i t y was n o t always
associated with highest p e r c e n t a g e sucrose. They a t t r i b u t e d t h e i r
resuits to be d u e to an i n t e r a c t i o n b e t w e e n g e n o t y p e s (as r e p r e sented by hybrids) a n d locations. P o w e r et al. (9) f o u n d t h a t

130

JOURNAL OF THE A. S. S. B. T

some genotypes m i g h t have a high n i t r a t e n i t r o g e n c o n t e n t in


the petioles a n d low total n i t r o g e n in t h e t h i n j u i c e as compared
with o t h e r genotypes. T h e same was f o u n d to hold t r u e for potassium. Some p o p u l a t i o n s were f o u n d to be h i g h in level of
pota ssium in the petioles a n d low in levels of p o t a s s i u m in the
thin juice as compared with other populations.
Materials and Design of the E x p e r i m e n t
T h e materials used in t h e study are as follows. T h e r e is a
total of 20 p o p u l a t i o n s in the e x p e r i m e n t . O n e is a commercial
variety, 4 are three-way hybrids, each c o m p o s e d of 3 inbreds,
a n d 15 a r e Fi h y b r i d s each c o m p o s e d of 2 i n b r e d s . T h e dates of
harvest are S e p t e m b e r 14, O c t o b e r 3, a n d O c t o b e r 16. T h e exp e r i m e n t was c o n d u c t e d in 1961. T h e characters s t u d i e d are
levels of total n i t r o g e n , p o t a s s i u m a n d s o d i u m in t h e petioles
a n d in the t h i n j u i c e . In t h e petioles the characters are expressed
as m i l l i g r a m s p e r 100 g r a m s a n d in the t h i n j u i c e as milligrams
p e r 100 milliliters of t h i n j u i c e e q u a t e d to a refractive dry substance of 10. T h e t h i n j u i c e was p r e p a r e d by T h e G r e a t Western
Sugar C o m p a n y by an oxalate m e t h o d s t a n d a r d w i t h them
[see (1) ]. In t h e process t h e n i t r a t e n i t r o g e n s are r e m o v e d . Flence
t h e total n i t r o g e n for the t h i n j u i c e does n o t i n c l u d e all the nit r o g e n o u s c o m p o u n d s f o u n d in t h e total n i t r o g e n analysis of the
petioles.
!
T h e design of the e x p e r i m e n t is a split p l o t with populations j
r a n d o m i z e d w i t h i n r e p l i c a t i o n s a n d dates of harvest randomized'
w i t h i n blocks. Each block is c o m p o s e d of 3 d a t e s of harvest andj
each d a t e of harvest has 2 r e p l i c a t i o n s with 20 p o p u l a t i o n s ran-:
d o m i z e d w i t h i n each r e p l i c a t i o n . T h e r e are 5 such blocks. Hence |
the design of the e x p e r i m e n t is a modified r a n d o m i z e d complete!
block.
!
Results
T h e F values calculated from an analysis of variance are listed
in T a b l e 1. A study of this table reveals t h a t t h e r e are significant
differences b e t w e e n p o p u l a t i o n s as regards all chemical characters
for b o t h t h e levels in t h e petioles a n d in t h e t h i n j u i c e . This is
T a b i c 1.The F values calculated from the analyses of variance for levels of total 1
nitrogen,
potassium,
and
sodium
in
the
petioles
and
in
the
thin
juice.
Total nitrogen
Variation
due to
Populations
Dates
P X D
1

Potassium

Petioles

Thin
juice

Petioles

6.85

41.84

48.87

1.22

1.20

1.33

Thin
juice
19.43
1.59

Sodium

Value of F at:

Petioles

Thin
juice

5%

1%

9.62
1.99
1.45

9.36
7.80
1.08

1.60
4.46
1.42

1.92
8.63
1.64

signifies that the error mean square is the larger.

x 52-407 F1
CMS x (52-430 X 52-407) F1
CMS X 52-430 F1
CMS x 52-407 F1
X 52-307 F1i

52-303
52-130
52-130
52-305
52-130

CMS x 52-307 F i ,
x 52-108 Ft
x 51 52D Fi
CMS x 51-520 Fi
x 51-505 Fi

52-305
52-305
52-130
52-305
52-305

CMS x 51-505 fi
C.MS x 51-453 Fi
y 51-310 Fi
CMS x 51-310 Fi
CMS x (52-130 X 54-346) Fi

52-305 CMS
52-305 CMS
52 305 CMS
51-505 x 52
A 50 -3
LSI) ai 5';;.
LSI) at 1";,
Awrajrc

x (52-130 x 54-520) Fi
x 3-1 Fi
x (54-458 X 34) Fi
407 Fi

nitrogen

Petioles

Thin
juice

Mg/100gm

Mg/100gm

Potassium

Sodium

Petioles

Thin
juice

Petioles

Thin
juice

Mg/100gm

Mg/100gm

Mg/100gm

Mg/100gm

1305.0
1370.7
1374.0
1345.0
3470.8

49.2
50.1
49.0
02.1
40.9

]0.2
24.2
20.5
17.0
18.0

00.3
09.!)
05.7
73.4
53.9

40.0
32.3
28.5
34.5
35.9

50.3
43.9
37.2
39.3
45.2

6
7
8
9
10

1489.8
113I.H
I-18S.7
1383.5
1330.8

52.3
44.8
57.0
05.0
41.0

27.8
21.1
13.8
17.9
18.1

02.2
00.4
00.0
05.2
44.7

32.2
30.9
35.3
31.3
32.9

41.8
30.8
30.5
37.8
30.9

11
12
13
14
15

1-131.7
1321.0
1278.2
1315.3
1310.2

45.0
04.9
37.4
42.0
44.5

21.1
10.4
14.2
20.7
20.5

48.2
71.9
41.9
53.1
53.0

30.9
32.0
32.3
32.7
30.4

30.0
39.4
38.0
35.9
37.3

10
17
18
19
20

14 42.0
1185.7
1401.5
1-I8H.2
1531.2

05.2
01.3
00.1
57.4
54.4

22.4
24.7
21.0
17.9
10.4

07.2
57.9
02.1
7'!.2
00.7

30.8
35.4
33.7
39.0
35.9

30.3
42.1
30.8
40.0
55.7

77.8
102.0

3.9
5.1

1.0
2.1

5.7
7.0

2.7

5.5
7.2

M03.5

52.0

20.0

00.0

33.7

39.0

1
2
3
4

1964

52-430
52-305
52-305
52-305
52-430

Entry
number

13. No. 2, JULY

Total
Population, LSD
and average

VOL.

Tabic 2.Means and the least significant differences for levels of total nitrogen, potassium, and sodium in the petioles and in the thin juice.

131

132

JOURNAL OF THE A. S. S. B. T.

n o t t r u e for dates of harvest (with t h e possible e x c e p t i o n of sod i u m in t h e t h i n juice) as the differences n o t e d b e t w e e n dates
of harvest can readily be e x p l a i n e d by chance. T h i s is also true
of the first o r d e r i n t e r a c t i o n of p o p u l a t i o n s X dates of harvest.
It may be c o n c l u d e d that t h e changes in levels of total nitrogen,
potassium a n d s o d i u m when harvested S e p t e m b e r 14, O c t o b e r 3,
a n d O c t o b e r 16 have little if any practical significance. F u r t h e r research is necessary to d e t e r m i n e w h e t h e r the c h a n g e in levels of
sodium for t h e different dates of harvest are o t h e r t h a n chance
deviations, as for the t h i n juice they were significant at the 5%
level, b u t just barely so. F o r that reason the different dates of
harvest will n o t be considered i n d i v i d u a l l y in this article.
T h e m e a n s a n d least significant differences for levels of total
n i t r o g e n , potassium, a n d s o d i u m in the petioles a n d in t h e thin
juice are listed in T a b l e 2. As has b e e n shown by P o w e r s et al.
(11) very little e n v i r o n m e n t a l variability is i n c l u d e d in the
differences b e t w e e n m e a n s of p o p u l a t i o n s , the differences noted
are p r e d o m i n a n t l y genetic. In this article the d a t a in T a b l e 2
have t h e i r greatest interest in the degrees of association between
lev els of a chemical in the petioles a n d the level of the same chemical in the t h i n j u i c e ; a n d the i n t e r a c t i o n s i n v o l v i n g levels of
the chemicals in the petioles as c o m p a r e d with levels of the chemicals in t h e t h i n j u i c e . T h e c o r r e l a t i o n coefficients will be considered first.
Associations
F r o m T a b l e 3 it can be seen that the genetic c o r r e l a t i o n between total n i t r o g e n in the petioles a n d in t h e t h i n j u i c e is 0.29,
b e t w e e n potassium in the petioles a n d in the t h i n j u i c e is 0.06,
a n d finally b e t w e e n s o d i u m in t h e petioles a n d in t h e t h i n juice
is 0.60. H e n c e the greatest p e r c e n t of the variability accounted
Table 3.Correlation coefficients for levels of
sodium in the petioles and in the thin juice.i
Total
nitrogen
Character and
material analysed
Total nitrogen
Petioles
T h i n juice
Potassium
Petioles
T h i n juice
Sodium
Petioles
T h i n juice

Thin
juice
0.29

Pot

total

nitrogen,

assium

potassium
Sodiu

Petioles

Thin
juice

Petioles

Thin
juice

0.20
0.03

0.22
0.77

0.38
_0.01

0.43
0.13

0.06

_0.44
0.21

_0.21
0.40

T h e approximate value of r at the 5% level

0.60

13, No.

2, JULY

133

1964

for by the c o r r e l a t i o n of a chemical in the petioles a n d in the


thin juice is 36 p e r c e n t a n d is for s o d i u m .
T h e strongest association (0.44) b e t w e e n chemicals in the
petioles involves potassium a n d sodium a n d h e r e only 19 p e r c e n t
of the genetic v a r i a t i o n of o n e is a c c o u n t e d far by the genetic
variation of the o t h e r . T h e greatest association between chemicals
in the thin j u i c e is 0.77 a n d involves total n i t r o g e n a n d potissium.
Here 59 p e r c e n t of the total v a r i a t i o n is covariation. T h e n e x t
strongest association for the thin j u i c e is 0.40 a n d is b e t w e e n
potassium a n d s o d i u m .
Further, from the c o r r e l a t i o n coefficients listed in T a b l e 3 it
can be d e t e r m i n e d that in no case is a chemical c h a r a c t e r in t h e
petioles closely associated with a n o t h e r chemical character in the
thin juice. T h e s e results show that by p r o p e r b r e e d i n g p r o c e d u r e s
it should be possible to r e c o m b i n e desirable levels of these chemical characters in the petioles with desirable levels of the same
characters in t h e t h i n j u i c e .
Interactions
T h e m e a n s s h o w i n g the i n t e r a c t i o n s are t a k e n from T a b l e 2 .
Means for levels of total n i t r o g e n s h o w i n g i n t e r a c t i o n s of
populations X materials analyzed are listed in T a b l e 4. T h e F 1
hybrid 52 430 X 54 346 has a low level of total n i t r o g e n in
both the petioles a n d in the t h i n juice. T h e F 1 h y b r i d 52 305
CMS X 54 458 has a low level of total n i t r o g e n in the petioles
and a high level of total n i t r o g e n in t h e t h i n j u i c e . T h e F 1 h y b r i d
(52 430 X 52 307) shows" t h e reverse in t h a t it has a h i g h
level of n i t r o g e n in the petioles a n d a low level of n i t r o g e n
in the thin juice. T h e c o m m e r c i a l variety A56-3 has the highest
level of total n i t r o g e n in the petioles a n d has an i n t e r m e d i a t e
level of total n i t r o g e n in the t h i n juice.
Tabic 4.Means for levels of total nitrogen showing interactions of populations
X materials analwed.

Population
52-430 x 51-340 Fi
52-305 C.MS x 54-458 Fi
52-t;50 x 52-307 F1
A56-3
LSD at 5%
LSD at l%

Petioles

Thin
juice

Mg/lOOgm

MglOOtm

1278.2
1321.0
1470.8
1531.2

37.4
64.9
40.9
54.4

77.8
102.6

3.9
5.1

134

JOURNAL, OF THE A. S. S. B. T.

T h e c o m p a r i s o n s for t h e F 1 h y b r i d s 52-430 X 54-346 and


52-305 C M S X 54-458 are n o t significant for total n i t r o g e n in
the petioles b u t are significant for total n i t r o g e n in t h e thin
juice. T h e c o m p a r i s o n s for the F 1 h y b r i d s 52-430 X 54-346 and
52-430 X 52-307 are statistically significant for the petioles but
n o t for the t h i n j u i c e . T h e c o m p a r i s o n s for the F 1 h y b r i d s 52305 C M S X 54-458 a n d 52-430 X 52-307 are significant for both
the petioles a n d the t h i n juice. T h e same is t r u e for t h e comparison i n v o l v i n g 52-305 C M S X 54-458 a n d A56-3. T h e comparison b e t w e e n the F 1 h y b r i d 52-430 X 52-307 a n d A56-3 are
n o t significant for the petioles b u t are statistically significant
for t h e t h i n j u i c e .
T h e s e results definitely show that some p o p u l a t i o n s at time
of harvest have h i g h e r levels of total n i t r o g e n in t h e petioles as
c o m p a r e d to o t h e r varieties a n d lower levels of total n i t r o g e n in
the t h i n j u i c e . T h e reverse is also t r u e some p o p u l a t i o n s have
high levels of total n i t r o g e n in the t h i n j u i c e a n d low levels of
total n i t r o g e n in the petioles as c o m p a r e d with o t h e r populations.
T h e s e results definitely show that for total n i t r o g e n t h e r e is an
i n t e r a c t i o n b e t w e e n genotypes a n d material analysed (petioles
a n d t h i n juice) . H e n c e by p r o p e r b r e e d i n g p r o c e d u r e s different
c o m b i n a t i o n s of levels of total n i t r o g e n in the petioles a n d in the
t h i n juice are a t t a i n a b l e .
M e a n s for levels of potassium s h o w i n g i n t e r a c t i o n s of populations X material analysed are listed in T a b l e 5. T h e F 1 hybrid
52-430 X 54-346 has low potassium in b o t h the petioles and in
t h e t h i n juice, whereas 52-305 C M S X 54-458 is low in potassium
in the petioles a n d high in the t h i n juice. T h e F 1 h y b r i d 52-305
C M S X 54-565 is high in potassium in t h e petioles a n d low in
t h e t h i n juice. T h e variety A56-3 is low in p o t a s s i u m in the
petioles a n d m o d e r a t e l y high in the t h i n j u i c e .
T a b l e 3.Means fcr levels of
material analysed.

potassium showing interactions of populations


Potassium

Population
52-4r.O x 51-3-46 Fi
52-303 CMS x 54-458 Fi
52-305 CMS x 34-565 Fi
Af.ii 3
LSD at 5%
LSD at 1%

Petioles

Thin
juice

Mg lOOgm

Mg. 100ml

14.2
16.4
24.1
16.4

41.9
71.9
48.2
60.7

1.6
2.1

57
7.6

Vol.

13, No. 2, J U L Y 1964

155

A g a i n t h e r e is an i n t e r a c t i o n b e t w e e n g e n o t y p e s a n d levels
of p o t a s s i u m in t h e p e t i o l e s as c o m p a r e d w i t h levels of p o t a s s i u m
in the thin juice. It is a p p a r e n t that genotypes can be o b t a i n e d
having different levels of p o t a s s i u m in t h e petioles a n d in t h e
t h i n j u i c e . T h a t is, p o p u l a t i o n s c a n b e b r e d t h a t h a v e d e s i r a b l e
levels o f p o t a s s i u m i n t h e p e t i o l e s a n d d e s i r a b l e l e v e l s o f p o t a s sium i n t h e t h i n j u i c e .
M e a n s for levels of s o d i u m s h o w i n g i n t e r a c t i o n s of p o p u l a tions X m a t e r i a l a n a l y s e d a r e l i s t e d i n T a b l e 6 . T h e F , h y b r i d
52-805 C M S X 5 4 - 5 0 5 i s l o w i n l e v e l s o f s o d i u m i n b o t h t h e p e t i oles a n d t h i n j u i c e . T h e l \ h y b r i d 5 2 - 3 0 5 C M S X 5 2 - 3 0 7 p o s s e s s e s a low l e v e l o f s o d i u m i n t h e p e t i o l e s a n d a h i g h l e v e l i n t h e
thin juice. T h e F 2 h y b r i d 52-430 X 52-408 h a s t h e h i g h e s t level
of s o d i u m in t h e petioles a n d a m o d e r a t e l y low level in t h e t h i n
juice. A5G-3 h a s a h i g h level o f s o d i u m i n b o t h t h e p e t i o l e s a n d
i n t h e t h i n j u i c e . I n fact i t i s s i g n i f i c a n t l y h i g h e r i n l e v e l o f sodium in the thin juice than any o t h e r p o p u l a t i o n listed in F a b l e
G. Also for s o d u m , as was t h e case for total n i t r o g e n a n d potassium, tliere i s a n i n t e r a c t i o n b e t w e e n g e n o t y p e s a n d levels o f
sodium in t h e petioles as c o m p a r e d w i t h levels of s o d i u m in t h e
thin j u i c e . I t f o l l o w s t h a t d i f f e r e n t c o m b i n a t i o n s o f l e v e l s o f sodium in the petioles a n d in the thin juice can be o b t a i n e d by
proper b r e e d i n g p r o c e d u r e s .
Table 6.Means
material analyzed.

levels

of

sodium

of

showing

populations

S o d ium

Population

Petioles

Thin
juice

Mg/lOOgm

Mg/100ml

30.9
32.2
36.9
35.9

52-305 CMS X 5-1-565 Fi


52-305 CMS x 52-307 Fi
52-130 x 52-408 Fi
A56-3

30.6
41. S
36.8
55.7

LSD at 5%
LSD at 1%

Discussion
T h e i n t e r a c t i o n s i n v o l v i n g g e n o t y p e s X m a t e r i a l analysed
(petioles a n d t h i n j u i c e ) h a v e s h o w n t h a t a t t i m e o f h a r v e s t
levels of t h e t h r e e c h e m i c a l s vary in t h e petioles a n d t h i n j u i c e
according to p o p t d a t i o n s . W h e n i n t e r p r e t i n g these findings it is
well to h a v e in m i n d t h a t t h e petioles a r e a p a r t of t h e tops of
the sugar b e e t a n d t h e t h i n j u i c e i s p r e p a r e d from t h e r o o t s .
Hence, it a p p e a r s t h a t at t i m e of harvest s o m e g e n o t y p e s h a v e t h e
higher levels of these c h e m i c a l s in t h e tops of t h e p l a n t , w h e r e a s

JOURNAL OF T H E A. S. S. B. T

136

for otlier genotypes the h i g h e r levels are f o u n d in the roots as


r e p r e s e n t e d by analysis of the thin juice. T h e s e findings are of
e x t r e m e i m p o r t a n c e to the beet sugar i n d u s t r y in that this shows
p o p u l a t i o n s can be b r e d t h a t will have t h e h i g h e r levels of these
t h r e e chemical characters in the tops of t h e p l a n t r a t h e r t h a n in
the roots. T h e s e t h r e e chemicals at h i g h e r levels have a decided
adverse effect on p e r c e n t a g e sucrose a n d p e r c e n t a g e apparent
p u r i t y (see Powers a n d Payne, 10).
H e n c e it is of i m p o r t a n c e to k n o w w h e t h e r t h e genotypes
t e n d i n g to have t h e h i g h e r levels of the t h r e e c h e m i c a l characters
in the petioles r a t h e r t h a n in the t h i n j u i c e , at t i m e of harvest,
have yielding ability. T h e results from the studies i n v o l v i n g these
t h r e e chemical characters a n d weight p e r root, p e r c e n t a g e sucrose
a n d p e r c e n t a g e a p p a r e n t p u r i t y are p r e s e n t e d in a n o t h e r article
(see Powers a n d Payne, 10).
Summary

1. P o p u l a t i o n s of sugar beets were f o u n d to differ in the relative levels of total n i t r o g e n , potassium, a n d s o d i u m in t h e petioles
as c o m p a r e d with levels of these same chemicals in the t h i n juice,
It is well to k e e p in m i n d that the petioles are p a r t of the tops
of t h e sugar beet p l a n t , whereas the t h i n j u i c e is p r e p a r e d from!
t h e roots.
2. T h e i n t e r a c t i o n s i n v o l v i n g g e n o t y p e s X m a t e r i a l s analysed;
(petioles or t h i n juice) have shown that, at t i m e of harvest,
h i g h e r levels of t h e t h r e e chemicals occur in e i t h e r the petioles,
or the thin juice, or in b o t h . Conversely, at t i m e of harvest, some
genotypes have h i g h e r levels of these t h r e e chemical characters
in t h e petioles associated w i t h lower levels in the t h i n juice.
3. T h i s latter finding is of e x t r e m e i m p o r t a n c e to the beet
sugar i n d u s t r y , because it shows t h a t p o p u l a t i o n s can be bred
t h a t will have t h e h i g h e r levels of these chemicals in t h e tops
(petioles) of t h e sugar beet r a t h e r t h a n in t h e roots ( t h i n juice)
T h e h i g h e r levels of these three chemicals in t h e t h i n j u i c e have
a decidedly adverse effect w i t h p e r c e n t a g e sucrose a n d on percentage a p p a r e n t p u r i t y (see Powers a n d Payne, 10).
L i t e r a t u r e Cited
(1)

BROWN, R O B E R T J. a n d R O B E R T F. SERRO.

1951. A m e t h o d for deter

ruination of t l r n juice puritv from individual m o t h e r beets. Proc |


Am. Soc. Sugar Beet T e c h n o l . 8 ( 2 ) : 271-278.
(2) CARRUTHERS, A., and J. F. T. OLDFIELD. I960. Methods for the assess;
merit of beet quality. Intern. Sugar J. 63: 72-74, 103-103, 137-139
(3) E M M E R T , E. M. 19.10. A method for the rapid determination of phos
p h a t e in fresh p l a n t tissue. Plant Phys. 5: 113-117.

V O L 13, N o . 2, J r i . v 1961

137

A}) E M M E R T - E. M. 1932. Field method of estimating nitrate, p h o s p h a t e


and potassium in plants. Plant Phvs. 7: 3 15-321.
(5) E M M E R T , E. M. 1933. New methods for the determination of the availability of nitrogen a n d p h o s p h o r u s to plants. J. Am. Soc. Agr.
27: 1-7.
(6 GARDNE.R. R O B F R T ami D.

ROBE.RTSON.

1935. T h e use of sugar beet

petioles as indicators of soil fertility needs. Colo. Agr. Exp. Sta.


T e c h . Bull. 14. 16 p p .
(7) HADDOCK. J. I... D. C. LINTON and R. E. HursT. 1930. Nitrogcn constituents associated with reduction of sucrose percentage and purity
of sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 9 ( 2 ) : 110-117.
(8)

O w e n . F. V., M Y R O N STOUT. A. M. M U R P H Y . C. H . S M I T H a n d G. K.

RYSER. I960. Interaction of c o m p o n e n t s of impurity a n d location in


hybrids from inbred lines of sugar beet*. J. Am. Soc. Sugar beet
T e c h n o l . 11 (1) : 37-13.
(9)

POWERS. LE ROY. W. R. SCHMFIH., W. T. FEDERER and M E R L E G. P A V N F .

1963. Chemical genetic and soils studies involving thirteen characters in sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 1 2 ( 5 ) :
393-118.
(10) POWERS. LE ROY and Merle. G. PAYNE.. 1964. Associations of levels
of total nitrogen, potassium, and sodium in petioles and thin
juice with weight of root per plot, percentage sucrose a n d percentage a p p a r e n t purity. J. Am. Soc. Sugar Beet T e c h n o ! . 1 3 ( 2 ) : 1383 50.
(11)

POWERS, L E R O Y , E. E. R E M M E N G A a n d

N.

S.

Urquhart.

In

process

of publication. T h e p a r t i t i o n i n g method of genetic analysis applied


to a study of weight per root a n d percentage sucrose in sugar beets
(Beta vulgaris I.)
(12) RORABAUGH. Guy a n d Lloyd) W. Norman. 1956. T h e effect of various
impurities on the crystallisation of sucrose. J. Am. Soc. Sugar Beet
T e c h n o l . 9 (3) : 238-252.
(13)

ROUNDS, H U G H G.. GEORGE. E. R U S H ,


PARRISH a n d FRANK X. R A W L I N G S .

DONAID
1958. A

L.
OldemeyeR.
C. P.
study a n d economic

appraisal of the effect of nitrogen fertilization and selected varieties


on the production and processing of sugar beets, f. Am. Sec. Sugar
Beet T e c h n o l . 10 (2) : 97-1 16.
(14)

(15)
(16)
(17)
(18)

RYSER. G. K.. M Y R O N STOUT. A. U L R I C H a n d F. V. O W E N . 1959. Some

chemical a n d physiological characteristics of inbred litres of sugar


beets. J. Am. Soc. Sugar Beet T e c h n o l . 1 0 ( 6 ) : 525 - 543
Toi-MAN, BION and R. C. JOHNSON. 1958. Effect of nitrogen on the
vielci a n d sucrose content of sugar beets. J. Am. Soc. Sugar Beet
T e c h n o l . 1 0 ( 3 ) : 251-257.
ULRICH, ALBERT. 1912. T h e relationship of nitrogen to the formation
of sugar in sugar beets.' Proc. Am. Soc. Sugar Beet T e c h n o l . 3: 66-80.
UI.RICII, ALBERT. 1948. Plant analysis as a guide to the nutrition of
sugar beets in California. Proc. Am. Soc. Sugar Beet T e c h n o l .
5: 364-377.
U L R I C I I , AI.BERT and F. J.
Hills.
1952. Petiole sampling of sugar
beet fields in relation to their- nitrogen, phosphorus, potassium and
sodium status. Proc. Am. Soc. Sugar Beet T e c h n o l . 7: 32-15.

Associations of Levels of T o t a l N i t r o g e n , Potassium,


and Sodium in Petioles and in Thin Juice with
W e i g h t of Roots Per Plot, Percentage
Sucrose and Percentage A p p a r e n t
Purity in Sugar Beets 1 4
LEROY

POWERS-

Received

AND

MERLE

for publication January

G.

PAYNE3

16, 1964

Studies c o n d u c t e d (see P a y n e et al.) 5 to d e t e r m i n e levels of


total n i t r o g e n , potassium, a n d s o d i u m in the petioles as compared
with levels of these same chemicals in the t h i n juice, at time of
harvest, have shown that t h e r e are i n t e r a c t i o n s of genotypes and
material analysed. It was f o u n d that some genotvpes, as compared
to others, t e n d e d to have h i g h e r levels of the t h r e e chemicals in
the petioles as c o m p a r e d with the t h i n juice of t h e sugar beet
(Beta vulgaris L.) . F o r o t h e r c o m p a r i s o n s t h e reverse was found
to be t r u e . T h e p u r p o s e of the study r e p o r t e d in this article is
to d e t e r m i n e relations b e t w e e n weight of roots per plot, percentage sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t v a n d levels of total
n i t r o g e n , potassium a n d s o d i u m in the petioles as c o m p a r e d with
levels of these same chemicals in t h e t h i n juice. Also the relations b e t w e e n levels of p h o s p h o r u s in t h e petioles w i t h weight
of roots p e r plot, p e r c e n t a g e sucrose a n d p e r c e n t a g e apparent
p u r i t y were studied. T h e petioles analysed were collected at time
of harvest a n d the t h i n j u i c e was p r e p a r e d from t h e roots harvested.
Literature R e v i e w
T h e studies on t h e associations of levels of total nitrogen.|
potassium, a n d s o d i u m in the petioles a n d in the t h i n juice with,
w e i g h t of roots p e r plot, p e r c e n t a g e sucrose, a n d percentage app
p a r e n t p u r i t y p r o v i d e i n f o r m a t i o n f u n d a m e n t a l to an under
1 Cooperative investigations of the Colorado Agricultural Experiment Station. the
Crops Research Division.' Agricultural Research Service. U. S. Department of Agriculture
ami the Beet Sugar Development Foundation. T h e Colorado State l*ni\ersitv gratefully
acknowledges financial support from the James G. Boswell Foundation administered by
the Agricultural Research Center of Stanford Research Institute, the National institutes;
of Health, the National Plant Food Institute and contract-research-funds [12-14-100 \2-U-]9'\
4549 (34) ] from the Agricultural Research Sen ice of the I". S. Department of Agriculture
Approved b\ the Colorado Agricultural Experiment Station for publication as Scientific
Series Article No. 885.
,
-Geneticist, Crops Research Division. Agricultural Research Service. U. S. Department
of Agriculture.
3
Professor of Chemistry. Colorado State University.
,
4 T h e writers are indebted to R. Ralph Wood of T h e Great Western Sugar Company
for obtaining thin juice samples bv an oxalate method standard with his company and
to the Western Data Processing Center at the University of California at Los Angeles
for use
cf the computing facilities for analysing data, Job No. 398.
5
Numbers in parentheses refer to literature cited.

. i.s, N o . 2, J c i v 1904

130

standing of c o m b i n i n g ability. especially for yield of sugar p e r


plot. T h e l i t e r a t u r e p e r t a i n i n g t o c o m b i n i n g ability i n sugar
beets is r a t h e r limited. T h i s p r o b a b l y is d u e to t h e fact that u n t i l
quite recently the n u m b e r of i n b r e d lines of sugar beets ayailable
for testing has been a n d , c o m p a r a t i v e l y speaking, still is r a t h e r
limited.
O l d e m e y e r (5) used a c o m m e r c i a l sugar beet variety a n d t h e
German red beet as topcross testers to d e t e r m i n e the general
combining ability of i n b r e d lines of sugar beets a n d c o n c l u d e d
that the Cierman red beet is satisfactory to test general c o m b i n i n g
ability for both yield a n d p e r c e n t a g e sucrose. Peterson a n d Dickenson (9) using the r e d - m a r k e r beet to test for g e n e r a l c o m b i n i n g
ability f o u n d that t h e single crosses p r o d u c i n g t h e most sugar
per acre were those whose p a r e n t s w e r e high in general combining ability w h e n tested by crossing will) the r e d - m a r k e r beet
and whose F, h y b r i d s e x h i b i t e d heterosis for p e r c e n t a g e sucrose.
O l d e m e y e r a n d R u s h (4) m a d e a very i n t e r e s t i n g study u s i n g
male-sterile testers. Seventeen self-fertile i n b r e d lines a n d o n e
open-pollinated variety of sugar beets were crossed to five cytoplasmic male-sterile tester lines. T h e h y b r i d s a n d t h e i r corresponding p a r e n t s w e r e g r o w n in a field test. T h e results of this
test showed that t h e r e are differences a m o n g the i n b r e d lines
for general c o m b i n i n g ability a n d that specific c o m b i n i n g ability
is i m p o r t a n t , p a r t i c u l a r l y in r e g a r d to yield. H e t e r o s i s a n d p h e n o typic d o m i n a n c e were found for b o t h yield a n d sucrose percentage. P a r e n t a l p e r f o r m a n c e showed little association with t h e
combining ability of t h e i r respective i n b r e d s . T h i s points o u t
the necessity of m a k i n g test crosses w h e n e v a l u a t i n g i n b r e d s . T h e
variance a t t r i b u t a b l e to t h e males a n d females is c o n s i d e r e d bv
them to be an i n d e x of that part of t h e over-all v a r i a t i o n a m o n g
the test crosses clue to t h e general c o m b i n i n g ability of t h e parents. T h e i n t e r a c t i o n v a r i a n c e ( m a l e X f e m a l e ) is considered an
index of t h a t p a r t of t h e over-all v a r i a t i o n due to specific combining ability. To stuclv the effect of specific c o m b i n i n g ability,
the m e a n s of the i n d i v i d u a l crosses w e r e adjusted by a d d i n g to.
or subtracting from t h e m , t h e d e v i a t i o n s of t h e m e a n s of all
respective crosses of each p a r e n t from t h e test averages.
H e l m e r i c k ct al. (3) e m p l o y i n g varietal crosses m a d e r a t h e r
extensive studies p e r t a i n i n g to heterosis a n d c o m b i n i n g ability.
They c o n c l u d e d t h a t r a t h e r substantial gains c o u l d be m a d e by
utilizing heterosis in the p r o d u c t i o n of beet sugar. T h e y also
studied the e n v i r o n m e n t a l a n d g e n e t i c variances a n d identifiable
Proportion of genetic deviates a n d p o i n t e d o u t the v a l u e of this
information in b r e e d i n g h y b r i d p o p u l a t i o n s of sugar beets.

140

J O U R N A L OF THE A. S. S. B. T.

P o w e r s e t a l . ( 1 0 ) c o n d u c t e d s t u d i e s w h i c h s h o w e d t h a t cert a i n p l a n t i n g a r r a n g e m e n t s i n i s o l a t i o n p l o t s c o n t a i n i n g t w o pare n t a l s o u r c e s r e s u l t e d i n a p p r o x i m a t e l y 6 8 p e r c e n t o f t h e progeny


b e i n g t h e r e s u l t o f c r o s s - f e r t i l i z a t i o n b e t w e e n s o u r c e s a n d that
p r o b a b l y t h e r e m a i n i n g 3 2 p e r c e n t o f t h e p r o g e n y r e s u l t e d from
c r o s s - f e r t i l i z a t i o n b e t w e e n p l a n t s w i t h i n s o u r c e s . O t h e r studies
( 1 1 , 12, 13) s h o w e d t h a t c e r t a i n i n b r e d s p r o d u c e d h y b r i d s that
e x h i b i t e d heterosis for p e r c e n t a g e sucrose a n d p e r c e n t a g e app a r e n t p u r i t y . I t i s e x p e c t e d t h a t h e t e r o s i s f o r w e i g h t p e r root
w o u l d b e o b t a i n e d . Such was f o u n d t o b e t h e case.
C h e m i c a l g e n e t i c s t u d i e s ( P o w e r s e t a l , 13) r e v e a l e d t h a t the
d o m i n a n c e p h e n o m e n a f o r t h e c h e m i c a l c h a r a c t e r s a s s o c i a t e d with
p e r c e n t a g e sucrose a n d p e r c e n t a g e a p p a r e n t p u r i t y are such as to
r e s u l t i n b o t h o f t h e s e c h a r a c t e r s e x h i b i t i n g h e t e r o s i s i n hybrids
b e t w e e n certain selected i n b r e d lines. T h i s indicates that by emp l o y i n g m e t h o d s o f b r e e d i n g d e s i g n e d t o u t i l i z e h e t e r o s i s , hybrid
p o p u l a t i o n s t h a t a r e s u p e r i o r i n w e i g h t p e r r o o t , p e r c e n t a g e suc r o s e , a n d p e r c e n t a g e a p p a r e n t p u r i t y t o t h o s e v a r i e t i e s n o w being
g r o w n f o r t h e p r o d u c t i o n o f b e e t s u g a r c a n b e b r e d . S o m e such
hybrid c o m b i n a t i o n s involving i n b r e d lines were obtained.
F o r m e t h o d s o f c h e m i c a l a n a l y s i s s e e P a y n e e t a l . ( 6 , 7 , ) and
f o r a r e v i e w o f t h e l i t e r a t u r e i n v o l v i n g t h e h e r i t a b i l i t y o f the
c h e m i c a l c h a r a c t e r s s e e P o w e r s e t a l . ( 1 1 , 12) a n d F i n k n e r e t

al. ( 2 ) .
Materials and Design of the E x p e r i m e n t
T h e materials used in t h e study a r e as follows: T h e r e is a
total of 20 p o p u l a t i o n s in t h e e x p e r i m e n t . O n e is a commercial
variety. 4 are three-way h y b r i d s , each c o m p o s e d of 3 inbreds, and
15 a r e F 1 h y b r i d s each c o m p o s e d of 2 i n b r e d s . T h e dates of harvest are S e p t e m b e r 14, O c t o b e r 3. a n d O c t o b e r 16. T h e experim e n t was c o n d u c t e d in 1961. T h e characters studied are weight
of roots per plot, p e r c e n t a g e sucrose, p e r c e n t a g e a p p a r e n t purity,
a n d levels of total n i t r o g e n , potassium a n d s o d i u m in the petioles
a n d in the t h i n juice, a n d levels of p h o s p h o r u s in t h e petioles
W e i s h t of roots p e r plot is expressed as kilograms, sucrose and,
p u r i t y as percentages, the levels of t h e chemical characters are
expressed as m i l l i g r a m s p e r 100 g r a m s in the petioles, and the
levels of the chemical characters a r e expressed as milligrams per
100 milliliters of t h i n juice e q u a t e d to a refractive dry substance
of 10 in the t h i n juice. T h e t h i n juice was p r e p a r e d by The
G r e a t W e s t e r n Sugar C o m p a n y bv an o x a l a t e m e t h o d standard
with t h e m [see ( 1 ) ] . In this process t h e n i t r a t e n i t r o g e n is removed. Hence t h e total n i t r o g e n for t h e t h i n juice does not
i n c l u d e all t h e n i t r o g e n o u s c o m p o u n d s f o u n d in t h e total nitrogen

VoL.

13,

No.

2,

JULY

1964

141

analysis of t h e petioles. H o w e v e r , as shown by Powers et al.


(12),
the association b e t w e e n total n i t r o g e n in the t h i n juice a n d the
press juice is e x t r e m e l y high, most of the variability of o n e b e i n g
accounted for by the v a r i a b i l i t y of t h e o t h e r .
T h e design of the e x p e r i m e n t is a split plot with p o p u l a t i o n s
randomized w i t h i n r e p l i c a t i o n s a n d dates of harvest r a n d o m i z e d
within blocks. Each block is c o m p o s e d of t h r e e d u e s of harvest
and each d a t e of harvest, has t w o r e p l i c a t i o n s with 20 p o p u l a t i o n s
randomized w i t h i n each r e p l i c a t i o n . T h e r e are five such blocks.
Hence, t h e design of t h e e x p e r i m e n t is a m o d i f i e d r a n d o m i z e d
complete block.
Results
T h e F values calculated from the analyses of v a r i a n c e for all
the characters are listed in T a b l e 1. F o r all characters, t h e r e are
significant differences b e t w e e n m e a n s of p o p u l a t i o n s . T h e r e are
significant differences b e t w e e n m e a n s of dates of harvest for percentage sucrose a n d possibly for levels of s o d i u m in t h e t h i n
juice. T h e i n t e r a c t i o n s having possible statistical significance are
for the characters p e r c e n t a g e sucrose, p e r c e n t a g e a p p a r e n t p u r i t y .
and levels of s o d i u m in the petioles. T h e i n t e r a c t i o n i n v o l v i n g
the levels of p h o s p h o r u s in t h e petioles is fairly well established
statistically. T h e data will be c o n s i d e r e d on the basis of t h e
average of all dates of harvest as t h e a m o u n t of t h e v a r i a b i l i t y
accounted for by the i n t e r a c t i o n s i n v o l v i n g dates of harvest is
small, c o m p a r a t i v e l y . T h e i n t e r a c t i o n s of greatest i m p o r t a n c e in
this article involve p o p u l a t i o n s , chemical c o n s t i t u e n t s , and materials analysed.
T h e m e a n s for weight p e r plot, p e r c e n t a g e sucrose, a n d percentage a p p a r e n t p u r i t y : for levels of total n i t r o o e n potassium,
sodium in t h e petioles a n d in the t h i n juice: a n d levels of phosphorns in the petioles are listed in T a b l e 2. Also, the least significant differences a n d t h e g r a n d averages are listed at t h e bottom of this table. P o w e r s et al. (13) have shown t h a t verv little
of the e n v i r o n m e n t a l v a r i a b i l i t y is i n c l u d e d in t h e differences
between the m e a n s of populations. H e n c e , the differences n o t e d
between m e a n s of p o p u l a t i o n s a r e p r e d o m i n a n t l y genetic. In this
article, t h e data in T a b l e 2 have t h e i r greatest imprest in t h e de-rees of association b e t w e e n t h e level of a c h e m i c a l in t h e petioles
and the level of t h e same chemical in t h e t h i n juice. t h e i r corresponding i n t e r a c t i o n s , a n d t h e association of t h e level of these
chemicals with t h e i m p o r t a n t a g r o n o m i c characters, w e i g h t of
roots p e r plot, p e r c e n t a g e sucrose, a n d p e r c e n t a g e a p p a r e n t puri t y . T h e associations a r e d e t e r m i n e d b y s t u d y i n g t h e s i m p l e correlation coefficients.

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JOURNAL OF THE A. S. S. B. T.

Associations
T h e simple c o r r e l a t i o n coefficients are listed i n T a b l e 3 .
W e i g h t p e r plot is positively associated with levels of total
n i t r o g e n a n d s o d i u m in the petioles a n d with levels of s o d i u m in
t h e thin juice. T h e association of weight p e r plot with levels of
potassium in the petioles is not statistically significant, only 4 percent of the variability b e i n g a c c o u n t e d for by covariation. The
association b e t w e e n p e r c e n t a g e sucrose a n d total n i t r o g e n in the
petioles is negative a n d 22 p e r c e n t of t h e v a r i a t i o n is covariation.
Likewise, t h e association b e t w e e n p e r c e n t a g e sucrose a n d level
of s o d i u m in th petioles is negative, a n d h e r e 18 p e r c e n t of the
v a r i a t i o n is covariation. In no case is t h e association b e t w e e n percentage a p p a r e n t p u r i t y a n d levels of total n i t r o g e n , potassium,
a n d s o d i u m in the petioles statistically significant.

W e i g h t p e r plot does n o t show any statistically significant association with levels of total n i t r o g e n or potassium in the thin
juice. H o w e v e r , levels of total n i t r o g e n in the thin juice and
p e r c e n t a g e a p p a r e n t p u r i t y are very closely associated and the
association is negative. Also levels of potassium in t h e thin juice
are r a t h e r closely associated with p e r c e n t a g e a p p a r e n t purity and
again the association is negative. W i t h this high a d e c r e e of association. it is n o t at all likely that the b r e e d e r can o b t a i n genotypes h a v i n g a high p u r i t y a n d a high level of total nitrogen in
t h e t h i n juice. T h e same associations h o l d for p e r c e n t a g e s u c r o s e
a n d levels of total n i t r o g e n a n d potassium in the t h i n juice but
t h e asociations are n o t nearly so p r o n o u n c e d . Likewise, percentage sucrose is r a t h e r strongly associated with levels of sodium in
t h e t h i n juice a n d t h e association is negative, 64 p e r c e n t of the|
variability b e i n g c o v a r i a t i o n .
i

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T h e s e results show t h a t at t i m e of harvest it is m u c h move


desirable to have the higher levels of total n i t r o g e n , potassium,
and sodium in the petioles r a t h e r t h a n in the t h i n j u i c e : as h e r e
they are positively associated with weight of roots per plot a n d
are not closely associated negatively with e i t h e r p e r c e n t a g e sucrose or p e r c e n t a g e a p p a r e n t p u r i t y . Levels of p h o s p h o r u s in t h e
petioles show little, or n o , association with weight per plot, b u t
the associations w i t h p e r c e n t a g e sucrose a n d p e r c e n t a g e a p p a r e n t
purity are statistically significant a n d positive. H o w e v e r , t h e
closeness of the association is n o t m a r k e d , t h e greatest a m o u n t
of the variability b e i n g covariation is 31 p e r c e n t .
T h e i n t e r a c t i o n s of w e i g h t p e r plot, p e r c e n t a g e sucrose, a n d
percentage a p p a r e n t p u r i t y with total n i t r o g e n in the petioles
and in the t h i n juice are d e p i c t e d by the m e a n listed in Table 4.
These p o p u l a t i o n are selected from T a b l e 2 because they show
that certain c o m b i n a t i o n s of characters can be o b t a i n e d . T h e
comparisons b e t w e e n the m e a n s of the F 1 h y b r i d 52-303 C M S
X 54-458 a n d t h e F 1 h y b r i d 52-4.0 X 52-408 show that an increase of total n i t r o g e n in t h e petioles a n d a decrease of total nitrogen in the t h i n juice are a c c o m p a n i e d by increases in weight
of roots p e r plot, p e r c e n t a g e sucrose, a n d p e r c e n t a g e a p p a r e n t
purity. T h e c o m p a r i s o n s of F, h y b r i d 52-430 X 52-408 with
A56-3 show t h a t f u r t h e r increases of total n i t r o g e n in t h e petioles
and in the t h i n juice are a c c o m p a n i e d by a f u r t h e r increase in
weight p e r p l o t a n d by d e c i d e d decreases in p e r c e n t a g e sucrose
and in p e r c e n t a p p a r e n t p u r i t y .

These c o m p a r i s o n s c o n f i r m t h e r e l a t i o n s shown by t h e correlation coefficients: n a m e l y , t h a t increases of total n i t r o g e n in


the petioles r a t h e r t h a n in t h e thin j u i c e can result in an increase
in all 3 of t h e i m p o r t a n t a g r o n o m i c characters, weight p e r plot,
percentage sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t y . T h e y f u r t h e r
show that an increase of total n i t r o g e n in the t h i n j u i c e does n o t

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J O U R N A L OF T H E A. S. S. B. T.

h a v e a n a d v e r s e r e l a t i o n w i t h w e i g h t o f r o o t s p e r p l o t b u t i t does
h a v e d e c i d e d l y a d v e r s e r e l a t i o n s w i t h p e r c e n t a g e s u c r o s e and
p e r c e n t a g e a p p a r e n t p u r i t y . H e n c e , w e i g h t o f r o o t s p e r plot,
p e r c e n t a g e s u c r o s e , a n d p e r c e n t a g e a p p a r e n t p u r i t y i n s o m e popu l a t i o n s a r e f a v o r a b l y a s s o c i a t e d w i t h t o t a l n i t r o g e n i n t h e petioles
b u t n o t w i t h t o t a l n i t r o g e n i n t h e t h i n j u i c e . H e n c e , t h e breeder
s h o u l d b e a b l e t o i n c r e a s e t h e s e t h r e e d e s i r a b l e a g r o n o m i c chara c t e r s b y b r e e d i n g g e n o t y p e s h a v i n g h i g h l e v e l s o f t o t a l nitrogen
i n t h e p e t i o l e s a t t i m e o f h a r v e s t . T h e s e r e s u l t s i n d i c a t e that!
h i g h e r levels of total n i t r o g e n in t h e petioles a r e associated with

h i g h e r yields a n d a r e n o t associated w i t h lower percentage sucrose a n d lower p e r c e n t a g e p u r i t y ; whereas h i g h e r levels of total


n i t r o g e n in the t h i n juice are n o t associated with h i g h e r yields
b u t are associated w i t h lower p e r c e n t a g e sucrose a n d lower percentage a p p a r e n t p u r i t y . T h e n , it seems as t h o u g h t h e plant
b r e e d e r can i m p r o v e b o t h yield a n d q u a l i t y by genetically con
t r o l l i n g t h e location, at t i m e of harvest, of the h i g h e r levels of
total n i t r o g e n ; that is, b r e e d i n g those genotypes h a v i n g higher
levels of this chemical in t h e petioles instead of the t h i n juice.
T h e m e a n s s h o w i n g the i n t e r a c t i o n s of w e i g h t p e r plot, percentage sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t y w i t h levels off
potassium in t h e petioles a n d in t h e t h i n j u i c e are listed in Table
5. T h e c o m p a r i s o n s i n v o l v i n g t h e two F 1 h y b r i d s 52-305 CMS X
54-458 a n d 52-430 X 52-408 show t h a t an increase in levels of
potassium in t h e petioles a n d a decrease in the levels of the potassium in t h e t h i n juice are a c c o m p a n i e d by m a t e r i a l increases
in w e i g h t p e r plot, p e r c e n t a g e sucrose, a n d p e r c e n t a g e apparent
p u r i t y . T h e c o m p a r i s o n s b e t w e e n p o p u l a t i o n s 52-430 X 52-408
a n d A56-3 show t h a t a decrease in the level of potassium in the
petioles a n d no c h a n g e in t h e level of p o t a s s i u m in t h e t h i n juice
are a c c o m p a n i e d by decreases in p e r c e n t a g e sucrose a n d percentage a p p a r e n t p u r i t y a n d a c o m p a r a t i v e l y small increase in weight
p e r plot. As was t h e case for levels of total n i t r o g e n , increased
levels of potassium in t h e petioles are associated with increased

VOL. 13, No. 2, JULY 1964

147

percentage sucrose a n d p e r c e n t a g e a p p a r e n t p u r i t y , whereas inlicases in levels of potassium in the t h i n j u i c e show the reverse
associations. A g a i n , if h i g h e r levels of p o t a s s i u m a r e essential
to those metabolic, processes c o n d u c i v e to h i g h e r yields, it is
more desirable to have these h i g h e r levels in t h e petioles at t i m e
of harvest r a t h e r t h a n in the t h i n j u i c e .
T h e m e a n s s h o w i n g t h e i n t e r a c t i o n s of w e i g h t per plot, percentage sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t y w i t h levels of
sodium in t h e petioles a n d in the t h i n j u i c e a r e listed in T a b l e
6. C o m p a r i n g the F 1 h y b r i d s 52-305 C M S X 54-458 a n d 52-430
X 52-408 it can be seen that an increase in levels of s o d i u m in
the petioles a n d no m a t e r i a l c h a n g e in levels of s o d i u m in t h e
thin juice a r e a c c o m p a n i e d by increases in weight of roots p e r
plot, p e r c e n t a g e sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t y . Comparing 52-430 X 52-408 a n d A56-3 no m a t e r i a l c h a n g e in levels
of sodium in the petioles a n d an increase in levels of s o d i u m in
the thin j u i c e are associated with decided decreases in p e r c e n t a g e
sucrose a n d p e r c e n t a g e a p p a r e n t p u r i t y a n d a m o d e r a t e increase
in weight of roots p e r plot. A g a i n , if h i g h e r levels of s o d i u m
are conducive to favorable m e t a b o l i c processes in the sugar beet
plant, it is p r e f e r a b l e to have the h i g h e r levels in the petioles
rather t h a n h a v i n g t h e h i g h e r levels in t h e t h i n j u i c e at t i m e
of harvest.

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T h e m e a n s s h o w i n g t h e i n t e r a c t i o n s o f w e i g h t p e r p l o t , percentage sucrose, a n d p e r c e n t a g e a p p a r e n t p u r i t y w i t h levels of


p h o s p h o r u s i n t h e p e t i o l e s a r e l i s t e d i n T a b l e 7 . F o r a l l three
p o p u l a t i o n s listed in this table, the h i g h e r levels of phosphorus
i n t h e p e t i o l e s a r e a s s o c i a t e d w i t h i n c r e a s e s i n w e i g h t o f r o o t s per
p l o t a n d t h e r e a r e n o c o n s i s t e n t r e l a t i o n s i n v o l v i n g percentage
s u c r o s e a n d p e r c e n t a g e a p p a r e n t p u r i t y . H o w e v e r , f r o m t h e correlation coefficients listed in T a b l e 3 it c a n be seen that, on an
a v e r a g e , t h e g e n e t i c v a r i a b i l i t y s h o w s t h e a s s o c i a t i o n between
w e i g h t o f r o o t s p e r p l o t a n d l e v e l s o f p h o s p h o r u s i n t h e petioles
t o b e n e g l i g i b l e : w h e r e a s , t h o s e i n v o l v i n g s u c r o s e a n d purity
a r e n o t m a r k e d b u t a r e s t a t i s t i c a l l y s i g n i f i c a n t a n d favorable,
being positive.
Discussion

and

Summary

(A) T h e associations b e t w e e n levels of total n i t r o g e n in the


t h i n j u i c e , p e r c e n t a g e sucrose, a n d p e r c e n t a g e a p p a r e n t purity
a r e negative a n d for p e r c e n t a g e a p p a r e n t p u r i t y is extremely
close. In fact, t h e association is so close (r = -0.95) as to practically p r e c l u d e the possibility of genetically c o m b i n i n g high
total n i t r o g e n in t h e t h i n j u i c e w i t h high p e r c e n t a g e apparent
p u r i t y . H o w e v e r , t h e association b e t w e e n high levels of total nit r o g e n in t h e petioles a n d h i g h p e r c e n t a g e a p p a r e n t purity is
m u c h lower, only 4 p e r c e n t of t h e variability b e i n g attributable
to covariance. M o r e o v e r , the h i g h e r levels of total n i t r o g e n in
t h e petioles a r e positively associated w i t h h i g h e r yields of roots
(weight of r o o t p e r plot) . T h i s indicates t h a t h i g h levels of
total n i t r o g e n in t h e petioles are c o n d u c i v e to g r e a t e r weight of
roots p e r plot. P e r c e n t a g e sucrose is adversely asociated with
h i g h e r levels of total n i t r o g e n in t h e petioles b u t t h e adverse
associations a r e n o t as m a r k e d as for h i g h e r levels of total nitrogen in t h e t h i n j u i c e . In fact, only 22 p e r c e n t of t h e variability
is covariance, i n d i c a t i n g t h a t genetically h i g h e r levels of total
n i t r o g e n in t h e petioles can be c o m b i n e d w i t h h i g h e r percentage
sucrose. T h e d a t a in T a b l e 4 for t h e F 1 h y b r i d 52-430 X 52-408
as p o i n t e d o u t in t h e discussion u n d e r results, show t h a t these
t w o c h e m i c a l c h a r a c t e r s can b e favorably r e c o m b i n e d .
(B) Essentially t h e same findings h o l d for levels of potassium,
s o d i u m , a n d p h o s p h o r u s . T h a t is, it is m u c h m o r e desirable to
have t h e h i g h e r levels of these chemicals in t h e petioles as con
trasted w i t h t h e t h i n j u i c e .
(C) F u r t h e r , it seems t h a t t h e h i g h e r levels of total nitrogen
potassium, a n d s o d i u m in t h e petioles a r e c o n d u c i v e , if not essential, t o t h e p r o d u c t i o n o f h i g h e r yields. T h e h i g h e r levels o f
p h o s p h o r u s in t h e petioles a r e associated w i t h h i g h e r percentage

VOL. 13, No. 2, JULY 1964

149

sucrose a n d h i g h e r p e r c e n t a g e a p p a r e n t p u r i t y . T h e r e l a t i o n i s
stronger for p e r c e n t a g e a p p a r e n t p u r i t y t h a n f o r p e r c e n t a g e
sucrose.
(D) F i n a l l y , t h e r e s e e m s t o b e n o r e a s o n w h y t h e m e t a b o l i c
r e q u i r e m e n t s for h i g h e r yields o f r o o t s , h i g h e r p e r c e n t a g e sucrose, a n d h i g h e r p e r c e n t a g e p u r i t y c a n n o t b e m e t b y p r o d u c i n g
and g r o w i n g g e n o t y p e s w h i c h , a t t h e t i m e o f h a r v e s t , t e n d t o
have t h e h i g h e r levels o f t o t a l n i t r o g e n , p o t a s s i u m , s o d i u m , a n d
phosphorus i n t h e p e t i o l e s r a t h e r t h a n i n t h e t h i n j u i c e .
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M E R L E G. PAYM,.

1963. Chemical genetic a n d soils studies involving thirteen characters


in sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 1 2 ( 5 ) : 393-448.
(14)

POWERS, L E R O Y , E. E. R E M M E N G A a n d

N . S. U R Q L H A R T .

In

process of

publication. T h e p a r t i t i o n i n g m e t h o d of genetic analysis applied


to a study of weight per root a n d percentage sucrose in sugar beets
(Beta vulgaris L.) .

Variation in Emergence of Sugar Beet Plants


Attributable to Different Drill Units
of the Same Planter
H.

L.

Received for publication

BUSH1
November

27,

1963

Stand c o u n t s in t h e u n t r e a t e d " c h e c k s " of a c o m p r e h e n s i v e


sugar beet seed t r e a t m e n t test in 1961 p r o v i d e d an o p p o r t u n i t y
for s t u d y i n g t h e v a r i a b i l i t y of i n i t i a l s t a n d s o b t a i n e d f r o m p l a n t ing with different u n i t s of t h e s a m e d r i l l .
T h e r e s u l t s for t w o m a k e s of d r i l l s ( d e s i g n a t e d A a n d B)
were r e p o r t e d in e n o u g h cases for s o m e statistical analysis of
the data to be m a d e . A s u m m a r i z a t i o n of t h e highest s t a n d c o u n t
compared w i t h t h e lowest is p r e s e n t e d in T a b l e 1 for t h e t w o
makes of d r i l l s , b o t h in c o m m o n usage. F e w d r i l l s h a d all u n i t s
producing s t a n d s w i t h i n t h e l i m i t s o f r a n d o m v a r i a t i o n ,
LSD = 11.
Table 1.Seedling counts of the row planted by a drill unit which had the highest
count and the lowest count in four 100-inch samples per row for two makes of drill.

LSD 5% pt. = 11 between units for each drill.

T h i s v a r i a t i o n of t h e r e s u l t a n t stands m a y reflect differences


between u n i t s as to seed m e t e r i n g , d e p t h of p l a n t i n g , p r e s s u r e
on press w h e e l s a n d u n i f o r m i t y of c o v e r i n g , to list a few. T h i s
variation i s e n l i g h t e n i n g b e c a u s e i t confirms t h e n e e d t o a c c o u n t
f o r drill u n i t v a r i a t i o n i n t h e d e s i g n o f field e x p e r i m e n t s i n w h i c h
Statistician-Agronomist, T h e Great Western Sugar Co., Exp. Sta., Longmont, Colorado.

152

J O U R N A L OF THE A. S, S. B. T.

c o m m e r c i a l m a c h i n e s a r e u s e d a n d i n w h i c h s e e d l i n g c o u n t s are
i n v o l v e d . F o r t h e f a r m e r , i t i n d i c a t e s t h a t a l l p a r t s o f t h e drill
w h i c h m i g h t affect u n i f o r m i t y o f s t a n d s m u s t b e c a r e f u l l y calibrated a n d adjusted before the drill is taken to the field. It is
d i f f i c u l t , i f n o t i m p o s s i b l e , t o p r o d u c e a u n i f o r m f i n a l b e e t population by m e c h a n i c a l m e a n s w h e n such large differences as to
s t a n d e x i s t i n t h e d i f f e r e n t r o w s . A h i g h e r d e g r e e o f precision
i n t h e c o n s t r u c t i o n o f t h e d r i l l , e s p e c i a l l y t h e m e t e r i n g device,
t o g e t h e r w i t h a n o v e r - a l l c o n s t r u c t i o n t o a c h i e v e a u n i f o r m sowi n g u n d e r v a r i a t i o n s i n s e e d b e d , i s i n d i c a t e d f o r d r i l l manufacturers.

Evaporator H e a t Transfer Coefficients


for Beet Sugar Solution
ALLISON

S.

Received for publication

CHANG1
January

20,

1964

E v a p o r a t o n plays o n e of t h e m o s t i m p o r t a n t roles in t h e s u g a r
refineries. T h e existing d a t a a v a i l a b l e to give a f u n c t i o n a l relationship b e t w e e n heat transfer coefficients a n d operating: pressures are s o m e w h a t l i m i t e d . K e r r (2) 2 o b t a i n e d t h e h e a t transfer
coefficients for t h i r t y - n i n e different e v a p o r a t o r s , d o u b l e - , triple-,
and quadruple-effect i n actual o p e r a t i o n . H e p l o t t e d h e a t transfer
coefficients against o p e r a t i n g pressures in p o u n d s gage a n d in
inches o f m e r c u r y v a c u u m . T h e p o i n t s s c a t t e r e d s o b a d l y t h a t i t
requires a careful j u d g m e n t to use t h e m .
Several E u r o p e a n t e c h n o l o g i s t s h a v e w o r k e d o u t t h e f o r m u l a s
which take i n t o c o n s i d e r a t i o n b r i x of t h e juice a n d t h e t e m p e r a ture of the h e a t i n g v a p o r . H o w e v e r , t h e h e a t transfer coefficients
calculated b y t h e f o r m u l a s d o n o t a g r e e w i t h t h e a c t u a l v a l u e s
obtained by h e a t b a l a n c e in t h i s i n v e s t i g a t i o n .
T h e d a t a r e p o r t e d h e r e w e r e o b t a i n e d from full size evaporators in actual o p e r a t i o n . H e a t transfer coefficients w e r e p l o t t e d
against solids i n s o l u t i o n r a t h e r t h a n o p e r a t i n g pressures. T h e
result showed a b e t t e r c o r r e l a t i o n b e t w e e n h e a t transfer coefficients a n d solids i n s o l u t i o n . T h e c u r v e s h o w n o n F i g u r e 1 i s
applicable t o b o t h s t a n d a r d s h o r t v e r t i c a l - t u b e e v a p o r a t o r s a n d
standard h o r i z o n t a l - t u b e e v a p o r a t o r s w i t h t u b e s i n n o r m a l cleanliness.
A f o r m u l a based on t h e c u r v e s h o w n on F i g u r e 1 was proposed to e s t i m a t e t h e h e a t t r a n s f e r coefficients for b e e t s u g a r
juice in t h e e v a p o r a t o r .
Apparatus
T h e a p p a r a t u s used i n this i n v e s t i g a t i o n w e r e q u i n t u p l e effect full size e v a p o r a t o r s a t F a c t o r i e s # 1 t o # 4 . T h e y w e r e
standard s h o r t v e r t i c a l - t u b e e v a p o r a t o r s w i t h c e n t e r d o w n t a k e .
Evaporators a t F a c t o r y # 5 w e r e h o r i z o n t a l - t u b e e v a p o r a t o r s except the first effect w h i c h was a s t a n d a r d v e r t i c a l - t u b e e v a p o r a t o r
with a c e n t e r d o w n t a k e .
T h e sizes of e v a p o r a t o r u s e d in t h i s i n v e s t i g a t i o n w e r e as
follows:
1

Chemical
Engineer, Engineering Department, T h e Amalgamated Sugar Company,
Ogden, Utah.
2
Numbers in parentheses refer to literature cited.

154

J O U R N A L OF T H E A. S. S. B. T.

Figure 1.Heat transfer coefficient for beet sugar solution.

Factory
1st

#1
Effect: T w o bodies
25,176 sq.ft., 1 1/4" O D , 14 ga. c o p p e r tubes,
11'-1/4" long.
10,750 sq.ft., 1 1/4" O D , 0.049" steel o u t s i d e , 0.035"
c o p p e r inside, bi-metal t u b e s , 9'-6 1/4" l o n g .
2 n d Effect: T h r e e bodies
16,300 sq.ft., 1 1/4" O D , 15 ga. c o p p e r tubes, 9'3/8" long.
15,400 sq.ft., 1 1/4" O D , 15 ga. c o p p e r tubes, 8'6 3/8" l o n g .
10,730 sq.ft., 1 1/4" O D , bi-metal t u b e s , 9'- 6 1/4"
long.
3 r d Effect: 6,900 sq.ft., 1 1/4" O D , 15 ga. c o p p e r tubes, 6'2 3/8" l o n g .
4 t h Effect: 6,630 sq.ft., 1 1/4" O D , 14 ga. c o p p e r tubes, 6'2 3/8" long.
5 t h Effect: 6,200 sq.ft., 1 1/4" O D , 14 ga. c o p p e r tubes, 5'6 3/8" l o n g .
Factory
#2
1st Effect: 16,744 sq.ft., 1 1/4" O D , 14 ga. c o p p e r tubes, 9'-

14" long.
2 n d Effect: T w o b o d i e s
15.217 sq.ft., 44 1 1/4" O D , 7 / 3 2 wall, steel tubes.
9' 1 4 " long.
5914 1 1/4" O D , 14 ga. c o p p e r t u b e s , 9'- 14" long.
6,385 sq. ft., 1 1/2" O D , 14 ga. c o p p e r tubes, 5'6 1/4" long.

VOL. 13, N o . 2, J U L Y 1064

155

3rd Effect: 10,032 sq.ft., 26 1 1/4" OD, 7/32" wall, steel tubes,
8'- 1/4" long.
4440 1 1/4" OD, 14 ga. copper tubes, 8'- 1/4" long.
4th Effect: 6,854 sq.ft., 1 1/4" OD, 14 ga. copper tubes, 6'1/4 " long.
5th Effect: 7,231 sq.ft., 1 1/4" OD, 14 ga. copper tubes, 6'4 1/4" long.
Factory #3
1st Effect: 16,744 sq.ft., 1 1/4" OD, 14 ga. copper tubes, 9'1/4" long.
2nd Effect: T w o bodies
11,132 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 6'7 3/4" long.
3,300 sq.ft., 2" OD, 14 ga. copper tubes, 4'- 1 1/4"
long.
3rd Effect: 8,614 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 6'7 3/4" long.
4th Effect: 6,058 sq.ft., 1 1/2" OD, 14 ga. copper tubes, 5'9 1/4" long.
5th Effect: 6,854 sq.ft., 1 1/4" OD, 14 ga. copper tubes, 6'1/4" long.
Factory #4
1st Effect: 15,843 sq.ft., 2,640 1 1/4" OD, 14 ga. copper tubes,
9'- 3/8" long.
2,106 2" OD. 14 ga. copper tubes, 9', 3/8" long.
2nd Effect: 8,474 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 7'0" long.
3rd Effect: 6,355 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 5'- 3"
long.
4th Effect: 4,540 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 3'- 9"
long.
5th Effect: 5,145 sq.ft., 1 1/4" OD, 15 ga. copper tubes, 4'- 3"
long.
Factory #5
1st Effect: 5,860 sq.ft., vertical-tube, 2" OD, 12 ga. copper,
5'- 5 1/2" long.
2nd Effect: 5,836 sq.ft., horizontal-tube, 1" OD, 16 ga. admiralty brass, 15'- 4" long.
3rd Effect: 3,945 sq.ft.; horizontal-tube, 1" OD, 16 ga. admiralty brass, 13'- 7" long.
4th Effect: Same as above.
5th Effect: Same as above.
Factory steam flow rates, dome pressure and temperature recorders were supplied by Taylor Instruments Company. T h i n
juice brix to 1st effect and thick juice brix from 5th effect were

156

J O U R N A L OF T H E A. S.

S. B. T

measured by laboratory refractometer. All intermediate brixes


of juice were measured by laboratory hydrometer. Quantities
of thin juice to 1st effect and thick juice from 5th effect at #2
and #4 factory for run #4 were measured by Foxboro, Model
No. 9650 C, Magnetic Flowmeter. Others were calculated by
overall factory heat and material balances. Juice inlet tempera
tures at 1st effect were measured by Model R, Dillon Dial Thermometers.

SOLIDS IN SOLUTION, BRIX

Figure 2.Boiling Point elevation for sugar solution.

SOLIDS IN SOLUTION, BRIX

F i g u r e 3.Specific h e a t of s u g a r solution t a k e n from laboratory data'

VOL.

13,

No.

2,

JULY

1064

157

General arrangement of evaporators for factories #3 and


#4 is shown on Figure 1. General arrangements of evaporators
and flash tanks for factories' #1 and #2 are shown on Figure 5.
General arrangement of evaporators for factory #5 is shown on
Figure 6.

FACTORIES 1 AND 2

Figure 4.Flow diagram (top) for factories #3 and # 4 .


Figure 5.Flow diagram (below) for factories #1 and #2.

Over-all Heat Transfer Coefficients


The heat transfer coefficients depend on the cleanliness of
surfaces on both the vapor and juice sides, on the metal of which
the tubes are made, on the ratio length and diameter of the
tubes, on the temperature difference between the vapor and
Juice side and, finally, on the brix of the juice and the temperature of juice.

158

JOURNAL OF THE A. S. S. B. T

FACTORY 3

Figure 5.Flow diagram for factory #5.

Dessin has w o r k e d o u t t h e following f o r m u l a w h i c h takes


i n t o c o n s i d e r a t i o n b r i x of t h e j u i c e leaving t h e e v a p o r a t o r and
t h e t e m p e r a t u r e o f t h e h e a t i n g steam o r v a p o r .
U = 960 (1OO B x ) ( t v 130)
16,000
T h e a b o v e f o r m u l a gives h i g h e r h e a t t r a n s f e r coefficients for 1st,
2 n d a n d 3 r d effect a n d l o w e r h e a t transfer coefficients for 4th
a n d 5 th effect u n d e r i n v e s t i g a t i o n .
Swedish t e c h n o l o g i s t s p r o p o s e t h e following f o r m u l a :
49.2 ( t j 32)
U
=
Bx
T h e above f o r m u l a gives l o w e r h e a t transfer coefficients for 4th
a n d 5 t h effect i n t h i s i n v e s t i g a t i o n .
MacDonald and Rodgers propose the following formula:
(tj 32)2

U = 55 100
j X j
T h e a b o v e f o r m u l a gives l o w e r h e a t t r a n s f e r coefficients for every
effect f r o m 1st to 5 t h b o d y in t h i s i n v e s t i g a t i o n .
D a t a s h o w n in T a b l e 1 w e r e 24 h o u r s average. D o m e pres
sures, d o m e t e m p e r a t u r e s , e x h a u s t steam pressures w e r e recorded
every h o u r o n t h e h o u r . J u i c e i n l e t t e m p e r a t u r e s , b r i x e s o f j u i c e
to a n d from each effect b e t w e e n 2 n d a n d 4 t h effect w e r e measured
a n d r e c o r d e d o n c e every t w o h o u r s . J u i c e a n d s t e a m flow rates
to 1st effect a n d t h i c k j u i c e f r o m 5 t h effect w e r e calculated by
overall h e a t a n d m a t e r i a l b a l a n c e t o w i t h i n 2 % accuracy f o r
every factory w i t h o u t m a g n e t i c flowmeter.

TABLE 1.

*1st flash tank in service only.

TABLE 1.(Continued)

TABLE 1.(Continued)

Atmospheric pressure
Factory No. 1: 13,59 psia, 27.63 "Hg.
Factory No. 3: 12.81 psia, 26.03 "Hg.
Foctory No. 5: 12.4 psia, 25.2 "Hg.

Factory No. 2: 13.41 psia, 27.33 "Hg.


Factory No. 4: 12.56 psia, 25.52 "Hg.

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JOURNAL OF THE A. S. S. B. T.

O v e r a l l h e a t transfer coefficients w e r e c a l c u l a t e d by
U=Q
and tabulated in T a b l e 2. T X A
Where

T = effective t e m p e r a t u r e difference, F
= a p p a r e n t t e m p e r a t u r e difference b.p.r., F.

A p p a r e n t t e m p e r a t u r e difference:
1st effect = e x h a u s t steam t e m p e r a t u r e
temperature, F.
2 n d effect = 1st v a p o r sat. t e m p e r a t u r e
temperature, F.
3rd effect = 2 n d v a p o r sat. t e m p e r a t u r e
temperature, F.
4 t h effect = 3rd v a p o r sat. t e m p e r a t u r e
temperature, F.
5th effect = 4 t h v a p o r sat. t e m p e r a t u r e
temperature, F.

1st v a p o r sat.

2 n d vapor sat.

3 r d vapor sat.
4 t h vapor sat.

5th vapor sat.

S a m p l e c a l c u l a t i o n s : Factory # 2 , R u n 4 .
T h i n j u i c e to 1st effect = 510,000 # / h r .
Solids in j u i c e = 67,800 # / h r . @ 13.3 b r i x .
S t e a m to 1st effect = 188,000 # / h r . @ 24.5 psig a n d 264 F.
1st Effect:
H e a t from steam = 188,000 (936) = 176,000,000 B t u / h r .
H e a t i n g j u i c e = 510,000 (245-223)
(0.925) = 10,350,000
Btu/hr.
A v a i l a b l e h e a t = 165,650,000 B t u / h r .
165,650,000
1st v a rp o r
=
= 174,500 # / h r .
949.5
Juice
Brix
510,000 # / h r .
T o process = 54,400 # / h r .
174,500 # / h v .
To 2 n d effect = 120,100 # / h r .
335,500 # / h r . 20.2
B . P . R . = 1.0F from F i g u r e 2
T = 264 244 1 19F
H e a t i n g surface = 16,744 sq.ft.
176,000,000
U = 19 X 16,744 = 553 Btu p e r hr. p e r sq ft. per F.

2nd Effect:
H e a t from v a p o r = 120,100 (949.5) = 114,000,000 Btu/hr.
J u i c e flash = 335,500 (245 230) 0.82 = 4,130,000 Btu/hr.
A v a i l a b l e h e a t = 118,130,000 B t u / h r .

164

JOURNAL OF THE A. S. S. B. T

E v a p o r a t o r h e a t transfer coefficients a r e o n e of t h e most


i m p o r t a n t d a t a for sugar refineries. W i t h t h e d a t a of h e a t transfer
coefficients it e n a b l e s t h e refineries to e s t i m a t e t h e required
h e a t i n g surface for c e r t a i n o p e r a t i o n by r e a r r a n g i n g t h e equation
to t h e following form.

Apparent temperature difference


1st effect: 1st vapor sat. temperature === exhaust steam tens
perature apparent temperature difference a
1st effect.
2nd effect: 2nd vapor sat. temperature = 1st vapor sat, tem
perature apparent temperature difference:
2nd effect.
3rd effect: 3rd vapor sat. temperature = 2nd vapor sat, tem
perature apparent temperature difference
2nd effect.
4th effect: 4th vapor sat. temperature 3rd vapor sat, ten
perature apparent temperature difference at
4 th effect.
5th effect: 5th vapor sat. temperature = 4th vapor sat. tem
perature apparent temperature difference at
5 th effect.

VOL.

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2,

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1964

165

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1964
TABLE 2.(Continued)

Btn per hour per sq.ft. per

F.

I t also e n a b l e s t h e refineries t o e s t i m a t e t h e v a p o r t e m p e r a ture a n d p r e s s u r e for e x i s t i n g h e a t i n g surface b y r e a r r a n g i n g


the e q u a t i o n to t h e following f o r m .

From the data obtained in this investigation, the following


formula is p r o p o s e d .

O v e r a l l h e a t t r a n s f e r coefficients c a l c u l a t e d b y t h e a b o v e
formula a g r e e w i t h t h e v a l u e s o b t a i n e d from F i g u r e 1 .
S u m m a r y a n d Conclusions
T h e r e s u l t of t h i s i n v e s t i g a t i o n s h o w e d a b e t t e r c o r r e l a t i o n
between h e a t t r a n s f e r coefficients a n d solids in s o l u t i o n .
T h e curve shown on Figure 1 is applicable to both standard
short v e r t i c a l - t u b e e v a p o r a t o r s a n d s t a n d a r d h o r i z o n t a l - t u b e e v a p orators w i t h t u b e s i n n o r m a l c l e a n l i n e s s p r o v i d e d t h a t t h e t u b e
size a n d o p e r a t i n g c o n d i t i o n s a r e w i t h i n t h e f o l l o w i n g r a n g e s :

168

JOURNAL OF THE A. S. S. B. T,

T u b e d a t a a n d o p e r a t i n g c o n d i t i o n s for h o r i z o n t a l - t u b e evapo r a t o r a r e s h o w n on pages 155-157 a n d T a b l e 1 at factory #5.


F i g u r e 1 a n d E q u a t i o n (1) m a y or m a y n o t be applicable
to l o n g - t u b e e v a p o r a t o r , since t h e d a t a w e r e n o t available and
no a t t e m p t was m a d e to investigate t h e overall h e a t transfer
coefficients for vertical l o n g - t u b e e v a p o r a t o r .
E q u a t i o n (1) or F i g u r e 1 will a i d t h e d e s i g n e r to design a
n e w e v a p o r a t o r or to e s t i m a t e t h e v a p o r pressure from the exist
ing e v a p o r a t o r .
Nomenclature

VOL.

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1964

Acknowledgments
C o o p e r a t i o n s from E n g i n e e r i n g D e p a r t m e n t , O p e r a t i n g Department a n d R e s e a r c h D e p a r t m e n t o f T h e A m a l g a m a t e d S u g a r
Company m a d e t h i s w o r k possible. M r . S. M. H e i n e r , C h i e f
Engineering, assisted in t h e i n v e s t i g a t i o n .
Literature Cited
(1) HONIG, P., "Principles of Sugar T e c h n o l o g y . "

1963.

Vol. I l l , Elsevier

Publishing C o m p a n y , New York.


(2) KERR, T r a n s . A m . Soc. Mech. Engrs., 38, 67 (1917).
(3) SPENCER, G. L. a n d G. P. M E A D E . 1945. " C a n e Sugar H a n d b o o k , " 8th
Edition, J o h n Wiley a n d Sons, Inc., N e w York.

Influence of Prolonged Association of Sugar-Beet


N e m a t o d e and T o m a t o on Intensity of Parasitism 1 I
ARNOLD E. STEELE2

Received for publication January 27, 1964

j
J

Riggs and Winstead (7)4 found that a small proportion of


populations of root-knot nematodes, Meloidogyne incognita, M.
incognita acrita, and M. arenaria, reproduced on root-knot-resistant Hawaii 5229 tomatoes. By three successive transfers of
these nematodes on this variety of tomato, populations to which
the tomato had very little or no resistance were obtained. There
was no change in virulence after maintenance of these populations on tobacco, okra, cucumber, or tomato for 9 months. The
authors considered that the change was not due to a physiological
effect of the resistant plants, but that the resistant plants merely
served to select out individuals capable of reproducing on the|
resistant variety.
Similar studies by Sauer and Giles (8) demonstrated that
the resistance of Hawaiian Experiment Station tomato lines to
M. javanica was broken by the selection of an aggressive strain
of M. javanico which could reproduce on resistant plants. Resistance-breaking biotypes of Heterodera rostochiensis and H.
schachtii have been reported by Jones (3) and Shepherd (9)
respectively.
I
Tomato is frequently included in rotations with sugar beets.
Jones (2) and Mulvey (5) reported instances where the beet
nematode did not develop to maturity on tomatoes grown in
infested soil. Reports by Raski (6) and Golden and Shafer
(1), however, indicate that certain varieties of tomato are suitable
hosts for the beet nematode.
In preliminary tests, populations of the sugar-beet nematode
were recovered from tomato and sugar beet previously inoculated
with cysts obtained from a single population. Reinoculation of|
these populations on tomato revealed that consistently fewer
adult females and larvae of the nematode were recovered from
tomatoes grown for 15, 30, or 60 days in soil containing cysts
the population from sugar beets than from tomato receiving
cysts of the population from tomato.
I

VOL- 13, No. 2, JULY 1964

171

T h e increased infectivity o f t h e tomato-selected p o p u l a t i o n s


appeared to h a v e r e s u l t e d from e i t h e r a r a p i d a d a p t a t i o n of t h e
parasite to t o m a t o or f r o m t h e selection of a race t h a t was p a t h o
genic to t o m a t o .
T h e q u e s t i o n o f w h e t h e r biological races o r a d a p t i v e m e c h anisms exist in Heterodera schachtii is of practical i m p o r t a n c e to
programs d e s i g n e d t o d e v e l o p effective c o n t r o l t h r o u g h c r o p p i n g
systems and t h r o u g h b r e e d i n g s u g a r beets for t o l e r a n c e to n e m a tode infections. C o n s e q u e n t l y , a s t u d y was u n d e r t a k e n to e x p l o r e
the host-parasite r e l a t i o n s of t o m a t o a n d t h e sugar-beet n e m a t o d e .
Materials and Methods
Cysts of Heterodera schachtii w e r e r e c o v e r e d from t o m a t o
plants used i n t h e p r e l i m i n a r y e x p e r i m e n t a n d d i v i d e d i n t o t w o
groups. O n e g r o u p was a d d e d to a flat of seedlings of A g g l e r
and Musser's P e a r s o n A-l
tomato
(Lycopersicon esculentum),
and the o t h e r g r o u p was a d d e d to a flat of s e e d l i n g of s u g a r b e e t
(Beta vulgaris c u l t i v a r U.S. 75). Cysts r e c o v e r e d from s u g a r beets
were added to a n o t h e r flat of s u g a r b e e t seedlings. In each case,
the seedlings w e r e s t a r t e d in sterilized s a n d a n d t r a n s p l a n t e d to
sterilized soil. M a t u r e cysts of each of t h e p o p u l a t i o n s w e r e recovered 120 days after i n o c u l a t i o n a n d d e s i g n a t e d as follows:
A . T o m a t o p o p u l a t i o n . O n t o m a t o 6 0 p l u s 120 days.
B . T o m a t o - b e e t p o p u l a t i o n . O n t o m a t o 6 0 days a n d o n s u g a r
beet 120 days.
C. Beet p o p u l a t i o n . On s u g a r b e e t 60 p l u s 120 days.
For the first e x p e r i m e n t t w e n t y - f o u r a l u m i n u m foil c y l i n d e r s
filled with sterilized soil w e r e p l a c e d in each of 3 flats. A single
tomato seedling t r a n s p l a n t a n d 15 sugar-beet n e m a t o d e cysts of
one of t h e t h r e e n e m a t o d e p o p u l a t i o n s w e r e t r a n s f e r r e d to each
cylinder. T h e flats w e r e p l a c e d in a r o w on a single b e n c h in
the greenhouse.
Six p l a n t s w e r e r e m o v e d from each flat 15, 30, a n d 60 days
after t r a n s p l a n t i n g . T h e r o o t s o f t h e p l a n t s w e r e w a s h e d a n d
weighed a n d t h e r o o t s a n d soil e x a m i n e d for cysts a n d w h i t e
female n e m a t o d e s .
After m a t u r e female n e m a t o d e s w e r e r e m o v e d , t h e r o o t s
were stained in a b o i l i n g s o l u t i o n of l a c t o p h e n o l a n d acid fuchsin
and destained in l a c t o p h e n o l , as d e s c r i b e d by M c B e t h . T a y l o r .
and Smith (4). R o o t s w e r e t h e n f r a g m e n t e d in a b l e n d e r a n d
the root d e b r i s was e x a m i n e d for larvae.
Sixty days after i n o c u l a t i o n , t h e r e m a i n i n g six t o m a t o p l a n t s
w e r e r e m o v e d f r o m each o f t h e t h r e e f l a t s , a n d r e p l a n t e d i n
individual two-gallon crocks. Sixty days l a t e r t h e p l a n t s w e r e
removed from crocks
a n d t h e r o o t s e x a m i n e d for w h i t e f e m a l e
nematodes. Soil f r o m i n d i v i d u a l crocks was t h o r o u g h l y m i x e d ,

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J O U R N A L OF T H E A. S. S. B. T.

w e i g h e d , a n d s a m p l e d , a n d t h e s a m p l e s w e r e w a s h e d , screened,
a n d e x a m i n e d f o r a d u l t f e m a l e s a n d c y s t s . C o u n t s o f nematodes
in v a r i o u s stages of d e v e l o p m e n t a r e r e c o r d e d in T a b l e 1.
A s e c o n d e x p e r i m e n t w a s d e s i g n e d t o d e t e r m i n e t h e increase
of two nematode populations on tomato. O n e population inc l u d e d c y s t s o b t a i n e d f r o m t o m a t o i n t h e p r e c e d i n g t e s t . The
o t h e r p o p u l a t i o n w a s i n s o i l f r o m a s u g a r - b e e t f i e l d located
a b o u t 20 miles from the source of the p o p u l a t i o n s described in
t h e p r e c e d i n g t e s t . B e e t s w e r e g r o w n i n t h e s o i l 1 2 0 d a y s and
t h e soil w a s h e d a n d s c r e e n e d t o r e c o v e r cysts. E i g h t - i n c h c l a y
p o t s w e r e filled w i t h s t e r i l i z e d soil.
E a c h o f 3 2 p o t s received
o n e P e a r s o n A - 1 t o m a t o s e e d l i n g t r a n s p l a n t a n d 1 5 nematode
c y s t s , s i x t e e n o f t h e s e p o t s r e c e i v e d c y s t s f r o m t o m a t o , a n d the
s e c o n d 1 6 p o t s r e c e i v e d c y s t s f r o m b e e t s . E a c h t r e a t m e n t (population)
w a s r e p l i c a t e d 4 t i m e s a n d i n c l u d e d 4 s a m p l e s (pots).
T h e p o t s w e r e a r r a n g e d i n a r a n d o m i z e d b l o c k - d e s i g n o n benches
i n a g r e e n h o u s e . S i x t y a n d n i n e t y d a y s a f t e r i n o c u l a t i o n , samples
w e r e r e m o v e d t o t h e l a b o r a t o r y w h e r e t o m a t o r o o t s a n d soil
w e r e p r o c e s s e d a s p r e v i o u s l y d e s c r i b e d f o r r e c o v e r y o f adult
f e m a l e n e m a t o d e s a n d exsts. D a t a f r o m t h i s test ( T a b l e 2)
w e r e a n a l y z e d f o r s t a t i s t i c a l s i g n i f i c a n c e b y t h e a n a l y s i s o f variance
method.
A t h i r d e x p e r i m e n t w a s d e s i g n e d t o d e t e r m i n e w h e t h e r popul a t i o n differences o b s e r v e d in t h e p r e v i o u s tests w o u l d be obt a i n e d w h e n l a r v a e r a t h e r t h a n c y s t s w e r e u s e d f o r inoculum.
T o m a t o o r b e e t s e e d l i n g s w e r e i n o c u l a t e d w i t h approximately
1,000 l a r v a e o f e a c h o f t h e t h r e e p o p u l a t i o n s u s e d i n t h e f i r s t
e x p e r i m e n t . E a c h o f 5 t r e a t m e n t s w e r e r e p l i c a t e d 4 t i m e s . The
p l a n t s w e r e a l l o w e d t o g r o w 9 0 d a y s i n a g r e e n h o u s e a f t e r which
a d u l t f e m a l e n e m a t o d e s a n d c y s t s w e r e r e c o v e r e d b y washing
a n d s c r e e n i n g p l a n t r o o t s a n d s o i l . C o u n t s o f a d u l t nematodes
are listed in T a b l e 3.
T h e f o u r t h e x p e r i m e n t w a s d e s i g n e d to d e t e r m i n e the effect
o f s u g a r - b e e t - r o o t d i f f u s a t e o n h a t c h i n g o f 2 4 0 c y s t s selected a t
r a n d o m f r o m e a c h o f t h e t w o n e m a t o d e p o p u l a t i o n s inoculated
o n t o m a t o i n t h e s e c o n d t e s t . E a c h c y s t g r o u p w a s exposed 6
w e e k s to t r e a t m e n t s of sugar-beet-root diffusate or tap water.
T r e a t m e n t s w e r e r e p l i c a t e d 4 t i m e s in i n d i v i d u a l Syracuse watch
glasses w h i c h c o n t a i n e d 30 cysts a n d 15 ml of t r e a t m e n t solution.
T h e dishes with contents were stored at r o o m temperatures in a
d a r k a e r a t e d c a b i n e t i n t h e l a b o r a t o r y . A t w e e k l y intervals, c y s t s
w e r e t r a n s f e r r e d t o c l e a n w a t c h g l a s s e s c o n t a i n i n g fresh t r e a t ment solutions and the emerged larvae were counted.

VOL-

13,

No.

2,

JULY

1964

173

Microscopic studies of second-stage larvae, a d u l t m a l e s a n d


females, a n d b r o w n cysts w e r e m a d e t o d e t e r m i n e w h e t h e r t h e
populations c o u l d b e m o r p h o l o g i c a l l y d i s t i n g u i s h e d . Five larvae
from five cysts of each p o p u l a t i o n w e r e collected a n d e x a m i n e d
on each of four s a m p l i n g dates.
Results
I n the f i r s t t h r e e e x p e r i m e n t s m o r e larvae a n d a d u l t s w e r e
recovered from t o m a t o e s i n o c u l a t e d w i t h cysts o b t a i n e d from
tomato t h a n from t o m a t o e s i n o c u l a t e d w i t h cysts o b t a i n e d from
sugar beets ( T a b l e s 1, 2, a n d 3). In t h e first e x p e r i m e n t . 22.4
times as m a n y female a d u l t s a n d cysts w e r e r e c o v e r e d after 120
days from t o m a t o e s i n o c u l a t e d with cysts o b t a i n e d from t o m a t o
as from t o m a t o e s i n o c u l a t e d w i t h cysts o b t a i n e d from s u g a r
beets. T h e second a n d t h i r d e x p e r i m e n t s gave c o m p a r a b l e
values, 27.4 a n d 17.1 t i m e s , respectively.
In the first experiment, the majority of nematodes had
apparently c o m p l e t e d t w o cycles of d e v e l o p m e n t in 120 days;
i.e. the a d u l t s w e r e p r o b a b l y offspring of a d u l t s p r e s e n t at a b o u t
6 0 days. T h e n u m b e r o f a d u l t s r e c o v e r e d from t o m a t o p l a n t s
120 days after i n o c u l a t i o n w i t h cysts o b t a i n e d from t o m a t o e s
(population A. T a b l e 1) r a n g e d from 1,618 to 5,401, w i t h an
average of 2,934 cysts p e r p l a n t . T h i s a m o u n t s to a 194.6-fold
increase i n cysts o v e r t h e o r i g i n a l n u m b e r i n o c u l a t e d c o m p a r e d
with 11.5-fold increase in t h e p o p u l a t i o n s from s u g a r b e e t s ( p o p u lation C, T a b l e 1).
T h e average n u m b e r s of l a r v a e e m e r g e d from 40 cysts of
tomato or b e e t p o p u l a t i o n s t r e a t e d 6 weeks w i t h beet-root diffusate were 5.546 a n d 4,954, respectively, w i t h a significant
difference o f 492 a t t h e 5 % level. T h e ' a v e r a g e n u m b e r s o f
larvae e m e r g e d in t a p w a t e r from cysts o b t a i n e d from b e e t or
tomato were 1,697 a n d 1,920, respectively.
Microscopic e x a m i n a t i o n s o f larvae, a d u l t m a l e s a n d females,
a n d b r o w n cysts revealed t h a t p o p u l a t i o n s from beet o r t o m a t o
differed in t h e a v e r a g e l e n g t h s of second-stage larvae a n d a v e r a g e
lengths of a d u l t males. L a r v a e from beet-selected p o p u l a t i o n s
averaged 456.6 m i c r o n s , w h e r e a s l a r v a e from tomato-selected
populations a v e r a g e d 438.5 m i c r o n s . A d u l t m a l e s from b e e t s
averaged 1.44 m m , w h i l e those from t o m a t o a v e r a g e d 1.21 m m .
These differences a r e n o t c o n s i d e r e d sufficient to s e p a r a t e t h e
populations as d i s t i n c t species at t h i s t i m e , since they m a y he
t h e r e s u l t o f e n v i r o n m e n t a l influence ( h o s t - d e t e r m i n e d variations) rather t h a n genetically transmitted characteristics.

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Table 1.Total numbers of Heterodera schachtii larvae adults and cysts recovered
from roots and soil of tomato examined 15, 30, 60, and 120 days after inoculation. 1

1
Figures given are the total number of nematodes removed from 6 samples.
2
Populations are represented as follows: A. Tomato population. B. Tomato-beet population.
C. Beet population.
3
Excluding cysts recovered from soil.

Table 2.Total number of adult females and cysts of Heterodera schachtii recovered
from tomato plants 60 and 90 days after inoculation with cysts.

1
Populations are represented as follows: ATomato population. CBeet population.
2
Figures are total numbers of nematodes recovered from 2 tomato plants sampled at
the indicated date.

Table 3.Total numbers of adult females and cysts of Heterodera Schachtii recovered
from beet or tomato plants 90 days after inoculation with larvae.

1
Populations are represented as follows: ATomato population. BTomato-beet
population. GBeet population.
2
Figures are total numbers of nematodes recovered from 5 tomato plants.

VOL.

13,

No.

2,

JULY

1964

175

Discussion and Conclusions


Reproduction of Heterodera schachtii on tomato was increased during a brief period of association of this host and
parasite. Although 3 years had lapsed between the first and
third tests of this study with the populations maintained on
their respective host plants in the meantime, the ratio of nematode progeny obtained from tomato inoculated with beet- of
tomato-selected populations were about the same. This indicates that prolonged association did not further increase the
reproductive rate. When a population obtained from tomato
was allowed to reproduce on sugar beet and the progeny reinoculated on tomato, the numbers of adult nematodes obtained
from the third transfer host were intermediate to the numbers
obtained from tomato inoculated with beet- or tomato-selected
populations (Table 1 and 3). The data suggest that reproduction
of tomato-selected populations on tomato was decreased during
the brief association with sugar beet. Association of the beet
nematode with tomato also appeared to reduce ability to reproduce on sugar beet (Table 3).
Since results were comparable regardless of whether cysts of
larvae were used as inoculum, failure of the nematodes to hatch
from the cysts can be eliminated as a possible explanation of
the differences in reproduction on sugar beets and tomatoes.
The fact that beet root diffusate had about the same effect on
both populations also supports this view. Results of all tests
strongly indicate that there was an "adaptation" to tomatoes.
The mechanism of the "adaptation" is of theoretical and
practical interest. Steiner (10) and others suggested that physiological adaptation may take place, but many of the instances
cited are now known to be due to confusion of nematode species
or to misinterpretation of field observations. Experiments with
populations derived from a single cyst might conclusively demonstrate whether results as found in these tests were due to "adaptation" or to selective development of taxa (races or subspecies)
present in the original inoculum.
From the practical standpoint, the existence of field populations of nematodes able to attack sugar beets and capable of
increasing in ability to attack tomatoes is of importance, since
it shows that tomatoes should not be used in rotations with sugar
beets, and also suggests that tomatoes grown on former sugarbeet fields may increase nematode populations. The development of sound rotation practices and effective breeding programs
for control of the sugar-beet nematode depend on an understanding of host-parasite relations. The possibility of an adaptive

176

J O U R N A L OF T H E A. S. S. B. T.

m e c h a n i s m in
Heterodera
schachtii c a l l s a t t e n t i o n
for a d d i t i o n a l i n f o r m a t i o n o n t h i s subject.

to

t h e need

Summary

R e p r o d u c t i o n o f t h e s u g a r - b e e t n e m a t o d e (Heterodera
schachtii S c h m i d t ) on t o m a t o (Lycopersicon
esculentum)
was
increased d u r i n g a brief p e r i o d of association of host a n d parasite.
P r o l o n g e d association d i d n o t f u r t h e r increase t h e nematodes
r e p r o d u c t i v e r a t e on t o m a t o . R e s u l t s of four s e p a r a t e green
house tests suggest t h a t an a d a p t a t i o n of t h e b e e t nematode to
t o m a t o o c c u r r e d . H o w e v e r , the possibility t h a t test results were
d u e to selective d e v e l o p m e n t of taxa has n o t b e e n eliminated.
Literature
(1)

GOLDEN,

A.

M.

and

THELMA

Cited

SHAFER.

1959.

Host-parasite

of various p l a n t s a n d t h e sugar-beet n e m a t o d e ,
U S D A P l a n t D i s . R e p t r . 4 3 (12) : 1 2 5 8 - 1 2 6 2 .

relationships

Heterodera

schachtii

(2)

J O N E S , F . G . W . 1 9 5 0 . O b s e r v a t i o n s o n t h e b e e t e e h v o r m a n d other
c y s t - f o r m i n g s p e c i e s o f Heterodera.
A n n . A p p l . Biol.
3 7 : 407-440.

(3)

J O N E S , F . G . W . 1 9 5 7 . R e s i s t a n c e - b r e a k i n g b i o t y p e s o f t h e p o t a t o root
e e l w o r m (Heterodera
rostochiensis
W o l l . ) . N e m a t o l o g i c a 2: 185-192,

(4)

M C B E T H , G. W., A.

L. T A Y L O R a n d A.

ing n e m a t o d e s in r o o t tisue.

T.

SMITH.

1941.

P r o c . H e l m . Soc. W a s h .

N o t e o n stain

8 (1) : 26.

(5)

M U L V E Y , R . H . 1 9 5 7 . S u s c e p t i b i l i t i e s o f c u l t i v a t e d a n d w e e d plants t o
the
sugar-beet
nematode,
Heterodera
schachtii
Schmidt,
1871, in
S o u t h w e s t e r n O n t a r i o . J . H e l m . 3 1 : 225-228.

(6)

R A S K I . D. J. 1952. On t h e host r a n g e of t h e s u g a r - b e e t nematode in


C a l i f o r n i a . U S D A P l a n t D i s . R e p t r . 3 6 : 5-7.

(7)

R I G G S . R . D . a n d N . W . W I N S T E A D . 1 9 5 9 . S t u d i e s o n t h e resistance i n
t o m a t o i n r o o t - k n o t n e m a t o d e s a n d o n t h e o c c u r r e n c e o f pathogenic
biotypes. P h y t o p a t h o l o g y 49: 716-724.

(8)

S A U E R , M . R . a n d J . E . G I L E S . 1 9 5 9 . F i e l d t r i a l w i t h a root-knot r e sistant t o m a t o variety.


I n v e s t i g a t i o n s R e s e a r c h S t a t i o n technical
p a p e r N o . 3.
C o m m o n w e a l t h S c i e n t i f i c a n d I n d u s t r i a l Research
O r g a n i z a t i o n . A u s t r a l i a , 1959.

(9)

S H E P H E R D , A U D R E Y M . 1959. T e s t i n g p o p u l a t i o n s o f b e e t e e l w o r m ,
Heterodera schachtii S c h m i d t f o r r e s i s t a n c e b r e a k i n g b i o t y p e s , using
t h e w i l d b e e t (Beta p a t e l l a r i s M o q . ) a s i n d i c a t o r . N a t u r e (London)
1 8 3 ( 4 6 6 8 ) : 1141-1142.
( 1 0 ) S T E I N E R , G . 1 9 2 5 . T h e p r o b l e m o f h o s t s e l e c t i o n a n d h o s t specializt i o n o f c e r t a i n p l a n t - i n f e s t i n g n e m a s a n d its a p p l i c a t i o n i n t h e
s t u d y o f n e m i c p e s t s . P h y t o p a t h . 1 5 (9) : 5 0 0 - 5 3 4 .

Longevity of Sugar Beet Seed1


D.

W.

ROBERTSON

AND

MILDRED

Received for publication

March

L.

THORNTON2

9, 1964

It is c o m m o n k n o w l e d g e t h a t t h e v i a b i l i t y of m o s t cereal
seeds r e m a i n s h i g h e r w h e n s t o r e d a t low h u m i d i t y t h a n w h e n
stored a t h i g h h u m i d i t y . T h e same r e l a t i o n s h i p h o l d s i n g e n e r a l
for sugar beet seed viability w h e n s t o r e d u n d e r s i m i l a r c o n d i t i o n s
of h u m i d i t y . H o w e v e r , few d a t a a r e a v a i l a b l e on t h e g e r m i n a tion p e r f o r m a n c e of processed ( s e g m e n t e d ) s u g a r b e e t seed w h e n
stored for an e x t e n d e d p e r i o d u n d e r c o n d i t i o n s of low h u m i d i t y .
T h e objective of this s t u d y was to d e t e r m i n e w h e t h e r or n o t
segmentation h a d a n y d e l e t e r i o u s effect on t h e g e r m i n a t i o n of
sugar beet seed after s t o r a g e in c l o t h sacks for v a r y i n g p e r i o d s
of years.
R e v i e w of Literature
In 1928 t w o lots of s u g a r b e e t seed w e r e p l a c e d in a commercial r e f r i g e r a t i o n storage h o u s e i n Salt L a k e C i t y . U t a h ( P a r k
and Owen) 3 . T h e t e m p e r a t u r e d u r i n g t h e test p e r i o d r a n g e d
from + 1 0 F to 10 F. G e r m i n a t i o n tests w e r e m a d e at intervals from 1928 to 1950. T h e g e r m i n a t i o n p e r c e n t in 1928 was
83.5, whereas in 1950 it was 75.0, or a d r o p of only 8.5 p e r c e n t .
Under n o r m a l storage c o n d i t i o n s , US N o . 1 s u g a r b e e t seed
showed 27 p e r c e n t g e r m i n a t i o n after storage for 19 years, as
compared w i t h t h e o r i g i n a l g e r m i n a t i o n of 80 p e r c e n t .
Materials and Methods
T h e sugar b e e t seed u s e d i n this i n v e s t i g a t i o n was o b t a i n e d
from T h e G r e a t W e s t e r n S u g a r C o m p a n y E x p e r i m e n t a l S t a t i o n ,
Longmont, C o l o r a d o . O n e lot e a c h c a m e from t h e c r o p years
of 1943, 1945 a n d 1946 a n d t w o lots from 1944. E a c h of these
five lots is r e p r e s e n t e d by t w o sub-lots. O n e sub-lot consisted
of whole seed, w h i l e t h e o t h e r sub-lot r e p r e s e n t e d t h e s a m e seed
i n segmented f o r m . T h e seeds w e r e s t o r e d i n t h e o r i g i n a l sacks
i n a dry, u n h e a t e d r o o m a t t h e C o l o r a d o A g r i c u l t u r a l E x p e r i ment Station, F o r t C o l l i n s , C o l o r a d o . E a c h year, from 1943 to
U 6 3 , samples w e r e d r a w n from t h e w h o l e a n d s e g m e n t e d seed
lots a n d g e r m i n a t i o n tests w e r e m a d e . B o t h ball a n d s p r o u t
counts were r e c o r d e d .
1

Contribution
of
the Department of Agronomy and the Colorado Seed Laboratory,
Colorado
Agricultural Experiment Station, Fort Collins, Colorado: Colorado Agricultural
2
Agronomist Station Scientific Series No. 934.
Colorado.
3
Park, D. A. and F. V. Oven. 1950. Viability of sugar beet seed held in cold storage
for 22 years. Proc. Sixth General Meeting, Am. Soc. Sugar Beet Technol.

Table 1.Germination percentages of whole and segmented sugar beet seed stored lor one to twenty years.

Table 2.Adjusted germination percentages assuming the original germination to be 100 percent.

J O U R N A L OF T H E A. S. S. B. T.

180

Experimental Results
A c t u a l g e r m i n a t i o n r e s u l t s a r e p r e s e n t e d in. T a b l e 1 . I n
o r d e r t o d e t e r m i n e t h e p e r c e n t a g e d e c r e a s e from t h e original
tests, all g e r m i n a t i o n s w e r e c o n v e r t e d to p e r c e n t a g e s ot the
o r i g i n a l g e r m i n a t i o n , t h a t is, t h e o r i g i n a l g e r m i n a t i o n was cons i d e r e d as 100 p e r c e n t ( T a b l e 2).
A statistical analysis of t h e first 17 years of t h e test (Table 1)
was m a d e t o d e t e r m i n e t h e significance o f t h e d a t a .
T h e statistical analysis s h o w e d (a) t h a t t h e r e was a significant
difference i n g e r m i n a t i o n b e t w e e n t h e w h o l e a n d segmented
seed ( T a b l e 3), (b) t h e r e was no significant difference between
years for t h e w h o l e seed, b u t a very significant difference in years
for t h e s e g m e n t e d seed. ( T a b l e s 4 & 5), (c) t h e r e was a significant
difference in s a m p l e s w i t h i n each t r e a t m e n t for t h e period of
t h e s t u d y , ( T a b l e 4 a n d 5).
T a b l e 3.Analysis of variance for both treatments. 4

4
Le Clerg, E. L., W. H. Leonard, and A. G. Clark.
Publishing Company.
Second edition.

1962 Field Plot Technique. Burgess

T a b l e 4Analysis of variance for whole seed.

Table 5.Analysis of variance for segmented seed.

VOL. 13, No. 2, JULY 1964

Figure 1.Percentage g e r m i n a t i o n of w h o l e a n d s e g m e n t e d seed


years for a period of 17 years.

181

by

T h e r e was a d r o p i n g e r m i n a t i o n w h e n t h e s e g m e n t e d seed
w a s c o m p a r e d t o t h e w h o l e seed. T h e difference i n t h e o r i g i n a l
g e r m i n a t i o n was 5, or a d e c r e a s e of 6.4 p e r c e n t . T h e final g e r m ination of t h e o n e lot of w h o l e seed t h a t was s t o r e d for 20 years
was 97 p e r c e n t of t h e o r i g i n a l g e r m i n a t i o n . At t h e e n d of 17
years, w h e n t h e a v e r a g e of all five s a m p l e s is c o n s i d e r e d , t h e r e
was n o d r o p i n g e r m i n a t i o n for t h e w h o l e seed. W i t h t h e segm e n t e d seed t h e r e was a s l i g h t d r o p i n t h e a v e r a g e g e r m i n a t i o n
after six years ( T a b l e 2, F i g u r e 1). A f t e r this d r o p , t h e g e r m i n a t i o n increased a n d h e l d fairly c o n s t a n t for t h e n e x t 6 years,
after w h i c h i t d r o p p e d t o 1 0 p e r c e n t b e l o w t h e o r i g i n a l g e r m i n a t i o n . T h e final g e r m i n a t i o n o f t h e o n e lot t h a t h a d b e e n s t o r e d
for 20 years was 88 p e r c e n t of t h e o r i g i n a l g e r m i n a t i o n . T h e s e
results i n d i c a t e t h a t s e g m e n t e d seed loses v i a b i l i t y to a g r e a t e r
extent t h a n does w h o l e seed w h e n s t o r e d f r o m six to 17 years.
In sugar beets, t h e " s e e d " n o r m a l l y is a b a l l w h i c h c o n t a i n s
several seeds; w h e n g e r m i n a t e d m o r e t h a n o n e s p r o u t m a y e m e r g e
from each seed b a l l . T a b l e 6 r e p r e s e n t s t h e a c t u a l n u m b e r of
sprouts p e r 100 seed balls for b o t h t h e w h o l e a n d s e g m e n t e d
seeds. I n t h e w h o l e seed t h e n u m b e r o f s p r o u t s r e m a i n e d a b o u t
the same for t h e e n t i r e p e r i o d of t h e test. In t h e s e g m e n t e d seed
the n u m b e r of s p r o u t s f o l l o w e d a p a t t e r n s i m i l a r to t h a t o b s e r v e d
for its g e r m i n a t i o n
p e r c e n t a g e s . At t h e e n d of t h e 17 y e a r s s t o r a g e
period it h a d d r o p p e d f r o m 106 to 92 or a d e c r e a s e of 13 p e r c e n t .
T h e c o r r e l a t i o n coefficient b e t w e e n g e r m i n a t i o n p e r c e n t a n d
n u m b e r of s p r o u t s was 0.74, w h i c h is significant at t h e 0.01 perc e n t point.
I n e x a m i n i n g T a b l e 7 , it. will b e seen t h a t a s t h e seeds b e c a m e
o l d e r the p e r c e n t a g e o f w e a k s p r o u t s d e c r e a s e d i n b o t h w h o l e a n d

Table 6.Number of sprouts per 100 seed balls.

Table 7.-Percent of weak sprouts per one hundred seed balls.

184

J O U R N A L OF T H E A. S.

S.

B. T.

s e g m e n t e d s e e d . I n t h e e a r l y y e a r s o f s t o r a g e t h e s e g m e n t e d seeds
h a d a h i g h e r p e r c e n t a g e o f w e a k s p r o u t s t h a n d i d t h e w h o l e seed.

Summary and Conclusions

W h o l e s u g a r b e e t seed a n d s e g m e n t e d seed f r o m e a c h o f f i v e )
lots w e r e s t o r e d in c o t t o n sacks in a d r y , u n h e a t e d r o o m at the
C o l o r a d o A g r i c u l t u r a l E x p e r i m e n t S t a t i o n at F o r t C o l l i n s from
1943 to 1963. A f t e r 17 years of s t o r a g e , t h e a v e r a g e germination
of t h e w h o l e seed w a s 79 p e r c e n t as c o m p a r e d w i t h 78 percent
for t h e o r i g i n a l g e r m i n a t i o n . T h e s e g m e n t e d seed d r o p p e d from
73 p e r c e n t o r i g i n a l g e r m i n a t i o n to 67 p e r c e n t g e r m i n a t i o n after
s t o r a g e for 17 years, a d e c r e a s e of 8 p e r c e n t .
W i t h t h e w h o l e seed t h e n u m b e r o f s p r o u t s p e r 100 balls
v a r i e d f r o m 136 to 159, b u t r e m a i n e d a b o u t t h e s a m e for the
17-year p e r i o d . T h e n u m b e r of s p r o u t s p e r 100 b a l l s was lower
for t h e s e g m e n t e d seed, v a r y i n g from 95 to 114, a n d dropping
off slightly in t h e last f o u r y e a r s of s t o r a g e . T h e percentage of
w e a k s p r o u t s d e c r e a s e d in t h e last f o u r y e a r s of s t o r a g e in both
w h o l e a n d s e g m e n t e d seed. T h e p e r c e n t a g e o f w e a k sprouts
was h i g h e r i n t h e s e g m e n t e d seed t h a n i n t h e w h o l e seed.

Sucrose Determination in Sugar Beets Using Paper


Chromatography and Spectrophotometry 1
SPIROS M . CONSTANTINIDES AND C . L . BEDFORD 2
Received

for publication February

19,

1964

T h e m e t h o d s g e n e r a l l y e m p l o y e d for t h e d e t e r m i n a t i o n o f
sucrose i n s u g a r b e e t s h a v e c h a n g e d very l i t t l e d u r i n g t h e past
thirty years. C o p p e r r e d u c t i o n a n d p o l a r i m e t r i c m e t h o d s h a v e
generally b e e n used. I n r e c e n t years c h r o m a t o g r a p h i c p r o c e d u r e s
have been d e v e l o p e d t h a t p r o v i d e e x c e l l e n t m e a n s of s e p a r a t i o n
of the sugars a n d e l i m i n a t e s o m e e r r o r s i n v o l v e d in t h e a b o v e
methods ( 1 , 2 , 7 , 12) 3 . T h e basic m e t h o d s o f p a p e r c h r o m a t o graphy of sugars g i v i n g s o l v e n t a n d spray r e a g e n t s h a v e b e e n
reviewed b y K o w k a b a n y (10). M a n y c o l o r i m e t r i c m e t h o d s for
quantitative analysis f r o m t h e c h r o m a t o g r a m s h a v e b e e n r e p o r t e d
(3, 5, 6, 8, 11, 13, 14). D u b o i s et al. (4) r e c e n t l y r e p o r t e d a
colorimetric m e t h o d for sugars b a s e d on t h e f o r m a t i o n of a yellow color b e t w e e n p h e n o l a n d t h e h y d r o x y f u r f u r a l s p r o d u c e d
from the sugars by t r e a t m e n t w i t h s u l f u r i c a c i d .
T h i s p a p e r r e p o r t s t h e a p p l i c a t i o n o f t h e D u b o i s m e t h o d for
the d e t e r m i n a t i o n of sucrose in s u g a r b e e t s .
Materials and Methods
Sugar b e e t s w e r e o b t a i n e d f r o m t h e C r o p Science D e p a r t ment in t h e Fall of 1963, w a s h e d a n d frozen at - 2 0 C , u n t i l u s e d .
Extraction of sugar. T h e frozen b e e t s w e r e t h i n l y sliced. O n e
hundred g r a m s a m p l e s w e r e e x t r a c t e d for 10 m i n u t e s in a W a r i n g
blendor w i t h 100 m l . o f b o i l i n g 9 5 p e r c e n t e t h a n o l . T h e b l e n d e d
material was q u a n t i t a t i v e l y t r a n s f e r r e d to a 1000 m l . v o l u m e t r i c
f l a s k with h o t 9 0 p e r c e n t e t h a n o l a n d h e l d i n a w a t e r b a t h a t
70C. for 3 0 m i n u t e s . I t was t h e n c o o l e d t o 2 0 C C . a n d m a d e t o
volume w i t h 9 0 p e r c e n t e t h a n o l . T h e l i q u i d was filtered t h r o u g h
E and D 512 filter p a p e r a n d 700 m l . of filtrate collected. T h e
nitrate was c o n c e n t r a t e d to a b o u t 50 m l . in a B u c h l e r flash
evaporator a t 3 8 C , m a d e u p t o 100 m l . w i t h w a t e r , a n d c e n t r i fuged at 2500 r p m for 20 m i n u t e s . T h e e x t r a c t n o t u s e d immediately for c h r o m a t o g r a p h y w a s s t o r e d at 1 7 C .
Paper chromatography.
T h e e x t r a c t was c h r o m a t o g r a p h e d
on W h a t m a n N o . 1 filter p a p e r , 24 X 60 c m . u s i n g t h e d e s c e n d i n g
Procedure. T w o m i c r o l i t e r s o f t h e s u g a r b e e t e x t r a c t w e r e s p o t t e d
on 4 places, 6 cm a p a r t a n d 3 c m . f r o m t h e e d g e of t h e p a p e r ,
on a line inches from the t o p of the paper. T h e chromato1
Journal
2

Article No. 3297, Agricultural Experiment Station.


Department of Food Science, Michigan State University. East Lansing.

186

JOURNAL OF THE A. S. S. B.T .

g r a m s w e r e d e v e l o p e d u s i n g n - b u t a n o l - a c e t i c a c i d - w a t e r (4:1:5)
for 60 h o u r s at 20-22C. in a c h r o m a t o g r a p h i c c a b i n e t (15). The
c h r o m a t o g r a m s w e r e d r i e d a t 2 5 C for a b o u t 1 2 h o u r s a n d the
t w o 6 c m . s i d e s t r i p s , e a c h c o n t a i n i n g o n e o r i g i n a l s p o t , w e r e cut
f r o m t h e p a p e r . T h e s e w e r e s p r a y e d w i t h b e n z i d i n e s p r a y (9)
a n d h e a t e d a t 1 0 5 C . for 10-15 m i n u t e s for c o l o r development
a n d l o c a t i o n o f t h e s u c r o s e . T h e s t r i p s w e r e t h e n m a t c h e d with
t h e c e n t e r p o r t i o n o f t h e p a p e r a n d t h e t w o s e c t i o n s correspondi n g t o t h e l o c a t i o n o f t h e s u c r o s e s p o t s w e r e c u t o u t . E a c h section
was c u t i n t o a b o u t 1 s q u a r e c e n t i m e t e r p i e c e s , p l a c e d in 50 ml.
b e a k e r s a n d c o v e r e d w i t h 1 0 m l . o f r e d i s t i l l e d w a t e r . T h e beakers
w e r e c o v e r e d a n d a l l o w e d t o s t a n d for 3 0 m i n u t e s w i t h occasional
s h a k i n g . T h e e l u a t e w a s filtered t h r o u g h glass w o o l t o remove
t h e c e l l u l o s e l i n t a n d t h e a m o u n t o f s u c r o s e was d e t e r m i n e d b y
t h e D u b o i s m e t h o d (4). A b l a n k p a p e r w a s d e v e l o p e d a n d e x t r a c t e d t o serve a s a c o n t r o l .
Colorimetric
procedure. T w o m l . of s u g a r s o l u t i o n were
t r a n s f e r r e d i n t o 1.5 X 18 c m . p y r e x t u b e s a n d 0.125 m l . 80 per
c e n t p h e n o l s o l u t i o n was a d d e d . T h e n 5 m l . o f concentrated
s u l f u r i c a c i d was r a p i d l y i n t r o d u c e d (10-20 s e c o n d s ) . T h e tubes
w e r e h e l d in a w a t e r b a t h at 25-30C. for 10-20 m i n u t e s for the
d e v e l o p m e n t o f t h e y e l l o w - o r a n g e c o l o r , c o o l e d a n d t h e absorba n c y m e a s u r e d a t 487 m w i t h a B e c k m a n D . U . spectrophotom e t e r , slit w i d t h .015 m m . T h e c o n c e n t r a t i o n o f sucrose was
d e t e r m i n e d from the standard curve.
Preparation of standard curve.
A s t a n d a r d s o l u t i o n of sucrose
a n d d i l u t i o n s w e r e m a d e t o c o n t a i n b e t w e e n 3 0 a n d 1 0 0 . per
2 m l . a t 1 0 g . i n t e r v a l s . T h e o p t i m u m p h e n o l concentration
w a s d e t e r m i n e d by a d d i n g .05, .100, .150, .200, .250 a n d .300 ml.
of t h e 80 p e r c e n t s o l u t i o n to 80 m i c r o g r a m s of sucrose. The
a b s o r p t i o n s p e c t r u m w a s d e t e r m i n e d w i t h t h e B e c k m a n D.U.
spectrophotometer.
Results a n d Discussion
T h e a b s o r p t i o n c u r v e for s u c r o s e s h o w s m a x i m u m absorption,
a t 487 m . ( F i g u r e 1). T h e o p t i m u m p h e n o l c o n c e n t r a t i o n f o r
m a x i m u m c o l o r f o r m a t i o n was 100 m g . (.125 m l . 8 0 percent
p h e n o l s o l u t i o n ) ( F i g u r e 2). T h e s t a n d a r d c u r v e for sucrose f o l l o w s B e e r ' s law i n t h e r a n g e o f c o n c e n t r a t i o n u s e d (Figure 3 ) .
Raffinose h a s t h e s a m e m a x i m u m a b s o r p t i o n a s sucrose, r e q u i r e s
t h e s a m e p h e n o l c o n c e n t r a t i o n for m a x i m u m c o l o r f o r m a t i o n
a n d t h e s t a n d a r d c u r v e follows B e e r ' s L a w ( F i g u r e s 1 , 2 , 4 ) .
Raffinose w a s n o t d e t e r m i n e d q u a n t i t a t i v e l y b e c a u s e very l i t t l e
was p r e s e n t . I t w o u l d h a v e r e q u i r e d t h e a p p l i c a t i o n o f m o r e

187

Vol 13, No. 2, JULY 1964

Than 50 microliters of the extract to the paper and because of


the high concentration of sucrose, the two sugar spots would
overlap.

450
WAVELENGTH

Figure I.Absorption curves of the phenolic condensation products


of sucrose and raffinose.

Milligrams Of Phenol

figure 2. - Effect of phenol concentration on the absorbancy of the


phenolic condensation products of sucrose and raffinose

J O U R N A L OF THE A. S. S. B. T

188

MICROGRAMS

OF

RAFf i N O S t

Figure 4.Standard curve for raffinose.

T h e results obtained using a pure 15 percent sucrose solu


tion are given in T a b l e 1. T h e recovery of.sucrose averaged
percent. T h e procedure was found sensitive and
accurate enough to determine quantitatively sucrose concentrations of 25 to 500 micrograms per 2 microliters.
T h e variations between replicate determinations of sucrose in
one sugar beet extract are given in T a b l e 2. T h e average sucrose
concentration was
percent. T h e results obtained were
0.5 to 1.0 percent lower than those obtained by the polarimetric
method.
3

Numbers in parentheses refer to literature cited.

VoL.

13, N o . 2, J U L Y 1964

189

Table I.Sucrose recovery from chromatogram by clutioun1.

Average and standard deviation


1

60.21.0

100.31.2

2 microliters containing 60 micrograms sucrose spotted.

Table 2.Variation of sucrose concentration within one sample of sugar beets'.


Sample
No.

Absorbancy

Sucrose
micrograms

Sucrose
%

4 microliters of beet extract spotted.

The recovery of sucrose added to the sugar beet samples is


presented in T a b l e 3. Five grams of sucrose were mixed thor|oughly with representative beet samples and the sucrose determined by the method given. T h e overall recovery of sucrose for
16 determinations was 5.0 = 0.2 grams representing an average
of
99
percent.

JOURNAL OF THE A. S. S. B. T
Table 3.Sucrose recovery from sugar beet extracts 1 .

4 microliters spotted.

T h e sucrose concentration in sugar beets may be calculated


by the following formula:

E
S
U
C

= micrograms or sucrose on the standard curve corresponding to the absorbancy.


= milliliters of water in which the p a p e r s a r e elated
(10 ml.).
= milliliters of sugar beet e x t r a c t after concentration
(100 ml.).
= milliliters of eluent used in DU (2 m l ) .
= microliters of sugar beet extract applied on the paper
(2 lambdas).
= grams of sugar beets which were vised (70 grams).

ties given are used.


From the results obtained in this study. it is concluded tha
the method described provides an accurate procedure for the
determination of sucrose in sugar beets.
Acknowledgments

Vol. 13, No. 2, July 1964 191

References
(1) BLOCK, R- J-,

R-

LeSTRANGE

and

G. ZWEIG.

1952.

Paper C h r o m a t o -

graphy. 'Academic Press Inc., N e w York. p p . 170-214.


(2) CASSIDY, FT. G. 1951. A d s o r p t i o n a n d C h r o m a t o g r a p h y . Interscience
Publishers, Inc., N e w York.
(3)

DIMLER,

R.

J.,

W.

C.

SCHAEFER,

C.

S.

WISE

Q u a n t i t a t i v e p a p e r c h r o m a t o g r a p h y of
charides. A n a l . C h e m . 24, 141 1-1414.
(4)

and

glucose

C.

E.

and

RIST.

its

1952-

oligosac-

DUBOIS, M., K. A. G I L L E S , J. K. H A M I L T O N , P. A. REISERS a n d F. S M I T H .

1956. Colorimetric m e t h o d for d e t e r m i n a t i o n of sugars a n d related


substances. A n a l . C h e m . 2S, 350-356.
(5)

(6)
(7)
(8)
(9)
(10)

(11)
(12)
(13)

(14)

FLOOD, A. E., J . K. J O N E S a n d E. L. H I R S T .

1947.

Q u a n t i t a t i v e estima-

tion of m i x t u r e s of sugars by t h e p a p e r c h r o m a t o g r a p h i c m e t h o d .
N a t u r e 160, 86-87.
H A W T H O R N E , J. R. 1947. Microestimation of sugars separated on
filter p a p e r c h r o m a t o g r a m s . N a t u r e 160, 714-715.
H E F T M A N , E.
1962.
Chromatography.
R e i n h o l d P u b l i s h i n g Corp.,
New York, p p . 502-533.
HIRST, E. L. a n d K. N. J O N E S . 1949. Q u a n t i t a t i v e analysis of mixtures
of sugars. C h e m i c a l Society J o u r n a l , p p . 1659-1662.
HORROCKS, H. T-I. 1949. P a p e r p a r t i t i o n c h r o m a t o g r a p h y of r e d u c i n g
sugars with b e n z i d i n e as a spray reagent. N a t u r e 164, 444-445.
KOWBABANY, G. N. 1954. P a p e r c h r o m a t o g r a p h y of carbohydrates a n d
related c o m p o u n d s . A d v a n c e s in C a r b o h y d r a t e C h e m i s t r y 9. 303353.
LAIDLAW, R. A. a n d S. G. R E I D . 1950. Extraction of sugars from the
p a p e r at r o o m t e m p e r a t u r e , N a t u r e 166, 476-478.
LEDERER, E. a n d M. LEDERER.
1953. Chromatography. Elsevier P u b lishing Corp., N e w York ( 1 9 5 3 ) .
MCCREADV, R. M. a n d E. A. M C C O M B . 1954. Q u a n t i t a t i v e determination of sugars on p a p e r c h r o m a t o g r a m by a reflectance m e t h o d ,
Anal. C h e m . 28, 1645-1646.
M C F A R R E N , E. F., K. B R A N D a n d H . R . BUTKOW SKI.

1951.

Quantitative

d e t e r m i n a t i o n of sugars of filter p a p e r c h r o m a t o g r a m s by direct


photometry., A n a l . C h e m . 23, 1146-1149.
(15) PARTRIDGE, S. M. 1948. Filter p a p e r p a r t i t i o n c h r o m a t o g r a p h y of
sugar, Biochem. J. 42, 238-48.

Beet Pulp in All-Barley Rations


W.
Received

A.

HARRIS1

for publication March 13, 1964

Introduction
T h e s u g a r b e e t i n d u s t r y h a s n e v e r h e s i t a t e d t o maintain
t h a t m o l a s s e s - d r i e d - p u l p for f a t t e n i n g l i v e s t o c k is j u s t as good
as g r a i n . T h i s h a s n e v e r b e e n a m i s g u i d e d loyalty, for dozens
of r e e d i n g tests c a n be p o i n t e d to a n d p r a c t i c a l e x p e r i e n c e s with
a v a r i e t y o f r a t i o n s , t h a t s u s t a i n e d a n d n u r t u r e d t h i s faith.
B u t t h e m a n w h o b u y s t h e feed d o e s n ' t d o s o o n faith alone
H e w a n t s p r o o f t h a t c o m p e t i t i v e m a t e r i a l s c a n ' t d o a b e t t e r jot
for h i m , a n d s o m e t i m e s i t i s n e c e s s a r y t o h a v e g o o d proof.
A b o u t five years a g o t h e s t r o n g p o s i t i o n of d r i e d p u l p , as;
ration c o m p o n e n t , was threatened. T h e "all-barley" ration was
b e i n g w i d e l y e x t o l l e d a n d w a s g a i n i n g in p o p u l a r i t y in some
areas.
B a r l e y a p p a r e n t l y has e n o u g h fiber in t h e h u l l to satisfy
r u m i n a l r e q u i r e m e n t s for r o u g h a g e , p r o v i d e d i t i s rolledto
m a i n t a i n a c o u r s e p h y s i c a l s t r u c t u r e r a t h e r t h a n ground. A
c o m m e r c i a l s u p p l e m e n t h a d t o b e u s e d , o f c o u r s e , t h a t supplied
l a c k i n g v i t a m i n s a n d m i n e r a l s ( a n d p r o t e i n s ) . U s u a l l y stilbestrol
was i n c o r p o r a t e d i n t h e s u p p l e m e n t t o h e l p t h i n g s a l o n g . But i t
w o r k e d . C a t t l e finished o u t w e l l , w i t h fast g a i n s a n d excellen
feed c o n v e r s i o n t o s h o w g o o d e c o n o m y o f g a i n .
If the m e t h o d were to g a i n widespread acceptance as it was
o r i g i n a l l y p r o m o t e d , b e e t p u l p would, lose p o s i t i o n in the feed
t r a d e s i m p l y by b e i n g i g n o r e d as a r a t i o n c o m p o n e n t . It became
necessary t o l e t o u r feeders k n o w t h a t p u l p c o u l d fit into t h i s
s c h e m e as w e l l as i n t o a n o r m a l f e e d i n g r e g i m e . At that time
t h e r e w a s no e x p e r i m e n t a l e v i d e n c e (as t h e r e is n o w ) to bac
up any recommendations that might be made.
Y e t p u l p l o o k e d l i k e a n a t u r a l c o m p o n e n t for a "non
r o u g h a g e r a t i o n . It is h i g h in c r u d e fiber. M o s t of it differs
from b a r l e y f i b e r i n b e i n g h i g h l y d i g e s t i b l e , b u t t h e amount
o f n o n - d i g e s t i b l e fiber i n b a r l e y a n d p u l p a r e n o t far apart. A n d
it is k n o w n that p u l p does have some r o u g h a g e value.
I t s e e m e d r e a s o n a b l e t o t h i n k t h a t p u l p w o u l d replace P a r t
o f t h e b a r l e y i n t h i s " a l l - b a r l e y " r a t i o n a n d a g a i n prove i t s e l f
to be a m o n e y - s a v e r .
F a i t h i n s u c h beliefs w a s p u t t o test o n N o v e m b e r 6 , 1 9 5 9
w i t h a feeding trial at the e x p e r i m e n t a l lots of Holly Sugar
Corporation in Torrington, Wyoming.
1

Research Chemist, Ffoliy Sugar Corporation, Colorado Springs, Colorado-

Vol. 13, No. 2, JULY 1964

193

Experimental
A laroge purchase of yearling whiteface cattle received at t h e
yards was placed on f u l l feed of alfalfa, d r i e d beet p u l p , and
cottonseed cake. G r a i n was g r a d u a l l y s u b s t i t u t e d for alfalfa a n d
pulp until all a n i m a l s w e r e receiving a b o u t 6 p o u n d s of g r a i n
at the end of t h r e e weeks.
At this t i m e , a n i m a l s t h a t a p p e a r e d to deviate far from
Average in conformity w e r e rejected. All o t h e r steers were individually weighed a n d d i v i d e d to the e x p e r i m e n t a l pens according to weight. T h u s any steer of a p a r t i c u l a r s t a r t i n g weight in
the experimental p e n h a d its c o u n t e r p a r t in t h e c o n t r o l p e n .
Each pen h a d 15 steers with an average weight of 793 p o u n d s .
T h e s u p p l e m e n t was c h a n g e d to 2 p o u n d s p e r head p e r day
If Purina Special 3 2 % Steer F a t e n a for b o t h pens. T h i s s u p p l i e d
10 mg of stilbestrol. H a y was g r a d u a l l y w i t h d r a w n a n d barley
Substituted d u r i n g t h e n e x t t h r e e weeks. At this t i m e t h e c o n t r o l
"all-barley" pen was receiving only rolled bailey a n d the supplement; the e x p e r i m e n t a l p e n was receiving 1 / 2 barley a n d 1 / 2
pulp with the s u p p l e m e n t . Feeds were n o t p r e m i x e d .
Monthly weights w e r e t a k e n u n t i l t h e e x p e r i m e n t was terminated, after 171 days. After an o v e r n i g h t stand, final weights
were taken on A p r i l 26, 1960, a n d a 4% p a p e r s h r i n k was a p p l i e d
to arrive at n e t weight figures. T h e cattle were slaughtered, on
|consignment, at Swift's p l a n t in Scottsbluff, N e b r a s k a .
Results
No off-feed or o t h e r difficulties w e r e seen on t h e feedlot.
One steer in the c o n t r o l p e n did show signs of f o u n d e r in t h e
latter weeks of t h e test.
A difference in t h e two r a t i o n s was e v i d e n t from start to
finish. Steers on the p u l p - b a r l e y c o m b i n a t i o n consistently ate 3/4
to 1 1 / 2 pounds m o r e feed daily t h a n d i d the all-barley cattle.
The result was a faster r a t e of gain all the way t h r o u g h .
Final results a r e s u m m a r i z e d in T a b l e 1. T h e p u l p - b a r l e y
pen showed h i g h e r feed c o n s u m p t i o n , faster gain, b e t t e r feed
utilization, lower dressing % , b e t t e r g r a d e a n d m o r e profit.
Actually an excess fill of t h e p u l p - b a r l e y cattle at t h e t e r m i n a l
weighing may have given t h e i n d i c a t i o n of p o o r e r dressing percent Adjustment for this difference w o u l d a m o u n t to 3.5 p o u n d s
p e r head. T h i s w o u l d i n d i c a t e t h e t r u e c o m p a r a t i v e gains perhaps should have been 2.46 a n d 2.62, a n d t h e conversion of feed
would be identical.
The results completely
justified confidence in beet p u l p . For
once again, and under conditions foreign to its normal use, beet
pulp proved that it is difficult indeed to find a ration that cannot
be improved by its addition.

194

J O U R N A L OF T H E A. S.
Table

1.Pulp-barley vs.

all-barley

Intial Weight
Final Net Weight (using 4% shrink)
Total Gain
Net Daily G a i n
Daily F e e d
Rolled barley
D r i e d Molasses Beet P u l p
3 2 % Special Steer F a t e n a
G r o u n d Alfalfa*
Salt
Mineral
Avg. Lbs. Feed Per Lb. Gain
Avg.** Feed Cost Per L b . Gain
Avg. Dressing %
G r a d e , N o . of Steers in
U. S. G o o d
U. S. C h o i c e
N e t Profit Per Steer Over Control

in

Avg.
Avg.
A\g.
Avg.
Avg.

"non-roughage**

* H a y l e d d u r i n g 1st t h r e e week.;,.
' - F e e d prices used; P u l p , S32.50/T; Rolled
s 2 5 / T ; S a i l , S 2 7 . 5 0 / T ; M i n e r a l , $ 5 - 2 0 / 10O.

Barley,

B. T

(control)
all-barley
ration

P e n 15
barley-pulp
ration

793.3
1213.4
420.1
2.4C

793.3
1245.4
452.1
2.64

17.57
_
1.99
0.41
0.022
0.035
8.10
16.19
63.64

>

5
10

S.

i attons.

1.75/100ir;

9.33
9.3?
1.99
0.65
0.022
0.022
8.04
15.44
b3.i5
3
12
S6.42

Fatena,

$86/T;

Hay,

Summary
F i f t e e n h e a d of 793 p o u n d H e r e f o r d steers w e r e fed 171 days
o n a n ' a l l - b a r l e y " r a t i o n a n d t h e i r p e r f o r m a n c e c o m p a r e d with
t h a t of 15 s i m i l a r steers fed w i t h 1:1 p u l p : b a r l e y .
T h e p u l p : b a r l e y c o m b i n a t i o n g a v e s u p e r i o r f e e d l o t perforin
a n c e a n d e q u a l or b e t t e r carcass q u a l i t y . N e t profit was in
c r e a s e d w i t h t h e p u l p - b a r l e y feed.

JOURNAL
of the
American Society of Sugar
Beet Xechnologists
Volume 13
Number 3
October 1964

Published

quarterly

by

A m e r i c a n Society of S u g a r Beet Xechnologists


Office of the Secretary
P . O . B o x 538
F o r t Collins, C o l o r a d o , U. S. A.

Subscription
$4.50
$5.00
$1.25
$1.40

per
per
per
per

year,
year,
copy,
copy,

prices:
domestic
foreign
domestic
foreign

M a d e in the U n i t e d States of America

TABLE OF CONTENTS

Author

Page

The Effect of Incorporation Methods on


Weed Control With Tillam in the Rocky
Mountain Region
_E. G. Eckrolh
E. M. Hoist
D. F. Peterson
Mathematical A p p r o a c h
to D e t e r m i n e
Vacuum Pan Boiling Time for Full Seeding Method Using Milled Fondant
Thin Film Ultra-Violet S t e r i l i z a t i o n of
Liquid Sugar, Using the Aquafine Sterilizer.

Paper Chromatographic Determination


Raffinose in Sugar Beet Molasses

The Effect of Simulated Hail Injuries


Yield and Sugar Content of Beets
Soil Temperature and Nitrogen
Yield and Phosphorus Uptake
Beets

195

Allison S. Chang

Robert S. Gaddie
Robert R. West
E. G. Bennison _

214

of
Fern Jantzef
A. L. Potter

.218

on
M.

M.

Afanasiev

225

Effect on
by Sugar

Losses Caused by Beet Mosaic Virus in California Grown Sugar Beets

An Evaluation of a Multiple Bed Dionizaon Process for Beet Sugar Recovery

5. Dubetz
G. C. Russell 238

R. J. Shepherd
F. J. Hills
D. H. Hall

-244

Frank

-252

X.

McGarvey

Methods of Loosening Tight Seed Caps in


lonogerm Seed to Improve Germination _ F. H, Peto

.281

The Effect of Incorporation Methods on W e e d


Control W i t h Tillani in the Rocky Mountain Region
E.

G.

ECKROTH,

E.

M.

Received

for

publication

H O L S T AND
February

E>.
19,

F.

PETERSON1

1964

X h e v a l u e o f i n c o r p o r a t i n g h e r b i c i d e s i n t o t h e soil h a s b e e n
thoroughly d e m o n s t r a t e d . It is i m p o r t a n t to d e t e r m i n e the most
efficient m e a n s o f i n c o r p o r a t i o n , t h e m o s t e c o n o m i c m e t h o d , t h e
most p r a c t i c a l , a n d t h a t m e t h o d w h i c h e n h a n c e s t o l e r a n c e o f
sugar b e e t s t o t h e c h e m i c a l . W e h a v e r e a c h e d a p o i n t i n s u g a r
beet p r o d u c t i o n w h e r e t h e u n c e r t a i n l a b o r p i c t u r e p o s e s a g r e a t
challenge to raising a c r o p of beets w i t h an a b s o l u t e m i n i m u m
o r n o h a n d l a b o r a t a l l . M a n y o f u s feel t h a t w e h a v e t h e
knowledge, m e c h a n i c a l e q u i p m e n t a n d herbicides t o d o the j o b .
M o s t R o c k y M o u n t a i n f a r m e r s a r e c o n s t a n t l y f a c e d w i t h rising costs a n d f o r t h e s m a l l e r a c r e a g e g r o w e r t h i s e q u i p m e n t
cost is s e r i o u s .
T h i s p r o j e c t w a s set u p t o c o m p a r e t h e i n e x p e n s i v e m e t h o d s
of incorporation such as the Sinner a n d Ridgecover with the
more expensive p o w e r driven Bye-Hoe a n d Eversman.
Considerable w o r k has b e e n d o n e w i t h n u m e r o u s different
incorporation devices b u t o n l y a l i m i t e d a m o u n t of i n f o r m a t i o n
r e g a r d i n g a c t u a l field c o m p a r i s o n s i s a v a i l a b l e ( 1 , 2 , 3) 2 .
Materials and Methods
Four i n c o r p o r a t i o n devices w e r e c o m p a r e d , using the herbicide T i l l a m ( p r o p y l e t h y l n - b u t y l t h i o l c a r b a m a t e S t a u f f e r C h e m ical C o m p a n y ) a t t w o d e p t h s a n d t w o r a t e s o f a p p l i c a t i o n . X h e
four i n c o r p o r a t i o n d e v i c e s r e p r e s e n t t w o b a s i c m e t h o d s o f incorporation, power-driven m i x i n g a n d layering. X h e Bye-Hoe
and E v e r s m a n t i l l e r a r e p o w e r d r i v e n m i x e r s a n d t h e R i d g e cover a n d S i n n e r X i l l e r ( a l s o c a l l e d R u s s - K e n ) r e p r e s e n t t h e
layering m e t h o d o f i n c o r p o r a t i o n . X h e B y e - H o e c o n t a i n s reshaped k n i v e s a n d t h e E v e r s m a n h a s p e g s w h i c h c o n s t i t u t e s t h e
primary difference b e t w e e n t h e t w o p o w e r d r i v e n tillers. X h e s e
two p r o v i d e c o m p l e t e m i x i n g o f t h e h e r b i c i d e a n d soil. X h e
Ridgecover m e t h o d a s u s e d i n t h e s e t e s t s u t i l i z e d d i s c s i n p a i r s
t o cover t h e h e r b i c i d e . X i l l a m w a s s p r a y e d a h e a d o f t h e discs a s
a 7 - i n c h b a n d w i t h t h e discs c o v e r i n g t h e s p r a y e d b a n d w i t h soil.
This forms a modified r i d g e on t o p of t h e h e r b i c i d e . X h e S i n n e r
t i l l e r m e t h o d a l s o p l a c e s t h e h e r b i c i d e i n a flat b a n d b e h i n d
a furrow opener with the Sinner blades covering the chemicaltreated b a n d s w i t h soil.
1

Associate Agronomists and Assistant Director of Agricultural Research Respectively,


Agricultural
Research Department, Holly Sugar Corporation, Sheridan, Wyoming,
2
numbers
in
parentheses
refer
to
literature
cited
*

196

J O U R N A L OF T H E

A.

S.

S.

B.

T.

VOL.

13,

No-

3,

OCTOBER

1964

197

taken from an a r e a S inches by 48 i n c h e s w i t h t h e b e e t r o w in


the c e n t e r . S u g a r b e e t c o u n t s i n c l u d e d 1 0 feet o f r o w . T h e p l a n t
p o p u l a t i o n w a s classified a s t o s u g a r b e e t s , b r o a d l e a v e d w e e d s ,
a n d grassy w e e d s , b u t t h e w e e d d a t a a r e p r e s e n t e d a s t o t a l w e e d s .
Results a n d Discussion
C o n s i d e r a b l e difficulty w a s e n c o u n t e r e d i n o b t a i n i n g t h e d e e p
d e p t h o f i n c o r p o r a t i o n i n t h e M o n t a n a tests d u e t o h i g h soil
m o i s t u r e c o n t e n t . T h i s m o i s t soil c o n d i t i o n s h o w e d t h e S i n n e r
to advantage. T h e Bye-Hoe, Eversman, a n d Ridgecover caused
the m o i s t soil t o b e e x p o s e d e v e n m o r e a n d t h i s r e s u l t e d i n t h e
p l a n t e r discs a n d d e p t h b a n d s p i c k i n g u p t h e t r e a t e d soil a n d
s o m e t i m e s l e a v i n g s e e d e x p o s e d a n d t r e a t e d soil n o l o n g e r i n
the b a n d . T h e S i n n e r r e a c t e d s o m e w h a t d i f f e r e n t l y a s t h e r e w a s
just e n o u g h d r y soil o n t h e s u r f a c e s o t h a t w i t h t h e f u r r o w o p e n e r
r u n n i n g s h a l l o w t h e S i n n e r b l a d e s h a d sufficient d r y soil t o c o v e r
the herbicide b a n d r e s u l t i n g i n b e t t e r p l a n t e r o p e r a t i o n . H o w ever, t h i s b e t t e r p l a n t i n g o p e r a t i o n was n o t r e f l e c t e d b y i m p r o v e d
weed c o n t r o l .
I t was possible t o o b t a i n t h e t w o i n c o r p o r a t i o n d e p t h s i n
Wyoming and Colorado. T h e two depths of incorporation in
t h e W y o m i n g a n d C o l o r a d o tests r e s u l t e d i n a n a v e r a g e o f 8
p e r c e n t m o r e w e e d c o n t r o l for t h e d e e p e r d e p t h w i t h v e r v l i t t l e
influence o n b e e t e m e r g e n c e . A s a r e s u l t o f t h e s m a l l d i f f e r e n c e
in weed control d u e to d e p t h of i n c o r p o r a t i o n , a n d o b t a i n i n g
only t h e s h a l l o w i n c o r p o r a t i o n d e p t h i n t h e M o n t a n a tests, t h e
data a s p r e s e n t e d i n t h e f o l l o w i n g t a b l e s a r e n o t s e p a r a t e d o n
the basis of d e p t h of i n c o r p o r a t i o n .
Table 2 presents the percent weed control a n d percent beet
stand r e d u c t i o n a s i n f l u e n c e d b v i n c o r p o r a t i o n m e t h o d s , n a m e l y
Bye-Hoe, E v e r s m a n , S i n n e r a n d R i d g e c o v e r .
All m e t h o d s o f i n c o r p o r a t i o n r e s u l t e d i n v e r y g o o d w e e d
control. T h e R i d g e c o v e r m e t h o d g a v e s l i g h t l y less w e e d c o n t r o l
than the o t h e r t h r e e m e t h o d s a n d was also t h e m o s t severe in
beet s t a n d r e d u c t i o n w i t h t h e o t h e r m e t h o d s b e i n g c o m p a r a b l e .
The beet stand r e d u c t i o n for R i d g e c o v e r is greatly influenced
b y t h e M o n t a n a t e s t s w h e r e b e c a u s e o f t h e v e r y m o i s t soil c o n ditions t h e R i d g e c o v e r m e t h o d b r o u g h t u p r i b b o n s o f m o i s t soil
resulting i n a v e r y p o o r s e e d b e d .
The increase of h e r b i c i d e r a t e from four p o u n d s to five pounds increased
w e e d c o n t r o l 6.6 p e r c e n t a g e p o i n t s a n d i n c r e a s e d b e e t s t a n d
reduction
5.8 p e r c e n t a g e p o i n t s . A l l i n c o r p o r a t i o n m e t h o d s
followed a similar p a t t e r n in weed control a n d beet
stand r e d u c t i o n w h e n t h e h e r b i c i d e rate was increased.

198

J O U R N A L OF T H E

A.

S.

S.

B. T.

I n o r d e r t o p r e s e n t s o m e w h a t m o r e d e t a i l e d i n f l u e n c e o f inc o r p o r a t i o n m e t h o d s o n w e e d c o n t r o l , T a b l e 3 p r e s e n t s data
for t h r e e l o c a t i o n s .
To i l l u s t r a t e t h e v a r i a t i o n of p e r f o r m a n c e by i n c o r p o r a t i o n
m e t h o d s a t different l o c a t i o n s , t h e S i n n e r gave t h e least weed
c o n t r o l a t S i d n e y b u t w a s best a t W o r l a n d a n d R i v e r t o n . H o w ever, i n o v e r a l l a v e r a g e w e e d c o n t r o l , all f o u r a r e very c o m p a r able, as shown in b o t h Table 2 a n d Table 3.
X a b l e 4 p r e s e n t s t h e effect of i n c o r p o r a t i o n m e t h o d s on sugar
beet emergence.
A l l f o u r m e t h o d s o f i n c o r p o r a t i o n r e s u l t e d i n r e d u c e d beet
e m e r g e n c e a s c o m p a r e d t o t h e c h e c k . T h e d a t a s h o w t h e power
d r i v e n i n c o r p o r a t i o n m e t h o d s t o a d v a n t a g e , w i t h a b o u t t w o more
p l a n t s e m e r g i n g in every 10 feet of r o w . This a d v a n t a g e could
n o t b e d i s t i n g u i s h e d i n t h e field b u t c o u n t s b r o u g h t o u t this
difference.
It is of i n t e r e s t to o b s e r v e f r o m t h e s e tests t h a t b e e t seedlings
w e r e affected w h e n w e e d c o n t r o l w a s o b t a i n e d . I n n o test was
t h e r e a n e x c e p t i o n t o t h i s d u e t o i n c o r p o r a t i o n m e t h o d . This
s e e d l i n g effect i s a n i n w a r d c u p p i n g a p p e a r a n c e o f t h e cotyled o n a r y leaves. W h e n c u p p i n g was s e v e r e , u s u a l l y o n l i g h t soils,
Table 2.The percent weed control and percent beet stand reduction due to the
listed incorporation methods at two herbicide rates in Holly Sugar's Rocky Mountain
area in 1963.**
Incorporation
method

Tillam
rate*

Percent
weed control

27.0

73.0

10.0

79.0

14.0

Eversman

0
4
5

18.0
76.5
79.5

5.0
7.5
15.5

Sinner Tiller

0
4
5

25.0
68.5
80.0

7.0
12.5
210

Ridgecover

0
4
5

15.0
68.5
74.5

13 0
12.5
25.5

Average of all
Incorporation
Methods

0
4
5

21.2 7.2
71.6
78.2

13.2
19.0

Bye-Hoe

Percent beet
stand reduction

Expressed as pounds of active ingredient on overall coverage but applied as a seven


inch band treatment.
* * Averages of 29 tests.

V O L . 13, N o . 3, O C T O B E R 1964

199

the power driven incorporation methods showed a lesser degree


of damage. In general Tillam gave very good over-all weed
control with beet seedling reaction being a temporary effect.
The following weed species were being controlled: Foxtails
(Setaria spp.), Barnyard grass (Echinocloa crusgalli) , W i l d oats
(Avena Fatua), broadleaved weeds, Pigweed (Amaranthus retroflexus), L a m b s q u a r t e r (Chenopodium album), a n d Nightshade
(Solarium spp).
Table 3.The percent weed control due to the listed incorporation methods at two
herbicide rates at three locations in 1963.
Incorporation methods

Table 4.The effect of different methods of incorporation on sugar beet emergence


with two rates of T i l l a m in the Rocky Mountain area in 1963.
Incorporation methods
Average beet emergence per 10 feet of row
Location
Sidney
Hardin
Worland
Riverton
Torrington

rate*

Bye-Hoe

Eversman

16

20

17

12

20

5
4
5
4
5
4
5
4

14
18
19
S3
30
22
21
32

19
20
20
33
28
21
20
34

17
18
18
34
32
17
16
30

12
15
13
30
30
18
18
36

20
20
20
37
36
25
23
33

Total Average

__

32

23.7

Sinner tiller Ridgecover

Cheek

33

28

32

31

24.8

22.7

216

26.5

Expressed as pounds of active ingredient on overall coverage b u t was a pplied in a


seven inch band.

200

J O U R N A L OF T H E

A.

S.

S.

B. T.

Summary
A n u m b e r of herbicide trials using Tillam were conducted
comparing four methods of incorporation, (Bye-Hoe, Eversman,
Sinner Tiller, and Ridgecover) in Holly's Montana, Wyoming
and Colorado areas.
A good sampling of factors influencing weed control was
obtained in covering a three state area. These factors were as
follows: differences in soil types, soil moisture content, seed
bed condition, air temperature, irrigation and rainfall, and
cultural practices.
T h e two depths of incorporation provided almost similar
beet emergence results with the deeper depth providing eight
percent more weed control. T h i s could not be observed without
counts.
Very good weed control was obtained with all four methods
of incorporation, all of them being equally effective.
All four methods of incorporation caused a reduction in sugar
beet emergence b u t this difference was usually very difficult to
observe except in a few tests. T h e emergence results gave the
Bye-Hoe and Eversman an advantage of 9 percent more seedlings
than the Sinner and Ridgecover method. T h i s may not be true
when using a herbicide with a greater range of beet tolerance.
From the data obtained in these tests, it indicates that the
inexpensive layering methods of incorporation will do as good
a j o b in weed control as the more expensive power driven incorporators in the Rocky Mountain area.
Literature Cited
(1)

C O S T E L , G . L., C. F . B E C K E R , a n d H . P . A L L E Y .

t h e A p p l i c a t i o n of H e r b i c i d e s to S u g a r Beets.
Sta. C i r c u l a r N o . 158.
(2)

1961.

E q u i p m e n t for

W y o m i n g Agr. Exp.

C O S T E L , G . L., E . C H A M B E R L A I N , C . B E C K E R a n d H . A L L E Y .

1962.

m e n t for t h e A p p l i c a t i o n of H e r b i c i d e s to S u g a r Beets.
Agr. E x p . Sta. C i r c u l a r N o . 179.

Equip-

Wyoming

(3) A N T O G N I N I , J. 1962. E x p e r i m e n t a l a n d c o m m e r c i a l results with tillam


for weed c o n t r o l in sugar beets. J. A m . Soc. Sugar Beet Technol
1 2 ( 2 ) : 94-99.

Mathematical Approach to Determine Vacuum Pan


Boiling Time for Full Seeding Method
Using Milled Fondant
ALLISON S. C H A N G 1
Received

for publication

February

19,

1964

Sucrose crystallization rate is a controlling factor in sugar end


operation. T h e r e are several methods available at the present
time for determining sucrose crystallization rate in the laboratory.
In fact many methods, proved to be satisfactory in the laboratory,
have failed to be applicable in actual full-scale operation. T h i s
is particularly true in pan boiling.
T h e purpose of this investigation was to develop a suitable
equation for use at the factory level for determination of pan
boiling time.
Equipment
T h e following e q u i p m e n t was used in this investigation:
(1). White calandria pan.
3,340 sq.ft. heating surface, 23 3/4" O D , 1677 b i m e t a l tubes,
0.065" steel outside, 0.042" copper inside, 3'3/8" long,
437 cu.ft. graining volume, 1750 cu.ft. strike volume,
15', 0" OD pan diameter, 13'6" OD carandria diameter,
75 hp circulator at 75 R P M , steam at 10 PSIG.
(2). Pan automatic controls.
Pan instruments were supplied b y T a y l o r I n s t r u m e n t
Company.
T h e automatic controls consisted of the following items:
(a) Consistency probe and controller.
(b) Level controller and transmitter.
(c) T e m p e r a t u r e controller and transmitter.
(d) Absolute pressure controller.
(3). George Lory design, Lasico crystalscope (pan microscope)
was inserted i n t o the pan just below the normal graining
level. Crystalscope micrometer has a scale which is 5 units
long. Each u n i t is divided into 20 subdivisions. Each subdivision is equivalent to 0.001 inches. T o t a l length of scale
is 0.1 inches.
(4). Polaroid camera.
(5). Complete ball mill to make the fondant.
1

Chemical Engineer, Engineering Department, The Amalgamated Sugar Company,


Ogden, Utah.

202

JOURNAL OF THE A. S. S. B. T.

Equations
R a t e of decrease in m o t h e r l i q u o r purity is determined by
the degree of super-saturation and purity; namely
dP
(1)

dt
=
k
P
(S-1)
Degree of super-saturation is the ratio of the sugar in solution
per 100 units water to the true solubility of sugar in that solution
per 100 units of water both at the same temperature. Purity is
the percentage r a t i o of sugar to total solids.

W h e n degree of super-saturation of m o t h e r liquor is equal


to 1, the seeded crystals will n e i t h e r dissolve nor grow in the
m o t h e r liquor. Therefore, the purity of l i q u o r will not increase
nor decrease with time. In this case the above equation becomes
dP
dt=0
If purity of m o t h e r liquor is zero, sugar content of mother
liquor is zero and will not change with time. Therefore,
dP
dt=0
dP
R e a r r a n g i n g E q u a t i o n (1) to
P

= k (S-1) dt

Integrate E q u a t i o n (2) between t = 0, P = P and t


P = P at constant super-saturation S.
I n P I n P = k ( S 1 ) t
Hence,
In P In P = + k (S l ) t
Solve for 1n P, In P = 1n P k (S 1) t
In P I n P
Solve for t,
t
=
k
(
S

1
)

(2)
= t,

(3)
(4)
(5)

At t = 0, E q u a t i o n (4) is reduced to 1n P = 1n P . Hence,


P =
P , purity of m o t h e r l i q u o r at graining is equal to the
purity of standard liquor.
If S approaches to infinite super-saturation; that is,
lim
In P I n P

t =

( S 1 ) = 0

T i m e r e q u i r e d to boil the p a n from initial purity P to final


purity P is zero.
If super-saturation of l i q u o r approaches to 1,
lim In P In P

t=s1k(S1)=
T i m e r e q u i r e d to reduce the m o t h e r l i q u o r purity from P
P is infinite.

VOL. 13, N o . 3, O C T O B E R 1964

203

From theoretical a n d actual standpoint, it is desirable to boil


the p a n a t h i g h e r s u p e r - s a t u r a t i o n . H o w e v e r , s u p e r - s a t u r a t i o n
above 1.5 s h o u l d b e a v o i d e d for full s e e d i n g m e t h o d , b e c a u s e
s u p e r - s a t u r a t i o n a b o v e 1.5 i s i n t h e L a b i l e z o n e w h i c h w i l l c a u s e
the f o r m a t i o n o f f i n e g r a i n a n d u n w a n t e d g r a i n size.
There is an o p t i m u m condition at which fine grain will not
form a n d t h e b o i l i n g t i m e i s m i n i m u m . T h i s c o n d i t i o n e x i s t s
a t s u p e r - s a t u r a t i o n o f 1.5 w h i c h i s t h e u p p e r l i m i t o f M e t a s t a b l e
zone. I n t h i s z o n e , t h e c r y s t a l w i l l g r o w b u t p r a c t i c a l l y n o n e w
crystal will f o r m .
E q u a t i o n (4) i s t o b e u s e d t o e s t i m a t e t h e f i n a l g r e e n p u r i t y
o r m o t h e r l i q u o r p u r i t y P for b o i l i n g t h e p a n f r o m i n i t i a l p u r i t y
P a n d c o n s t a n t s u p e r - s a t u r a t i o n S for t o t a l b o i l i n g t i m e t .
E q u a t i o n (5) i s t o b e u s e d t o e s t i m a t e t h e b o i l i n g t i m e f o r
boiling t h e p a n f r o m i n i t i a l p u r i t y P t o final g r e e n p u r i t y P
at c o n s t a n t s u p e r - s a t u r a t i o n S.
I t i s r a t h e r difficult t o b o i l t h e p a n a t c o n s t a n t s u p e r - s a t u r a tion w i t h o u t p r o p e r a n d r e l i a b l e i n s t r u m e n t a t i o n s . F o r t u n a t e l y ,
Taylor I n s t r u m e n t C o m p a n y has d e v e l o p e d a t e m p e r a t u r e measuring device w h i c h is q u i t e u n i q u e in p a n i n s t r u m e n t a t i o n s .
T h e temperature m e a s u r i n g e l e m e n t is called T r a n s a i r e T e m perature Capillary B u l b w h i c h was inserted in t h e v a p o r line
between p a n a n d c o n d e n s e r . T h i s t e m p e r a t u r e s e n s i n g b u l b i s
q u i t e sensitive, m e a s u r i n g a n y s m a l l c h a n g e i n t h e t e m p e r a t u r e
of super-heated v a p o r leaving the surface. T h i s t e m p e r a t u r e
change i s a m p l i f i e d a n d r e c o r d e d o n a r e c o r d e r c a l i b r a t e d f o r
20 C s p a n o v e r f u l l c h a r t . W i t h t h i s t e m p e r a t u r e - m e a s u r i n g
i n s t r u m e n t a n d p r o p e r r e c o r d i n g c h a r t s h o w n o n F i g u r e 1 together w i t h a c c u r a t e c o n s t a n t s u p e r - s a t u r a t i o n c u r v e s f o r f i l l m a s s
as shown on Figures 2, 3 a n d 4 for 6" Hg abs., 7" Hg abs. a n d
8 " H g abs. r e s p e c t i v e l y , i t i s p o s s i b l e t o b o i l t h e p a n a t c o n s t a n t
super-saturation.

Figure 1.

204

JOURNAL OF THE A. S. S. B. T,

Figure 2.Constant super-saturation lines.

Figure 3.Constant super-saturation lines.

Figure 4.Constant super-saturation lines.

VOL.

13,

No.

3,

OCTOBER

1964

205

Figure 5 . T y p i c a l p a n b o i l i n g at constant super-saturation.

O n F i g u r e 5 , l i n e A B s h o w s t h e t y p i c a l b o i l i n g cycle. P o i n t
A indicates the initial j u i c e p u r i t y a n d t e m p e r a t u r e at g r a i n i n g .
Point B indicates t h e final m o t h e r l i q u o r or g r e e n p u r i t y a n d
temperature. L i n e AB is on the constant super-saturation of
1.35. D u r i n g b o i l i n g p e r i o d A B , s u g a r f r o m t h e m o t h e r l i q u o r
i s d e p o s i t i n g o n t h e s u r f a c e o f e x i s t i n g s u g a r crystals a t v e r y
rapid r a t e . T h e r e f o r e , t h e p u r i t y o f m o t h e r l i q u o r i s d e c r e a s i n g
with t i m e a l t h o u g h t h e p a n i s still f e e d i n g w i t h fresh h i g h p u r i t y
standard l i q u o r . H e n c e , r a i s i n g t h e f i l l m a s s t e m p e r a t u r e f r o m
point A to p o i n t B will n o t raise t h e super-saturation p r o v i d e d
that a b s o l u t e p r e s s u r e o f t h e p a n r e m a i n e d t h e s a m e .
B o i l i n g a Strike
T h e strike was i n i t i a t e d by t u r n i n g a switch w h i c h o p e n e d
feed a n d c o n d e n s e r v a l v e s . W h e n j u i c e l e v e l r e a c h e d t h e g r a i n ing v o l u m e , s t e a m w a s t u r n e d o n t o c o n c e n t r a t e t h e l i q u o r a n d
the level m a i n t a i n e d c o n s t a n t . A n a l a r m s o u n d e d w h e n s u p e r saturation r e a c h e d t o p r e s e t v a l u e o f 1.35. T h e n t h e p a n w a s
seeded w i t h 2 0 0 c.c. o f m i l l e d f o n d a n t . A s t h e b o i l i n g p r o c e e d e d ,
fillmass t e m p e r a t u r e t e n d e d t o r i s e a b o v e t h e set p o i n t w h i c h
was 1.35 s u p e r - s a t u r a t i o n , t h e s t e a m f l o w w a s t h r o t t l e d t o p r e v e n t
exceeding t h a t l i m i t . A s t h e p a n c a m e t o g e t h e r , t h e i n c r e a s i n g
tightness o p e n e d t h e f e e d v a l v e . T h e c o m b i n a t i o n o f f e e d a n d
increased crystal a r e a s t e n d e d t o c a u s e t h e s u p e r - s a t u r a t i o n t o
f a l l away f r o m 1.35. T h e r e f o r e , t h e s t e a m v a l v e w a s o p e n e d t o
m a x i m u m t o k e e p t h e s u p e r - s a t u r a t i o n c o n s t a n t a t 1.35. B o i l i n g
p r o c e e d e d u n t i l t h e p a n r e a c h e d t h e m a x i m u m set l e v e l , t h e
f e e d valve was t h r o t t l e d t o p r e v e n t f u r t h e r r i s e a n d t h e p a n

206

JOURNAL OF THE A.

S.

S.

B. T.

b e g a n t o b r i x u p . A t t h i s t i m e t h e f i l l m a s s t e m p e r a t u r e started
t o rise a b o v e t h e set p o i n t , t h e s t e a m f l o w w a s t h r o t t l e d again
t o a v o i d excess t e m p e r a t u r e rise a n d t o h o l d t h e super-saturation
c o n s t a n t a t 1.35 u n t i l t h e p a n was d r o p p e d .
E s t i m a t i o n of C o n s t a n t K
K n o w i n g t h e p u r i t i e s o f s t a n d a r d l i q u o r a n d h i g h g r e e n , the
b o i l i n g t i m e f r o m g r a i n i n g t o f i n i s h i n g t h e s t r i k e a n d t h e cons t a n t s u p e r - s a t u r a t i o n a t w h i c h t h e p a n i s b e i n g b o i l e d , the
c o n s t a n t k c a n b e c a l c u l a t e d f r o m E q u a t i o n (3).
1n P 1n P = k ( S 1 ) t
1n
P 1n P
Solve for k,
k
=
(
S

1
)
t
(6)
For the experimental boiling shown on Figure 1:
initial standard liquor purity
=
94.0
final h i g h g r e e n p u r i t y
=
86.2
super-saturation
=
1.35
boiling time
= 120 m i n u t e s
absolute pressure
=
7" Hg abs.
k c a n be e s t i m a t e d f r o m E q u a t i o n (6).
k = 1n 94.0 1n 86.2
=
0.08662 = 0.08662 = 0.002062
(1.35 1) 120
0.35 (120)
42
D u r i n g t h e b o i l i n g t i m e o f 120 m i n u t e s , t e m p e r a t u r e o f fillmass was c o n t r o l l e d so t h a t s u p e r - s a t u r a t i o n of fillmass not to
e x c e e d o r d r o p b e l o w 1.35. T h i s w a s v e r y difficult t o accomplish
b e c a u s e t h e l o c a t i o n of 1.35 s u p e r - s a t u r a t i o n was n o t clearly
k n o w n f r o m t h e r e c o r d i n g c h a r t s h o w n o n F i g u r e 1 . Therefore,
after t h e s t r i k e w a s c o m p l e t e d , t h e t e m p e r a t u r e c h a r t was exa m i n e d for i n t e r m e d i a t e p o i n t s t o d e t e r m i n e i f t h e e n t i r e boili n g was c o n d u c t e d at c o n s t a n t s u p e r - s a t u r a t i o n of 1.35. This
was d o n e b y c o m p u t i n g t h e i n t e r m e d i a t e m o t h e r l i q u o r purity
P 1 u s i n g t h e k v a l u e o b t a i n e d a b o v e , t i m e t after g r a i n i n g and
c o n s t a n t s u p e r - s a t u r a t i o n of 1.35. P 1 was l o c a t e d on F i g u r e 3 at
1.35 s u p e r - s a t u r a t i o n a n d t h e c o r r e s p o n d i n g f i l l m a s s temperature
was r e a d . T h e r e s u l t i n g f i l l m a s s t e m p e r a t u r e w a s t h e same a s
t h e t e m p e r a t u r e r e c o r d e d o n F i g u r e 1 . H e n c e , t h e p a n was
b o i l i n g at c o n s t a n t s u p e r - s a t u r a t i o n of 1.35.
For the experimental boiling:
k = 0.002062, P = 9 4 . 0 , t = 30 m i n u t e s a f t e r g r a i n i n g ,
s u p e r - s a t u r a t i o n = 1.35.
S u b s t i t u t i n g t h e a b o v e d a t a i n t o E q u a t i o n (4):
1n P 1 = 1n 94 0.002062 (1.35 1) 30
= 6.84588 0.002062 (0.35) 30
= 6.84588 0 . 0 2 1 6 5 6.82423
H e n c e , P 1 = 91.9 p u r i t y .

VOL. 13, N o . 3, O C T O B E R 1964

207

Fillmass t e m p e r a t u r e of 7 4 . 8 C w a s o b t a i n e d by l o c a t i n g P 1 of
91.9 o n F i g u r e 3 . F i l l m a s s t e m p e r a t u r e o f 9.8 c h a r t u n i t s , w h i c h
was 74.8 C , w a s o b t a i n e d b y l o c a t i n g a p o i n t 3 0 m i n u t e s a f t e r
graining on F i g u r e 1. Both t e m p e r a t u r e s shown above were
exactly t h e s a m e , t h e r e f o r e , t h e p a n w a s b o i l i n g a t c o n s t a n t s u p e r s a t u r a t i o n of 1.35 f r o m t = 0 m i n . to t = 30 m i n . T h e b o i l i n g
curve was c h e c k e d a t 15. m i n u t e i n t e r v a l s i n t h e s a m e m a n n e r .
I t was f o u n d t h a t for b o i l i n g p e r i o d o f 2 h o u r s a n d 1 C
temperature rise b e t w e e n g r a i n i n g a n d d r o p p i n g t h e p a n , t h e
entire b o i l i n g w a s c o n d u c t e d a t c o n s t a n t s u p e r - s a t u r a t i o n p r o vided t h a t t h e l i n e b e t w e e n i n i t i a l a n d f i n a l p o i n t s w a s fairly
straight a n d t h e a b s o l u t e p r e s s u r e r e m a i n e d c o n s t a n t .
T o verify t h e v a l u e o f k c a l c u l a t e d f r o m E q u a t i o n (6), t h e
following p r o c e d u r e w a s u s e d :
U s i n g P o l a r o i d c a m e r a , series
o f p i c t u r e s w e r e t a k e n f r o m p a n m i c r o s c o p e a t 15, 20, 2 5 , 4 0 a n d
110 m i n u t e s f r o m g r a i n i n g a s s h o w n o n F i g u r e s 6 , 7 , 8 , 9 a n d
10. T h e v o l u m e o f fillmass i n t h e p a n w a s r e c o r d e d f o r e a c h
picture t a k e n . W i t h t h i s d a t a , t o g e t h e r w i t h t h e k n o w n q u a n t i t y
of milled fondant used, it was possible to calculate t h e constant k.
Milled fondant was p r e p a r e d by a d d i n g o n e p o u n d of granulated s u g a r w i t h a l i t e r ( 1 0 0 0 c.c.) o f i s o - p r o p y l a l c o h o l i n t h e
mill jar. T h e m i l l i n g w a s c o n t i n u e d f o r e x a c t l y 2 4 h o u r s . I t
was f o u n d t h a t t i m e s o f m i l l i n g i n excess o f 2 4 h o u r s d i d n ' t
accomplish a n y f i n e r g r i n d i n g b u t t i m e s o f less t h a n 2 3 h o u r s
were f o u n d t o p r o d u c e i n t e r i o r seed.
F o r m i l l i n g t i m e o f 2 4 h o u r s , 1 c.c. o f f o n d a n t c o n t a i n s 2.5
X 10 9 crystals. T h e a v e r a g e c r y s t a l size h a s b e e n o b s e r v e d t o b e
4.5 m i c r o n s or 0 . 0 0 0 1 7 7 " .
F o r e x p e r i m e n t a l b o i l i n g 2 0 0 c.c. o f m i l l e d f o n d a n t w a s u s e d
to grain t h e p a n . T o t a l n u m b e r of c r y s t a l s in 2 0 0 c.c. = 5 X 10 1 1
crystals. F r o m F i g u r e 6 , a v e r a g e g r a i n size w a s 0 . 0 0 6 " a t 1 5
m i n u t e s after g r a i n i n g .
V o l u m e o f f i l l m a s s a t 1 5 m i n u t e s a f t e r g r a i n i n g w a s 4 3 7 cu.ft.
Purity of fillmass = 9 4 . 0 . S u p e r - s a t u r a t i o n = 1.35 (a) 7 4 . 5 C
and 7" H g a b s .
S u p e r - s a t u r a t i o n = 1.35

s u g a r i n g m . p e r 100 g m . w a t e r
3 4 4 g m . s u g a r p e r 100 g m . w a t e r
S u g a r i n l i q u o r = 4 6 4 g m . p e r 100 g m . w a t e r
dry substance = 464 = 494 gm.
0.94
Dry substance + W a t e r

594 gm.

208

JOURNAL OF THE A. S. S. B. T.

RDS (Refractometer Dry Substance) = 494 X 100 = 83.2


594
Density at 83.2 RDS = 89.4#/cu.ft.
W h i t e fillmass = 437 X 89.4 = 39,068 lb., Dry substance =
39,068 X .832 = 32,505 lb.
Sugar = 32,505 X 0.94 = 30,555 lb.
Volume of Crystal = 0.7 (6.0 X 10 -3 ) 3 = 1.52 X 10-7 in 3 / crystal
T o t a l crystal volume = 5 X 1011 X 1-52 X 10 -7 = 76,000
in 3 = 44.0 cu.ft.
Density of sugar = 99.15#/cu.ft. Crystal weight = 99 15 X
44 = 4,363 lb.
30,555 4,363
Mother liquor purity = 32,505 4,363 X 100 = 93.0
Substituting P i = 93.0, P = 94.0, super-saturation S= 1.35
1n 94 - 1n 93 0.0107
t = 15 into Equation (6). k = (1.35 - 1) 15 = 5.25 = 0.002038
T h e value of k calculated above checked fairly well with the
value calculated by using initial and final mother liquor purities
Therefore, k = 0.002062 is sufficiently accurate to be used for
the estimation of boiling time and final high green purity.
Changing in mean dimension of crystal with time was not
linear as shown on Figure 11. Figure 11 was obtained from
Figures 6, 7, 8, 9, 10 and 11. Mean dimension of crystal at one
h o u r after graining war 0.013" which d i d n ' t agree with the
observed value of 0.016" by Ziegler of Taylor Instrument Co.
His value of 0.016" at one h o u r after graining was based on
linear crystal growth of 0.004" per 15 minutes.
Substituting k = 0.002062 into Equation (4),
In P = In P 0.002062 (S 1) t
.
(7)
is obtained. If 1n P is plotted against time t, a straight line
with a slope of - 0 . 0 0 2 0 6 2 (S - 1) is obtained as shown on
Figure 12.

Figure 6.15 minutes after


graining. 437 cu. ft.

Figure 7.20 minutes after graining. 580


cu. ft.

Figure 8.25 minutes after graining. 700


cu. ft.

Figure 9.-40 minutes after graining. 900


cu. ft.

Figure 10.110 minutes after graining.


1650 cu. ft.

JOURNAL OF THE A.

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S.

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210

Figure 11.Mean crystal dimension vs. boiling time.

Figure 12.LN P vs. time based on e q u a t i o n 7.

Final high green purity can be estimated by knowing the


amount of milled fondant used and the screen analysis of sugar
produced. For the experimental boiling 200 c.c of milled
fondant was used to grain the pan and 0.01b MA sugar was
obtained.
Volume of 0.016 MA crystal = 0.7 a3 = 0.7(1.6 X 10-2)3 =
2.87 X 10-6 in3/crystal
Number of crystal in 200 c.c. milled fondant = 200 X 2.5 X
109 crystals
Total crystal volume - 5 X 1011 X 2.87 X 1 0 - 6 = 1,435,000
in3 = 830.44 cu.ft.
Density of sugar = 99.15#/cu.ft.
Weight of crystallized sugar = 830.44 X 99.15 = 82,338 lb.
Strike volume = 1750 cu.ft. @ 91.4 RDS, 94.04#/cu.ft., 94.0
purity

VOL. 13, No. 3, OCTOBER 1964

211

Refractometer Dry Substance


150,417 l b .
Sugar in fillmass
High green purity
Crystallization
High green p u r i t y calculated above checked well w i t h the actual
value o f 86.2 w h i c h w a s d e t e r m i n e d i n t h e l a b o r a t o r y . S u c r o s e
crystallization c a l c u l a t e d a b o v e also c h e c k e d w i t h 6 0 % crystallization o b t a i n e d i n t h e l a b o r a t o r y .
A m o u n t o f m i l l e d f o n d a n t r e q u i r e d t o o b t a i n specified M A
and high green p u r i t y was s h o w n on t h e following e x a m p l e :
Basis:

1000 cu.ft. o f f i l l m a s s
94.0 purity, 91.4 R D S ,
t o o b t a i n 0.017 M A s u g a r a n d 86.0 h i g h
g r e e n p u r i t y . T h e p a n t o b e b o i l e d a t c o n s t a n t 1.4
super-saturation.

From E q u a t i o n
===== 108 m i n u t e s
RDS in fillmass
Sugar in fillmass
Let
of sugar to be crystallized

Volume of sugar to be crystallized


855,844 c u . i n .

1 h o u r 48 m i n u t e s

J O U R N A L OF T H E A.

212

S.

S.

B. T.

A m o u n t of milled fondant required


T h e r e f o r e , 99.6 c.c. of m i l l e d f o n d a n t for 1000 ft 3 strike is req u i r e d t o seed t h e p a n i n o r d e r t o o b t a i n 0.017 M A s u g a r and
86.0 h i g h g r e e n p u r i t y for total b o i l i n g t i m e of 1 h o u r 48 minutes
at 1.4 c o n s t a n t s u p e r - s a t u r a t i o n .
Summary
V a c u u m p a n b o i l i n g t i m e can be accurately e s t i m a t e d by
p r o p e r a p p l i c a t i o n of a u t o m a t i c c o n t r o l s a n d E q u a t i o n (7) prov i d e d t h a t t h e b o i l i n g is to be c o n d u c t e d at c o n s t a n t supers a t u r a t i o n a n d c o n s t a n t pressure.
F o r t h e given i n i t i a l s t a n d a r d l i q u o r a n d final g r e e n purities,
b o i l i n g t i m e c a n b e greatly r e d u c e d b y b o i l i n g t h e p a n a t higher
s u p e r - s a t u r a t i o n p r o v i d e d t h a t t h e s u p e r - s a t u r a t i o n does n o t exceed 1.5 w h i c h is t h e u p p e r l i m i t of M e t a s t a b l e zone. If t h e pan
is to be b o i l e d a b o v e 1.5 s u p e r - s a t u r a t i o n , m u c h of t h e fine grain
f o r m e d d u r i n g b o i l i n g will be so fine t h a t t h e y will n o t be ret a i n e d o n t h e centrifugal screen. they m e r e l y g o t h r o u g h the
screen a n d a r e r e m e l t e d i n t h e h i g h g r e e n s y r u p . T h u s , t h e high
g r e e n p u r i t y for t h e p a n b o i l i n g a b o v e 1.5 s u p e r - s a t u r a t i o n will
n o t b e t h e s a m e a s t h e p u r i t y e s t i m a t e d b y E q u a t i o n (7). Equat i o n (7) is a p p l i c a b l e to t h e c o n d i t i o n s w h e r e t h e sugar from
t h e m o t h e r l i q u o r is d e p o s i t e d on t h e e x i s t i n g sugar crystals and
i s r e t a i n e d o n t h e centrifugal screen.
H i g h g r e e n p u r i t y also can b e e s t i m a t e d b y u s i n g Equation
(7), if i n i t i a l t h i c k j u i c e p u r i t y , b o i l i n g t i m e a n d c o n s t a n t supersaturation at which the pan is to be boiled are known.
C o n s t a n t k m a y vary from factory to factory d u e to different
c o n s t i t u e n t s in t h e n o n - s u g a r a n d different s t e a m pressure at
c a l a n d r i a section of p a n . T h e r e f o r e , c o n s t a n t k is to be determ i n e d for each factory.

P
P
S
k
t
a

Nomenclature
= P u r i t y of m o t h e r l i q u o r
= Initial standard liquor purity
= Super-saturation
=
C o n s t a n t to be d e t e r m i n e d from E q u a t i o n (6)
=
Boiling time
=
M e a n d i m e n s i o n of s u g a r crystal

Vol.

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OCTOBER

1964

213

Acknowledgment
Cooperation from Engineering Department, Research Department and Operating Department made this work possible. Mr.
J. G. Ziegler, Taylor Instrument Company, and Mr. Julian
Johnson, Manager of Research Laboratory, T h e Amalgamated
Sugar Company have contributed to this investigation.
Literature Cited
GILLETT, EUGENE C. 1948. Low grade sugar crystallization. California a n d
Hawaiian Sugar Refining Corp., ltd., Crockett, California.
HONIG, PIETER. 1959. Principle of Sugar Technology. Vol. II, Elsevier P u b lishing Company, New York.
MCGINNIS, R. A. 1951. Beet-Sugar Technology. Reinhold Publishing Corp.,
New York.
ZIEGLER, J. G. 1963. Experiments in vacuum pan control. J. Am. Soc. Sugar
Beet Technol. 12 (6) : 462-467.

Thin Film Ultra-Violet Sterilization


of Liquid Sugar, Using the Aquafine Sterilizer1
ROBERT S. GADDIE, ROBERT R. W E S T , AND E. G. BENNISON 2
Received for publication March

21,

1964

Applying the thin film penetration theory of ultra-violet sterilization, considerable success in the sterilization of water has
been demonstrated the past few years. Using this knowledge the
Aquafine Corporation has designed equipment to pass a film of
syrup over an ultra-violet source under extremely turbulent conditions so that theoretically a thin film condition was presented
to the radiation for each particle of syrup. T h e equipment used
in the experiments detailed in this report was the Model G-4
Aquafine Liquid Sugar Sterilizer. It contains 4 Westinghouse
germicidal lamps 4-G36-T6L, wave length 2537 angstroms.
At the West Jordan plant there was available a 4000-gallon
cone-bottom liquid-sugar tank. T h e outlet at the bottom of the
cone was piped with
inch pipe to a positive displacement
(Viking) p u m p which pumped through the sterilizer, up over
the top of and back into the tank. Circulation was at the rate
of 20 gallons per minute. A sample cock was placed on the
suction line to the p u m p and another on the discharge line from
the sterilizer. Yeast and mold counts were made according to
the method of the A.B.C.B.
Investigation No. 1
One thousand two hundred gallons of liquid sucrose, 66.5
Brix were placed in the tank and seeded with a variety of yeasts
normally found in liquid sugar. T h e mixture was stirred thoroughly and the yeast count determined to be 900 per 10 grams
solids. T h e circulation through the sterilizer was started, and

VOL. 13, N o . 3, OCTOBER 1964

215

samples were t a k e n for p l a t i n g each 2 h o u r s from t h e i n t ak e


of the p u m p a n d from t h e discharge of the sterilizer. Results
are shown in T a b l e 1.
Investigation N o . 2
T h e same 1200 gallons of l i q u i d sucrose were used in the
second e x p e r i m e n t except t h a t this t i m e they were seeded with
a variety of m o l d s c o m m o n l y f o u n d in l i q u i d sugar. T h e m o l d
count after t h o r o u g h m i x i n g was 6500 p e r 10 g r a m s solids, a n d
it was estimated t h a t 1 or
of this c o u n t was yeasts. Circulation was at 20 gallons p e r m i n u t e , a n d samples were taken at
0, 2, 4, 12, a n d 20 h o u r s . R e s u l t s are shown in T a b l e 2.
Table 2.Mold counts, circulated liquid sucrose, 20 GPM, single sterilizer.

Average kill per pass approximately 40%.

E x p e r i m e n t N o . 2 confirms w h a t has b e e n f o u n d in previous


laboratory investigations, t h a t m o l d spores a r e considerably m o r e
resistant to ultra-violet r a d i a t i o n t h a n yeasts.
Investigation N o . 3
As a result of t h e first t w o investigations, a n o t h e r Aquafine
G-4 u n i t was p u r c h a s e d a n d p i p e d in series w i t h t h e first o n e .
Another batch of 1200 gallons of l i q u i d sucrose was p u m p e d
to the t a n k a n d seeded w i t h sufficient yeasts to grossly c o n t a m i nate it. After s t i r r i n g , t h e yeast c o u n t was d e t e r m i n e d to be
Table 3.Yeast counts, circulated liquid sucrose 20 GPM, two sterilisers in series.
Yeasts per 10 gm solids
elapsed,
hours

Before
sterilizer

After
sterilizer

216

JOURNAL OF THE A. S. S. B. T.

14,400 p e r 10 g r a m s solids w i t h p e r h a p s 1 or 2% of these being


m o l d s . T h e l i q u i d was a g a i n c i r c u l a t e d a t 2 0 gallons p e r m i n u t e
e x c e p t t h a t t h i s t i m e it passed t h r o u g h 2 sterilizers in series.
Samples before a n d after t h e sterilizers w e r e t a k e n each hour
for 8 consecutive h o u r s . R e s u l t s a r e s h o w n in T a b l e 3.
L i q u i d Sucrose Corn Syrup B l e n d s
F r o m t h e s t a n d p o i n t of t h e c o n t r o l of m i c r o - o r g a n i s m s , the
m o s t difficult of t h e l i q u i d s w e e t e n e r s to p r o d u c e , transport,
a n d store a r e t h e l i q u i d s u g a r c o r n s y r u p b l e n d s . If they are
s u b j e c t e d to sterilizing t e m p e r a t u r e s for a n y l e n g t h of t i m e , they
d a r k e n b a d l y . T h i n film r e g e n e r a t i v e h e a t pasteurizers are
effective b u t to get a practical t h r o u g h - p u t capacity, a r e ex
pensive. T h e ultra-violet sterilizers a r e e c o n o m i c a l , easy to install, a n d w i t h t h e a i d of hoses, t h e y m a y be m a d e p o r t a b l e . In
view of this, it was d e c i d e d to try these u n i t s on b l e n d s to
d e t e r m i n e if o n e pass at 20 gallons p e r m i n u t e w o u l d reduce
t h e yeast a n d m o l d c o u n t t o s o m e practical c o n c e n t r a t i o n . Two
A q u a f i n e G-4 u n i t s w e r e p u r c h a s e d for t h e T o p p e n i s h plant.
H e r e all types of b l e n d s are p r o d u c e d , usually in 4500-gallon
batches. I n i n s t a l l i n g t h e t w o u n i t s b e c a u s e t h e p i p i n g was
simpler, t h e factory installed t h e m in parallel on t h e t h e o r y that
20 gallons p e r m i n u t e from t h e p u m p g i v i n g 10 gallons per
m i n u t e t h r o u g h each sterilizer w o u l d effect t h e s a m e kill as
if they h a d b e e n c o n n e c t e d in series. T h i s d i d n o t w o r k out as
e x p e c t e d as s h o w n in T a b l e 4 for several r e p r e s e n t a t i v e batches.
Table 4.Yeast and mold counts, 75-25 liquid sucrose corn syrup Mend, two sterilizers
in parallel, 10 GPM through each, single pass.
Before sterilizers

After sterilizers

Yeasts

Molds

Yeasts

Molds

632
728
872
320
834

156
54
60
40
110

510
400
440
272
688

162
22
8
36
74

Table 5.Yeast and mold counts, 75-25 liquid sucrosecorn syrup blend, two sterilizers
In series, 20 GPM, single pass.
Before sterilizers

After sterilizers

Yeasts

Molds

Yeasts

752
484
472
664
872
468

120
10
36
12
8
6

92
36
0
68
148
54

Molds
52
0
0
8
0
4

VOL.

13,

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3,

OCTOBER

1964

217

T h e two u n i t s w e r e t h e n p i p e d i n series w i t h results shown


in T a b l e 5 for several r e p r e s e n t a t i v e b a t c h e s .
Summary
In the first t h r e e investigations t h e l i q u i d sucrose was circulated o u t o f t h e t a n k , t h r o u g h t h e sterilizer, a n d back i n t o t h e
tank. T h e o r e t i c a l l y , t h e m i c r o - o r g a n i s m c o u n t i n t h e t a n k will
be zero only after infinite c i r c u l a t i o n t i m e . H o w e v e r , results
indicate t h a t t h e f o l l o w i n g c o n c l u s i o n s m a y b e safely d r a w n .
At a c i r c u l a t i o n r a t e of 20 G P M t h r o u g h a single G-4 A q u a fine u n i t o p e r a t i n g at n o r m a l r a d i a t i o n level, 6 v o l u m e t h r o u g h puts of t h e 1200 gallons of l i q u i d sugar killed sufficient yeasts
in a heavily c o n t a m i n a t e d l i q u i d sucrose to b r i n g it w i t h i n
Bottler's S t a n d a r d s for yeasts.
At a c i r c u l a t i o n r a t e of 20 G P M t h r o u g h a single G-4 A q u a fine u n i t , t h e 1200 gallons of l i q u i d sucrose grossly c o n t a m i n a t e d
with m o l d s r e q u i r e d 1 2 t o 2 0 v o l u m e t h r o u g h - p u t s t o b r i n g
it within t h e l i m i t s of B o t t l e r ' s S t a n d a r d s for m o l d s .
At a p u m p i n g r a t e of 20 G P M of grossly yeast c o n t a m i n a t e d
liquid sucrose t h r o u g h t w o A q u a f i n e G-4 u n i t s in series, a single
pass will p r o d u c e an effluent m e e t i n g B o t t l e r ' s S t a n d a r d s for
yeasts.
It was c o n c l u d e d from t h e r e s u l t s s h o w n in T a b l e s 4 a n d 5
on liquid sucrosecorn syrup blends that 10 G P M did not
result in sufficient t u r b u l e n c e t h r o u g h t h e sterilizer to p r o d u c e
the t h i n film c o n d i t i o n necessary for ultra-violet r a d i a t i o n sterilization. At 20 G P M a n d w i t h t w o sterilizers in series, sufficient
kill of m i c r o - o r g a n i s m s c o u l d be o b t a i n e d in a single pass to
produce a b l e n d m i c r o b i o l o g i c a l l y s u i t a b l e for most b l e n d users.

Paper Chromatographic Determination of Raffinose


in Sugar Beet Molasses
F E R N JANTZEF A N D A. L. POTTER 1
Received for publication April

6,

1964

A c c u r a t e k n o w l e d g e of t h e raffinose c o n t e n t of molasses is
i m p o r t a n t t o sugar b e e t g r o w e r s a n d processors. C a r r u t h e r s and
co-workers (4) 2 h a v e r e v i e w e d m e t h o d s c o m m o n l y u s e d for the
d e t e r m i n a t i o n o f raffinose a n d d e s c r i b e d t h e i r l i m i t a t i o n s .
P o l a r i m e t r i c m e t h o d s a r e m o s t w i d e l y u s e d i n t h e sugar beet
i n d u s t r y . O p t i c a l l y active s u b s t a n c e s o t h e r t h a n t h e common
sugars r e d u c e t h e r e l i a b i l i t y o f these m e t h o d s .
P a p e r c h r o m a t o g r a p h i c m e t h o d s a r e m o r e specific a n d more
r e l i a b l e . A l b o n a n d Gross (1) d e t e r m i n e d raffinose directly,
de W h a l l e y (5) h y d r o l y z e d raffinose to m e l i b i o s e w i t h invertase
b e f o r e c h r o m a t o g r a p h y to e l i m i n a t e i n t e r f e r e n c e of kestoses and
s l o w - m o v i n g c o l o r e d c o m p o u n d s . T h e s e m e t h o d s i n v o l v e visual
c o m p a r i s o n of t h e c o l o r d e v e l o p e d from spots of raffinose or
m e l i b i o s e w i t h t h a t from a series of s t a n d a r d s . T h u s , they are
l i m i t e d b y subjective e r r o r s . B e v e n u e a n d W i l l i a m s ' method
(3) is s i m i l a r to t h a t of de W h a l l e y e x c e p t t h a t t h e y measure
t h e d e v e l o p e d c h r o m a t o g r a p h i c spots of m e l i b i o s e by reflected
light. R e p r o d u c i b i l i t y of these m e t h o d s is a p p r o x i m a t e l y 10%.
W e i d e n h a g e n a n d Schiweck (9) o b t a i n e d b e t t e r chromata
g r a m s by clarifying t h e molasses w i t h l e a d a c e t a t e a n d then de
s a l t i n g w i t h i o n - e x c h a n g e resins.
T h e y r e p o r t e d incomplete
recovery of sugars from t h e resins a n d u s e d a c o r r e c t i o n factor.
Several m e t h o d s (6, 7, 8) i n v o l v e p h o t o m e t r i c measurements
after e l u t i o n of t h e raffinose- or m e l i b i o s e - c o n t a i n i n g zones of
p a p e r c h r o m a t o g r a m s . T h e s e m e t h o d s c a n give very good precision. H o w e v e r , e l u t i o n m a y n o t r e c o v e r all t h e sugar.
W e r e p o r t a p a p e r c h r o m a t o g r a p h i c m e t h o d for determining
raffinose d i r e c t l y . It avoids i o n - e x c h a n g e resins, invertase hydrolysis, or e l u t i o n . T h e d i l u t e d molasses, clarified by lead
a c e t a t e , is s p o t t e d on a c h r o m a t o g r a p h i c sheet. T h e sugars are
s e p a r a t e d by d e s c e n d i n g c h r o m a t o g r a p h y , a n d t h e raffinose spots
a r e d e v e l o p e d w i t h p-anisidine. T h e r e s u l t i n g yellow color i s
m e a s u r e d w i t h a c o l o r difference m e t e r or a t r a n s m i s s i o n densitometer.
T h i s r e p o r t d e s c r i b e s a s e c o n d clarification p r o c e d u r e , which
r e m o v e s m o r e c o l o r e d m a t e r i a l s . A f t e r clarification with l e a d
1
Western Regional Research Laboratory, Western Utilization Research and Develop-,
ment Division, Agricultural Research Service, U. S. Department of Agriculture, Albany
California.
2
Numbers in parentheses refer to literature cited.

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acetate, the molasses solution is washed through a cellulose


column with
ethanol. Both procedures gave good chromatograms with the samples analyzed, that is, raffinose appeared as
a compact spot separated from other color-producing compounds.
If chromatography of the lead-treated molasses does not separate
raffinose from colored materials, the second procedure may be
preferred.
Experimental
Apparatus.Gardner Automatic Color Difference M e t e r ,
Model AC-2 (Gardner Laboratory, Inc., Bethesda, Md.). This
instrument measures colors in terms of percent reflectance Rd
and chromaticity coordinates, a (red, plus; green, minus) and
b (yellow, plus; blue, minus) (2).
Photovolt Transmission Densitometer (Electronic Photometer
Model 501-A with Transmission Density Unit Model 52-C).
Reagents.Neutral Lead Acetate. Saturated aqueous solution.
Raffinose Standard
anhydrous raffinose). Dilute
3.54 g raffinose pentahydrate to 100 ml with water. Prepare
working standards from this stock solution. The stock and standard solutions may be preserved by adding a few crystals of
thymol and storing in a refrigerator.
Chromatographic Solvent. w-Propanol-ethyl a c e t a t e - w a t e r
(7:1:2 by volume) (1).
p-Anisidine. Suspend 20 g p-anisidine in about 400 ml absolute ethanol. Dissolve by adding 100 ml concentrated hydrochloric acid. Dilute to 1000 ml with absolute ethanol.
Chromatographic Paper. Whatman No. 1, chromatographic
grade paper
Whatman Standard Grade Cellulose Powder.
Sample Treatment.Heat the molasses with occasional stirring in a water bath at 100 C or in an oven at 110 C until
it is homogeneous. Clarify the sample by one of the methods
A or B below.
A. Clarification with Lead Acetate. Transfer a 5-g sample to
a 25 ml volumetric flask with water. Add 1 ml lead acetate solulon. This quantity precipitates interfering substances of most
samples. Mix and dilute to volume. If foam hinders dilution
to volume, add one drop of amyl alcohol. Let stand about 30
minutes, then filter using Eaton-Dikeman No. 509 folded filter
paper.
B. Clarification with Lead
Acetate
and
Cellulose
Column.
to dilute a 5-g sample of molasses and I ml lead acetate solution
to 10.0 ml with water. Let stand about 30 minutes, filter.

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JOURNAL OF THE A. S. S. B. T.

P r e p a r e t h e c o l u m n , 3 0 m m i n d i a m e t e r , w i t h a fritted glass
disc, by p o u r i n g in a s l u r r y of cellulose p o w d e r in absolute
e t h a n o l . Stir a n d let settle a s t h e l i q u i d f l o w s . T h e f i n a l volume
of settled cellulose is 120 m l .
R u n 4.0 m l o f t h e f i l t r a t e i n t o t h e c o l u m n . R i n s e t h e sides
of t h e c o l u m n a b o v e t h e cellulose w i t h s m a l l p o r t i o n s of 80%
e t h a n o l . T h e n a d d 8 0 % e t h a n o l c o n t i n u o u s l y t o t h e t o p o f the
c o l u m n . D i s c a r d t h e first 75 ml of effluent. C o l l e c t t h e next
250 ml of effluent. E v a p o r a t e it on a s t e a m b a t h to a few ml
a n d t h e n d i l u t e to 10.0 m l . Q u a l i t a t i v e tests s h o w e d t h a t all the
p - a n i s i d i n e - p o s i t i v e sugars w e r e i n t h i s fraction.
Paper
Chromatography.Place
10.0 liter-spots of t h e solut i o n s on a c h r o m a t o g r a p h i c sheet at 1.5 i n c h i n t e r v a l s . Apply
on e a c h sheet f o u r spots of molasses s o l u t i o n , t h r e e spots of a
raffinose s t a n d a r d of l o w e r c o n c e n t r a t i o n , f o u r spots of inter
m e d i a t e c o n c e n t r a t i o n , a n d t h r e e spots o f h i g h e r concentration.
A r r a n g e t h e spots i n g r o u p s s o t h a t t h e spots o f a n y o n e solution
a r e a b o u t e q u a l l y d i s t r i b u t e d across t h e sheet. U s e raffinose
standards varying f
r
o
m
M
o
s
t samples fell
in a s m a l l e r r a n g e , 1.5
D i l u t e t h e molasses
s o l u t i o n s if necessary so t h a t t h e raffinose c o n c e n t r a t i o n is near
that of the intermediate standard.
S e p a r a t e t h e sugars by d e s c e n d i n g c h r o m a t o g r a p h y . Spot test
p a p e r strips w i t h a molasses s o l u t i o n . C h r o m a t o g r a p h t h e strips
w i t h t h e sheets a n d d e v e l o p t h e strips a t i n t e r v a l s t o determine
w h e t h e r raffinose is well s e p a r a t e d f r o m o t h e r sugars. A b o u t 64
h o u r s is u s u a l l y r e q u i r e d to s e p a r a t e raffinose f r o m sugars (probably kestoses) t h a t i n t e r f e r e w i t h its m e a s u r e m e n t . W h e n chroma t o g r a p h y i s f i n i s h e d , a i r d r y t h e sheets.
D i p each sheet by d r a w i n g it t h r o u g h a t r o u g h containing
p - a n i s i d i n e r e a g e n t . L e t t h e s h e e t d r y 15 m i n u t e s in a fume
h o o d . H e a t in a forced a i r o v e n at 70 C for 2 m i n u t e s . After
a l l o w i n g it to s t a n d for a few m i n u t e s at r o o m temperature,
m e a s u r e t h e c o l o r of t h e d e v e l o p e d raffinose spots w i t h the color
difference m e t e r o r t h e t r a n s m i s s i o n d e n s i t o m e t e r .
Measurement
with
Color Difference Meter.-Operate the ins t r u m e n t a c c o r d i n g t o t h e m a n u f a c t u r e r ' s i n s t r u c t i o n s . Standardize t h e c o l o r difference m e t e r w i t h a " y e l l o w " c a l i b r a t e d enameled
standard
o b t a i n e d from the
m a n u f a c t u r e r . P o s i t i o n t h e s h e e t so t h a t a raffinose s p o t is centered
o v e r t h e a p e r t u r e (1.5 i n c h d i a m e t e r ) . P l a c e t h e e n a m e l e d
standard on the paper to hold it flat. O b t a i n readings of
a n d b. M o v e t h e p a p e r slightly w h i l e b a l a n c i n g t h e 6
aticity c o o r d i n a t e t o assure t h e m a x i m u m r e a d i n g . T h e
c o l o r fades w h e n e x p o s e d t o t h e l i g h t o f t h e i n s t r u m e n t ,

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color of each spot must be measured after an equal time of


exposure. Record the b reading of each spot at 25 seconds of
exposure.
For each sheet, plot the average b readings of each standard
solution against the raffinose concentration. T h e relation between b reading and raffinose concentration is shown in Figure
1. Average the b readings for the sample on the same sheet and
determine the amount of raffinose from this curve.

Figure 1.Relation between b chromaticity coordinate and raffinose


concentration.

Measurement with
Transmission Densitometer.-Place the
sheet over the aperture
densitometer. Using a
that the optical density reads zero (100% transmission) on a
blank area of the chromatographic sheet in the region of the
raffinose spots and infinity (zero percent transmission) on the
most dense area of a developed sucrose spot. Measure the maximum optical density of each raffinose spot. Determine the amount
of raffinose of the sample from the standard curve for the same
sheet.

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JOURNAL OF THE A. S. S. B. T.

Results and Discussion


Replicate determinations of about 1% raffinose in molasses
by the methods presented in this report were within the following limits: If the color of the p-anisidine-rafnnose spots was
measured with the color difference meter, the results were within about five percent of the mean. If the transmission densitometer was used, the results were within about ten percent.
T w o experiments were carried out to evaluate the accuracy
of the paper chromatographic method. First, molasses samples
were analyzed before and after the addition of known amounts
of raffinose. Second, a sample of molasses was fermented until
free of sucrose and raffinose. Sucrose and known amounts of
raffinose were then added, and the raffinose was determined.
Table 1 summarizes the results of these experiments. T h e data
show that the method correctly determined the amounts of raffinose within experimental error.
Table 1.Paper chromatographic determination of raffinose with known amounts of
raffinose added.
Sample

Percent raffinose
Initially found 1
Added

Found 1

Molasses 1
2

1.13
1.08

0.17
0.36

1.26
1.46

Fermented molasses 2

0.00
0.00

1.00
1.30

0.99
1.34

1
Paper chromatographic method.
2
Free of raffinose and sucrose by treatment with Bakers' yeast. Sucrose was added after
treatment to imitate original molasses (60% sucrose).

Some samples were analyzed to compare the two clarification


procedures. Each method produced good chromatograms, and
the raffinose values obtained by both agreed within experimental
error. T h e simple treatment with lead acetate (A) was satisfactory for the samples encountered. T h e cellulose column procedure (B) removed more colored materials from the molasses
solutions. If chromatography of the lead-treated molasses does
not separate raffinose from colored compounds, the cellulose
column procedure may be required.
Solutions of raffinose alone and of raffinose containing sucrose
in amounts equivalent to those in molasses, were chromatographed. T h e raffinose spots gave the same values whether
sucrose was present or not. We concluded that it is not necessary
to add sucrose to the raffinose standards.
Several reagents, including p-anisidine, orcinol, napthoresorcinol, and a-napthol were tested as developers of the raffinose

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spots. p-Anisidine is sensitive, produced the most stable color,


and gave consistent results.
F a i n t spots of p-anisidine-positive sugars, probably kestoses,
usually a p p e a r e d between raffinose and sucrose on chromatograms
of the samples analyzed. Chromatography for about 64 hours
removed t h e m far e n o u g h from the raffinose that they did not
interfere. C a r r u t h e r s et al. (4) reported that 1-kestose and
neokestose can be chromatographically separated from raffinose,
but that the mobility of 6-kestose is very similar to that of raffinose. In addition, they reported that only traces of 6-kestose are
present in beet molasses. Therefore, we assume that, if 6-kestose
was in the raffinose area, the color developed from it contributed
only negligibly.
T a b l e 2 lists raffinose contents determined from the same
samples by polarimetric methods and by the paper chromatographic m e t h o d . T h e s e data show a large variation in raffinose
content of t h e sugar beet molasses as determined by polarimetric
methods. If it is assumed that the paper chromatographic method
gives the t r u e a m o u n t of raffinose, polarimetric methods may err
by a factor of three or four.
Table 2.Comparison of raffinose content of suger beet molasses determined by polarimetric methods and by paper chromatography.
Sample
1

2
3
4
5

Percent raffinose
Polarimetric1
Paper chromatographic
1.44
1.19
1.08
0.25
0.54

1.20
1.18
1.21
1.01
1.13

1
The values by polarimetric methods were determined at the two sugar beet plants where
the samples were produced.

Summary
A p a p e r c h r o m a t o g r a p h i c determination of raffinose in sugar
beet molasses was developed. T h e treatment of the sample before
chromatography is simple. T w o clarification procedures are reported: A. t r e a t m e n t with lead acetate; B. treatment with lead
acetate followed by washing the sugars through a cellulose powder
column with 8 0 % ethanol. Each procedure gave the same raffinose values. T h e sugars were separated by descending chromatography. p-Anisidine was chosen to develop the color of the raffinose spots because it is as sensitive as other reagents tested and
produces m o r e stable color. T h e intensity of the color of the
spots was m e a s u r e d with a color difference meter or a transmission
densitometer.

224

J O U R N A L OF T H E A.

S.

S. B. T.

Replicate results were within about 5% of the mean when


the color difference meter was used, and within about 10%
when the transmission densitometer was used.
Acknowledgment
T h e authors are indebted to R. M. McCready for samples
of sugar beet molasses and yeast-treated molasses, and to H. C.
Lukens for assistance in using the Gardner Color Difference
Meter.
Reference to a company or product name does not imply
approval or recommendation of the product by the U. S. Department of Agriculture to the exclusion of others that may be
suitable.
Literature Cited
(1)

ALBON, N., a n d D. GROSS. 1952. T h e chromatographic determination


of raffinose in raw sugars. Analyst. 77: 410-412.

(2)

AMERICAN SOCIETY FOR T E S T I N G MATERIALS.

1955.

T e n t a t i v e method

of test for color difference using the H u n t e r Color Difference Meter.


Book of A S T M Standards, Part 8, p p . 1043-1046.
(3)

BEVENUE,

ARTHUR,

and

KENNETH

T.

WILLIAMS.

1958.

Quantitative

d e t e r m i n a t i o n of raffinose or melibiose by p a p e r chromatography.


Arch. Biochem. Biophys. 73: 291-295.
(4)

CARRUTHERS, A., J. V. D U T T O N , J. F. T. O L D F I E L D , C. W. E L L I O T T , R. K.

H E A N E Y a n d H. J. T E A G U E . 15th May, 1963. Estimation of Sugars


in Beet Molasses. Sixteenth A n n u a l T e c h n i c a l Conference of the
British Sugar C o r p o r a t i o n Limited.
(5) DE W H A L L E Y , H. C. S. 1952. Raffinose in molasses a n d low beet products.
I n t . Sugar J. 54: 158.
(6) GOL'DFARB, R. I., a n d P. L. DANILENKO. 1960. Quantitative determination of the raffinose content of molasses by p a p e r chromatography.
Sakharnaya promyshlennost. 34 (5) : 21-23. (Abstr.) Int. Sugar J.
1960, 62: 262.
(7)

SCHNEIDER, F., A. E M M E R I C H , C . R E I C H E L a n d H . R O T H E R .

1959.

Uber

die papierchromatographische Bestimmung von Raffinose. ZuckerBeihefte 3 ( 4 ) : 95-102. (Abstr.) I n t . Sugar J. 1959, 6 1 : 317.
(8)

T A U F E L , K., H . R U T T L O F F a n d A. T A U F E L .

1961.

Z u r Mikroanalytik

d e r Raffinose. N a h r u n g . 5: 353-361.
(9)

W E I D E N H A G E N , R., a n d H . SCHIWECK.

Raffinosebestimmung in Melasse.

1959.

Papierchromatographische

Zeit. Zuckerind.

9: 443-445.

The Effect- of Simulated Hail Injuries


on Yield and Sugar Content of Beets1
M.

M.

AFANASIEV 2

Received for publication March 30, 1964

Introduction
Sugar beets g r o w n in M o n t a n a represent a very important
cash crop. E n v i r o n m e n t a l conditions, soil type and general farm
practices are favorable in M o n t a n a for raising good quality crops
of sugar beets with high sugar content. Most of the sugar beets
in M o n t a n a are grown east of the Continental Divide. T h i s
area in M o n t a n a is sometimes subject to hail storms which cause,
in some seasons, extensive damage to sugar beets and to other
crops. In general, there is very little information regarding the
effect of hail injury on the yield and sugar content of sugar
beets.
In o r d e r to d e t e r m i n e the a m o u n t of damage caused by hail
to sugar beets, a limited a m o u n t of work was conducted in Montana d u r i n g 1946-1949 (6,7) 3 on this subject. However, a more
extensive investigation of this problem was undertaken in 1957
and was c o n t i n u e d t h r o u g h 1962. Preliminary results for the
first three years of this study were published earlier (1). T h i s
paper contains a s u m m a r y of the results of six years of investigation.
Materials and Methods
In this study, the p l a n was to investigate hail damage throughout the w h o l e g r o w i n g season, beginning soon after beets recovered from t h i n n i n g a n d t e r m i n a t i n g as close as possibe to
harvest. In previous studies (6,7) simulated hail damage to
beets was investigated mainly d u r i n g the middle portion of the
growing season.
T h r e e e x p e r i m e n t s were conducted d u r i n g this study:
E x p e r i m e n t 1: Simulated hail injury was inflicted on sugar
beets seven times t h r o u g h o u t the season, and each set of plots
was damaged only once.
E x p e r i m e n t 2: Beets were subjected to two consecutive defoliations d u r i n g t h e m i d d l e of the growing season.
E x p e r i m e n t 3: A comparison was made of the effect of detonation of beets with scissors a n d wooden sticks.
1
Contribution from Montana State College, Agricultural Experiment Station, Bozeman,
Montana. Paper No. 650, Journal Series.
Department of Botany and Bacteriology, Montana Agricultural Experiment Station,
Bozeman,
Montana.
3
Numbers in parentheses refer to literature cited.
2

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JOURNAL OF THE A. S. S. B. T.

All these experiments were conducted at t h e H u n t l e y Branch


Station which is located in t h e Yellowstone Valley of Montana,
a b o u t 20 miles east of Billings. A p p r o x i m a t e l y t w o acres of
beets were used in these studies each year. Beets were grown
in two-year rotations w i t h corn. Soil for corn was usually fertilized with m a n u r e (12-16 tons) a n d s u p p l e m e n t e d in t h e spring
with 400 p o u n d s of 15-20-0 fertilizer p e r acre. Soil for sugar
beets also was fertilized with m a n u r e in the fall and, during
most of the years, 200 p o u n d s of 0-45-0 fertilizer a n d some
nitrogen were added in the spring. Sugar beets were planted
in rows, 24 or 22 inches apart, a n d the usual care was given to
them d u r i n g t h e growing season.
In simulating hail damage in the first e x p e r i m e n t a n d wherever it was applied in other tests, the following procedure was
used:
1. Sugar beets were injured seven times d u r i n g the growing
season. T h e first injury was m a d e in the m i d d l e of J u n e and
subsequent injuries followed at a b o u t 15-day intervals until
the m i d d l e of September.
2. On each date 25, 50, 75 or 1 0 0 % of the beet foliage on
each beet p l a n t in different plots was destroyed. Each leaf blade
was cut separately with scissors to remove an area appropriate
for t h e chosen degree of injury. Approximately one third of
the leaves on each beet p l a n t in every t r e a t m e n t was cut crosswise, one t h r i d lengthwise, a n d one t h i r d diagonally.
3. Each plot of beets consisted either of four 21.8-foot rows
spaced 24 inches apart or 23.6 foot rows spaced 22 inches apart
(each linear 21.8 or 23.6-foot row, hereinafter called a "row",
is equivalent to 1/1000 of an acre). All four rows of beets in
a plot were subjected to this t r e a t m e n t ; however, at harvest
time, d a t a were taken from only the two m i d d l e rows. Each
t r e a t m e n t was applied to four replications of randomized plots
on every injury date. F o u r u n i n j u r e d plots were left as checks
for each d a t e . Beets grown in the defoliated plots were later
c o m p a r e d to beets in the check plots.
4. D u r i n g the last week of September of each year, beets
were harvested a n d counted, weights of tops a n d roots were
d e t e r m i n e d for each plot, a n d sugar analyses were made on
samples taken from each plot.
Effect

of

First Experiment
Simulated Hail Injury on

Sugar

Beets

Results o the First Experiment


In this discussion all weights and yields are average values

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227

DATE OF INJURY

Figure 1.The effect of various degrees of defoliation at different


dates on the relative top weights of sugar beets at harvest (1957-1962).

for the appropriate treatment and injury date for the six years
of the experiment.
T h e tops of the check beets were almost always heavier than
the tops of beets subjected to injury (Figure 1). The 25, 50
and 75% defoliated beets had about the same top weights, and
were only slightly below those of the check beets for the 0rst
five injuries including the one made in the middle of August.
During September however, weights of the tops of 25% defoliated
beets resembled the checks, while the tops of beets with 50 and
75% injuries showed lower weights. The tops of 75% defoliated
beets weighed less than those with 50% defoliation and the
top weights of plants subjected to complete defoliation were
much lower than for beets with smaller degrees of injuries, and
this gradually decreased with later defoliations.
The greatest reduction in weight of the tops of beets with
25% defoliation occurred in plants injured in the middle of
August and this weight was about 10% less than the check. The
greatest losses for 50 and 7 5 % treatments occurred when injuries
were made during September with losses of about 16 and 20%,
espectively. Completely defoliated beets showed great losses in
top weights during the latter part of the season, with the greatest
reduction of 7 3 % occurring during September.
Sugar beets with 25%defoliation,
when c o m p a r e d with
the checks, showed (Figure 2) only a slight decrease in yield,
with a maximum reduction of about 5% when injury was made
during the first part of August. The yields of plants subjected
to 50 and 75% defoliation were more or less similar. However,

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Figure 2.Relative yields of sugar beets subjected to various degrees


of defoliation on different dates (1957-1962).

beets with 7 5 % injury showed a slightly greater reduction in


yield than those with 50% defoliation. T h e greater losses in
yield occurred during the July-August period and were equal
to 9 and 10%, respectively, for the 50 and 7 5 % injuries.
Complete defoliation of beets on the first five dates of injury
considerably reduced the yields. T h e greatest losses due to these
treatments occurred with injuries made in the beginning of
August and amounted to about 3 1 % .
Yield losses of all beets corresponded quite closely to a given
degree of defoliation for all the injuries including the midAugust treatment. However, defoliations made during the first
part of September had, in general, only a slight effect on the
reduction of the yield of beets for all degrees of injuries and
especially for the 25 and 50% defoliated beets. T h e last defoliations, made in the middle of September, had practically
no effect on the yield of beets.
T h e yield of sugar was calculated on the basis of sugar production per acre. T h e content of sugar was not determined in
beets in 1959, so results are given only for 5 years of study.
T h e amount of sugar produced by beets with all degrees of
defoliation was about the same, and was quite similar to the
check plants for the first three injuries (Figure 3). Beginning
with August injuries, variation is evident in the amount of sugar
produced by beets subjected to different degrees of defoliation.
T h e yields of sugar in beets with 25 and 50% defoliations during the remainder of the season were very much like the check

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Figure 3.Relative yields of sugar (on acre basis) in beets subjected


to various degrees of defoliation on different dates (1957-1962).

beets, showing only a slight reduction of from 3 to 4% of sugar


for injuries m a d e in the middle of August. Beets with 7 5 %
defoliation showed some reduction in yield of sugar for both
August injuries, with the greatest loss in the one made d u r i n g
the middle of August which was 7.5% below the check. Practically no r e d u c t i o n in sugar occurred for 7 5 % defoliation made
in September. Losses in yield of sugar in completely defoliated
plants d u r i n g August and September ranged from about 5 to
18%, with only a b o u t 5% for the last date of injury. T h e maxim u m loss in sugar occurred in completely defoliated beets in
the m i d d l e of August.
It appears that 25, 50 a n d 7 5 % defoliations throughout the
season, with the exception of 7 5 % injury in the middle of August,
had little effect on sugar production at harvest time. Beets with
100% injury showed considerable loss in sugar for mid-August
and early September defoliations.
Discussion and Conclusions
T h e results show that sugar beet plants can rapidly restore
their leaves destroyed by defoliation. However, the data also
indicate that recovery was almost never complete. T h e same
results were o b t a i n e d with potatoes in Idaho when more than
2 5 % of the foliage was lost (12). T h e average top weights of
injured beets, for all degrees of defoliation, including 7 5 % ,
showed that these injuries had only a slight effect in reducing
weight of beet tops. Even with 7 5 % defoliation the greatest
reduction in weight of beet tops was equal to only 2 0 % Completely defoliated beets showed considerable reduction in top

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weights. All later defoliations, with the exception of 25% injury,


had a greater effect in reducing the weight of beet tops than did
early treatments. These results are quite similar to those obtained for the first three years of this investigation (1) and for
beet defoliation studies in Canada (5).
The 25% defoliation had only a very slight depressing effect
on the yield of beets per acre with the greatest reduction of
about 5% for early August treatments. Similar results were obtained by other investigators working with beets (2, 5) and
other crops. Work in Texas showed (4) that cotton plants were
not markedly affected by the removal of one third to two thirds
of their leaves. Studies with potatoes (12) and beans (14) in
Idaho showed that early foliage losses up to 25% had only a
small effect in the reduction of yield of these crops. Slightly
greater reduction in yield of beets occurred for 50 and 75%
defoliations and were equal to about 9 and 10%, respectively.
Beets completely defoliated showed a substantial reduction in
yield, with the greatest weight reduction of about 3 1 % for early
August treatment. Very little reduction in the yields of beets
occurred for all injuries made in early September and practically
no reduction for all the mid-September defoliations.
It appears that slight defoliations do not have too much
detrimental effect on the continuous growth of beet roots. A
possible explanation for this situation is that a beet plant develops more leaves then it needs for normal growth of a root.
However, in spite of the apparent excess of leaves which beet
plants have, it appears that not all leaves have the same value.
Indications are (13) that the lower, shaded leaves probably have
less photosynthetic activity than those fully exposed to light.
Early defoliations had much less effect on the reduction of
the yield of beets than those made during the middle of the
season. This undoubtedly was due to the fact that beets, injured
early in the season, had more time to recover than those defoliated
later. The same results were obtained by other investigators
working with potatoes (8,11,12), beans (10,14), cotton (4), flax
(3) and tobacco (9).
All September defoliations and particularly those made during the middle of the month, had little influence on the yield
of beets. This undoubtedly was due to the slow rate of beet
growth during this period of the season. Results from six years
of experimentation appear to give a more uniform picture of
the effect of various degrees of defoliation on the reduction of
the yield of beets than those for only three years of study (1).
Average results for six years of defoliation studies showed greater
beet losses than those for only three years.

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Very little reduction in the yield of sugar took place for 25


and 5 0 % defoliations. Beets with 7 5 % injury, in the middle
of August, showed a slight reduction in the yield of sugar. Complete defoliation resulted in a moderate loss in the yield of sugar
for injuries made d u r i n g the middle of August and the first part
of September. However, even here losses were below 2 0 % . Results from six years of studies showed smaller losses in the yield
of sugar for 75 a n d 100% defoliations than those obtained for
the first three years of investigation (1).
Second Experiment
Effect of Two Consecutive Defoliations on Sugar Beets
In the previous study, the effect of only one defoliation on
the growth a n d yield of sugar beets was investigated. D u r i n g
1960 an e x p e r i m e n t was conducted on the effect on beets of two
consecutive defoliations. In this study the same size plots and
procedures were used as in the previous test.
T h e first defoliation in this experiment was made on July 15,
and the second on August 2. On July 15, 7 5 % of the foliage
was removed from beets grown in 20 plots and 100% from beets
grown in a n o t h e r 20 plots. In addition, two sets of four check
plots were left u n i n j u r e d for 75 and 100% defoliations. On
August 2 the second injury was made. Each group of 20 plots
in which 75 or 100% of foliage was previously removed, was
divided i n t o five sub-groups consisting of four plots each. Twentyfive, 50, 75 a n d 100% defoliation was made on sugar beets grown
in the above m e n t i o n e d groups of plots. F o u r plots each of
beets with 75 a n d 100% defoliation made on July 15 were not
injured on August 2 and were used as checks for the first defoliation. All beet plots in this test were randomized.
Results of the Second Experiment
Figure 4 presents the results of this test and indicates an
average percentage of weights of tops, yield of beets and sugar
for four replications of each treatment.
Weights of beet tops with only one 7 5 % defoliation and also
those with an additional 2 5 % injury were the same and were
about 1 2 % below the check. T h e s e reductions were similar to
those for identical degrees of injuries d u r i n g the same period
in the first e x p e r i m e n t . Apparently an additional 2 5 % iniury
did not have m u c h effect on beets after they had sustained 7 5 %
defoliation from the first treatment. Weights of beet tops with
50 and 7 5 % defoliations in the second treatment were similar
and showed only a moderate reduction in weight (22%) as
compared to the checks. A complete defoliation on the second
date produced a p r o n o u n c e d reduction (59%) in weight of tops.

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Figure 4.The effect of various degrees of defoliation, made at two


consecutive dates, on the relative weight of tops, yield of beet roots and
yield of sugar on an acre basis (1960).

Beets with only one complete defoliation showed a moderate


reduction in weight of tops (22%), which was a somewhat
smaller reduction than that resulting from similar treatments
of the first experiment. However, beets with subsequent 25,
50 and 75% defoliations showed a substantial and quite a similar
reduction in top weights (37 to 45%). These results indicate
that lesser injuries made on the second date produced a much
greater reduction in weight of beet tops after the first complete
defoliation than after 75% defoliation. Weights of beet tops,
exposed to two complete defoliations, were quite similar to
those exposed to 75 and 100% injury.
Only a very slight reduction (7.0%) in the yield of beets
resulted from one 75% defoliation. Yields of beets defoliated a
second time at 25, 50 or 75% were more or less similar and
they varied from 22 to 28% below the check beets. Beets with
25% defoliation showed a slightly smaller reduction in the
yield than those with 50 or 75% injury. Beets completely defoliated from the second injury had 46% loss in yield.
Beets with only one complete d e f o l i a t i o n showed 33%
reduction in yield when compared to the checks. Losses in
yield of beets, defoliated a second time at 25, 50 and 75%, were
only slightly below those with only one complete defoliation
and showed a slight, but gradual increase in their losses with
greater degrees of defoliation (from 45 to 50%). Beets completely defoliated twice had 64% reduction in yield.
Since the percentage of sugar in all these beets was about
the same, the yield of sugar followed about the same pattern
as the yield of roots.
Discussion and Conclusions
These results showed that 75% removal of beet foliage in
the middle of July depressed the yield of beets only slightly

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(7.0%). T h i s reduction was very similar to the one for a similar


treatment in the first experiment (10.0%). Subsequent defoliation of beets at 25, 50 and 7 5 % , made two weeks later, produced
another slight reduction in the yield of beets. These results
emphasize the p o i n t stressed in the first experiment, that removal up to a n d including 7 5 % of beet leaves had only a slight
depressing effect on the yield of roots and sugar in beets. Beets
subjected to a n o t h e r 100% defoliation, showed a substantial
reduction in yield.
Beets with only one 100% defoliation in the middle of July,
showed 3 3 % r e d u c t i o n in yield, which was also quite comparable
to losses in a similar treatment of the first erperiment (29%).
As compared to a single 7 5 % defoliation, these beets suffered
a further 2 6 % reduction in yield. These results show that complete defoliation is very detrimental to beets and greatly affects
the yield of beets. Subsequent defoliations of 25, 50 and 7 5 %
showed only a slight additional reduction in yield of beets. It
appears that the first complete defoliation had a much greater
effect on the yield of beets than had any subsequent injuries,
even for those of 75% Beets with two complete defoliations,
showed a great reduction in yield (64% of the check). T h e
yield of these beets was about 19% lower than those in which
75 and 1 0 0 % of the foliage was consecutively removed.
T h e s e results showed that if beets are subjected to one complete defoliation, and later to lesser ones, not exceeding 7 5 %
injury, no p r o n o u n c e d reduction in the yield of beets resulted
from the second defoliation. These results are in agreement
with those obtained in the work with cotton (4).
Experiment 3
Comparison of Defoliation of Sugar Beets
Made with Scissors and Wooden Sticks
All defoliations in the preceding experiments with simulated
hail injuries to sugar beets were made with scissors. During
these investigations the question was asked whether this type
of injury is comparable to damage incurred bv natural hail. In
previous work with simulated hail injury with field beans in
Massachusetts (10) it was reoorted that damage of similar intensity p r o d u c e d the same end results, whether by hand clipping
or bv machine-blown ice, wind and water.
To investigate w h e t h e r the removal of leaves with scissors
is comparable to injuries inflicted on plants bv some other means
more similar to hail stones, experiments were conducted in which
one set of beet plots was defoliated with scissors and the others
were iniured bv beating the foliage with wooden sticks. D u r i n g
1962, two parallel defoliations of beets were made with wooden

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Figure 5.The effect of various degrees of defoliation m a d e with


wooden sticks a n d scissors on the relative weight of tops, yield of beet
roots a n d yield of sugar on an acre basis (1962).

sticks and with scissors four times during the season: July 2,
17, August 1 and 15. The same size of plots and the same procedure were used here as in the previous tests.
Results of the Third Experiment
Figure 5 illustrates the results of this test and presents the
average percentages of weights of tops, yield of beets and sugar
for four replications of each treatment for all the above mentioned dates of defoliation. Injuries made with wooden sticks
produced a gradual reduction in the final weight of beet tops,
reducing them from 4% in beets with 25% injury to 35% for
completely defoliated beets as compared to the checks. Tops
of beets defoliated with scissors at 25, 50 and 75% intensity,
showed more or less the same or only slight reductions in their
top weights in comparison to the checks. Weights of beet tops
with 25% defoliation with scissors were quite similar to those
defoliated with sticks; however, tops of beets with 50 and 75%
injury with scissors weighed considerably more than those damaged with sticks. Weights of tops completely defoliated with
scissors were similar to the same injured with sticks.
Yields of beets in plots defoliated with sticks and scissors
were quite comparable for all degrees of injuries. However,
beets injured with sticks, with the exception of completely defoliated plants, showed slightly lower yields than those injured
with scissors. These reductions were equal to 5.1, 2.0 and 3.7%.
respectively, for 25, 50 and 75% defoliations. The yield of 100%
defoliated beets with sticks was 3.3% higher than in those injured with scissors.

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T h e average yield of sugar in beets, defoliated with sticks


and scissors, showed a proportionate decrease in yield of sugar
with greater degrees of defoliation for both types of injuries.
Beets defoliated with sticks showed moderately lower yields of
sugar t h a n those injured with scissors. These decreases were
equal to 11.9, 7.6 a n d 5.8%, respectively, for 25, 50 and 7 5 %
injuries. T h e yields of sugar in completely defoliated beets for
both types of injuries were a b o u t the same.
Discussion and Conclusions
It appears that the greatest difference between plants defoliated with sticks a n d scissors was shown by weight of beet
tops in 50 a n d especially in 7 5 % injured beets. Beets damaged
with sticks showed greater reductions in tops than those defoliated with scissors. T h e a m o u n t of sugar harvested from beets
in plots subjected to the first three degrees of injuries with sticks
were also lower t h a n from those injured with scissors.
In m a k i n g defoliations with scissors, clean cuts of leaves were
made w i t h o u t m u c h abrasion or injury to the remaining part
of the leaves a n d petioles or to very small leaves. In defoliations
made with sticks, a considerable a m o u n t of injury and numerous
small w o u n d s were inflicted on many leaves and also on petioles.
W h e n a b o u t the same a m o u n t of leaf tissue was removed in
both types of defoliation, beets injured with sticks, undoubtedly
sustained m a n y small wounds which, to a large extent, were not
made in beets cut with scissors. W o u n d s inflicted on petioles
with stick injury u n d o u b t e d l y also injured the vascular tissue.
It is believed that injury made with sticks required a longer
time for the beets to repair, and in this process they needed
more time a n d greater amounts of sugar as a source of energy.
Hail studies conducted with potatoes in Maine showed (8)
that injury to vascular tissue in the stem, caused greater damage
than w h e n a large a m o u n t of leaf tissue was removed. Work
with simulated hail damage to flax in Idaho (3) also showed
that mechanical injuries to the stems led to a greater reduction
in yield t h a n injuries to, or even removal of the leaves. T h e same
situation appears to be evident here.
T h e yield of beets subjected to both types of defoliation were
about the same, with slightly higher tonnage for those injured
with scissors. Beets completely defoliated with sticks yielded
slightly m o r e t h a n those whose leaves were removed with scissors.
It is believed t h a t w h e n beet leaves were destroyed with sticks,
small a m o u n t s of leaf tissues were left attached to the crowns
of the beets which allowed some photosynthetic activity immediately after t h e injury. T h i s was not the case in the beets when
all leaves were c u t off by scissors. T h e same phenomenon was
observed for cotton in T e x a s (4).

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In spite of slight variations in the results, both methods of


defoliation showed about the same reductions in yield and can
be used for estimating natural hail damage.
Summary
A number of experiments were conducted over a six-year
period to simulate hail damage and to evaluate the losses which
resulted from these injuries.
Defoliations of sugar beets up to and including 75% had
only a slight effect in reducing weight of beet tops. The greatest
losses in weight of tops were equal to 10, 16 and 20%, respectively, for 25, 50 and 75% defoliations. Beets completely defoliated starting with August treatments, showed great losses in
top weights for all injuries.
Maximum reduction in the yield of beets for all degrees
of defoliations occurred during the middle of the season and
were equal to 5, 9, 10 and 3 1 % , respectively, for 25, 50, 75 and
100% defoliations. It is apparent that only completely defoliated
beets suffer serious loss in yield. All September injuries had
little effect on the yield.
Very little reduction (3 to 4%) in yield of sugar took place
for 25 and 50% defoliations. The greatest losses in the yield of
sugar for 75 and 100% defoliated beets were equal respectively,
to 7.5 and 18% of sugar.
Beets which sustained 75% defoliation on the first date (July
15) showed additional losses in beet tops with amounts 0, 22,
25 and 59%, respectively, for 25, 50, 75 and 100% injuries for
the second defoliation (August 2). Similar losses for beets with
the first complete defoliation were equal to 45, 37, 37 and 61%,
respectively, for 25, 50, 75 and 100% of the second injuries.
These results showed that all second defoliations, except a complete one, produced a much smaller effect on the reduction of
weight of beet tops after the first 75, than after 100% defoliations.
The losses in the yield of beets, defoliated the first time at
75 and the second time at 25, 50 and 75% were more or less
similar (22 to 25%). The losses in the yield of beets for similar
injuries, following 100% defoliation, were only slightly below
losses due to only one complete defoliation. Beets injured once
by 75 and the second time by 100% had a 46% reduction in
the yield, and those twice completely defoliated lost 64%.
Yield of sugar in all these beets was quite comparable to
the yield of beets.
These results showed that if beets were subjected to one
75 or 100% defoliation and later on to a lesser one, not exceeding

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7 5 % , no p r o n o u n c e d r e d u c t i o n in the yield of beets results


from t h e second defoliation.
In c o m p a r a t i v e defoliations m a d e with scissors a n d sticks,
weights of tops of beets in 25 a n d 100% injured beets were q u i t e
similar for b o t h m e t h o d s of defoliations. However, tops of beets
in 50 a n d 7 5 % defoliations with scissors weighed more than
those i n j u r e d w i t h sticks.
Yields of beets were q u i t e comparable for both types of defoliations a n d all degrees of injuries.
Yield of sugar in beets for the first three degrees of injuries
made w i t h sticks were lower t h a n in those defoliated with
scissors. T h e yield of sugar in completely defoliated beets was
a b o u t t h e same for b o t h types of injuries.
Literature Cited
(1) AFANASIEV, M. M., S. D. LYDA and I. K. MILLS.

1960. Simulated hail

injury to sugar beets. J. Am. Soc. Sugar Beet Technol. 11: 196-200.
(2)

(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)

JONES,

F.

G.

W.,

R.

A.

DUNNING

and

K.

P.

HUMPHRIES.

1955.

The

effect of defoliation and loss of stand upon yield of sugar beets.


Ann. Appl. Biol. 43: 63-70.
KLAGES, K. H. W. 1933. T h e effect of simulated hail injuries on flax.
J. Am. Soc. Agron. 25: 534-540.
LANE, H. C. 1959. Simulated hail damage experiments in cotton.
Texas Agr. Exp. Sta. Bull. 934.
LILLY, C. E. and A. M. HARPER. 1962. Effect of defoliation and reduction of stand on yield of sugar beets in southern Alberta. J. Am.
Soc. Sugar Beet Technol. 12: 192-199.
MORRIS, H. E. 1948. Simulated hail damage to sugar beets. Proc. Am.
Soc. Sugar Beet Technol. pp. 358-362.
MORRIS, H. E. Simulated hail damage to sugar beets 1948-49. 1950.
Proc. Am. Soc. Sugar Beet Technol. pp. 302-304.
MURPHY, H. J. and M. J. GOVEN. 1962. The effect of simulated hail
damage on yield and quality of potatoes in Maine. Maine Agr.
Exp. Sta. Bull. 607.
POINTER, J. P. and W, C. WOLTZ. 1956. Investigations of hail damage
to tobacco. N. C. Agr. Exp. Sta. Tech. Bull. 123.
SNYDER, G. B. and Louis MICHELSON. 1958. T h e effect of simulated
hail damage on field beans. Mass. Agr. Exp. Sta. Bull. 506.

(11)

SPARKS, W. C., G. W. WOODBURY and F. H. TAKATORI.

(12)

TAKATORI, F. H., W . C. SPARKS and G. W. WOODBURY.

1957.

Esti-

mating hail injury in potatoes. Idaho Agr. Exp. Sta. Bull. 274.
1952. A study

of simulated hail injury on potatoes. Idaho Agr. Exp. Station Res.


Bull. 22.
(13) WATSON, D. J. 1958. T h e dependance of net assimilation rate on leafarea index. Ann. Bot. 22: 37-54.
(14) WOODBURY, G. W . a n d MARSHALL L E BARON.

1959.

A study of simu-

lated hail injury in beans. Idaho Agr. Exp. Sta. Bull. 322.

Soil Temperature and Nitrogen Effects on Yield and


Phosphorus Uptake by Sugar Beets1
S. DUBETZ AND G. C. RUSSELL 2
Received for publication April 13, 1964

About 45,000 acres of sugar beets are planted annually in


southern Alberta. Growers have been applying 100 lb of 11-48-0
fertilizer per acre, and the need for additional nitrogen has been
reported previously (8)3. Beets are planted from the second
week in April to the middle of May. The relatively cool temperatures that may be encountered in spring together with the increased use of nitrogen fertilizer merits the study of the effect of
these factors on the early growth of the plant.
Review of Literature
There is very little published information available on the
effect of soil temperature on the growth of sugar beets. The
percentage emergence of sugar beet seedlings at soil temperatures
of 13, 18, and 24 C was significantly increased (6) over that
at 6 C, and the speed of emergence increased as the temperature
increased. Nielsen et al. (10) found that the yield of roots and
foliage of lucerne increased with increase in temperature to at
least 19.4 C. The phosphorus content of the roots and foliage
tended to increase with increasing temperature. Oats produced
higher yields (11) of grain and straw when soil temperature
was increased from 41 to 67 F (19.4 C). There was a trend
toward increased concentration of phosphorus in the oat plants
with increasing temperature. Low soil temperature depressed
the growth of corn seedlings (9), and the percentage phosphorus
and total phosphorus were also lower at the low temperature.
In his review of the effects of nitrogen on the availability
of phosphorus to plants, Grunes (13) separates them into biological and chemical effects. Some of the biological effects include stimulation of root and, top growth, thereby increasing
absorbing capacity and phosphorus uptake. The chemical effects
are related to alterations of the phosphorus solubility in the
soil. Rennie and Soper (14) reported an increase in fertilizer
phosphorus absorption at a very early stage of growth of cereal
crops when nitrogen in the ammonium form was mixed with
the phosphorus carrier. Caldwell (4), working with corn, also
found that ammonium forms of nitrogen had to be intimately
1
Contribution from the Soils Section, Canada Agriculture Research Station, Lethbridge.
Alberta.
2
Agronomist and Head, Soils Section, respectively.
3
Numbers in parentheses refer to literature cited.

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mixed with phosphorus before phosphorus absorption by t h e


plant occurred. He stated that the N to P 2 O 5 ratio had to be 1:4
or n a r r o w e r for significant u p t a k e of phosphorus to take place
and suggested that the effects of nitrogen on phosphorus absorption were b r o u g h t a b o u t by chemical interactions.
Balba (2) f o u n d that nitrogen increased yields but had
almost no effect on the phosphorus percentage of onion plants.
He suggested that nitrogen enabled the onion roots to "forage"
for m o r e phosphorus, hence the percentage phosphorus tended
to r e m a i n constant, a n d that the a m o u n t of phosphorus taken
up by the p l a n t was p r o p o r t i o n a l to t h e yield.
T h e purpose of the experiments reported in this paper was
to study t h e effect of t e m p e r a t u r e and the time of application
of n i t r o g e n fertilizer on the growth and phosphorus uptake of
sugar beets.
Materials and Methods
T h e e x p e r i m e n t was conducted in a greenhouse and consisted of o n e soil type, four soil temperatures, five fertilizer
treatments, a n d two harvest dates.
T h e soil used was a C h i n silt loam (3), which is the pred o m i n a n t soil type on which sugar beets are grown in southern
Alberta. T e n p o u n d s of soil were placed in 1-gallon plastic pots.
T h e pots were placed in constant temperature tanks similar to
those described by Cooper et al. (5). T h e temperatures used in
the tanks were 7, 12, 19, a n d 27 C.
Sugar beet seeds (Alberta blend) were germinated in flats
containing compost soil. W h e n the seedlings were 41/2 weeks old
they were transplanted, three to a pot in duplicate pots. Prior
to transplanting, all of the pots received triple superphosphate
fertilizer (0-43-0) at the rate of 40 lb P 2 0 5 per acre. A m m o n i u m
nitrate fertilizer (331/2-0-0) at the rate of 40 lb N per acre was
applied to o n e set of pots at transplanting time and to three
other series of pots at 2-week intervals for 6 weeks.
T h e soil surface was insulated with a 1/2-inch layer of beaded
polystyrene. C o p p e r constantan thermocouples set at 1/2-inch
depth in t h e soil a n d connected to a recorder showed that the
temperature at this d e p t h varied 2 C. T h e temperature in
the air-conditioned w i n g of the greenhouse was 21 C 3.
W a t e r was a d d e d to m a i n t a i n the soil water in the u p p e r
half of the available range as determined by periodic weighing.
T h e first harvest of sugar beets, from half the pots, was 8
weeks after t r a n s p l a n t i n g w h e n the plants were 121/2 weeks old,
and the second harvest was taken 10 weeks after transplanting.
The e x p e r i m e n t was repeated two m o r e times to permit statistical
analysis with t h r e e replications in time.

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JOURNAL OF THE A. S. S. B. T.

After harvest the tops and roots from each pot were dried
at 180 F in a forced draft oven and weighed. T h e phosphorus
content of the plant material was determined colorimetrically
(15) after preparation by the wet digestion method with perchloric acid (13).
At the time of each harvest the available phosphorus content of the soil in each pot was determined by the sodium bicarbonate method (1). Golden (7) found that the control of
temperature at which soil extractions were made was highly
desirable. Therefore, freshly prepared solutions were kept in
the respective temperature tanks for several hours, and the extraction was then carried out in the tanks.
Results and Discussion
As the soil temperature increased from 7 to 27 C, the
dry weights of sugar beet tops increased significantly (Table 1).
Table 1.Effect of four soil temperatures on sugar beet seedlings and on extractable
soil phosphorus, and significant differences1 between means (means per pot calculated from
three replications and two harvests).

T h e yields of roots increased significantly with temperature increases to 19 C only. T h e optimum temperature for root growth
appears to be around 20 C. While the percentage of phosphorus
in the beet tops was not significantly affected by soil temperature, there was a tendency for it to decrease in the roots with
increasing soil temperature. Total phosphorus uptake by tops
and roots increased with increasing soil temperature in
proportion to increased plant material. T h e amount of available phos-

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p h o r u s e x t r a c t e d from the soil was n o t significantly affected by


soil t e m p e r a t u r e . T h i s can be partially a t t r i b u t e d to the increased g r o w t h a n d , therefore, increased total uptake as temperature increased.
Table 2.Effect of phosphorus and nitrogen applied at successive dates on sugar beet
seedlings and on extractable soil phosphorus, and signficant differences1 between means
(means per pot calculated from three replications and two harvests)

Root yields were highest when nitrogen was applied either


at transplanting time or 2 weeks later (Table 2). The percentage
of phosphorus in the roots was higher from the treatment that
received the last application of nitrogen than from either of
the two treatments that received the earliest application of nitrogen. T h e first application of nitrogen resulted in roots that
were higher in total phosphorus than those of the other treatments. These results agree with those of Balba (2), who found
that nitrogen increased the total uptake of phosphorus as a result
of increased plant growth. There was no evidence of an apparent
increase in the solubility of phosphorus as indicated by the
percentage of phosphorus in sugar beets grown in the presence
of nitrogen.
N i t r o g e n at 40 lb p e r acre at any of the four dates of application resulted in yields of beet tops a n d total phosphorus content
of tops t h a t w e r e h i g h e r t h a n from the treatment that d i d not

242

J O U R N A L OF T H E A.

S.

S.

B.

T.

receive any nitrogen. T h e percentage of phosphorus in the tops


was higher when no nitrogen was used, and there did not appear
to be any pattern among treatments that received nitrogen.
T h e lower extraction of available soil phosphorus from the
pots that received nitrogen further illustrates that total uptake
of phosphorus was increased with nitrogen fertilizer.
In roots and tops the dry matter weights and total phosphorus
were higher while the percentage phosphorus was lower between
dates of harvest. There were no significant temperature-fertilizer
interactions.
In general, these results showed that as the soil temperature
increased from 7 to 27 C the dry matter and total phosphorus
increased in both roots and tops of sugar beet seedlings. T h e
early applications of nitrogen resulted in the highest dry matter
and phosphorus content of seedling roots. T i m e of nitrogen
application had no effect on seedling beet tops. T h e results
would indicate that early availability of nitrogen to the seedling would promote a more rapid root growth.
Literature Cited
(1)

ATKINSON, H. J., G. R. G I L E S , A. J. M A C L E A N , a n d J. R. W R I G H T .

1958.

Chemical m e t h o d s of soil analysis. Contr. N o . 169, Chem. Div.,


Science Service, C a n . D e p t . Agr., Ottawa.
(2) BALBA, A. M O N E M . 1960. Phosphorus a n d nitrogen uptake as expressed by a Mitscherlich-type equation. T r a n s . 7th I n t e r n . Cong.
Soil Sci. 3: 154-160.
(3)

BOWSER, W . E., T . W . PETERS, a n d A. A.

KJEARSGAARD.

1963.

Soil

Survey St. Mary a n d Milk Rivers Development Project. Univ. of


Alta. Bull. SS-5.
(4) CALDWELL, A. C. 1960. T h e influence of various nitrogen carriers on
the availability of fertilizer phosphorus to plants. T r a n s . 7th Intern.
Cong. Soil Sci. 3: 517-525.
(5)

(6)

COOPER,

W.

KALBFLEISCH.

1960. Note on an a p p a r a t u s for controlling soil


C a n . J. Soil Sci. 40: 105-107.

D.

J.,

K.

F.

NIELSEN,

J.

W.

WHITE,

and

temperatures.

DUBETZ, S., G. C. RUSSELL, a n d D . T . ANDERSON.

1962.

Effect of soil

t e m p e r a t u r e on seedling emergence. C a n . J. P l a n t Sci. 42: 481-487.


(7) GOLDEN, LARON E. 1962. Effect of temperature of extractant on P.
K, Ca, a n d Mg removal from different soil types. Soil Sci. 93:
154-160.
(8) H I L L , K. W., a n d S. DUBETZ. 1952. Fertilizer side-dressing studies on
sugar beets in s o u t h e r n Alberta. Proc. Am. Soc. Sugar Beet Technol.
7: 207-211.

V O L . 13, N o . 3, OCTOBER 1964

243

(9) KETCHESON, J. W. 1957. Some effects of soil temperature on phosphorus requirements of young corn plants in the greenhouse. Can.
J. Soil Sci. 37: 41-47.
(10)

NIELSEN, K. F., R. L. HALSTEAD, A. J.

M A C L E A N , R. M. HOLMES, and

S. J. BOURGET. 1960. Effects of soil temperature on the growth and


chemical composition of lucerne. Proc. 8th Intern. Grassland Cong.
p p . 287-292.
(11)

(12)

(13)
(14)
(15)

NIELSEN, K. F., R. L. HALSTEAD, A. J.

MACLEAN,

R.

M.

HOLMES, and

S. J. BOURGET. 1960. T h e influence of soil temperature on the


growth and mineral composition of oats. Can. J. Soil Sci. 40:
255-263.
N O R M A N , A. G. (Ed.) 1959. Advances in Agronomy, Vol. 11: Grunes,
D. L. Effect of nitrogen on the availability of soil and fertilizer
phosphorus to plants. Academic Press, Inc., New York, N. Y.
PIPER, C. S. 1953. Soil and plant analysis. Inter-science Publishers,
Inc., New York, N. Y.
R E N N I E , D. A., and R. J. SOPER. 1958. T h e effect of nitrogen additions
on fertilizer-phosphorus availability. II. J. Soil Sci. 9: 155-167.
W A R D , G. M., and F. B. JOHNSTON. 1962. Chemical methods of plant
analysis. Can. Dept. Agr. Publ. 1064.

Losses Caused by Beet Mosaic Virus in California


Grown Sugar Beets
R. J. SHEPHERD, F. J. HILLS AND D. H. H A L L 1
Received for publication

April

13,

1964

Introduction
T h e beet mosaic virus is widespread in many California beetgrowing areas. T h e virus is particularly common in areas where
large acreages of sugar beets have been overwintered successively
for a number of years and where early plantings are made near
overwintered fields. In some areas early spring plantings have
been observed with almost 100% infection by early summer.
Generally mild strains of the beet mosaic virus seem to predominate in most areas, although the virus occurs as a number
of strains which differ in severity. Plants, following infection
with mild strains of the virus, usually show a flush of severely
mottled and distorted foliage accompanied by some stunting,
but soon recover. Chronically diseased plants generally show
little signs of disease except for mottling and some blistering
on the younger center leaves. Because of the temporary nature
of severe symptoms and its sporadic occurrence in most areas,
the virus has generally been assumed to be of little economic
importance.
Little, if any, data have been collected on the effect of the
beet mosaic virus on beet sugar yields under field conditions
in the U. S. Several reports from Europe on the effect of the
virus on yields of sugar beet suggest it may be economically
important in some cases but in general these reports have indicated the damage is minor in comparison with that associated
with
the beet yellows virus. In England, Watson and Watson
(S)2 found mosaic decreased sugar yields 10 to 20%, an economically important loss. Beet yellows under the same conditions
reduced the sugar yield 50%. Liidecke and Neeb (5) reported
mosaic reduced the yields of beet roots, foliage and sugar 6, 10,
and 9%, respectively, in Germany. In similar experiments beet
yellows caused losses about 9-fold greater than mosaic alone. In
plants infected with both yellows and mosaic, the effects of the
two were additive. Wiesner (9) obtained similar results: mosaic
reduced sugar yields 6 to 10% whereas yellows alone caused
losses of 35-55 percent. Again, in doubly infected plants the
effects of the viruses were additive.
1 Assistant Plant Pathologist, Extension Agronomist, and Extension Plant Pathologist'
University of California, Davis.
2 Numbers in parentheses refer to literature cited.

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T h e s e r e p o r t s suggest losses d u e to mosaic are probably m i n o r


in c o m p a r i s o n w i t h beet yellows b u t apparently little a t t e m p t
has b e e n m a d e to assess t h e effect of mosaic in the U. S. T h i s
lack of e x p e r i m e n t a t i o n has probably b e e n d u e to t h e difficulty
in m a i n t a i n i n g disease-free control plots in areas where t h e a p h i d
vectors a r e active t h r o u g h o u t t h e growing season. U n d e r conditions in w h i c h t h e virus is rapidly spread i n t o the control
p l a n t i n g it is difficult to m a k e an accurate assessment of the
effect of t h e v i r u s on yield. In the i n l a n d valleys of California,
however, a p h i d n u m b e r s d r o p to a very low level d u r i n g the
s u m m e r m o n t h s a n d little secondary spread of the virus occurs
d u r i n g this p e r i o d . Dr. W. H. Lange a n d his colleagues in the
D e p a r t m e n t of E n t o m o l o g y at the University of California, Davis,
have shown that following the massive spring flights of the green
peach a p h i d (Myzus persicae Sulz.), the most i m p o r t a n t vector
of beet mosaic virus in the San J o a q u i n a n d Sacramento valleys
of California, t h e i r n u m b e r s decrease rapidly d u r i n g May a n d
are m a i n t a i n e d at a very low level t h r o u g h o u t the s u m m e r
months. T h i s absence of a p h i d activity has allowed an evaluation of t h e effect of mosaic u n d e r field conditions in which very
little spread of t h e virus occurred u n t i l several m o n t h s after its
i n t r o d u c t i o n i n t o treated plots. T h e results of tests to d e t e r m i n e
the effect of b e e t mosaic on the root yields and sucrose content
of sugar b e e t d u r i n g t h e last 2 years are r e p o r t e d herein.

Materials and Methods


R e p l i c a t e d field trials with beet mosaic a n d the 2 yellows
viruses were m a d e at Davis in 1962 a n d 1963. T h e effects of each
virus a l o n e a n d in various combinations of 2 or more were
compared w i t h u n i n o c u l a t e d plots using late-planted beets of
the variety Spreckels Sugar 202H. In the 1962 tests beets were
planted May 8, irrigated up on May 14 a n d inoculated with
the viruses on J u n e 20-21, a b o u t 1 week after thinning, when
the plants w e r e in t h e 6- to 8-leaf stage. T h e s e plants were sidedressed w i t h sufficient a m m o n i u m sulfate at t h i n n i n g time to
give 200 lb n i t r o g e n p e r acre. All three viruses in all possible
combinations w e r e i n c l u d e d in this test. In the 1963 e x p e r i m e n t
the plots were p l a n t e d later, on J u n e 7, d u e to the late aphid
flights, a n d i n o c u l a t e d on July 20 to 23 when the plants were
in the 8- to 12-leaf stage. In these tests an additional variable,
nitrogen fertilization, necessitated eliminating most of the virus
combinations; each virus alone plus only the western yellowsmosaic c o m b i n a t i o n was included. In each experiment the various
treatments w e r e r a n d o m i z e d in 5 to 6 replicated blocks.

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T h e beets were grown in beds on 40-inch centers with 2


rows o plants on each bed a n d the plants t h i n n e d to 8 inches
within the row. Each plot consisted of 4 beds 60 feet in length.
Only the center 30 feet of the two middle beds was inoculated
in each case. T h i s gave an u n t r e a t e d buffer zone 2 beds wide
with an in-the-row distance of 30 feet between different treatments. In the 1962 experiment 6 replications of each treatment
were included at low (no additional) and high (200 lb per
acre) nitrogen levels. T h e nitrogen was applied beneath the
row before planting as a m m o n i u m sulfate granules.
A relatively mild strain of the beet mosaic virus was used
for the tests. T h i s isolate was obtained from a naturally infected
field near Davis a n d was passed t h r o u g h 2 local lesion transfers
on Chenopodium capitatum L. (Asch.) before use. Stock cultures
of the virus were m a i n t a i n e d in Nicotiana clevelandii Gray.
I n o c u l u m in sufficient quantity for the field inoculations was
b u i l t up in peas, Pisum sativum L., variety Dwarf Telephone.
Infected peas were collected about 2 weeks after mechanical
inoculation when showing incipient necrotic streak symptoms,
homogenized in 10 volumes of 0.05 M phosphate buffer, pH 7.5,
per g of tissue and held in an ice bath u n t i l used shortly thereafter in field inoculations. T h e homogenate was used to mechanically inoculate 2 to 3 of the lower leaves of the beet seedlings
using a small cheesecloth pad moistened in t h e inoculum. A
small quantity of 600 mesh c o r u n d u m was added to 50-60 ml
portions of the homogenate in a small wide-mouthed container.
T h i s was stirred into suspension each time t h e cloth pad was
dipped to renew the inoculum.
T h e beet yellows a n d western yellows viruses were introduced
into the field plots by the use of green peach aphids as described
by Bennett, et al. (2).
A severe vein-clearing strain, Bennett's strain 5 (1), of the
beet yellows virus was used d u r i n g b o t h years' trials. T h i s isolate
was transferred to a large n u m b e r of N e w Zealand spinach plants
(Tetragonia expansa T h u n b . ) on which aphids, reared on radish
(Raphanus sativus L.. variety W h i t e Icicle), w e r e transferred
a b o u t 24 hours before the inoculations were made. T h e s e plants
were transported to the field a n d pieces of leaf containing about
10 aphids were clipped off a n d d r o p p e d i n t o the crown of each
sugar beet plant to be inoculated. T h e western yellows virus
was inoculated by a similar procedure except that the aphids
were reared on healthy radish a n d transferred to infected radish
about 48 h o u r s before use. T h e m o r e severe Spence field strain
of the virus was used for the 1963 trials 8 . An aerial spray of

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systox was a p p l i e d to kill t h e a p h i d s 2 days after t h e inoculations


were c o m p l e t e d . T h e plots were harvested o n O c t o b e r 23, 1962,
a n d D e c e m b e r 9, 1963. T h e c e n t e r 25 ft of t h e four rows of
each 30 ft p l o t i n o c u l a t e d , was harvested. T h e beets were lifted
by h a n d u s i n g a 2-pronged fork, t h e excess soil r e m o v e d a n d
the tops c u t off for separate weighing. T w o 10-beet samples were
taken p e r p l o t for sucrose a n d tare d e t e r m i n a t i o n s .
Results
T h e m e c h a n i c a l i n o c u l a t i o n o f beet mosaic gave b e t t e r t h a n
90 p e r c e n t infection w i t h this virus w i t h t h e exception of 2 plots
in the 1963 trials. C o u n t s of yellowed plants m a d e 4 to 6 weeks
after i n o c u l a t i o n in b o t h t h e 1962 a n d 1963 e x p e r i m e n t s showed
that t h e a m o u n t of infection w i t h beet yellows, with the exception of a single p l o t in t h e 1963 test with 5 4 % , r a n g e d from
7 9 t o 9 8 % w i t h a n average o f a b o u t 8 5 % . I n t h e 1962 tests n o
symptoms w e r e o b t a i n e d w i t h t h e beet western yellows virus
a l t h o u g h at least some of the plants were f o u n d to be infected
as d e m o n s t r a t e d by transfer of t h e virus to Capsella bursa-pastoris
(L.) M e d i k . a n d by yield decreases. Similarly, t h e accuracy of
t h e estimates of infection d u r i n g the 1963 tests, based on c o u n t s
of p l a n t s s h o w i n g yellowed o l d e r leaves, is q u e s t i o n a b l e . In t h e
1963 e x p e r i m e n t a p p a r e n t infection with western yellows r a n g e d
from 57 to 8 9 % w i t h a m e a n of 7 0 % ; however, t h e n u m b e r of
plants s h o w i n g yellowed foliage varied m a r k e d l y with n i t r o g e n
fertilization. F e w e r p l a n t s w e r e obviously yellowed at t h e h i g h e r
n i t r o g e n level suggesting t h a t visual symptoms may indicate an
erroneously low level of infection.
Plots infected w i t h beet yellows were clearly o u t l i n e d by the
color difference 2 m o n t h s after infection a n d s h a r p lines of
d e m a r k a t i o n w e r e m a i n t a i n e d u n t i l some secondary spread of
yellows o c c u r r e d i n t o adjacent buffer rows in t h e fall.
The effect of the beet mosaic and yellows viruses on top
growth.The effect of t h e viruses on t o p growth of sugar beets in
the 1962 a n d 1963 tests is shown in T a b l e 1. T h e lower t o p
yields o b t a i n e d in 1963 were p r o b a b l y d u e to the slowdown in
growth w i t h t h e cold w e a t h e r p r e c e d i n g t h e later harvest a n d
to t h e loss of m a n y o l d e r leaves which were killed by Cercospora
leaf spot in N o v e m b e r . T h e beet mosaic viruses r e d u c e d the t o p
yields from 28.0 to 23.6 tons p e r acre in 1962 a n d h a d a similar
effect in 1963 at t h e l o w e r n i t r o g e n level ( T a b l e 1). Tn the
1963 trial, t h e s t u n t i n g effect of mosaic on t o p g r o w t h was visually
Dr.C.W.
Bennett of the U. S. Agricultural Research at Salinas. California, kindly
provided inoculum of the western yellows virus for the 1965 experiment.

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JOURNAL OF THE A. S. S. B. T.

Table 1.Effect of the beet mosaic virus alone and in combination with the beet yellows
and western yellows viruses on the top growth of sugar beet in 1962, and at high and low
nitrogen levels in 1963.
Top yield, tons/acre; fresh weight

apparent at the lower nitrogen level b u t not at the high nitrogen


level. T o p growth with the mosaic and western yellows viruses
combined was not significantly different from mosaic alone in
either test regardless of nitrogen level, thus demonstrating the
more m a r k e d effect of mosaic on top growth. T h e western yellows
virus alone did not cause any decrease in top growth in either
test. T h e beet yellows virus, however, caused serious stunting in
the 1962 trial and at the low nitrogen level in the 1963 experiment.
T h e stunting effects of mosaic and beet yellows viruses together were additive as indicated by the loss of 11 tons per acre
in top growth in the 1962 test. Beet yellows alone, however,
had no a p p a r e n t effect on t o p growth at the higher nitrogen level
in the 1963 e x p e r i m e n t and in this case, at least, the effect
seemed to be less than with mosaic alone ( T a b l e 1). Similarly,
beet yellows symptoms at the high nitrogen level were partially
masked.
The effect of the beet mosaic and yellows viruses on root
yield.The reductions in root yields as a result of infection
with the various viruses are shown in T a b l e s 2 a n d 3. Mosaic
reduced yields in 1962 from 34.9 to 31.5 tons / a c r e for an average
loss in tonnage of 9 . 7 % thus demonstrating that even a mild
strain of the virus can cause significant losses in yield. The
losses in root yield in the 1963 test with mosaic were scarcely
significant at t h e 5% level b u t showed a 5.9 ton per acre decrease
as a result of infection. Although the same strain of virus was
used for these tests, the losses in yield were less probably because
of the greater size of the plants at the time of infection. No
reduction in the sucrose content of the roots from mosaic infected plants occurred in either test (Tables 2 a n d 3). No

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Table 2.-Effect off the beet mosaic virus alone and in combination with the beet yellows
and western yellows virus on the root yields and sucrose percentage of sugar beet in 1962.
Virus inoculation

Root yield
Tons/Acre

Loss
Tons/Acre

34.9
31.5
29.8
19.1

3.4
51

15.8

14.6
45.3

17.3
32.8
22.6
22.8

17.6

50.4

2.1

6.0

12.3
12.1

35.2
34.7

Control
Beet mosaic alone
Beet mosaic plus western yellows
Beet mosaic plus beet yellows
Beet mosaic plus beet yellows
and western yellows
Western yellows alone
Beet yellows alone
Beet yellows plus western yellows
LSD, 5 %

Percent
loss

Percent
sucrose
13.4
13.5
13.8
13.8

9.7

13.7
13-5
13.7
13.7
n.s.

2.3

Table 3.Effect of the beet mosaic virus, beet yellows and western yellows virus on the
root yields of sugar beet in 1963, at high and low nitrogen levels.
Virus inoculation
Control
Beet mosaic alone
Beet mosaic plus western yellows
Western yellows alone
Beet yellows alone
LSD, 5 %

Root yield-Tons/acre
LowN1
High N 2
32.0
30.3
25.4
27.2
26.5

32.8
30.7
27.4
29.0
25.4

2.9

2.9

Loss3
Tons/acre
5.9

18.5
13.3
19.8

Percent
sucrose
12.4
12-2
11.2
11.8
12.2

% loss in
sugar yield
7.7

26.6
17.9
21.6

0.4

No added nitrogen.
2
200 lb of nitrogen/acre added in the form of ammonium sulfate.
3
Since the losses at the high and low nitrogen levels were not significantly different the
values were averaged for these figures.

synergistic effect w i t h a n y of t h e virus c o m b i n a t i o n s was indicated. T h e losses i n r o o t yield d u e t o mosaic with e i t h e r o r


both of t h e yellows viruses w e r e additive. T h u s in the 1962 test
where b e e t m o s a i c caused an average loss of 9 . 7 % a n d western
yellows 6 . 0 % , t h e t w o viruses t o g e t h e r caused a 14.6% r e d u c t i o n
in yield. In t h i s e x p e r i m e n t t h e beet yellows virus caused 35.2
a n d 4 5 . 3 % r e d u c t i o n s i n yield a l o n e a n d i n c o m b i n a t i o n with
mosaic ( T a b l e 2); all t h r e e viruses caused a 5 0 . 4 % loss in yield,
showing t h a t t h e c o m b i n a t i o n effect in each case was additive.
Similar results w e r e o b t a i n e d w i t h mosaic a n d western yellows,
the only v i r u s c o m b i n a t i o n tested, i n t h e 1963 e x p e r i m e n t ( T a b l e
H i g h levels of nitrogen fertilization, a l t h o u g h showing an
a p p a r e n t effect on t o p g r o w t h of infected plants, d i d n o t change
the effect of v i r u s on r o o t yields in the 1963 tests ( T a b l e 3).
R o o t yields w e r e n o t significantly different at the t w o n i t r o g e n
levels in e i t h e r t h e c o n t r o l plots or with a n y of t h e viruses.
Observation as to t h e effect of n i t r o g e n fertilization on t o p
growth, as well as of analyses for N O - - N in petioles collected
August 5, S e p t e m b e r 16, a n d D e c e m b e r 9, indicated that most

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JOURNAL OF THE A. S. S. B. T.

plants at the "low" nitrogen level were only slightly deficient


in nitrogen. T h i s is b o r n e o u t by the failure of roots to respond
in growth to the fertilizer. T h u s the 1963 experiment did not
provide the wide differences in nitrogen fertility that would
have been desirable for assaying the effects of virus infection.
Neither mosaic n o r beet yellows had any noticeable effect
on the sucrose content of roots from infected plants in either
experiment (Tables 2 and 3). T h e western yellows virus reduced
the sucrose content from 12.4 to 11.8% in the 1963 trial; the
virus had a similar effect on sucrose percentage in combination
with the beet mosaic virus ( T a b l e 3).
Discussion
T h e s e tests indicate that losses caused by the beet mosaic
virus in the California sugar beet crop as a whole may be more
or less negligible compared with the losses to be expected with
the 2 yellows viruses. T h e mosaic virus is primarily restricted
to areas with overwintered beets a n d thus is not as widespread
or as commonly encountered as the western yellows virus. The
beet yellows virus, though similarly restricted, has a more severe
effect on root yields t h a n mosaic or western yellows. It is probably the singly most i m p o r t a n t virus economically in areas where
it occurs with any great abundance. T h e losses obtained with
the 2 yellows viruses in these experiments were comparable to
those reported by Bennett et al. (2) and Duffus (3).
Losses in yield with beet mosaic will vary with the virulence
of the virus strain a n d the age of plants at the time of infection.
Although considerably more virulent strains of the virus are
known, these do not appear to be very c o m m o n in California.
In areas where strains of mosaic similar in virulence to the
one used for these experiments occur with sufficient abundance,
the losses probably range from 5 to 1 0 % . T h e s e losses could
probably be minimized by isolation of the overwintered acreage
as suggested by the results of P o u n d (7) a n d Duffus (4) or by
the development and use of suitably resistant varieties.
Summary
A strain of the beet mosaic virus was used in replicated
field plots to determine its effect on sugar beet yield. T h e virus
reduced top growth slightly a n d root yields 9 . 7 % a n d 5.9%.
respectively, in 1962 and 1963. T h e virus h a d no effect on the
percentage sucrose in infected roots.
In similar trials the western yellows virus had no measurable
effect on top growth b u t reduced r o o t yields 6.0% a n d 11 - 8 %
in 1962 a n d 1963, respectively. In the latter experiment the

V O L . 13, N o . 3, OCTOBER 1964

251

virus also caused a decrease from 12.4 to 11.8% in the sucrose


content of infected roots. Under the same conditions the beet
yellows virus caused severe stunting of top growth, except under
conditions of high nitrogen fertilization, and reduced root yields
35.2 percent and 19.8 percent, respectively, in the 1962 and 1963
tests. In tests to determine the result of infection with combinations of the 3 viruses, the effects were additive in each case.
L i t e r a t u r e Cited
(1) B E N N E T T , C. W. 1960. Sugar beet yellows disease in the United States.
U. S. D e p t . Agriculture T e c h . Bull. 1218.
(2)

BENNETT, C. W . , C. P R I C E , a n d J . S. MCFARLANE.

1957.

Effects of virus

yellows on sugar beet with a consideration of some of the factors


involved in changes produced by the disease. J. Am. Soc. Sugar
Beet T e c h n o l . 9: 479-494.
(3) D U F F U S , J. E. 1961. Economic significance of beet western yellows
(radish yellows) on sugar beet. Phytopathology 51: 605-607.
(4) D U F F U S , J. E. 1963. Incidence of beet virus diseases in relation to overwintering beet fields. P l a n t Disease Reptr. 47: 428-431.
(5) LUDECKE, H., a n d O. N E E B .
1956. Ertrag u n d Beschaffenheit der
Z u c k e r r i b e bei kombinierter Infektion mit Vergilbungs - u n d Mosaikvirus. Z. Zuckerind., N.S. 6: 630-633.
(6)

NIKOLIC,

V.,

D.

CAMPRAG, a n d

I.

MATIC.

1958.

Ispitivanje

stetnosti

mozaika na industriskoj Secernoj Repi. Zasht. Bilja (Plant Prot.,


B e o g r a d ) , 1958, 46: 69-73. (Rev. Appl. Mycol. 39:65)
(7) POUD, G. S. 1947. Beet mosaic in the Pacific Northwest. J. of Agr.
Research 75: 31-41.
(8)

WATSON, D . J.,

a n d M A R I O N A. W A T S O N .

1953.

C o m p a r a t i v e physio-

logical studies on the growth of field crops. III. T h e effect of infection with beet yellows a n d beet mosaic viruses on the growth
a n d yield of the sugar beet root crop. Am. Appl. Biol. 40: 1-37.
(9) WIESNER, K. 1959. D e r Einfluss einer Rubenmosaik einer Rubenvergilbungs u n d einer Mischinfecktion beider Virosen auf Entwicklung, E r t r a g u n d technologischen W e r t der Zuckerrube. Zucker 12:
266-274.

S O H -}- MX(ionic n o n sugars)--* Resin -SO3M-J- HX


Resin
Loading
A;nion a Exchange Resin
HX
SO H N
4-HMXS 0 iphia,

VOL.

13,

No.

3,

OCTOBER

1964

253

O n l y t w o f u n d a m e n t a l modifications in processing will be


i n t r o d u c e d h e r e . I n s t e a d of a single cation exchange c o l u m n in
the i n i t i a l stage, t h e j u i c e is to be passed t h r o u g h t w o c o l u m n s
o f c a t i o n e x c h a n g e resin. T h e f i r s t i s t o pick u p t h e inorganic
cations. T h e s e c o n d i s t o pick u p t h e a m i n o acids t h u s relieving
the a n i o n e x c h a n g e resins of t h a t role. Secondly a high capacity
weak base a n i o n e x c h a n g e r e s i n will be used to take advantage
of g r e a t e r r e g e n e r a t i o n economics t h a n were realized w h e n strong
base resins or l o w e r capacity weak base a n i o n exchange resins
were used.
M o d e r n i o n e x c h a n g e resins have b e e n developed h a v i n g h i g h
capacity a n d i m p r o v e d physical a n d chemical resistance to sugar
liquors. W h i l e i n p r i n c i p l e , they p e r f o r m the same chemical
changes as those f o u n d in earlier resins, t h e n e w resins are
k n o w n to p e r f o r m these e x c h a n g e reactions with greater efficiency.
In this p a p e r , t h e p r o p e r t i e s of commercially available ion exchange resins a r e o u t l i n e d a n d some of t h e i r properties are
described i n c o n n e c t i o n w i t h t h e i r use i n sugar j u i c e t r e a t m e n t .
C a t i o n E x c h a n g e r Selection
In this process, t h e c a t i o n exchange resin operates in the
hydrogen circle to c o n v e r t non-sugar salts to their free acids. T h e
process is s o m e w h a t m o r e involved however t h a n simple exchange
a n d n e u t r a l i z a t i o n d u e to t h e n a t u r e of t h e non-sugars in the
sugar l i q u o r s . F i g u r e 1 shows a typical b r e a k t h r o u g h curve for
a 14 B r i x second c a r b o n a t i o n j u i c e of 9 1 % p u r i t y . T h i s juice
has 135 g r a m s p e r l i t e r of sucrose a n d 10.8 grams of non-sugars.
If it is a s s u m e d t h a t 1/2 t h e non-sugar salts are inorganic salts
having an e q u i v a l e n t w e i g h t of 100, t h e concentration of salts
will b e 0.108 e q u i v a l e n t / l i t e r o r a b o u t 5400 p p m a s C a C O 3 . T h e
equivalent w e i g h t of t h e non-sugars has b e e n d e t e r m i n e d from
a variety of tests w i t h A m b e r l i t e IRA-68, a n d this will be discussed i n t h e section o n a n i o n exchangers.
It is a p p a r e n t from F i g u r e 1 that the inorganic cations are
present as salts of weak acids since a large a m o u n t of buffering
exists in t h e system. T h e t i t r a t i o n curves for a strong acid ( H C 1 )
and a typical weak acid (acetic) are p l o t t e d to d e m o n s t r a t e this
buffering effect in a q u a l i t a t i v e sense. T h e s e curves are calculated based o n t h e a s s u m p t i o n t h a t actual b r e a k t h r o u g h for
ion e x c h a n g e o c c u r r e d at 15 b e d volumes, a figure reached by
using a c o l u m n capacity of 1.6 e q / l i t e r w h i c h is typical for a fully
regenerated b e d of A m b e r l i t e IR-120.
Capacity 1.60 = 15 bed volumes
0.108

254

JOURNAL OF THE A. S. S.

Figure 1.Hydrogen cycle treatment.


brix, 91% purity.

juice.

14

Figure 2.-Hydrogen cycle treatment, 2nd carbonation juice.


brix, Ambrelite IR-120H.

14#

10

15

20

2nd carbonation

B. T.

25

SO

BED VOLUMES

VOL. 13, N o . 3, OCTOBER 1964

255

In F i g u r e 2, t h r e e curves have b e e n p l o t t e d showing the effect


o f p u r i t y o n t h e effluent p H c u r v e . T h e curves have b e e n based
o n l a b o r a t o r y results o n v a r i o u s juices. Possible variation d u e
to changes in t h e ash to total non-sugar r a t i o c a n n o t be predicted
w i t h o u t e x t e n s i v e d a t a o n t h i s subject. T h e extensive buffering
evident i n p H b r e a k t h r o u g h curves i s d u e i n a large degree t o
the a m i n o acids w h i c h a r e p r e s e n t i n t h e juice. G l u t a m i c acid,
p y r r o l i d o n e c a r b o x y l i c acid (PCA), aspartic acid, t y r o s i n e ,
leucine, a n d isoleucine a r e usually found. T h e q u a n t i t y o f these
materials in some typical b e e t l i q u o r s has b e e n previously rep o r t e d (3, 6).
T h e a m i n o a c i d c o n t e n t of a typical b e e t sugar j u i c e a m o u n t s
to 15-20% of t h e total non-sugars a n d represents 30-40% of t h e
organic non-sugars. It is e v i d e n t t h a t these r e p r e s e n t major
constituents in t h e l i q u o r . T h e i r recovery is w o r t h y of consideration a n d t h e i r p r o p e r t i e s will certainly have an i m p o r t a n t bearing on t h e e n d p o i n t of t h e cation exchange cycle.
In this s t u d y it has b e e n considered desirable to operate the
cation e x c h a n g e system in w h a t is called a merry-go-round operation. T h i s p e r m i t s t h e recovery of t h e a m i n o acid fractions at
the same t i m e p e r m i t t i n g full l o a d i n g of t h e cation exchange
resin. F u r t h e r m o r e , it p r o v i d e s for t h e a c c u m u l a t i o n of the
a m i n o acids o n t h e c a t i o n exchange resin, r a t h e r t h a n r e q u i r i n g
a d d i t i o n a l v o l u m e s of t h e m o r e expensive a n i o n exchange resin.
T h i s m e r r y - g o - r o u n d system has as its a i m t h e a c c u m u l a t i o n
of t h e n i t r o g e n o u s fraction on o n e bed. It is necessary to balance
the cycle so t h a t t h e b e d w h i c h receives t h e a m i n o acids can
operate u n t i l it is well l o a d e d w i t h the a m i n o acid fraction. At
any given m o m e n t , t w o c o l u m n s of cation exchange resin are
operated i n series w i t h t h e a m i n o acids b e i n g a c c u m u l a t e d o n
the second c o l u m n . A q u e s t i o n to be answered is h o w m u c h a m i n o
acid can b e l o a d e d o n t h e second c o l u m n before t h e effluent p H
rises a b o v e 3.5 at w h i c h p o i n t t h e g l u t a m i c acid will begin to
leak. To discuss this, c o n s i d e r first an " a v e r a g e " beet sugar l i q u o r
which is a s s u m e d to be as follows:
Table 1.Composition of 91 purity beet liquor.
Component
Sucrose
Inorganic non-sugars
Organic non-sugars
Betaine
Amino acids
Nitrogeneous fraction/ash

Grams/liter
135
5.42
5.42
1.30
1.41

Equivalent Weight
50 (averaged
119
147
_

Eq/liter
0.1085
0.0109
0.0096
0.19

T h e s e figures i n d i c a t e that the ion loading d u e to amino


acids is about 2 0 % of the inorganic non-sugars. For calculation

256

JOURNAL OF THE A. S. S.

B. T.

purposes, it should be assumed t h a t the exchange of a m i n o acids


a n d other nitrogenous materials follow the ratios in T a b l e 1,
although experimental evaluation will be r e q u i r e d for a firm
recommendation.
An examination of the pH curve in Figure 1 shows t h e importance of careful operation to insure full utilization of the
cation a n d anion exchange resins. To prevent d u m p i n g of amino
acids, it is necessary to stop the loading before the effluent pH
rises above 3.5. T h e c o l u m n which collects the a m i n o acids can
be used in connection with 2 or more other columns since the
columns picking up the inorganic ions will u n d o u b t e d l y be exhausted before the column picking up the a m i n o acids. Two
examples of merry-go-round systems of this type, o n e involving
three columns, the other a total of four cation exchange columns
are shown in this paper.
Three
column
merry-go-round
T h e following description will serve to outline a recovery
procedure based on three columns in the cation set.

M E R R Y - G O - R O U N D FLOW DIAGRAM
T h e stream of second carbonation juice will pass through
c o l u m n A, then t h r o u g h c o l u m n B, b o t h of which will load until
the pH of the effluent of A raises to 3.8-4.0. C o l u m n B now
contains a b o u t 2 0 % of its exchange sites in combination with
a m i n o acids a n d betaine. It is also likely that other organic
non-sugars will be held b u t it is n o t k n o w n in exactly what
portion. T h i s area requires m o r e study. O n c e the columns have
reached this condition, the A c o l u m n can be d r o p p e d off for
regenerations. T h e C c o l u m n is started up a n d the flow is now
t h r o u g h C to B. T h i s process continues u n t i l the pH of C
effluent reaches 3.8-4.0. C o l u m n B now is a b o u t 4 0 % loaded
with a m i n o acids. C is t h e n regenerated a n d A operated through
B u n t i l a similar b r e a k t h r o u g h occurs. R e g e n e r a t i o n of V can
now be accomplished while loading in a cycle of low flow rate

VOL.

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3,

OCTOBER

1964

257

continues on G. Once A or B is regenerated, either one can be


used to accumulate the amino acids on the next cycle.
Four column merry-go-round
With a three column merry-go-round system it is necessary
to operate an occasional cycle at low flow to keep the system
from getting out of phase. It is likely that a fourth column would
greatly assist in this matter. It is suggested that a four column
merry-go-round should be used in cases where continuous operation with amino acid recovery is practiced.
T h e ultimate recovery of the amino acids is involved in the
elution of column B with a base. In Europe 10% ammonium
hydroxide or NaCl is used for this step. T h e recovery of amino
acids from beet sugar plants is practiced in Europe as a byproduct
industry. T h e effluent mixture is heated with lime and the
ammonia recovered after separation of the insoluble calcium
glutamate. T h e precipitates are collected at a central factory
where the various fractions are purified and recrystallized for sale
as food supplements.
Regeneration
efficiency
T h e expenditure of acid for regeneration is a major factor in
the operation of the cation exchanger. Relatively little work has
been done on this subject in the sugar industry but certain effects
can be estimated accurately in regard to acid consumption. The
exchange is largely hydrogen for potassium and in monovalent
exchange, leakage can be expected which will affect materially
the shape of the breakthrough curve. These factors have been
evaluated for Amberlite IR-120 in Figure 3 which shows the
relationship between capacity (bed volumes) and acid useage.
Figure 4 gives an estimate of the monovalent cation leakage as
a function or regeneration level.
Figure 4 shows breakthrough curves for various purity liquors
being treated
with Amberlite IR-120 regenerated at 10 lbs.
H 2 S0 4 /ft 3 . These curves are quite typical and the values check
reasonably well with the estimated leakage of about 8% in the
flat range prior to breakthrough.
Sucrose Losses
Since the major purpose of ion exchange treatment of beet
juice is the recovery of sucrose, any losses of sucrose in the process
would cause much concern. In the cation exchange treatment
there are two possible sources of sugar loss. The most significant
are losses due to displacement and other mechanical handling
operations. T h e second most important loss is involved with
catalytic inversion of the sucrose. Careful consideration shows
that these causes for sugar loss are not sufficient reason for rejecting ion exchange treatment.

258

JOURNAL OF THE A.

S.

S.

B. T.

lbs. H 2 S0 4 /ft?

Figure 3.Relationship between capacity and regeneration level for


second carbonation juice.

BED VOLUMES

Figure 4.Potassium leakage as a function of regeneration level and


juice purity. Amberlite IR-120.

Losses D u e to Displacement
In arriving at the loss of sugar due to the displacement processes, estimates must be made on the basis of emperical field
data due to the variables of design and operating conditions.
In general, the displacement will be a function of bed void
volume, sugar liquor density, liquor viscosity, and resin bead
porosity. In addition, liquid heights in the column, Row rates

VOL.

13,

No.

3,

OCTOBER

1964

259

and under-drain volume will play an important part in the displacement process. From a practical standpoint, there are two
problems, namely, the added cost of evaporating diluted juice
and the loss of sugar in sweet waters resulting from continued
rinsing.
T h e idealized displacement involving no kinetic considerations would be a sharp step function which would occur when
the displacement had filled the bed and underdrain voids. Sucrose
molecules which have diffused into the resinous structure reduce
this sharpening effect and require time to diffuse out at the end
of the displacement. These effects result in a round frontal displacement with a tailing off at the end of the displacement.
T h e concentration of the sugar liquor plays an important
role during "sweetening on". A thick liquor will tend to diffuse
down the column under the influence of gravity. This will result
in an irregular displacement within the bed. Displacement of a
heavy liquor by water in a "sweetening off" step may present a
drag problem due to viscosity but it is possible to correct this
by using hot water. Flow rate can play an important part in
these matters. Generally, flow should be as slow as practical but
the improvement
in displacement is usually not considerable below about 1 / 2 gpm/ft 3 .

BED VOLUMES

Figure 5.Displacement curves, plant studies.

In Figure 5 are drawn displacement curves for a typical operation in a beet sugar factory. These curves indicate the influence
of flow rate but do not represent an optimum situation in regard
to this important operating function. Detailed studies of the
variables have been carried out.
T h e displacement step in juice treatment is an important
one from an economic standpoint. It is apparent that "sweeten-

260

JOURNAL OF THE A. S. S. B. T,

ing o n " results in a slight overall dilution when d i l u t e juices are


involved. T h i s is not serious in average purity liquors b u t could
become i m p o r t a n t for low purity liquors where r u n s will be
short. Losses are more i m p o r t a n t d u r i n g the "sweetening off"
step where a considerable tailing may result. T h e s e liquors may
be used for m a k e u p water to t h e diffusion station a n d for the
water used for slaking the lime as shown in a later section.
T h e d i l u t i o n of typical t h i n juice as a function of purity has
been estimated for a typical bed displacement step in T a b l e 2.
It is assumed that displacement is accomplished at 1 / 4 gpm/ft 3 .
a n d that the l i q u o r is of constant concentration. No estimate of
e q u i p m e n t voids are allowed for in this estimate.
Table 2.Displacement dilution of sugar liquor, Flow Rate1/4 gpm/ft. 3 .
Purity

%
91
88
84

Concentration Temperature
F
Brix
14
14
14

50
50
50

Bed Volume
Total bed volumes
treated
(with dilution)
juice net
16
11
8

18.3
13.3
10.7

Dilution

%
12.8
18.5
25.0

Although a n u m b e r of factors in this study show that ion


exchange treatment is most economical w h e n one is working
with low purity juices, in this instance the advantage of working
with high purity Hquors becomes apparent. It is expected that
the dilution water can be r e t u r n e d to the diffusion plant. Therefore, the values recorded above do not represent additional actual
evaporator load. T h e detailed operation of this phase of the
p l a n t will be estimated in the sections on economics and plant
design.
Invert Formation
Inversion d u r i n g cation exchange t r e a t m e n t can be broken
d o w n into two separate b u t related processes designated here as
heterogeneous a n d homogeneous inversion. T h e inversion of
sucrose to fructose a n d glucose is catalyzed by hydrogen ion. The
exchange sites when converted to the hydrogen form provide a
source of highly acidic materials which give a typical heterogeneous catalytic reaction, when the sucrose diffuse into the
resin beads. D u r i n g the exchange process, hydrogen ions are also
liberated from the resin. T h e s e ions e n t e r the solution where
they are available to act as homogeneous catalysts for the inversion.
Existing data on several cation exchangers have been evaluated
a n d inversion estimated as a function of contact time and temperature. T h e s e values are based on the assumption that pu re

VOL. 13, No. 3, OCTOBER 1964

261

F i g u r e 6 . I n v e r s i o n w i t h A m b e r l i t e XE-100 as function of t e m p e r a t u r e
and contact time.

Figure 7.Inversion with Amberlite IR-120 as function of temperature


and contact time.

262

JOURNAL OF THE A. S. S. B. T.

Figure 8.Inversion with Amberlite 200 or IR-124 as function of


temperature and contact time.

sugar solutions aie being treated so that only the heterogeneous


reaction is involved.
T h e curves in Figures 6, 7, a n d 8 show the a m o u n t of inversion
which will occur when a p u r e sugar solution at a certain temperat u r e contacts a bed of cation exchanger for a definite period of
time. It is assumed here that a 20 Brix l i q u o r is being treated.
For most thin juices it is expected that Figures 6-8 will give a
reasonable description. Less invert w o u l d be expected if a more
dilute juice were used and, of course, a greater a m o u n t would
invert if the liquor were of appreciably higher concentration. A
p l o t of field data for which, unfortunately, the contact time is
u n k n o w n , is drawn in each graph to show that the calculated
values are of the same o r d e r of m a g n i t u d e as those from the
field. T h e flow rates are expressed as contact times.
gpm/ft 8 =

to
of
by
as

7 48
:
, . s
contact time (min.)
Since the concentration of hydrogen ion is related directly
the rate of inversion, it is necessary to measure t h e amount
time that the cation effluent will exist before neutralization
t h e a n i o n exchanger. In Figure 9 data from Sadter are plotted
summarized by Payne (5), which enables inversion due to

VOL.

13,

No.

3, OCTOBER

1964

Figure 9.Homogeneous inversion. Data of Sadter Honig, Principles


of Sugar Xechnology. Volume 1page 448.

homogeneous catalysis to be estimated. While the amount of inversion by homogeneous catalysis is not as great as that caused
by a fully regenerated bed, it must be taken into account.
T h e graphs developed for inversion can be used quite effectively to calculate inversion for a variety of process conditions.
There are four process variables which deserve consideration.
These are: (1) EfiEect of temperature in a standard two bed
system; (2) Effect of flow rate in a standard two bed system; (3)
Effect of a merry-go-round operation and (4) Effect of regeneration levels. These situations are examined in the following
sections.
Effect of temperature
A standard operation with a 90% purity liquor at 15 Brix
which will contact a single
bed of cation exchanger at 4 minute
contact time (1.87 gpm/ft 3 ) will be assumed. The acid generated
will pass on to a bed of anion exchange resin. In Table 3, the
amount of inversion estimated from Figures 6-9 is listed for the
various resins at several temperatures. A sample calculation
follo w s :

JOURNAL OF THE A.

264

S.

S.

B. T.

Amount due to homogeneous catalysis


At the start of the r u n the bed is completely in the acid form
a n d there is free space equal to the volume of juice in the cation
exchange column above the anion exchange resin. T h e pH
generated is 2.1 (Figure 2).
T i m e through cation bed = 4 m i n u t e s
T i m e in piping and above anion exchange bed = 4 minutes
T e m p e r a t u r e = 0 C (using Figure 9)
% sucrose converted = 8 (0.02) = 0.00267% inverted
60
Since the cation exchanger will be exhausted at a b o u t the
same rate (geometrically) as the anion exchanger, the void displacement will remain relatively constant as the r u n progresses
so that the inversion d u e to homogeneous catalysis will remain
roughly constant t h r o u g h o u t the r u n .
Amount due to heterogeneous catalysis
Using Amberlite IR-120 and the contact time of four minutes,
T a b l e 3.Summary of invert estimated as function of contact time.
% I n v e r t b a s e d o n s u g a r solids
h e t e r g e n o u s catalysis
Temperature
~0G

20C

40C

"
60C

80C

"

Position
on run
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter
Start
Middle
3/4 quarter

Contact Homogeneous Amberlite Amberlite


IR-120
Time
catalysis
XE-100
8
"
"
2
"
"
8
"
"
2
"
"
8
"
"
2
*
"
8
"
"
2
"
'*
8
**
"
2
"
"

0.006
"
"
0.005
"
"
0.056
"
'*
0.014
"
"
0.62
"
0.155
*'
"
6.66
"
"
1.66
"
"
66.4
"
"
16.6
"
*

21.5
5.80
1.55
1.52
0.26
0.085
35.0
14.0
5.4
5.30
1.45
0.60
53.0
28.0
16.0
16.0
5.50
3.40
75.0
55.0
39.0
39
22
15
100.0
iOO.O
90.0
90.0
65.0
53.0

0.25

0.12
0.075
0.072
0.027
0.016
1.80
0.70
0.50
0.50
0.19
0.11
9.0
4.5
2.6
2.6
1.10
0.53
34.0
18.0
9.9
9.8
4.40
2.50
100.0
64.0
54.0
SS.0
16.0
4.0

Amberlite
IR-124
O.073
0.045
0.026
0.026
0.015
0.009
0.33
0.20
0.90
0.092
0.053
0.030
1.30
0.75
0.29
0.28
0.16
0.09
3.8
2.2
0.76
0.76
0.42
0.23
10.0
5.6
1.8
1.8
0.95
O.54

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265

the heterogeneous catalyzed inversion for various parts of a run


can be estimated by reference to Figure 7.
Start of r u n Contact time (acid resin) = 4 minutes
Inversion == 0.12%
Middle of r u n Contact time (acid resin) = 2 minutes
Inversion = 0.075%
3/4 of r u n Contact time (acid resin) = 1 minute
Inversion = 0.026%
In T a b l e 3, the effect of contact time is examined in the
same manner for a variety of resins, temperatures and contact
times.
Effect of merry-go-round operation
It becomes apparent that the time during which suerar liquor
is in contact with regenerated cation exchanger, is very important
in so far as invert formation is involved. For example, the merrygo-round operation would result in an increased time that the
liquor would exist in contact with acid resin. If one were to
compare the invert formation in a conventional two bed system
vs. a merry-go-round operation containing two cation exchansrer
beds and a double anion exchanger, we would expect the contact
time to be doubled.
T h e effect of a merry-go-round system on invert has been
examined for a four minute contact with Amberlite IR-120, a
conventional, commercially available cation exchanger of average
crosslinkage. These estimates are given in Table 4.
Table 4.Effect of merry-go-round operation on hetergeneous catalyzed invert formation, 90% purity - 15 Brix Amberlite IR-120H.
Temperature
C
0
0
20
20
40
40

Contact Convention
Minutes
4
8
4
8
4
8

Invert
0.07
0.125
0.40
0.75
2J!
4 4
-

These figures show that the merry-go-round operation will


roughly double the invert formation over that experienced for a
conventional two bed system.
On the other hand, numerous studies show that Amberlite
200 and Amberlite IR-124 give less than half the inversion noted
for Amberlite IR-120. At high temperatures the homogeneous
catalytic reaction will play an increasingly important role so that
inversion will tend to increase at a rate greater than that indicated above. Variations in the process can be examined by the
methods developed in the previous sections.

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JOURNAL OF THE A. S. S. B. T.

Effect of regeneration level


T h e effect of regeneration level on the a m o u n t of invert
formed is of interest. Theoretically, the effect will d e p e n d on
t h e regeneration efficiency of the acid which in t u r n is a measure
of the a m o u n t of hydrogen form available for catalytic inversion.
T h i s may be translated to the contact time that the l i q u o r has
with the hydrogen resin. T h e curves used in this report are
based on highly regenerated resin (20 lbs sulfuric acid/ft 3 ).
A contact time corrected for regeneration efficiency can be
employed to predict the a m o u n t of invert formation. T h e calculation of corrected contact times is illustrated in T a b l e 5.
Table 5.Effect of cation exchanger regeneration efficiency on invert formations
Amberlite 1R-120, 90% purity - 15 Brixes.
Time, minutes
Regeneration Level
lbs H2S04/fts
2.5
5
10
15
20
25

Full

% Saturation
Capacity

Initial contact
(saturation)

31
49
72
76
81
83
100

4
4
4
4
4
4
4

Corrected contact
time
1.24
1.98
2.88
S.04
3.25
3.3
4.0

T h e s e concepts can be used to derive a curve for the performance of Amberlite IR-120 when operated at 5 lbs/ft 3 using
a contact time of 6 minutes. T h e t e m p e r a t u r e for the run is
15C.
Fully regenerated bed:contact time = 6 m i n u t e s
1.98 X 6 4
So at 5 lbs H 2 S 0 4 / f t 3 , Contact time =
~
= 2.9 m i n u t e s contact at
the start of the run.
Heterogeneous inversion = 0 . 2 8 % of sucrose solids.
H o m o g e n e o u s Catalysis 0.07(6/60) 0 . 0 0 7 % of sucrose
solids.
T o t a l = 0 . 2 8 % heterogeneous inversion of sugar in liquor.
5 0 % point: % heterogeneous inversion = 0.14%
% homogeneous inversion = 0.007
T o t a l = 0.147% of sugar solids
7 5 % point: % heterogeneous inversion = 0.07
% homogeneous inversion = 0.007
T o t a l .= 0.077% of sugar solids.
T h e results of such calculations have been plotted and comp a r e d with actual measurements in Figure 10.

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267

**' KUN

Figure 10.Comparison of calculated invert formation with experimental values.

T h e results of these estimates for inversion show that the


calculated values are fairly consistent with those obtained by
experimentation. T h e calculation methods give a somewhat
higher value b u t these can easily be accounted for by particle
size variation, failure of the displacement scheme due to channeling, etc.
Anion Exchange Resins for Sugar Treatment
Several anion exchange resins have been studied for sugar
processing over the years. Amberlite IRA-68 has been found
to be a very suitable resin for this application due to its high
capacity for non-sugars and its ability to decolorize the liquors
to a high degree. Its high loading is a chief virtue but its stability
against fouling is also important. Also Amberlite IRA-68 is more
resistant to thermal degradation than strong base anion exchange
resins. Since this resin shows some instability at very high temperatures, it is necessary to examine this variable in regard to
resin life.
In the following sections it will be assumed that Amberlite
IRA-68 is used after a merry-go-round operation on Amberlite
IR-120. T h e average cation endpoint will be assumed to be
below pH 2.6. It will be assumed that the anion exchanger is
regenerated with ammonia but that the ammonia is recovered
with lime. Column studies have been employed to define capacity
and effluent quality as a function of juice purity.

268

JOURNAL OF THE A. S. S,

B. T.

B E D VOLUMES

Figure 11.Amberlite IRA-68 effluent characteristics. Juice of 81.4%


initial purity.

BED V O L U M E S

Figure 12.Amberlite IRA-68 effluent characteristics.


initial purity.

Juice of 88.2

Capacity as a Function of Purity


T h e capacity values of A m b e r l i t e IRA-68 have been related
to the characteristics of the sugar juice. It is n o t the intention
to develop this m a t t e r in detail at this t i m e b u t a few examples
of a typical operation will be used to illustrate a relationship
between influent purity a n d capacity.

13,

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1964

269

BED VOLUMES

Figure 13.History of the effluent from Amberlite IRA-68 in deionization of a 9 1 % purity juice loading 19.2 lbs/ft 3 . *w / / * * w c t .

% PURITY

(|NFLUF*,TX

Figure 14.Effect of juice purity on capacity of Amberlite IRA-68


with merry-go-round operation of Amberlite IR-120 beds.

In Figure 11 we show a typical run on a low purity juice


(81.4% purity) and Figure 12 shows a run on 8 8 % purity juice.
T h e most interesting results were obtained on a typical 9 1 %
purity beet j u i c e in France and these results are summarized
in Figure 13.
T h e utilization of these results in a general manner is required if an estimate of the capacity of Amberlite IRA-68 is to

270

JOURNAL OF THE A. S. S. B. T.

be o b t a i n e d for average sugar liquors. Fo r this purpose the


accumulative curves for purity were plotted a n d values picked
off to construct a plot of bed volumes treated to a purity endp o i n t as a function of influent purity. W i t h i n the purity ranges
available the curves are plotted in Figure 14. T h e curves have
been extended toward 100% purity by extrapolation.
T h e capacity of A m b e r l i t e IRA-68 is a b o u t 1.70 equivalents/
liter. T h i s figure will p e r m i t an estimate of the capacity provided
the equivalent weight is known. Unfortunately, the average
equivalent weight of non-sugars vary considerably so that a study
is r e q u i r e d to fix the equivalent weight.
T h e results shown in Figures 11-13 can be used to calculate
equivalent weights for these particular r u n s and this is shown
in T a b l e 6. T h e s e values are a p p r o x i m a t e d from the loading
data a n d t h e composition of the liquors. T h e values compare
favorably with those obtained by an accurate analytical procedure.
Table 6.Equivalent weight estimates for typical beet liquors, Amberlite IRA-68.
%
Purity
_____
88-2
91

Capacity
Equivalent capacity
lbs. non-sugar/ft3
lbs. Eq/ft:i
24
~
0.106
17
0.106
19.2
0.106

Equivalent weight
lbs. or grams
226~
loO
11

Regeneration Efficiency
T h e regeneration of low molecular weight acids from Amberlite IRA-68 is not difficult. Complete regeneration of the sites
results from a contact with an equivalent a m o u n t of ammonium
hydroxide or caustic. For a fully exhausted bed (1.7 meq/liter)
a regeneration level of 1.75 e q / l i t e r is suggested. In English units,
the r e q u i r e m e n t becomes 4.37 lbs N a O H / f t 3 or 1.86 lbs NH 3 /ft 3 .
T h e best r e c o m m e n d a t i o n is to regenerate with ammonia and
then to recover the regenerant with lime. Caustic is normally
n o t employed d u e to its cost.
Effluent Characteristics
An e x a m i n a t i o n of the effluent curves shows a gradual leakage of non-sugars which increases progressively u n t i l breakthrough
occurs. It should be recognized that t h e curves shown in Figures
11-13 represent almost complete b r e a k t h r o u g h . T h e cycles were
so operated to obtain complete utilization of the resin. T h e pro
cedure results in a 5 0 % reduction in non-sugars w h e n calculated
on an accumulative basis.
A consideration of the m a g n i t u d e of the capacity values in
terms of b e d volumes treated w o u l d show that accumulative
purity will n o t change sufficiently to be used for control p u r

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poses. It a p p e a r s t h a t a b r e a k t h r o u g h p o i n t based on effluent


pH is t h e m o s t s u i t a b l e o n e for a sugar a p p l i c a t i o n .
Brief m e n t i o n s h o u l d b e m a d e c o n c e r n i n g t h e non-sugars
which a r e l e a k i n g t h r o u g h t h e bed. It is expected that very
weakly i o n i z e d a c i d s a n d n o n - p o l a r non-sugars w o u l d pass t h r o u g h
the b e d w i t h o u t b e i n g r e t a i n e d b y i o n e x c h a n g e mechanisms.
W h i l e it is p o s s i b l e t h a t some of these materials will be held
b y a b s o r p t i o n , n o a l l o w a n c e h a s b e e n m a d e for such process.
It is likely t h a t t h e slight leakage, almost from t h e b e g i n n i n g of
the r u n s , h a s b e e n c a u s e d b y large m o l e c u l a r weight acids a n d
n o n - p o l a r s u b s t a n c e s . T h e exact n a t u r e o f these c o m p o n e n t s has
not b e e n p r o p e r l y e s t a b l i s h e d a n d t h e i r effect o n t h e sugar application h a s n o t b e e n m e a s u r e d i n these studies.
S o m e i n t e r e s t i n g w o r k o n l i q u o r s c o n t a i n i n g large a m o u n t s
of i n v e r t has s h o w n t h a t these acidic sugars are h e l d at least to
some d e g r e e on p a r t i a l l y e x h a u s t e d b e d s of A m b e r l i t e IRA-68.
T h i s i n t e r e s t i n g discovery has n o t b e e n considered i n this p a p e r
b u t m a y b e useful i n r e m o v i n g i n v e r t from high quality liquors.
T h e r m a l Stability of the A n i o n Exchanger
N o p r o b l e m i s e x p e c t e d i n r e g a r d t o t h e t h e r m a l stability o f
A m b e r l i t e I R A - 6 8 i n t h e cold m u l t i p l e b e d process. However,
the i m p o r t a n c e of t h i s p r o p e r t y c a n n o t be overlooked a n d the
q u e s t i o n o f o p t i m u m o p e r a t i n g t e m p e r a t u r e r e q u i r e s a n answer.
In g e n e r a l , it is e x p e c t e d t h a t t h e process t e m p e r a t u r e will be
fixed by t h e f o r m a t i o n of i n v e r t a n d n o t by resin instability.
C o n s i d e r a b l e i n f o r m a t i o n has b e e n collected o n t h e t h e r m a l
stability of A m b e r l i t e I R A - 6 8 . T h e m e c h a n i s m of t h e r m a l degrad a t i o n follows first o r d e r r e a c t i o n kinetics. T h i s result was useful
i n e x t r a p o l a t i o n t o t h e low t e m p e r a t u r e values. T h e m a g n i t u d e
of t h e t e m p e r a t u r e effect on resin half life is summarized in
T a b l e 7.
T a b l e 7.Thermal stability of Amberlite IRA-68, Half Life values for IRA-68.
Half/time
Days
50

Comment
measured

Anion E x c h a n g e R e s i n L i f e
It must be emphasized that thermal life may n o t be related
directly to actual life except at high temperatures. It is not ex-

272

JOURNAL, OF THE A. S. S. B. T.

pected that Amberlite IRA-68 will last 60,000 days (165 years)
if operated at 1 5 C O t h e r factors will be involved such as oxidation of the resin, fouling with organic m a t t e r a n d operational
attrition. Practice has indicated an average life of a n i o n exchangers at 1 X 10 6 gallons of treated liquor p e r cubic foot of
resin. A m o r e conservative figure will be used in the later
economic evaluation.
Selection of Process a n d G e n e r a l Assumptions
T h e process that has been discussed is one which involves
efficient removal of cations from second carbonation juice by
passing it t h r o u g h a cation exchange system utilizing Amberlite
IR-120 (H). T h e acidified juice is then passed t h r o u g h Amberlite IRA-68 for the removal of anions a n d color bodies. Since
Amberlite IRA-68 is a weakly basic a n i o n exchanger, it is not
surprising that the best loading is accomplished in a feed liquor
which has been converted completely in t h e cation exchange installation. Constancy of effluent composition from the cation
exchange components can best be accomplished in a feed liquor
which has been converted completely in the cation exchange
installation, and this can best be accomplished by a merry-gor o u n d operation of the units. T h i s procedure will also help in
the recovery of a m i n o acids.
T h e r e are several places where this process can be performed
in t h e sugar production flow sheet. It is believed, however, that
best performance would occur after the second carbonation step.
T h i s selection of feed liquor would insure that t h e juice is of
good quality, well established in regard to colloids, a n d yet not
too viscous. It is undesirable to heat liquors b e i n g passed through
a m u l t i p l e b e d system of this type a n d consequently, dilute
liquors are desirable. Generally, it is undesirable to force heavy
liquors through ion exchange beds u n d e r pressure. By the way,
pressure d r o p in an ion exchange b e d is a function of flow rate,
resin particle size, a n d viscosity. T h e pressure d r o p increases almost linearly with the viscosity of the flowing l i q u o r .
An examination of the c u m u l a t i v e p u r i t y curves on the
treated juice versus the instantantaneous effluent curves clearly
shows that considerable difficulty will result if blending of the
effluent was n o t employed to deliver a treated l i q u o r of constant
purity to the evaporator. To avoid the p r o b l e m of boiling high
purity unbuffered liquors which w o u l d degrade rapidly in fne
evaporators, one might employ a surge tank to allow the mixing
of t h e effluent from t h e a n i o n exchange installation or one might
b l e n d in some u n t r e a t e d juice.

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1964

F o r t h e p u r p o s e s of t h e c a l c u l a t i o n given in this paper, it


shall be a s s u m e d t h a t t h e t e m p e r a t u r e is c o n t r o l l e d at 15-20C
a n d t h a t t h e flow r a t e is 2 g p m / f t 3 . U n d e r these conditions, inversion w i l l be of t h e o r d e r of 0.2 - 1.0% d e p e n d i n g on the
r e g e n e r a t i o n level of t h e c a t i o n e x c h a n g e r a n d t h e m e t h o d of
o p e r a t i o n . X h e i n f o r m a t i o n o n t h e i n v e r t f o r m a t i o n has been
d e r i v e d f r o m t h e o r e t i c a l considerations a n d d a t a o b t a i n e d i n
the field a n d in t h e l a b o r a t o r y . F o r t h e p u r p o s e s of this calculation, a figure of 1.0% a p p e a r s to be conservative for b o t h invert
a n d m e c h a n i c a l losses.
D e v e l o p m e n t of the Process
T h e multiple bed
calculations b a s e d o n
be w i t h a 6,000 t o n
p u r i t y . T h e d e t a i l s of

process will be e v a l u a t e d by a series of


typical plants. T h e initial evaluation will
p e r day p l a n t h a v i n g a j u i c e of variable
t h e c a l c u l a t i o n s a r e as follows (4):.

Q u a n t i t y of b e e t s p e r day = 6,000 t o n s
Composition

14.97% sugar
0 . 3 9 % sugar
0 . 0 2 % sugar
14.97-0.41 =
juice.

o n beets
o n beets p u l p loss.
on beets lime, flume loss.
1 4 . 5 6 % sugar on beets in t h e

Solids in j u i c e = 14.0 B r i x
P u r i t y = 8 5 . 5 % b a s e d on solids
T o n s of s u g a r in j u i c e / d a y 6000 X 0.1456 874 tons
T o n s solids in j u i c e / d a y =

874
g^g

= 1021

tons

T o n s t h i n j u i c e / d a y - j ^ 7300 tons 1.66 X 10*


gallons
Flow

1 6 6 X 10 6
24 X 60

u 5 0 saiions/minute
s

W e i g h t in n o n - s u g a r s - 1021
X 10 5 l b s / d a y

- 874 - 147 t o n s / d a y = 2.94

JOURNAL OF THE A. S. S. B. T.

274

Ion exchange plant


A typical but not optimized system has been used as outlined
in the diagram.

S C H E M A T I C DIAGRAM OF ION E X C H A N G E S Y S T E M

Cation exchange system


Cation Exchange ResinAmberlite IR-120 in six beds.
Capacity8.5 bed volumes/cycle on a single bed based on
laboratory data or say an average of 16.5 bed
volumes on each primary bed in a merry-go-round
operation. Assume each primary bed is used 6
times/day.
Volume
of resin required/day.
Volume per Bed
schedule.
Flow Rate
Amberlite
installed.
Regenerant Requirement
Anion exchange system
Anion Exchange ResinAmberlite IRA-68 in six beds.
Capacity22 bed volumes based on laboratory data.
Volume
IRA-68 required in 24 hr day.
Volume per Bed
Amberlite IRA-68 Inventory
Regenerant Requirement

VOL.

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275

Capital and Depreciation Costs


T h e cost of ion exchange equipment varies roughly as the
resin bed volume according to the empirical0 . 7formula
cost = 200 (Bed Volume)
In addition to the initial outlay, installation cost is usually about
equal to the equipment cost less resin. Depreciation of installed
equipment can be figured at 5 years under normal conditions
although an actual life of 20 years is not unusual for well maintained installations.
T h e depreciation of resin cost is a difficult matter to calculate.
T h e experience with cation exchangers would lead to the conclusions that treated volumes of the order of
gallons per
cubic foot would be a reasonable value. Field knowledge of anion
exchanger stability would indicate a life ranging between 500,000
and 1,000,000 gallons per cubic foot. These figures can be used
to arrive at a life for these components; 4,070 days for Amberlite
IR-120 and 3,030 days for Amberlite IRA-68. However, this
intuitively seems to be too long and a five year (100 days/operating year) depreciation figure on the resin and on the equipment
will be utilized. This appears conservative on the basis of field
experience.
Labor
requirement
T h e labor required to operate a given plant will vary with
the amount of automation used and with the composition of
the beet juice being processed. A technically trained person will
be required for about
of an 8 hour shift. The shift group
would be usually 2 men with some mechanical help.
Other
considerations
An evaluation of the deionization process requires examination of its effect on auxiliary factors including cooling, sweet
water quantities, waste treatment, and steam requirements. These
matters are reviewed in the following sections.
Cooling costs
T h e need for the cooling of juice is an important requirement
due to invert formation in the cation exchanger bed and also
possible thermal instability of the anion exchanger. Cooling
costs are much a function of plant location and availability of
large quantities of cooling water. Since heat recovery is usually
practiced, it is necessary to balance the capital cost of heat exchange surface with the cost per B T U lost in the process. T h e
cost of invert formation increases rapidly with temperature and
becomes the major consideration.
.
T h e magnitude of these factors for a typical installation has
an important influence on the selection of the cation exchanger.
Since invert formation increases with operating temperature and

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JOURNAL OF THE A. S. S. B. T.

decreases with increasing crosslinkage of the cation exchanger,


the o p t i m u m operating t e m p e r a t u r e can be increased by the
p r o p e r selection of exchanger. For example, A m b e r l i t e IR-120
shows a m i n i m u m cost value at 16C. while Amberlite IR-124
gives an o p t i m u m cost at 30 C.
Steam
requirement
Dilution of juice d u r i n g displacement will occur a n d is much
d e p e n d e n t u p o n the purity of the juice. It would be expected
that the deionized juice would evaporate efficiently w i t h o u t scale
formation so that the steam useage d u e to dilution would be
counterbalanced by improved efficiency in the evaporator. It is
also unlikely that all the juice would be processed so that the
actual difference in steam r e q u i r e m e n t m u s t be evaluated for
each plant.
Sweet water utilization
T h e ion exchange process will result in varying amounts of
sweet water production d e p e n d i n g on the juice purity. The
utilization of this sweet water in the diffuser a n d for lime water
make up becomes an i m p o r t a n t problem in the reduction of
sugar losses. T h e following calculation will a t t e m p t to establish
how m u c h sweet water can be handled in a 6,000 t o n / d a y factory
operating with ion exchange on beet liquors of varying purities.
T h e following scheme will be used for the calculation

SWEET WATER RE-USE SYSTEM

In practice 50-75% r e t u r n from presses is used with a water


r e q u i r e m e n t of 1.0 lbs w a t e r / l b beets. It should be possible to
substitute sweet water from the ion exchange p l a n t for the press
water without h u r t i n g purity while the press water can be used
for lime preparation. We could also use high levels since the
ion exchange sweet water does not contain as m u c h non-sugars
as the press waters.
6,000 ton/day plantwater balance (lime plant)
Fresh W a t e r = 6,000 ( 0 . 2 5 ) = 1 , 5 0 0 tons/day
Sweet W a t e r a n d Press W a t e r = 6,000 ( 0 . 7 5 ) = 4 , 5 0 0 tons/day

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Lime Feed
Lime Salts
In a 3 0 % Slurry
Water Required, Lime System == 350 tons/day
Water from ion exchange
T h e amount of sweet water will depend on the success in the
design and operation of the equipment. It is likely that the
sweet water will not exceed about 4 bed volumes for each bed
volume in operation. For a 6,000 ton/day plant on 85.5% purity,
the amount of sweet water will be:
391 tons of sweet water
For the other purities under consideration, the sweet water
generated by ion exchange becomes:

It is evident from these estimates that the sweet waters produced by ion exchange can be returned either as lime slurry
or as part of the pulp water returned to the diffuser.
Ion exchange system waste disposal
Since it is likely that amino acid recovery cannot be justified
at the present time due to economic reasons, the spent regenerant
from the ion exchange plant will require treatment as a waste.
An indication of the actual amount of waste produced by ion
exchange is of interest in the adaptation of the process to an
existing factory. T h e 6,000 ton per day mill will be used operating on a 85.5% purity juice as the basis for calculations.

Cation regenerant
About 50 tons of sulfuric acid will be used in regeneration
of which about 50% is converted to salts during regeneration.

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JOURNAL OF THE A. S. S. B. T .

V o l u m e of spent regenerant will be a b o u t five times the total


cation resin volume r e q u i r e d for a day's operation.
13,500 x 5 x 7.46 === 505,000 gallons

Anion
regenerant
A b o u t 9 tons of N H 3 are r e q u i r e d and this in t u r n is treated
with lime to yield 15 tons of a spent lime slurry containing a
m i x t u r e of non-sugars. T h e non-sugar loaded on A m b e r l i t e IRA68 a m o u n t s to a b o u t 16 lbs/ft 3 .
Load
Waste
T h e spent regenerant would have a volume a b o u t five times the
total anion exchange resin volume r e q u i r e d for a day's operation.
10,000 x 5 x 7.46 = 374,000 gallons
Concentration of Solid
T h e B O D r e q u i r e m e n t s of the ion exchange waste would be
expected to be similar to that found in a Steffen process. The
ion exchange, waste a m o u n t s to a b o u t 150 gallons per ton of
beets while values of a b o u t 120 gallons per ton are reported
for Steffen's process by McGinnis (4). T h e B O D would probably be a b o u t
of the total a m o u n t found in the process
water a n d would be concentrated in a b o u t
of the total
waste effluent. Backwash water from the beds would be used
in the flumes a n d condensers a n d as such would n o t be considered as a dirtct waste related to the ion exchange process.
Economic Considerations
T h e economic evaluation of this deionization process requires detailed knowledge on the particular mill involved. A
j u d g e m e n t on profitability requires an estimate on the future
m a r k e t price of refined sugar a n d molasses d u r i n g the period
that t h e plant is b e i n g amortized. Certain general values can be
reached however which will serve as an i n d e x for the economic
evaluation of the process.
T h e purity of t h e juice a n d t h e mill size are important influences on t h e r e q u i r e d capital investment. F o r example, a
6000 ton per day mill treating of juice of 8 2 % purity will cost
a b o u t $860,000 or 143 dollars p e r ton of capacity. Capital cost
is also related to non-sugar removal
of non-sugar removed/day. F o r a

VOL.

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3,

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1964

279

increase of 1% in purity would decrease the capital investment


by $35,000. A proportional reduction would be found for smaller
plants.
T h e cost of removal for non-sugars varies from 2.8 to 8.6
cents/lb for a 6000 ton per day mill treating juices having purity
ranges of 82 to 9 6 % T h e cost rises sharply above 90% purity
and the operation becomes unprofitable at a refined sugar price
of 7 cents per lb and a molasses price of 3 cents/lb. For small
plants the cost of removal is somewhat higher but follows the
same pattern.
T h e operating cost breaks down roughly according to the
following distribution: depreciation (41%), chemicals and water
(49%), fuel (5%), and labor (5%) . T h e contribution of labor
would increase for small plants since it is almost as difficult to
operate a 1500 ton per day mill as a 6000 ton/day mill. As expected the contribution of fixed charges is very high and closely
related to the method of cost accounting used in the mill. The
ion exchange resin contributes about 30-35% of the total capital
cost and its depreciation represents about 14-15% of the total
daily cost when depreciated on a 5 year-100 day/year basis.
Comparison with the Steffen Process
Steffen's process has long been used to increase the extraction
of sugar from beets. While it is a simplification to state that
the ion exchange process is a substitution for the Steffen's House,
it is important to compare the two processes from an economic
standpoint. T h e following table gives a comparison for a 6,000
ton/day beet plant which would feed a Steffen's House rated at
400 tons molasses/day.

280

JOURNAL OF THE A. S. S. B. T.

cost p e r 100 lbs of sugar recovered indicate a significant advantage for t h e ion exchange process.
Conclusions
M o d e r n ion exchange resins have been developed which have
affected materially the economics of a two b e d deionization
process. U n d e r proper conditions it is possible to show a significant gain in sugar recovery at a capital cost of a b o u t 100-125
dollars/ton of beets processed. T h e capital write-off period not
including taxes was in range of 120-200 operating days. The
cost for non-sugar removal varied from 4-5 c e n t s / l b depending
on juice purity a n d p l a n t size.
A comparison with the Steffen H o u s e process was q u i t e favorable to ion exchange w h e n compared at the same depreciation
rate. T h e waste problems were n o t aggrevated a n d the sweet
water utilization was n o t unreasonable. Cooling water would
be a serious p r o b l e m which can be controlled at least in part by
p r o p e r section of the cation exchanger.
Acknowledgements
T h e a u t h o r is grateful to m e m b e r s of Society M1NOC, a
subsidiary of the R o h m & H a a s Company for collecting data on
field operation of the two b e d system; to Tapan O r g a n o for providing valuable field data on invert formation; to Dr. M. Andrus
of R o h m & Haas Company for valuable advice, direct assistance
a n d m u c h editorial help, a n d finally to m a n y individuals in the
American Beet Sugar Industry w h o have given critical advice and
stimulating discussion w h e n this paper was in its early stages.
References
(1) ASSALINI, G. and G. BRANDOLI. 1961.
sugar refining. I. Purification of beet
Sugar Beet Technol. 11: 341-349.
(2) ASSALINI, G. and G. BRANDOLI. 1961.
sugar refining. II. Purification of sugar
Beet Technol. 11: 349-357.
(3)

CARRUTHERS,

A.,

J.

V.

DUTTON,

J.

F.

T.

Ion exchange process for beet


difiusion juice. J. Am. Soc.
Ion exchange process for beet
solutions. J. Am. Soc. Sugar
OLDFIELD,

M.

SHORE and H.

TEAGUE. 1960. Juice composition in relation to factory performance.


T h i r t e e n t h Annual Technical Conference, British Sugar Corpora
tion, Ltd.
(4) M C G I N N I S , R. A. 1951. Beet Sugar Technology, Reinhold Publishing
Corporation, New York, N.Y.
(5) PAYNE, G. W. 1953. T h e physical and technical conditions in sugar
manufacture, Chapter 11. In Honig's Principles of Sugar Technology
Vol I, Elsevier, Amsterdam, Netherlands.
(6)

STARK, J. B., M. M. SANDOMIRE and R. M.

MCCREADY.

A study of the

relation of purity and non-sugars in molasses, Western Utilization


Research Branch Albany, Cal., Agricultural Research Service U. S.
Department of Agriculture.

Methods of Loosening T i g h t Seed Caps in Monogerm


Seed to Improve Germination
F.

H.

PETO 1

Received for publication February

24, 1964

T h e germination of monogerm seed produced in British


Columbia under good conditions is frequently lower than germination of multigerm seed found in the same sample. This fact
is illustrated by the data from a greenhouse soil emergence test
on monogerm and multigerm seed from the same seed sample of
CS36, Contract 3. T h e germ count of the multigerm was determined by examination and the emergence results of spaced seeds
are shown in T a b l e 1. T h e monogerm emergence on a single-germ
basis was 32.5% as compared to 44.8% for the multigerm. T h e
comparison on a seed ball basis was 32.5% for the monogerm as
compared to 69.5% for the multigerm. T h e cause for differences
in emergence obviously lies in the actual physical or physiological
differences as between the multigerm and monogerm. The present
report has to do mainly with the physical problem of tight seed
caps and methods of loosening them.

282

JOURNAL OF THE A. S. S. B. T.

ing germination. Sodium hydroxide a n d sodium hypochlorite


were also effective in loosening the seed caps, b u t depressed
germination.
Enzyme
Treatments
It was reasoned that if the pectins and hemicelluloses were
the cementing substances, then the enzymes hemicellulase and
pectinase should loosen the seed caps a n d improve germination.
T r e a t m e n t s reported in T a b l e 2 show that hemicellulase was the
more effective and gave responses over a very wide range of concentration. W h i l e enzymes could be economically used to increase laboratory germinations, it was felt that the acid treatments might prove to be more effective a n d likely m u c h cheaper
to use on a commercial scale.
Table 2.Effect of enzyme treatments on germination of monogerm seed.

Alternate Soaking and Drying


W h i l e soaking seed has long been employed to remove materials toxic to germination, the possibility of soaking and drying,
thus loosening the seed caps, appeared worthy of investigation.
T h e results of one test are shown in T a b l e 3. In this test the
germination was improved 1 7 % a n d 1 5 % by three a n d four one
h o u r soakings followed by drying u n d e r a heat l a m p . T h e treatments are, however, too t i m e c o n s u m i n g to have commercial
possibilities.
Table 3.Effect of alternate soaking and drying on germination of monogerm seed.

Laboratory
Scale Acid
Treatments
In a preliminary test on CS36, Contract 3, it was found that
a two-hour treatment with 3% H C L , followed by two-hour wash
ing before placing in the germinator, raised the germination from
5 7 % for u n t r e a t e d seed to 7 7 % for the H C L treated seed

VOL.

13,

No.

3,

OCTOBER

1964

283

Five different lots of seed were treated with 3% HCL for two
hours, and washed for two hours without subsequent drying with
the results as shown in Table 4. This acid treatment improved
germination 2 4 % to 2 7 % on treatments where germination was
between 4 1 % and 50%, but gave inconclusive results on lots
where untreated germination was already in the 74% to 78%
range.
Table 4.Effect of 3% HC1 treatments.
Seed
CS
CS
CS
CS
CS
CS
CS
CS
CS
CS

36-3
36-3
41-4
41-4
36-5
36-5
33-11
33-11
40-23
40-23

Material treatment

HC1 3 %
H2O

HC1 3%
H2O

HC1 3 %
H2O

HCI 3%
H2O

HC1 3 %

Hours treated

Hours wash

2
2
2
2
2
2
2
2
2

% Germination
47

2
2
2
2

71
50
77
41
65
78
71
74
77

A test was designed to determine whether washing was essential after treatment with 3% acid. Seed that was washed after
treatment gave 2 9 % improvement in germination, whereas, seed
that was not washed after acid treatment gave 28% and 32%
improvement. Therefore, washing may not be essential, but
washed seed should be less damaging on paper bags during shipment and storage, and less corrosive on seed drills and would,
therefore, be preferred for commercial use.
Phosphoric acid treatments were also tried giving in some
cases, improvement comparable to 3% HCL, but the optimum
concentration was somewhat higher than for HCL and the results were more variable.
Neutralization of either H 3 PO 4 or HCL after treatment by
accurate titration caused no damage but in other tests, where the
seed was neutralized by excess N H 3 gas, germination was severely
reduced. T h i s might have been anticipated since it has been reported that naturally occurring ammonia compounds inhibit
germination.
A further test to determine the optimum ratio of seed to acid
solution is shown in Table 5. T h e ratio of 3 seed to 1 acid by
weight indicated insufficient acid, whereas, an acid soak of onepart seed to two-parts acid resulted in less benefit than the intermediate ratios. These results lead to the opinion that percolation
with excess acid might be a practical commercial treatment. This
should insure that all surfaces are exposed to acid action without
damage by too deep a penetration from an excess soaking in acid.

284

JOURNAL OF THE A. S. S. B. T
Table 5.Seed-acid ratio of 8% add to seed (CS 36, contract 3).

Seed-acid
ratio

Hours treated

3:1
2:1
1:1
1:2

2
2
2
2

Hours drying
3
3
3
3

% Germination

Abnormals

76
85
91
76

1/2
1/2
1/2
1/2

3
2
0
l

Percolation with Muriatic Acid

F u r t h e r acid treatments were r u n to provide leads on com


mercial treatment methods. T h e s e were designed to suit operating
conditions in the L a d n e r seed cleaning plant where t h e seed is
h a n d l e d a n d stored in 45-bushel plywood boxes. T h e plant is
also e q u i p p e d with a New H o l l a n d Model 733 Drying Bin which
has a 10,000-pound capacity. T h e present tentative plan is to
p u t screen bottoms in about n i n e storage boxes, fill these to
within 8 to 10 inches of the t o p with processed seed. T h e boxes
will then be flooded with 3% muriatic acid from an overhead
tank. T h e seed will be allowed to drain for two hours and then
the excess acid will be flushed away with water. T h e seed will
t h e n be elevated i n t o the drier which must be filled to capacity
to permit the drier to operate.
In order to o b t a i n indications of the effectiveness of the above
method, a series of treatments were r u n using a 4" tile, three feet
long, as a percolator chamber. T h i s tile was stood upright on a
Buckner funnel a n d six p o u n d s of seed was percolated with 12
p o u n d s of 3% muriatic acid. T h e t r e a t m e n t period was two hours
a n d t h e excess acid was washed o u t with three changes of water,
T h e seed was then dried with a small-scale forced-air drier at
temperatures of a b o u t 105F to a p p r o x i m a t e conditions in the
commercial drier. T h e results of these tests are shown in Table
6. T h e results were q u i t e consistent a n d gave an average improve
merit in germination of 2 1 % a n d 2 0 % for the two- and three
h o u r t r e a t m e n t period, respectively.
Table 6.Effect of percolation with muriatic acid.
Seed
CS 36-3
CS 36-3
CS 36-3
CS 41-4
CS 41-4
CS 41-4
CS 36-5
CS 36-5
CS 56-5

Treatment
material

Seed-add
ratio

Draining No. of
hours
washings

H2O

26% Mur
26% Mur

1:2
1:2

2
3

1:2
1:2

2
3

1:2
1:2

2
3

HsO

3% Mur
3% Mur
H2O

3% Mur
3% Mur

0
3
3
0
3
3
0
3
3

Hours
drying % Germination
0
2-3
2-3
0
2-3
2-3
0
2-3
2-3

68
88
80
57
80
80
56
76
8O

VOL.

13,

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3,

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1964

285

Discussion
T h e e v i d e n c e p r e s e n t e d i n this p a p e r indicates that o n e o f
the i m p o r t a n t l i m i t i n g factors in t h e g e r m i n a t i o n of certain lots
of m o n o g e r m seed g r o w n in t h e V a n c o u v e r area is the tightness
of t h e seed c a p . S n y d e r (2) r e p o r t e d t h a t w h e n cloned seedb e a r i n g p l a n t s w e r e g r o w n a t m e a n t e m p e r a t u r e s o f 66 a n d 76
F, a g r e a t e r p r o p o r t i o n of loose seed caps o c c u r r e d at t h e h i g h e r
t e m p e r a t u r e . Since J u l y a n d A u g u s t m e a n t e m p e r a t u r e s i n Vancouver a v e r a g e 6 3 F a n d t h e m e a n at P h o e n i x is p r o b a b l y above
76 for t h i s p e r i o d , it is p r e d i c t a b l e t h a t t i g h t caps should be a
c o m p l a i n t in B r i t i s h C o l u m b i a a n d loose caps s h o u l d be a problem in P h o e n i x . C l i m a t i c c o n d i t i o n s at Salem m o r e closely resemble V a n c o u v e r t h a n P h o e n i x a n d t w o p o o r g e r m i n a t i o n lots
from O r e g o n w h i c h w e r e given t h e acid t r e a t m e n t were i m p r o v e d
2 3 % i n g e r m i n a t i o n w h i c h indicates t h a t t h e tight seed cap problem also exists i n O r e g o n .
T i g h t seed c a p s h a v e n e v e r b e e n recognized as a p r o b l e m in
m u l t i g e r m seed, b u t t h i s m a y be because t h e i r seed caps are
n a t u r a l l y t h i n n e r as was f o u n d in t h e o n e case e x a m i n e d .
W h i l e a c i d t r e a t m e n t s of m o n o g e r m seed may prove practical
a n d e c o n o m i c a l on a c o m m e r c i a l scale, it s h o u l d be possible to
b r e e d s t r a i n s w i t h t h i n n e r a n d looser seed caps. I n b r e d lines
now o n h a n d w i l l b e g r o w n i n British C o l u m b i a t o see w h e t h e r
sufficient useful v a r i a t i o n exists.
Conclusions
1. M o n o g e r m seeds g r o w n in t h e V a n c o u v e r area have b e e n
s h o w n t o possess t h i c k e r a n d t i g h t e r seed caps t h a n m u l t i g e r m
seed f r o m t h e s a m e s a m p l e . T h e s e thick tight seed caps a p p e a r
t o b e t h e m a i n r e a s o n t h a t m o n o g e r m has a lower percentage
g e r m i n a t i o n t h a n m u l t i g e r m g r o w n u n d e r the same conditions.
2. G e r m i n a t i o n of m o n o g e r m seeds h a v i n g tight seed caps was
greatly i m p r o v e d by c h i p p i n g off p a r t of t h e cap, by altern a t e s o a k i n g i n w a t e r a n d d r y i n g , b y enzyme t r e a t m e n t s a n d
treatments with 3 % H C L .
3 . S e m i - c o m m e r c i a l t r e a t m e n t s b y percolation with 3 % m u r i a t i c
w e r e successful i n b r i n g i n g g e r m i n a t i o n u p t o acceptable
levels. A c i d t r e a t m e n t s on a c o m m e r c i a l scale presents no
difficulties, b u t t h e d r y i n g of soaked seed in large quantities
will be slow a n d s o m e w h a t risky.

286

JOURNAL OF THE A. S. S. B. T.

Acknowledgments
T h e a u t h o r wishes to acknowledge the c o n t r i b u t i o n of G. M.
Guccione on the effect of alternate soaking a n d drying, of F. R.
Low a n d A. Schmand for the contributions to the seed treatment
a n d germination experiments, a n d of Sam C. C a m p b e l l for supplying low-germinating samples of Oregon-grown seed.
Literature Cited
(1) LACKEY, C. F. 1948. Chemical loosening of seed caps in relation to
germination of sugar beet seed. Proc. Am. Soc. Sugar Beet Techno].
V: 66-69.
(2) SNYDER, F. W. 1959. Influence of the seed ball on speed of germination
of sugar beet seeds. J. Am. Soc. Sugar Beet Technol. 10(6) : 513-520.

JOURNAL
of the
American Society of Sugar
Beet Technologists
Volume 13
Number 4
January 1965

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado, U. S. A.

Subscription
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Made in the United States of America

TABLE OF CONTENTS
Author
Instrumentation
Corporation

within

the

British

Page

Sugar
J.

E. A. Rich..

.287

O p e r a t i o n of juice softeners in a beet sugar


factory
-L. P. Orleans
W. A. Harris
L. W. Norman
H. W. Keller...
Sugar beet p r o d u c t i o n in Michigan as affected
by crop sequence a n d fertility levels
-L. S. Robertson
R. L. Cook
CD.
Piper
R. H. Dowdy
J. F. Davis
T h e effects of five legume crops on soil
p o p u l a t i o n s of t h e sugar beet nematode
(Heterodera
schachtii
Schmidt)
Arnold E. Steele
Charles Price
Historical highlights in sugar beet harvest
mechanization
.Austin A. Armer..
Some effects of ionizing r a d i a t i o n on the meta-Myron Stout
bolism a n d g r o w t h of sugar beets
John D. Spikes
Impact effects on g e r m i n a t i o n a n d seedling
vigor of sugar beets

On the n a t u r e of h a t c h i n g of Heterodera
schachtii.
I I . N a t u r a l sources of hatching s t i m u l a n t s
T h e p e r f o r m a n c e of sugar beet selections
m a d e for p u r i t y a n d chloride at three levels
of n i t r o g e n fertility

mosaic

virus

in

304

-314
.318

333

O. R. Kunze
F. W. Snyder
C. W. Hall

341

D. R. Viglierchio
P. K. Yu

354

C. W. Doxtator
R. E. Finkner
R. H. Helmerick
P. C. Hanzas

Resistance t o the sugar beet n e m a t o d e


(Heterodera schachtii) in
F
tetraploid
hybrids b e t w e e n Beta vulgaris a n d Beta
..Helen
Savitsky
patellaris
Charles Price
Occurence of t h e alfalfa
sugar beet in California

296

R. J. Shepherd
D. H. Hall
D. E. Purcifull

362

370

374

Instrumentation W i t h i n the British Sugar Corporation


J.

E.

A.

RICH1

Received for publication February 28, 1964

I n t h e e a r l y years after t h e war, i n s t r u m e n t a t i o n w i t h i n the


British S u g a r C o r p o r a t i o n was very sparse, a n d consisted mainly
o f i n d i c a t i n g a n d r e c o r d i n g t h e r m o m e t e r s , pressure gauges, a n d
steam f l o w m e t e r s w i t h a n occasional j u i c e m e t e r . T h e r e were
also a few i n d u s t r i a l p H m e t e r s used m a i n l y o n 2 n d carbonation
a n d t h i n j u i c e , b u t these w e r e a n y t h i n g b u t reliable.
All factory e q u i p m e n t was old a n d consisted mainly of a series
of b a t c h processes w h i c h d i d n o t l e n d themselves to a u t o m a t i c
control.
In t h e early 1950's B.S.C. e m b a r k e d u p o n a large reconstruct i o n p r o g r a m to m o d e r n i z e its factories. Each year one or m o r e
factories w e r e c h o s e n t o replace old batch plants with m o d e r n ,
c o n t i n u o u s m a c h i n e r y . Beet-end e q u i p m e n t was the first to be
r e p l a c e d , a n d w i t h c o n t i n u o u s diffusion a n d c o n t i n u o u s carbonation, t h e o p p o r t u n i t y arose t o increase the i n s t r u m e n t a t i o n .
I n s t r u m e n t a t i o n at t h a t t i m e was t h e prerogative of t h e W o r k s
M a n a g e r ( P l a n t S u p e r i n t e n d e n t ) at each factory with guidance
from his T e c h n i c a l Supervisor, a n d t h e Chief Engineer, b u t t h e r e
was n o over-all i n s t r u m e n t a t i o n p r o g r a m . I t was n o t u n t i l 1955
that a C o n t r o l E n g i n e e r i n g : D e p a r t m e n t was formed with t h e
p u r p o s e of d e s i g n i n g , specifying a n d c o o r d i n a t i n g the instrumentation t h r o u g h o u t the Corporation. T h e Works Manager
still h a d t h e final w o r d , b u t was n o w advised by a d e p a r t m e n t
w h i c h q u i c k l y amassed a w e a l t h of experience a n d know-how.
T h u s , w h e n a n e w p r o j e c t was p r o g r a m e d , t h e W o r k s M a n a g e r
w o u l d discuss t h e i n s t r u m e n t a t i o n with the C o n t r o l E n g i n e e r
w h o c o u l d a d v i s e o n p r o v e n practices a t o t h e r factories, a n d
w o u l d t h e n e n g i n e e r t h e scheme at C e n t r a l Offices.
T h e choice of the instrument manufacturer would depend
to a l a r g e e x t e n t u p o n c o m p e t i t i v e q u o t a t i o n s , b u t with a view
to s t a n d a r d i z e as m u c h as possible at a p a r t i c u l a r factory. T h i s
p r e s e n t e d n o p r o b l e m s w i t h t h e f i r s t schemes a s all the i m p o r t a n t
i n s t r u m e n t s w e r e n e w , b u t i n later years t h e aim was t o k e e p
the n u m b e r of manufacturers represented to a m i n i m u m , and
still a l l o w c o m p e t i t i o n . It was felt t h a t standardization taken to
t h e u l t i m a t e o f o n e m a n u f a c t u r e r only, w o u l d b e d e t r i m e n t a l t o
t h e factory, p a r t i c u l a r l y w i t h r e g a r d to service. In a d d i t i o n , some
pieces of e q u i p m e n t w e r e considered to be b e t t e r m a d e , or in1

Instrument Engineer, Holly Sugar Corporation. Colorado Springs, Colorado.

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JOURNAL OF THE A. S. S. B. T.

deed, only m a d e by certain manufacturers a n d these could be


specified irrespective of the major supplier.
O n c e the over-all scheme h a d been settled between the Works
Manager a n d the Control Engineering D e p a r t m e n t , the latter
w o u l d produce a flow schematic showing all the control loops
a n d an outline drawing for the panel, if required. T h e y would
also accurately specify each instrument or valve so that all quotations would be truly comparable. Over a period of years several
standard panel shapes were evolved, together with very complete
specifications. T h e s e were found to be of real value, as panel
prices often varied by as m u c h as 2 0 0 % between manufacturers,
whereas the i n s t r u m e n t prices were very close.
Orders would be placed with the selected manufacturer as
early as possible, preferably in the August or September, twelve
m o n t h s before the e q u i p m e n t was d u e to be commissioned. Extended delivery times dictated this, b u t it also allowed the dep a r t m e n t to concentrate on the more immediate problems of
commissioning the e q u i p m e n t just installed.
W h e n e v e r time permitted, and always with large installations,
a reasonably comprehensive " m a n u a l " would be produced, describing in simple terms the operation a n d function of all the
instruments being provided. T h i s " m a n u a l " was produced primarily for the factory operating personnel, b u t also proved its
worth within the d e p a r t m e n t both as a guide (if m o r e than one
new installation was being commissioned) a n d as a reference
when designing new i n s t r u m e n t schemes.
P r i o r to the start of the campaign the d e p a r t m e n t would
check the installation and supervise a t h o r o u g h test of all new
instruments a n d control panels. T h i s test would be carried out
d u r i n g the precampaign steam a n d water trials so that whenever
possible the control functions could be checked. If possible, a
service engineer from the i n s t r u m e n t m a n u f a c t u r e r would be
present d u r i n g these tests as he was in a good positon to obtain
any spares or extra items if they were urgently needed.
At the start of the campaign one or more engineers from
the d e p a r t m e n t would be present to oversee the start-up and
set up the controller functions. Sometimes this r e q u i r e d being
on site for a week or longer, b u t usually the p l a n t would be
operating efficiently w i t h i n two or three days.
Setting t h e controller
important job and under
T h e usual procedure was
t h e factory could operate

functions was u n d o u b t e d l y the most


start-up conditions this was not easy.
to set them sufficientlv close so that
adequately a n d t h e n adjust them, say

VOL. 13, No. 4, JANUARY 1965

289

two w e e k s l a t e r , w h e n t h e factory h a d settled d o w n a n d was


r u n n i n g at capacity.
C a r e f u l r e c o r d s w e r e k e p t of all c o n t r o l l e r settings a n d these
were of g r e a t assistance w h e n c o m m i s s i o n i n g similar e q u i p m e n t
in o t h e r factories.
R e c o r d s w e r e also k e p t at each factory of t h e c o n t r o l l e r o u t p u t
pressures for k n o w n c o n d i t i o n s . F o r instance, w i t h a magnetic
f l o w m e t e r a n d v a l v e c o n t r o l l i n g t h e m i l k o f l i m e f l o w t o lst
c a r b o n a t i o n , t h e c o n t r o l l e r o u t p u t w o u l d b e n o t e d for a b o u t three
definite flows. If, at a n y t i m e , t h e flowmeter failed, t h e valve
position c o u l d be set to give a k n o w n flow, w i t h o u t completely
d i s r u p t i n g t h e c a r b o n a t i o n station.
W i t h t h e i n c r e a s e of i n s t r u m e n t a t i o n w i t h i n the factories
came t h e n e e d for p r o p e r l y t r a i n e d i n s t r u m e n t mechanics. I n
the early days, i n s t r u m e n t m a i n t e n a n c e h a d b e e n looked after
by t h e l a b o r a t o r y , b u t this is no longer possible. As factory engin e e r i n g w a s a l r e a d y split b e t w e e n mechanical a n d electrical, i t
was d e c i d e d t h a t t h e i n s t r u m e n t mechanics should b e supervised
by t h e W o r k s E l e c t r i c i a n . T h i s system has w o r k e d well, especially
as m a n y of t h e p r e s e n t day i n s t r u m e n t m e n were once electricians.
I n o r d e r t o e n c o u r a g e m e n n o t t o consider i n s t r u m e n t work a s
a d e a d - e n d j o b , p r o m o t i o n was possible e i t h e r in the electrical
or processing fields. T h u s a skilled m e c h a n i c with a good knowledge of t h e process c o u l d b e c o m e a beet- or sugar-end foreman
and eventually a shift-superintendent.
T r a i n i n g was c a r r i e d o u t b o t h i n t h e factory a n d a t schools
r u n b y t h e i n s t r u m e n t m a n u f a c t u r e r s . O n several occasions i t
was possible to a r r a n g e special courses for B.S.C. personnel only.
In this way, o n l y those i n s t r u m e n t s of p a r t i c u l a r interest w o u l d
b e covered, a n d m e c h a n i c s from several factories h a d t h e opport u n i t y t o m e e t a n d discuss p r o b l e m s a m o n g themselves.
T h e f i r s t b i g i n s t r u m e n t scheme installed b y B.S.C. was a t
W i s s i n g t o n in 1957 ( l ) 2 N o t only was this t h e first factory to
have over-all c o n t r o l from t h e b e e t washers to t h e thick juice
tank, b u t it was also t h e first t i m e t h a t electronic controllers were
used on a l a r g e scale by B.S.C. M u c h could be w r i t t e n on t h e
pros a n d c o n s of p n e u m a t i c s versus electronics, b u t suffice to
say in t h i s i n s t a n c e t h a t E v e r s h e d & Vignoles was t h e only British
c o m p a n y w h o a t t h a t t i m e c o u l d m e e t t h e c o m p l e t e specification.
T h e r e c o n s t r u c t i o n a t W i s s i n g t o n involved p r i m a r i l y t h e
r e p l a c e m e n t of a diffusion b a t t e r y by a 5.4 m e t r e R . T . diffuser.
T h i s was t h e first t i m e t h a t B.S.C. h a d installed such a diffuser
2

Numbers in parentheses refer to literature cited.

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on the site of the battery it replaced, a n d the o p p o r t u n i t y was


taken to revise m u c h of the beet-end layout. T h e reconstruction
was completed in u n d e r seven m o n t h s and the instrumentation
was installed by factory personnel in less t h a n three weeks.
T h i s plant was now c o n t i n u o u s from the slicers to t h e thick
juice tank, with an absolute m i n i m u m of intermediate receiving
or waiting tanks, and the revised layout reduced the manpower
r e q u i r e m e n t to a m i n i m u m . O n e aspect of this was that only
one m a n was r e q u i r e d to attend both 1st a n d 2 n d carbonation
a n d the evaporators. In view of this it was impossible to justify
the great expense of m a k i n g the evaporators completely automatic.
Several simple electric analogue computers were installed in
the over-all scheme, a n d one of the most interesting applications
was to continuously display the a m o u n t of solids present at the
evaporators (4). T h u s , the supervisor knew w h e t h e r the inlet
sugar quantity was matched with the outflow.
In order to simplify the measurements across the evaporators,
the t h i n juice Brix was assumed to be constant. U n d e r these
conditions only three variables had to be measured, namely thin
juice flow, thick juice flow a n d thick juice Brix.
T h e signals representing thick juice flow (F,) a n d its specific
gravity (C) were fed i n t o the first of two simple computers,
solving the following equation which is a measure of the mass
of the solids leading the evaporators.
M = 0.1157F 1 C
1.124F1
(1)
T h e o u t p u t (M) from this computer, together with the signal
representing thin juice flow (F 2 ) were fed into the second comp u t e r solving this new equation, to give the mass flow difference
across the evaporators.
T = M - F 2 + 15
(2)
Equations 1 a n d 2 were derived as follows:
Assuming all solids as sugar, the mass flow across the evaporator may be written as:
f1b1 = f 2 b 2
(3)
where f 1 = thick juice flow in l b / m i n
f 2 = t h i n juice flow in l b / m i n
b 1 = thick juice Brix
b 2 = t h i n juice Brix
T h e thick juice specific gravity was measured by a Rotameter
densitometer which transmitted 20 30 ma over t h e range 1.20
- 1.45.

VOL. 13, N o . 4, J A N U A R Y 1965

291

W i t h i n t h e r a n g e of sp. gr. 1.20 t h r u 1.45, B r i x equals approximately [ (175 X sp. gr.) ] 165, so e q u a t i o n (3) can therefore be
writ-ten:

T h e t h i n j u i c e B r i x (b 2 ) was assumed t o b e 13.5.


T h e t h i n j u i c e f l o w t r a n s m i t t e r was r a n g e d 0-4480 l b / m i n
a n d gave 0-30 ma for this r a n g e . (F 2 ).
T h e t h i c k j u i c e f l o w t r a n s m i t t e r was r a n g e d 0-1600 l b / m i n
a n d gave 0-30 ma for this r a n g e (F 1 ).
E q u a t i o n (4) t h e n b e c o m e s :

If t h e mass flow across t h e evaporators was in balance, t h e n


F 2 = M a n d T = 15 m a . T h i s o u t p u t ( T ) was recorded on a
0 30 ma r e c o r d e r a n d was n o r m a l l y at mid-scale. If t h e mass
e n t e r i n g e x c e e d e d t h a t leaving, t h e r e c o r d e r fell below mid-scale
a n d vice-versa.
A l t h o u g h t h e t h i n juice B r i x was assumed to be 13.5, the
e q u a t i o n s c o u l d be modified to allow for any Brix, thus e q u a t i o n
(5) c o u l d be r e - w r i t t e n as:
K F 2 = 0.1157F 1 C 1.124F 1 .
.................(6)
w h e r e t h e coefficient (K) r e p r e s e n t s t h e difference between 13.5
a n d t h e a c t u a l B r i x . T h i s coefficient could b e a d d e d t o the comp u t e r by p l a c i n g a p o t e n t i o m e t e r across the c o m p u t e r coil receiving t h e t h i n j u i c e flow signal, t h u s p r o v i d i n g a simple m a n u a l
c o r r e c t i o n for t h i n j u i c e B r i x .
A n o t h e r e x a m p l e of t h e use of simple c o m p u t e r s , or comp u t i n g relays is t h a t used at t h e King's L y n n factory to m o n i t o r
t h e flow of j u i c e i n t o a n d t h r o u g h the purification p l a n t . It is
n o w u n i v e r s a l l y a c c e p t e d t h a t a really steady flow of raw j u i c e
is a p r e r e q u i s i t e of success at first c a r b o n a t i o n using s i m u l t a n e o u s
lime a n d gas a d d i t i o n . E v e n in t h e best factories t h e r e comes a
t i m e w h e n t h e r a w j u i c e f l o w gets o u t o f step w i t h t h e t h i n juice
f l o w o r b e e t slice r a t e . T h e following description shows t h e
scheme u s e d t o p r o v i d e a u t o m a t i c r e g u l a t i o n t o k e e p the f l o w
rates i n step w i t h o u t v i o l e n t f l u c t u a t i o n s .
T h e i n s t a l l a t i o n a t King's L v n n used two simple p n e u m a t i c
c o m p u t e r s , a n d t h r e e pressure selectors. It is shown schematically
in F i g u r e 1.

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JOURNAL OF THE A. S. S. B. T.

Figure 1.Juice purification schematic at King's Lynn.

T h e pressure selectors were simple devices utilizing a pneumatic balanced diaphragm and an o u t p u t relay. T h e y could be
arranged to select a n d pass either the higher or the lower of two
pressures.
T h e primary controller was the raw juice flow controller.
U n d e r n o r m a l factory conditions the desired value of this controller was set by the shift s u p e r i n t e n d e n t to an estimated maxim u m . T h e simple computers automatically reduced this desired
value if any one of three tank levels moved outside n o r m a l limits.
T h e three levels were those of the t h i n juice, the second carbonation, a n d the raw juice tanks. T h e latter was also used as a preliming tank a n d was fitted with a level controller operating a
valve on the inlet to it.
A l t h o u g h the tanks were different sizes, they were fitted with
transmitters ranged to give 3 to 15 psi signals over their normal
level ranges. T h e thin juice tank was 6 ft deep and if the level
rose above 4 ft, the raw juice flow was reduced. T h e carbonation
tank transmitter operated over the u p p e r 8 ft of the tank; if the
level rose i n t o the u p p e r half of this range, the raw juice flow
was also reduced. T h e raw juice tank was 10 ft deep and if the
level fell below 3 ft-4 in, t h e n the raw juice flow was again reduced. In addition, whenever any of these off-normal level zones
were entered, an audible alarm attracted the attention of the
supervisor.

VOL

13, No. 4, JANUARY 1965

293

T h e t h i n j u i c e level signal was fed i n t o a h i g h pressure selector, t o g e t h e r w i t h a p r e s e t 11 psi signal derived from a pressure
regulator. T h i s selecter w o u l d therefore n o r m a l l y pass 11 psi
a n d w o u l d o n l y pass a h i g h e r signal if the t a n k level rose above
4 ft.
T h e r a w j u i c e level signal was fed i n t o a low pressure selector,
together w i t h a pre-set 7 psi. T h i s selector w o u l d therefore n o r m ally pass 7 psi a n d w o u l d only pass a lower signal if t h e tank level
fell b e l o w 3 ft-4 i n .
T h e signals f r o m these t w o selectors were fed i n t o the first
computer, solving the equation:
Po = (A-C) R K, w h e r e R = 1.5 a n d K = + 15.
T h e o u t p u t of t h i s c o m p u t e r was n o r m a l l y 9 psi, b u t w o u l d
r e d u c e to 3 psi if t h e t h i n j u i c e level rose to 6 ft, or t h e r a w juice
level fell to zero. T h e o u t p u t from this c o m p u t e r was fed i n t o
the second c o m p u t e r , t o g e t h e r w i t h the o u t p u t from a second
high p r e s s u r e selector. T h e i n p u t s t o this were the c a r b o n a t i o n
tank level signal a n d a p r e s e t 9 psi. T h e o u t p u t was therefore
n o r m a l l y 9 psi r i s i n g to 15 psi, as t h e c a r b o n a t i o n tank level rose
above n o r m a l .
T h e s e c o n d c o m p u t e r was also solving the e q u a t i o n :
Po = (A-C) R K, b u t w i t h R = 2 a n d K = + 1 5 .
W i t h t w o n o r m a l i n p u t s of 9 psi, this c o m p u t e r ' s o u t p u t was
15 psi, b u t if t h e c a r b o n a t i o n t a n k level rose, or the o u t p u t from
the first c o m p u t e r fell, t h e n t h e o u t p u t from this second comp u t e r w o u l d fall to 3 psi.
T h e o u t p u t from t h e second c o m p u t e r was c o u p l e d t o t h e
adjustable p n e u m a t i c set p o i n t of t h e raw juice flow controller,
a n d b e i n g n o r m a l l y 15 psi, allowed t h e flow to be set m a n u a l l y
at any d e s i r e d v a l u e . If a n y of t h e t h r e e levels changed from
their n o r m a l state, t h e n t h e o u t p u t o f t h e second c o m p u t e r w o u l d
slowly r e d u c e to 3 psi, a n d this w o u l d automatically r e d u c e t h e
desired v a l u e s e t t i n g of t h e r a w j u i c e flow.
T h e successful c o n t r o l of m i l k of l i m e density in t h e B.S.C.
dates back to 1955 (2) w h e n t h e c o n t i n u o u s density t r a n s m i t t e r
conceived at S p a l d i n g factory was commercially developed. T h i s
i n s t r u m e n t i s n o w i n w i d e use t h r o u g h o u t E u r o p e a n d consists
basically of a b a l a n c e d 1" stainless steel U - t u b e , fitted w i t h e i t h e r
a n electric o r p n e u m a t i c force balance transmitter. T h e t u b e i s
mounted horizontally a n d the liquid to be measured flows through
i t B y k e e p i n g t h e t u b e horizontal, changes i n l i q u i d velocity
do n o t affect t h e t r a n s m i t t e d o u t p u t .
T h e s t a n d a r d i n s t r u m e n t has a span of 0.2 specific gravity,
and for m i l k o f l i m e m e a s u r e m e n t t h e n o r m a l r a n g e w o u l d b e

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1.00-1.20 sp. gr. Special models with spans down to 0.05 sp. gr.
a n d fitted with t e m p e r a t u r e compensation are manufactured, but
the standard instrument has a certain a m o u n t of i n h e r e n t temp e r a t u r e compensation d u e to the expansion of the U-tube, and
is adequate for most sugar factory purposes.
Apart from being totally enclosed a n d therefore clean, this
instrument has the added advantage that it can be positively
checked. It is supplied with weights proportional to the sp. gr. to
be measured and these can be h u n g on the U-tube for direct
calibration.
Milk of lime is produced at B.S.C. factories by slaking uncrushed b u r n t lime with 8 to 9 Brix sweet water in conventional
rotary d r u m slakers. T h e milk of lime is fed from the slaker to
a "thick lime" tank via a vibrating screen to remove the fines.
T h i s tank is the key to the automatic control of b o t h the limekiln a n d the slaker, a n d must be sized correctly d u e to the inter
m i t t e n t discharge from the slaker (3).
T h e level in the thick lime tank is used to control the burnt
lime and sweet water feeds to the slaker. T h e level in the limekiln is controlled by a gamma-switch so that as b u r n t lime is
d r a w n from the bottom, the level is automatically maintained.
Coke a n d limestone are added in a p r e s e t b u t adjustable ratio
via a completely automatic load-cell weigher a n d skip hoist.
T h e thick lime tank level controls are arranged to give a
working zone in the tank. T h u s the level will fall to 4 0 % before
the feeds to the slaker are started, a n d will rise to 8 0 % before the
feeds are stepped. T h e sweet water feed to the slaker is manually
adjusted so that the milk of lime p r o d u c e d is slightly heavier
t h a n that r e q u i r e d by the process.
T h e milk of lime is p u m p e d from the thick lime tank to a
constant head tank above 1st carbonation a n d overflows back.
It is sampled immediately after the p u m p a n d the density is
regulated by a control valve a d d i n g sweet water i n t o the suction
of the p u m p . A position alarm is usually fitted so that the main
slaking valve may be adjusted if the dosing valve shuts or goes
wide open. W i t h this type of control it is possible to regulate the
milk of lime density to within 0.5 Brix.
Acknowledgement
T h e a u t h o r wishes to thank Mr. J. C. Macdonald, O.B.E.
Technical Director of the British Sugar Corporation Ltd for
his permission to publish the data contained in this paper.

VOL. 13, N o . 4 , J A N U A R Y 1965

2 9 5

References
(1) A N O N .

1958. Sugar from beet. T h e Engineer. 51: 878-882.

(2) B O O T H , P. M.
191-192.

1955. C o n s t a n t density milk of lime. Int. Sugar J- 57:

(3) W I T H E R S , R. M. J. a n d J. E. A. R I C H . 1960. Control engineering applied


to the British Sugar Corporation. 13th Tech. Conf. British Sugar
Corporation, Ltd. London.
(4) W I T H E R S , R. M. J. a n d J. E. A. R I C H . 1962. T h e application of simple
c o m p u t e r s to process control. 15th T e c h . Conf. British Sugar Corporation, L t d . L o n d o n .

Operation of Juice Softeners in a Beet Sugar Factory


L. P. ORLEANS, W. A. HARRIS, L. W. N O R M A N , H. W.
Received for publication April 14, 1964

KELLER 1

Introduction
A continual degradation in quality of beets grown in the
Imperial Valley of Southern California has caused a gradual
increase in lime salts concentration of the sugar juices produced
from these beets. T h i s increase in lime salts concentration from
. 0 4 5 % C a O on thin juice solids in 1956 to .084% in 1961, has
made it increasingly difficult to concentrate these juices in the
evaporators because of scale formed on the heating surfaces.
T h e use of ion exchange resins to soften sugar containing juices
has proven successful in E u r o p e a n sugar mills and preliminary
softening trials using juices produced at Holly's Carlton mill,
indicated that this procedure should be economically feasible.
L a b o r a t o r y Tests
Laboratory tests were conducted in 1961 to determine the
best operating conditions for softening of evaporator thin juice.
T h e s e tests indicated that by using IllcoC-211W resin, a maxim u m loading of 1.89 lbs/cu ft as CaCO 3 , or 1.06 lbs CaO could
be expected by the time we reached a leakage of 4 2 % of the
hardness in the juice being treated. T h e average leakage for the
complete cycle would be 1 3 % at this point. Figure 1 gives the
percent of total hardness which leaked t h r o u g h i n t o the effluent
from the softening trials. T h e flow rate established by these
tests was 8 gpm per sq ft of bed cross section. (Used for water
softening).

Figure 1.Percent hardness leakage vs. t h r o u g h p u t .


1 Research Engineer, Research Chemist, General Chemist, Holly Sugar Corporation
Colorado Springs, Coloralo; Assistant Research Director, Illinois Water Treatment, Rock
ford, Illinois, respectively.

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13, N o . 4, J A N U A R Y 1965

297

From these preliminary tests the following equipment sizes


were established. T h e 1,200 gpm flow rate dictated a 150 sq ft
bed cross section in order to hold the average flow to 8 gpm per
sq ft. T h e column diameters were set at 10 feet with two columns
operating in parallel. We need one additional column being
regenerated while two are in service. The minimum bed depth
is 4 ft, making a total volume for each column of 315 cu ft. The
height of the column was set at 11 feet to allow enough free
board for the addition of another 158 cu ft of resin if needed.
Table 1 indicates the service cycle to be expected using a bed
volumn of 315 cu ft.
T a b l e 1.Expected service cycles.
% CaO on
dry substance
0.04
0.06
0.08
0.10
0.12
0.14

Throughput
to 42% Leakage
gallons/cu. ft.
1810
1210
910
730
600
520

Service
time
hours

_____
15.8
10.6
7.9
6.4
5.3
4.5

T h e preliminary tests indicated the desirability of reclaiming


brine for reuse in the following regeneration cycle. The volume
of brine to be reclaimed is 4.5 gallons per cu ft of resin with
8.5% brine concentration. A 10% fresh brine solution is used
for regeneration, at the rate of 7.5 pounds per cu ft of resin.
Equipment
T h e final arrangement of the equipment included three
columns 120 inches in diameter by 132 inches high with the
necessary headers and valve arrangements to facilitate automatic
operation of the units by a timer controlled panel. The columns
have top, center and bottom distributors. The bottom distributor
is covered by a quartz bed which supports the resin bed. T h e
center distributor is 12 inches above the top of the settled resin
bed. T h e top distributor is approximately 3 feet below the top
of the column and is used for the backwashing outlet. T h e
softener installation also includes a brine reclaim pump and
a horizontally mounted brine reclaim tank 78 inch diameter by
120 inches long. T h e softener columns and brine reclaim tank
are protected with a baked epoxy lining. T h e control panel includes a flow meter for each unit with a totalizing integrator
which sounds an alarm when a preset gallonage has passed
through the unit. T h e panel also contains a series of timers to
control the regeneration cycle and the necessary interlocks to

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298

prevent two columns from being p u t i n t o the regeneration cycle


at the same time. T h e complete regeneration cycle is listed in
T a b l e 2.
Table 2.Regeneration cycle.
Flow,
Sequence
Juice blowdown
Sweeten off
Sweeten off rinse
Backwash
Backwash blowdown
Reclaim brine
Fresh brine
Slow rinse
Fast rinse

gpm
600
600
600
450
600
120
120
120
600

Time,
minutes
1
5
5
20
6
12
22
15
10

Total
gallons
600
3000
3000
9000
3600
1440
2640
1800
6000

T h e juice blowdown is accomplished by stopping the juice


flow to the c o l u m n a n d allowing air pressure to blow the juice
from the c o l u m n u n t i l the level has d r o p p e d to t h e surface of
the bed. T h i s decreases the d i l u t i o n of juice w i t h sweetening off
water. Once the level of juice reaches the t o p of the resin bed,
the rinse water is t u r n e d on a n d continues to force the juice
from the c o l u m n back to the s u l p h u r tower for further processing. Sweetening off is c o n t i n u e d u n t i l t h e sugar concentration drops to approximately 2 % . T h e n e x t step is the sweeten
off rinse to finish removing all the sugar from the resin. (This
step was o m i t t e d from the process the second year, to decrease
the water r e q u i r e m e n t s for regeneration, a n d the backwash followed immediately after the sweetening off.)
T h e backwash leaves the c o l u m n t h r o u g h the t o p distributor
so the c o l u m n is completely full of water at the e n d of the backwash a n d this water is blown d o w n t h r o u g h t h e bed and out the
b o t t o m distributor by i n t r o d u c i n g air i n t o the column. This
blowdown continues u n t i l the water level reaches the center
distributor.
T h e reclaim b r i n e is t h e n i n t r o d u c e d t h r o u g h the bottom
distributor a n d flows u p w a r d t h r o u g h the b e d a n d o u t the center
distributor. T h e reclaim b r i n e is followed immediately by the
fresh brine, a n d this fresh b r i n e is reclaimed for t h e next regeneration cycle. T h e fresh b r i n e is followed by a slow rinse up-flow
t h r o u g h the b e d to force all t h e b r i n e t h r o u g h the resin. Once
the slow rinse has b e e n completed, the flow is reversed and the
fast rinse completely washes the r e m a i n i n g b r i n e from the resin
using a down-flow. T h e fast rinse completes t h e regeneration
cycle a n d t h e c o l u m n is ready to be sweetened on at the same
t i m e as the n e x t c o l u m n to be regenerated is sweetened off.

VOL.

13,

No.

4, J A N U A R Y

1965

299

Difficulties of Operation
Screens r e q u i r e c l e a n w a t e r which was n o t available, a n d the
screens w e r e c o n t i n u a l l y p l u g g e d from t h e inside. T h e pressure
r u p t u r e d t h e screens. The resin loss was h i g h , so the screens
were r e m o v e d f r o m t h e c e n t e r d i s t r i b u t o r s a n d t h e control circuits c h a n g e d t o b l o w d o w n the c o l u m n , following t h e backwashing o p e r a t i o n , t h r o u g h t h e bed a n d o u t t h e b o t t o m dist r i b u t o r . T h i s left o n l y t h e b r i n e a n d a slow rinse flowing out
t h r o u g h t h e u n s c r e e n e d c e n t e r distributors. Flow rates should
be low e n o u g h so t h a t t h e t o p of the bed w o u l d be well below
the d i s t r i b u t o r level. T h e only resin loss s h o u l d be caused by
m a l f u n c t i o n of t h e u n i t s , a n d to p r o v i d e for this possibility, a
resin t r a p was b u i l t in t h e flume c a r r y i n g waste from the units.
D i a p h r a g m valve o p e r a t o r s caused considerable t r o u b l e . Diap h r a g m t r a v e l w a s t o o s h o r t to allow full o p e n i n g of valves.
O p e r a t o r d i a p h r a g m s w e r e flat a n d p u l l e d o u t of r e t a i n i n g flanges.
T h e o p e r a t o r s h a v e b e e n c h a n g e d t o allow full travel a n d m o l d e d
d i a p h r a g m s h a v e r e p l a c e d t h e flat ones originally installed.
Excessive p r e s s u r e d r o p was e n c o u n t e r e d across the beds
d u r i n g t h e first c a m p a i g n . A fairly heavy layer of calcium carb o n a t e was b e i n g d e p o s i t e d on t o p of t h e b e d w h i l e juice was
flowing d o w n w a r d t h r o u g h t h e resin. It was impossible to
r e m o v e t h i s d e p o s i t b y backwashing, a n d the units were treated
with h y d r o c h l o r i c acid t o r e m o v e t h e calcium carbonate. T h e
source of this c a r b o n a t e was n o t located u n t i l the e n d of campaign w h e n m a i n t e n a n c e w o r k e r s discovered t h a t a by-pass cock
h a d a s h e a r e d p i n w h i c h , u n f o r t u n a t e l y h a d sheared with the
cock in a slightly o p e n position, a n d allowed a small stream of
second c a r b j u i c e to by-pass t h e filters.
I t was i m p o s s i b l e t o h o l d t h e j u i c e a t t h e p r o p e r level i n the
c o l u m n s b e c a u s e of t h e great change in pressure d r o p across the
bed from t h e b e g i n n i n g to t h e e n d of t h e cycle. It was necessary
to m a n u a l l y a d j u s t t h e level in t h e c o l u m n before starting the
r e g e n e r a t i o n cycle, in o r d e r to p r e v e n t excessive sweetening-off
losses. A n e w level c o n t r o l system was installed which could
c o m p e n s a t e for a large c h a n g e in pressure d r o p across the u n i t s
d u r i n g t h e j u i c e cycle.
C o n t r o l of p a r a l l e l flows t h r o u g h t h e u n i t s was impossible
by u s i n g a h a n d set valve, a n d c o n t r o l was accomplished by
utilizing a level c o n t r o l on t h e feed tank cascading to flow
controllers on each u n i t to balance the flows.
It is i m p o s s i b l e to tell w h a t r a t e of b r i n e a d d i t i o n is b e i n g
used d u r i n g t h e r e g e n e r a t i o n step a n d b r i n e c o n s u m p t i o n has
been c o n s i d e r a b l y a b o v e t h a t r e c o m m e n d e d for this application.

300

JOURNAL OF THE A. S. S. B. T.

Figure 2-Lime salts comparisons for treated and untreated juice.

Additional flow meters are b e i n g installed to correct this situation.


O p e r a t i n g Results
Figure 2 shows the variation in lime salts from the beginning
to the end of a campaign. T h e second carb juice contained
. 0 3 % C a O on solids at the b e g i n n i n g of campaign, and made
an erratic climb to more t h a n . 1 5 % as the campaign drew to a
close. T h e lime salts in the juice from t h e softeners was held
between . 0 1 5 % a n d . 0 3 % u n t i l the last few days of campaign
when it climbed as high as . 0 5 5 % . F i g u r e 2 shows the lime
salts in the effluent from the softeners as well as the lime salts
in the t h i n juice to the evaporators. T h e difference in these two
values is accounted for by a stream of juice which by-passes the
softeners to control t h e m i n i m u m softness of the juice going
to the evaporators. A completely softened juice is corrosive and
d u r i n g t h e early p a r t of campaign, the juice softening was carried too far a n d considerable corrosion was experienced in the
pipe lines to the evaporators a n d also in the first two effects of
the evaporators. A greater percentage of juice should have bypassed the softeners d u r i n g the first p a r t of the campaign.
Figure 3 shows the actual resin loading o b t a i n e d throughout
the campaign. T h e curve also indicates t h e m a x i m u m loading
to be expected with this resin a n d d u r i n g t h e earlier part or
campaign we were considerably below this expected loadingT h e service cycles were m u c h too short for t h e hardness of the
j u i c e a n d regenerations were b e i n g started m u c h sooner than
necessary. W h e n it was realized t h a t t h e juice was being oversoftened, the cycles were e x t e n d e d a n d b e t t e r loadings were
obtained.

VOL.

13, N o . 4, J A N U A R Y

1965

301

Figure 3.Resin loading.

T e s t s c o n d u c t e d o n t h e resin after the f i r s t year's operation


showed t h a t t h e capacity of t h e resin h a d decreased 4 . 3 % .
Analysis after t h e second year's use indicated capacity had decreased f u r t h e r to o n l y 7 9 . 6 % of t h e original capacity. T h i s
m e a n s t h a t t h e m a x i m u m l o a d i n g to be expected at the end of
the second c a m p a i g n w o u l d be 0.841 p o u n d s of C a O per cu ft
of resin i n s t e a d of 1.057 as i n d i c a t e d on F i g u r e 3.
T h e s t u d i e s also i n d i c a t e d t h a t the resin could b e cleaned
up a n d r e s t o r e d to 1 0 0 % of t h e original capacity by treating with
caustic soda a n d S o d i u m H y p o c h l o r i t e . Screen analysis showed
that very l i t t l e if a n y c h a n g e in particle size h a d taken place.
F i g u r e 4 gives a typical c u r v e for sweetening on a n d for
sweetening off t h e c o l u m n s . T h e sugar lost d u r i n g sweetening
on a m o u n t s to 25 p o u n d s p e r cycle a n d t h e loss d u r i n g sweetening off will be a p p r o x i m a t e l y 700 p o u n d s per cycle. T o t a l sugar
losses in t h e j u i c e softeners will vary from 0.06% of the sugar
in t h e j u i c e t r e a t e d p e r cycle at t h e b e g i n n i n g of campaign to
0 . 3 2 % a t t h e e n d o f c a m p a i g n w i t h t h e m u c h shorter service
cycles. W i t h 4 8 8 r e g e n e r a t i o n cycles, the sugar loss averaged
0 1 4 % for t h e c a m p a i g n .
D i l u t i o n d u r i n g s w e e t e n i n g off a m o u n t s to 1400 gallons a n d
d u r i n g s w e e t e n i n g on d i l u t i o n is 600 gallons for a total of 2,000
gallons p e r cycle. T h i s a m o u n t of w a t e r is n o t noticeable in the
861,000 g a l l o n s of j u i c e p e r cycle at t h e b e g i n n i n g of campaign.
T h e r e was n o m e a s u r a b l e difference i n the campaign averages
for b r i x of j u i c e g o i n g to t h e softeners a n d j u i c e going to the
evaporators.

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JOURNAL OF THE A. S. S. B. T.

Figure 4.Sweetening on a n d off curves.

D u r i n g the 1961 campaign (before softeners were installed)


evaporator bodies were boiled o u t 83 times while boilouts were
only necessary on 41 bodies d u r i n g the 1963 campaign. The
total tons of beets sliced d u r i n g the 1963 campaign was 17%
higher than the 1961 campaign. Basing costs on 1963 prices
a n d adjusting for differences in beets processed, the boilout
chemicals cost $5,560 less for 1963 t h a n for 1961.
T h e salt used for regeneration cost $15,100 for the 1963 campaign. T h e r e were 488 regenerations performed d u r i n g the 1963
campaign using a total of 1,200 tons of salt. T h i s salt consumption
gives a regeneration level of 15.6 p o u n d s per cubic foot of resin
regenerated. T h e r e c o m m e n d e d level is 7.5 p o u n d s per cubic
foot, a n d steps have been taken to remedy this excessive usage
of salt. Closer attention to length of cycles a n d increased loading of resin will further reduce salt c o n s u m p t i o n to less than
half of t h e 1963 figure.
T h e average slicing rate increased from 5,404 tons per day
for 1961 to 6,070 for 1963. T h i s increase of 1 2 % in slicing rate
m e a n t 14 less days in the 1963 campaign. O t h e r changes in the
mill operation u n d o u b t e d l y c o n t r i b u t e d to this higher slice rate,
b u t it is felt that a great p o r t i o n of t h e increase can be attributed
to the b e t t e r performance of the evaporators operating on
softened juice.
Summary
T h e juice softeners installed at Holly Sugar Corporation's
Carlton Mill reduced t h e lime salts in t h e t h i n juice from an
average of .066 to . 0 2 9 % of total solids in t h e juice. Evaporator

VOL.

13, N o . 4, JANUARY

1965

303

boilout requirements were decreased 65%. The decrease in cost


of boilout chemicals was only one-third of the cost of salt required for the softeners, but with better control of the brine
flows in the future, these costs should very nearly balance each
other. Slicing rate was increased 12% and, although all of
this increase cannot be attributed to the softer evaporator thin
juice, it is felt that enough savings were realized in the shorter
campaign and less labor required for evaporator boilouts to
justify the costs involved in the juice softener installation.

Sugar Beet Production in Michigan as Affected by


Crop Sequence and Fertility Levels1
L.

S.

ROBERTSON,

R.

L. C O O K ,
AND J. F.

C. D. P I P E R ,
DAVIS 2

R.

H.

DOWDY

Received for publication May 4, 1964

T h e original field layout for this e x p e r i m e n t was described


by Cook, et al. in 1945 (l) 3 . T h i s is the fourth successive five year
summary of sugar beet yields from the Ferden F a r m rotation
experiment (2,6,7). T h e yields that are in this r e p o r t are almost
double those produced d u r i n g the first five years in this experiment.
T h e higher production level is considered to be related to
changes that have been made in the experiment, and to a refined appreciation of the conditions which affect sugar beet
growth, as well as to a better u n d e r s t a n d i n g of methods and
tools that can be used to create a suitable e n v i r o n m e n t for sugar
beets.
T h e purpose of this paper is to discuss changes that have
been made in crop sequence and m a n a g e m e n t of the experiment, to discuss the effect of the changes a n d treatments upon
sugar beet yields, and to r e p o r t yields a n d observations from
a continuous sugar beet e x p e r i m e n t and a beet-bean strip crop
rotation experiment.
Characteristics

of

the

Soil

T h e soil u p o n which these e x p e r i m e n t s are located is a


relatively young h u m i c gley. As such this soil was developed
u n d e r a naturally poor drainage situation. T h e soil is relatively
high in organic matter a n d therefore dark colored. T h e soil is
classed as a Sims sandy clay loam. It contains 25 to 28% clay
and 50 to 5 8 % sand. T h e field is tile d r a i n e d every four rods
and is bordered by open ditches on three sides.
T h e structure of the soil is n o t as stable as desired (8)
Because of this, p o o r aeration at times is evident, especially
d u r i n g wet seasons (4). Periodically, the general outline and
shape of the beet roots suggests a soil that has relatively Poor
structure.
1
Authorizel for publication bv the Director as Journal Article No.. 3360 of the.Michigan Agricultural Experiment Station. East Lansing. T h e financial assistance of the Farmers
and2 Manufacturers Beet Sugar Association is gratefully acknowledged.
. lv
Professors, Instructor, Assistant Instructor, and Professor of Soil Science, respectively
* Numbers in parentheses refer to literature cited.

VOL. 13, N o . 4, JANUARY 1965

305

Tillage
Methods
T h e m i n i m u m tillage principle was p u t into effect in 1951
(3). On t h e sugar beet plots, this involves moldboard plowing
in late O c t o b e r or early November. Plowing depth averages
a p p r o x i m a t e l y 10 inches. No other tillage is practiced in the
fall. In t h e spring, a h a r r o w is used immediately prior to planting the sugar beets. T h e drill that has been used on these experiments has spring-loaded press wheels. T h e planting time fertilizer
is placed o n e i n c h to the side and two inches below the seed.
Systems of Farming
Seven systems of farming are evaluated in one experiment.
T h e original c r o p sequence plan included a legume sod crop
for hay in five of t h e seven systems (1). By 1951, it was evident
that clover-timothy h a d a less desirable effect than alfalfa-brome
u p o n the yield of o t h e r crops in the rotation, (7). Therefore,
in this o n e r o t a t i o n , alfalfa-brome was substituted for the clovertimothy c r o p .
D u r i n g t h e course of the research, marked changes occurred
in the sugar b e e t g r o w i n g area of Michigan. T h e n u m b e r of
farms h a v i n g livestock decreased rapidly while the proportion
of cash c r o p farms increased.
By 1958, it seemed desirable to alter some of the rotations
so that t h e systems of farming would realistically evaluate that
which m i g h t occur as a result of using more intense cash crop
systems. In a d d i t i o n , soybeans, a relatively new crop in the
Saginaw Valley, is p r o v i n g to be a valuable one which fits well
with sugar beet p r o d u c t i o n .
T h e systems of farming have been n u m b e r e d to facilitate
discussion. T h e crops a n d the sequence that have been grown
since 1958, w i t h t h e exception that is noted, in each of the rotations a r e as follows:
R o t a t i o n 1: alfalfa-brome, alfalfa-brome, b e a n s 4 , s u g a r
beets, barley
R o t a t i o n 2: sweet clover (oats), sugar beets, corn (GM),
beans, wheat
R o t a t i o n 3: beans, sugar beets, corn (GM), s o y b e a n s ,
wheat (GM)
R o t a t i o n 4: alfalfa-brome, corn, sugar beets, beans, wheat
R o t a t i o n 5: sweet clover, oats, beans, sugar beets, soybeans, wheat
R o t a t i o n 6: beans, wheat, corn, sugar beets, barley
R o t a t i o n 7: beans, wheat (GM), soybeans, sugar beets,
c o r n (GM)
_____
* Changed from corn to beans in 1962.

JOURNAL OF THE A. S. S. B. T.

306

In these studies, the word beans refers to the navy beans or


dry white pea beans. Barley in Rotations 1 a n d 6 is a springplanted crop, decreasing in importance in the sugar beet growing area. Sweet clover appears in Rotations 2 a n d 5 a n d is harvested for seed. Spring oats in Rotations 2 a n d 5 are planted as
a winter cover crop in August on the sweet clover areas. The
" G M " refers to a m i x t u r e of equal parts of alfalfa, mammoth,
J u n e , a n d alsike clover which is used as a green m a n u r e crop.
Each of the rotations is evaluated on the basis of two fertility
levels. W h e n the experiment was initiated in 1940, 200 pounds
of 2-16-8 was used for sugar beets on the low fertility side. This
represented the rate used by the average sugar beet grower at
that time. Five h u n d r e d pounds of the same fertilizer was used
on the high fertility side. By 1951, it was evident that more
than 500 p o u n d s of fertilizer might economically be feasible.
Therefore, t h e rates used on the low side (200 pounds) were
changed to 1000 pounds. T h u s , the previous low became the
new high.
A n o t h e r change was made in 1962 because there was a lack
of a definite crop yield response between the high and low
fertility plots. In addition, soil tests did n o t reflect a great change
in value from the use of the higher fertilizer rates. T h e rate on
the high fertility side was doubled, b r i n g i n g the a m o u n t to 2,000
pounds of 2-16-8 used on sugar beets. D u r i n g this time, commercial fertilizer became more concentrated so that 5-20-10 is
now used instead of 2-16-8. In other words, now 1,600 pounds
of 5-20-10 is used which supplies the same phosphate and potash
as 2,000 p o u n d s of 2-16-8.
In regard to the use of supplementary nitrogen, each of the
plots is subdivided so that one half of each plot is treated with
an extra 40 p o u n d s at t h i n n i n g time. A m m o n i u m nitrate has
been used most frequently although in some instances urea was
the source of the supplemental nitrogen.
Plot

Design

Basically, the e x p e r i m e n t has a split, split plot design (5).


Each crop in each rotation is grown each year. T h e treatments
are replicated 4 times. Each rotation is split a n d data are collected from 2 fertility levels. T h e fertility levels are subdivided,
a n d 1/2 of each plot receives s u p p l e m e n t a r y nitrogen. In other
words, the entire e x p e r i m e n t contains 560 plots, of which 132.
are devoted to sugar beets each year. P l a n t i n g is done with a
6-row commercial planter. Harvesting is accomplished mechanically with a modified one row machine.

VOL. 13, N o . 4, JANUARY 1965

307

Results and Discussion


In a d d i t i o n to t r e a t m e n t s , weather and timeliness of operations associated w i t h w e a t h e r affect the yield of crops (5). T h e
data in T a b l e 1 s u m m a r i z e t h e seasonal a n d a n n u a l precipitation. T h e l o n g t i m e average for the Ferden farm is approximately 30 inches. T h e r e f o r e , the sugar beet yields that are reported r e p r e s e n t those p r o d u c e d with only 60 to 7 0 % normal
p r e c i p i t a t i o n d u r i n g t h r e e of five years.
Table 1.Seasonal and annual precipitation on the Ferden Farm for 1959 through
1963.
Inches of precipitation

April
May

June
July
August
September
October
Total for season
Total for year

1959

I960

Year
1961

1962

1963

3.48
3.43
2.64
4.06
1.81
2.53
3.53
21.48
29.89

2.62
2.76
4.18
0.96
3.33
1.77
1.24
16.86
23.43

4.67
1.48
4.19
4.25
6.93
3.58
1.91
27.01
34.79

0.95
2.89
3.58
1.77
2.43
1.86
1.92
15.40
20.16

1.49
2.62
2.26
3.59
2.63
1.11
0.74
14.44
20.02

W h i l e t h e yield d a t a t h a t are r e p o r t e d were analyzed statistically, t h e analysis of variance data are n o t included in this
s u m m a r y . T h i s was to simplify the tables a n d to expedite discussion of t h e results. T h e only instance where treatments did
not cause a statistically significant difference in yield at the 5%
level o c c u r r e d w h e r e s u p p l e m e n t a l nitrogen was used in 1963.
In general, a n y differences in rotations greater than 9% are
significant. T h e same figures for levels of fertility a n d nitrogen
are 6 a n d 8% respectively.
T h e yields of sugar beets p r o d u c e d w i t h o u t supplemental
n i t r o g e n a n d w i t h 400 p o u n d s p e r acre of 5-20-10 fertilizer are
shown i n T a b l e 2 . T h e highest yields were produced d u r i n g
those t w o years w i t h average or m o r e than normal rainfall.
T h e h i g h e s t average yield was p r o d u c e d in R o t a t i o n 5. In
this r o t a t i o n , w h i c h c o n t a i n s sweet clover, the beets follow
beans. T h i s historically has b e e n a good sequence of crops.
W h e r e alfalfa precedes corn, the yield of sugar beets after
corn ( R o t a t i o n 4) was a p p r o x i m a t e l y the same as after beans
( R o t a t i o n 1).
T h e r a n g e in yield from o n e year to another was as great
in R o t a t i o n 5 as in a n y of t h e rotations. Most of the yield increase reflected in t h e final average occurred d u r i n g 1959 a n d
in 1961, w h i c h w e r e the wettest years. T h i s suggests that the

308

JOURNAL OF THE A. S. S. B. T.

Table 2.Sugar beet yields as affected by seven systems of crop sequence. (Low
fertility level - no supplementary nitrogen.)

""

T o n s o f s u g a r beets p e r acre
Rotation
no.

2
S
4
5
6
7

Crop sequence*

1959

A, A, Be, SB, Ba
Sw1 S B , C 2 , B e , W
Be, SB, C 2 , S, W i
A , C, SB, B e , W
Sw 1 , B e , S B , S, W
B e , W , C, S B , B a
Be, W 2 , S, SB, C2
Mean
L. S. D .

21-2
19.3
19.4
20.2
21.9
16.7
17.3
19.4
1.8

Year
1961

1962

1963

Range

Mean

13.9
17.6
18.0
15.5
14.6
20.4
18.0
16.2
24.0
16.0
14.7
16.2
18.4
14.6
18.7
15.3
1.4
1.6

13.3
13.9
11.5
13.4
16.2
11.3
13.9
13.4
3.2

16.8
11-2
12.9
16.6
15.2
15.0
14.8
14.6
1.6

7.9
8.1
8.9
6.8
8.8
5.4
4.5

16.6
15.6
15.8
16.9
18.7
15.8
15.8

1960

*A = alfalfa b r o m e h a y , Be = b e a n s , SB = s u g a r b e e t s , Ba = b a r l e y ,
Sw = sweet clover, C = c o r n , W = w i n t e r w h e a t , S = s o y b e a n s .
O a t cover c r o p s e e d e d a f t e r sweet clover.
A m i x t u r e of s m a l l s e e d e d l e g u m e s p l a n t e d in t h e c r o p for g r e e n m a n u r e purposes.

opportunity for high yields with adequate moisture is greater


in this rotation than in some of the others.
Rotations 6 a n d 7 historically have been relatively low yielding rotations. T h e range in yields in these rotations from year
to year was m u c h lower than in any of the other systems. As
will be shown, nitrogen problems are m o r e evident in these
rotations. Erickson a n d Van Doren demonstrated that soil
structure problems as measured by oxygen diffusion rates developed u n d e r these two systems, especially d u r i n g wet years (4),
T h e lowest average yields occurred in R o t a t i o n 2, the other
rotation that contained sweet clover. T h e low average yields
reflects strongly the low yield of only 11.2 tens per acre produced
in 1963. T h e effect of two exceedingly dry seasons in succession
probably accounts for this situation. D r o u g h t may have been
more severe because two to three tons per acre of dry matter as
sweet clover straw was plowed the fall before beets were planted.
As might be expected, the response from the use of 40 pounds
of supplemental nitrogen sidedressed to the beets at thinning
time varied from one year to another. T h e data in T a b l e 3 show
the increase or a p p a r e n t yield decrease associated with the use
of supplemental nitrogen in the various rotations on the low
fertility plots. T h e greatest average yield increase occurred in
Rotations 3 a n d 6 although the increase was very inconsistent
from one year to the next. T h e nitrogen was least effective in
Rotations 1 and 5. T h e s e are the two highest yielding rotations.
T h e data in T a b l e 4 show the yearly effects of using more
than 400 p o u n d s of 5-20-10 fertilizer on the sugar beets in each
of the rotations. In i n t e r p r e t i n g these data, one should keep

VOL.

13,

No.

4,

JANUARY

1965

309

Table 3.Yearly changes in yield of sugar beets as affected bv the use of 40 pounds
of supplemental nitrogen in seven systems of crop sequence. (Low fertility level - with
supplementary nitrogen.)
Change in yield - tons per acre
1959

1960

Year
1961

1962

1963

Mean

0.1
0.1
+1.4
+ 1.2
0.7
+2.3
+2.9
+1.0

+0.4
0.9
+0.9
0.7
+0.8
1.0
+0.8
+0.0

+0.5
+2.2
+0.6
+2.5
+0.3
+4.8
0.4
+ 1.5

+0.3
+ 1.5
+2.6
+ 1.1
+ 1.0
+3.4
+ 1.7
+ 1.7

+0.0
+0.7
+1.5
+0.9
+0.3
+ 1-7
+0.9

0.5

0.4

1.1
+0.6
+2.2
+0.2
+0.3
0.8
0.7
+0.1
N.S.

tion
Crop sequence*
A, A, Be, SB, Ba
Sw1, SB, C 2 , Be, W
Be, SB, C 2 , S, W1
A, C, SB, Be, W
Sw1, Be, SB, S, W
Be, W,2 C, SB, Ba2
Be, W , S, SB, C
Mean
L. S. D.

1
2
3
4
5
6
7

0.5

0.4

*A =: alfalfa brome hay, Be = beans, SB = sugar beets, Ba = barley,


Sw = sweet clover, C = corn, W = winter wheat, S = soybeans.
1
Oat cover crop seeded after sweet clover.
- A mixture of small seeded legumes planted in the crop for green manure purposes.
Table 4.Yearly changes in yield of sugar beets as affected by the use of extra
fertilizer in seven systems of farming. (High fertility level - no supplemental nitrogen.)
Change in yield - tons per acre

Rotation
no.
1
2
3
4
5
6
7

Crop sequence*
A, A, Be, SB, Ba
Sw1. SB, C2 2 , Be, 1W
Be, SB, C , S, W
A, 1 C, SB, Be, W
Sw , Be, SB, S, W
Be, W, C, SB, Ba
Be, W 2 , S, SB, C 2
Mean
L. S. D.

1959
+ 1.8
+2.1
+ 14
0.2
+0.6
+0.3
+2.5
+ 1-2
1.5

1960

Year
1961

1962

1963

Mean

+ 1.5
0.6
+2.4
0.3
0.4
3.0
+ 1.6
+0.17

+2.6
+ 1.0
+2.6
+ 11
+0.1
0.7
+2.0
+ 1.2

+3.4
+2.1
+4.7
2.2
+ 1.6
+2.8
+ 1.8
+2.0

+0.2
+5.9
+3.3
+2.1
+2.9
+ 1.5
+2.7

+ 1.9
+2.1
+2-9
+0.1
+1.0
+0.2
+2-1

0.4

0.6

0.6

+2.7
0.8

*A = alfalfa brome hay, Be = beans, SB = sugar beets, Ba = barley,


Sw = sweet clover, C '= corn, W = winter wheat, S = soybeans.
1
Oat cover crop seeded after sweet clover.
2
A mixture of small seeded legumes planted in the crop for green manure purposes.

in m i n d t h a t in 1959, 1960, a n d 1961, the rate of fertilizer used,


800 p o u n d s of 5-20-10, was twice that used on the low fertility
plots a n d t h a t in 1962 a n d 1963 the rates used, 1,600 pounds of
5-20-10, was e q u a l to four times that used on the low fertility
plots.
T h e average increase in yield from the use of 1,600 pounds
was g r e a t e r t h a n t h e increase in yield from the 800 p o u n d rate.
T h i s o c c u r r e d d e s p i t e t h e fact t h a t 1962 and 1963 were exceptionally d r y years, t h e driest of the five year period.
T h e a m o u n t of increase in yield varied with the rotation.
O n e of t h e lowest average increases was obtained in the cash
crop r o t a t i o n w h i c h h a d n o green m a n u r e o r cover crops (Rotation 6). T h i s m i g h t be expected in this situation where soil

310

JOURNAL OF THE A. S. S. B. T.

aeration may be a limiting factor as was suggested by Erickson


and Van D o r e n (4).
T h e average yield increase obtained in R o t a t i o n 4 was also
insignificant a m o u n t i n g to only one-tenth of a ton. An explanation for this situation is n o t readily apparent.
T h e data shown in T a b l e 5 indicate the yearly effect of sidedressing the sugar beets with 40 p o u n d s of nitrogen on the high
fertility plots. T h e difference in average response on all rotations from year to year was n o t great, being equal to only fivetenths of a ton per acre. T h e average response in Rotation 6
was greatest, being equal to 2.7 tons. T h e increase in yield was
not as large in this rotation in 1962 a n d 1963 because in these
years more planting time fertilizer was used which necessarily
means more nitrogen was used. T h e extra nitrogen used in
1962 a n d 1963 in the planting time fertilizer was equal to 40
pounds, the same a m o u n t as was used as a sidedressing.
T h e data in T a b l e 6 are the sugar beet yields that were produced in each of the rotations with the high fertilizer rate and
with the use of 40 p o u n d s of sidedressed nitrogen.
T h e highest average yields, 20.9 tons per acre, were produced
in Rotation 5, the rotation in which beets followed beans which
were preceded by sweet clover. T h e lowest yields were produced
in Rotation 6, the cash crop rotation w i t h o u t green manures or
cover crops.
T h e greatest range in yield from one year to another was
also obtained in Rotation 5 which suggests that with adequate
moisture the opportunity for satisfactory yields is greatest in
this rotation. T h e range in yields in R o t a t i o n 6 a n d 3 was approximately one half of those o b t a i n e d in Rotation 5. This
suggests that some factor other than rotation, planting time, or
fertilizer, is limiting the plants' growth.
Continuous
Sugar Beets
Because sugar beets are a high value cash crop, it would be
desirable to grow the crop year after year. T h i s , however, is not
practical because disease or pest problems frequently develop
To determine specificallv the k i n d of problems that develop, a
continuous sugar beet plot was established in 1959. T h e yields
that were obtained are 26.0, 16.5. 14.8, 13.0, a n d 17.4 tons per
acre respectively for the years 1959 t h r o u g h 1963.
T h e continuous decrease in yield for each of the seasons up
to 1963 suggests why continuous sugar beets is not practical
It is not immediately evident why the yields were down because
there were no clear-cut symptoms of pest or disease damage
except in 1961 when leaf blight (Cercospora beticola) was severe.

VOL. 13, N o . 4, J A N U A R Y 1965

311

Table 5Yearly changes in yield of sugar beets as affected by the use of extra
nitrogen in seven systems of crop sequence. (High fertility level - with supplementary
nitrogen.)
Change ini yield - tons per acre
Rotation
Crop sequence*

1959

I960

Year"
1961

1962

1963

Mean

Be, SB, Ba
sw 1 , SB, C 2 , Be, W
Be, SB, C 2 , S, W 1
A, C, SB, Be, W
bw1, Be, SB, S, W
Be, W, C, SB, Ba
Be, W 2 , S, SB, C 2
Mean
L. S. D.

_____

+0.8
+0.3
+0.1
+0.8
+2.7
+3.1
+0.1
+ 1.1
0.51

+0.5~
+ 1.6
+0.1
+ 1.1
+ 17
+4.4
+ 1.4
+ 1.5
0.46

+0.6
+0.8
+ 1.8
+0.7
+0.6
+2.4
+ 1.0
+ 1.1
0.38

+2.7
+2.3
+1.1
+0.4
+0.3
+0.5
+0.1
+ 1.1

+0.9
+ 1.0
+0.6
+ 1.1
+ 1.3
+2.7
+0.7

no.
A,A,

1
2
3
4
5
6
7

0.1
+0.1
+2.4
+ 1.3
+3.1
+0.7
+ 1-0
0.48

*A = alfalfa brome hay, Be beans, SB = sugar beets, Ba = barley,


Sw = sweet clover, C = corn, W = winter wheat, S = soybeans.
Oat cover crop seeded after sweet clover.
A mixture of small seeded legumes planted in the crop for green manure purposes.

1
2

Table 6.Sugar beet yields as affected by seven systems of crop sequence. (High
fertility level - with supplementary nitrogen.)
Tons of sugar beets per acre
Crop sequence*

1959

1960

Year
1961

1962

1963 Range Mean

A, A, Be, SB, Ba
sw 1 , SB, C2, Be, W
Be, SB, C 2 , S, W 1
A, 1 C, SB, Be, W
hw , Be, SB, S, W
Be, W, C, SB, Ba
Be, W 2 , S, SB, C 2
Mean
L. S. D.

22.8
21.3
20.9
22.4
23.8
20.1
20.5
21.7

16.2
15.2
17.7
16.7
18.3
16.3
16.3
16.6

20.7
20.6
22.3
21.0
25.8
18.8
21.8
21.6

17.3
16.8
18.0
16.5
18.4
16.5
16.7
17.2

19.7
19.4
17.3
18.7
18.4
17.0
17.6
18.4

1.8

1.4

1.6

3.2

1.6

tion
no.
1
2
3
4
5
6
7

6.6
6.1
3.8
5.9
7.4
3.8
4.2
5.4

19.3
18.7
19.1
19.1
20.9
17.7
18.6
19.1

*A = alfalfa brome hay, Be = beans, SB = sugar beets, Ba = barley,


Sw = sweet clover, C corn, W _ winter wheat, S = soybeans.
1
Oat cover crop seeded after sweet clover.
2
A mixture of small seeded legumes planted in the crop for green manure purposes.

The disease was not evident on other experiments in the same


field where beets were grown in rotation with other crops.
Sugar Beet - Bean Strip Cropping
Four-year average yields where sugar beets and beans were
grown in alternate strips are shown in Table 7. In this experiment, each strip was six rows wide. The rows ran east and west.
T h e rows of sugar beets adjacent to the beans averaged 21.1
tons per acre while the inside rows averaged only 17.0 tons. The
picture was different with the beans. Border rows yielded six
bushels per acre less than did those not bordering beets. The
practice might still be worthwhile though, because four tons
of beets are worth considerably more than six bushels of beans.

312

JOURNAL OF THE A. S. S. B. T.
Table 7.The effect of a beet-bean strip cropping plan upon yields.
Beets tons/acre

Year
1960
1961
1962
1963
Mean
% increase
% decrease

Beans bu/acre

Inside
rows

Outside
rows

Inside
rows

17.0
16.3
17.2
17.3
17.0

24.4
20.4
19.0
20.7

28.1
36.4
27.2
40.3
33.0

211

Outside
rows
18.9
32.1
20.2
36.8
27.0

24.1
18.1

Summary
T h e data in this paper represent the fourth successive fiveyear summary of the sugar beet yields produced in the Ferden
tarm rotation experiment. T h i s replicated field experiment is
located in the Saginaw Valley region of Michigan a n d involves
seven systems of farming, two fertility levels a n d the use of
supplementary nitrogen applied as a sidedressing.
On the basis of the data shown in this paper, the following
statements can be made:
1) T h e yields of sugar beets after corn were approximately
the same as after beans, providing the corn was preceded by
alfalfa.
2) T h e lowest sugar beet yields were produced in a cash
crop rotation that did not include a green m a n u r e or cover
crop immediately previous to the sugar beets.
3) Preceding sugar beets with sweet clover resulted in yields
approximately two tons per acre less than with a bean crop
between the sweet clover a n d the sugar beets.
4) T h e highest average of sugar beet yields was produced
in Rotation 5 where sugar beets followed beans which were preceded by sweet clover.
5) T h e use of more t h a n 400 p o u n d s of 5-20-10 fertilizer did
not consistently increase sugar beet yields to levels above those
produced with this rate.
6) T h e greatest yield response from the use of sidedressed
nitrogen was produced in the rotation that had no green manure
or cover crop.
7) T h e use of an extra 40 p o u n d s of sidedressed nitrogen
caused increases in yield ranging from 0 to 1.7 tons per acre on
the low fertility plots. T h i s was less t h a n on the high fertility
plots, despite the fact that the high fertility plots received significantly m o r e nitrogen in the p l a n t i n g time fertilizer.

VOL.

IS, N O . 4, J A N U A R Y

1965

313

8) T h e u s e of an e x t r a 40 p o u n d s of sidedressed n i t r o g e n
caused increases in y i e l d r a n g i n g from 0.5 to 2.7 tons p e r acre
on the h i g h fertility plots.
9) T h e yields of s u g a r beets were increased, b u t b e a n yields
were d e c r e a s e d w h e n a l t e r n a t e strips of six rows of beans a n d
six r o w s of b e e t s w e r e p l a n t e d in an east a n d west direction,
side by side.
10) G r o w i n g s u g a r b e e t s year after year on the same land
m a y n o t be d e s i r a b l e because leaf b l i g h t caused a r e d u c t i o n in
yield o n e y e a r o u t of four.
L i t e r a t u r e Cited
(1)

COOK, R . L., C. E. M I L L A R , a n d L. S. ROBERTSON.

1945.

A crop rota-

tion field layout with an illustration of the statistics involved in


c o m b i n i n g several years data. Soil Sci. Soc. Amer. Proc. 10: 213-218.
(2)

COOK, R . L., C. E. M I L L A R , a n d L. S. ROBERTSON.

in seven Michigan systems of crop rotation.


Beet T e c h n o l . 4: 73-87.
(3)

COOK,

R.

L.,

H.

F.

MCCOLLY,

L.

S.

1946.

Sugar beets

Proc. Am. Soc. Sugar

ROBERTSON,

and

C.

M.

HANSEN.

1958. M i n i m u m tillagesave money, water, soil, with. Mich. State


Univ. C o o p . Ext. Serv. Bull. 352.
(4) ERICKSON, A. E., a n d D. M. VAN DOREN. T h e relation of plant growth
a n d yield to soil oxygen availability. Trans. 7th Int. Congr. of Soil
Sci. 3: 428-434.
(5) GUTTAY, J. R., a n d R. L. COOK. 1953. T h e effect of crop sequence and
early s u m m e r t e m p e r a t u r e on the response of sugar beets to supplem e n t a l nitrogen. Quart. Bull. Mich. Agr. Exp. Sta. 35: 482-488.
(6)

G U T T A Y , J . R., R . L. COOK, a n d L. S. ROBERTSON.

1958.

Sugar beet

p r o d u c t i o n in Michigan as affected by cropping sequence and fertility level. J. Am. Soc. Sugar Beet Technol. 10: 65-75.
(7)

ROBERTSON, L. S., R. L. COOK, P. J. ROOD, and L. M. T U R K .

1952.

Ten

years results from the Ferden rotation and crop sequence experim e n t . Proc. Am. Soc. Sugar Beet Technol. 7: 172-179.
(8) ROBERTSON, L. S. 1952. A study of the effect of seven systems of cropp i n g u p o n yields a n d soil structure. Proc. Am. Soc. Sugar Beet
T e c h n o l . 7: 256-264.

The Effects of Five Legume Crops on Soil Populations


of the Sugar Beet Nematode
(Heterodera schachtii Schmidt) 1
ARNOLD E. STEELE AND CHARLES P R I C E 2
Received for publication May 11, 1964

In studies of rotation systems on n e m a t o d e infested land,


Johnson a n d Wheatley (2) 3 found that inclusion of beans in the
rotation greatly increased yields of sugar beets. Golden and
Shafer (1) demonstrated that Navy beans had a stimulatory,
trap crop effect on larvae of the beet nematode and suggested
that beans might have some practical value in rotation systems
for the control of this pest. T h i s suggested trials to determine
the magnitude of the trap crop effect of certain legumes.
Jones (3, 4 and 5) found that difficulties such as uneven
distribution of nematodes, soil type variations, a n d contamination were overcome when small plots were used instead of field
plots. His plots (termed microplots by the author) were constructed of slotted concrete posts a n d paving stones joined together with bitumastic, measured 2 feet 4 inches square and
two feet deep. Mai (6) reported successful use of 8 X 5 feet
plots b o u n d e d by 12 inch redwood boards for research involving
the golden nematode, Heterodera rostochiensis.
Materials and Methods
Twenty-five microplots, measuring 4 feet square and 3 feet
deep, constructed of 3/4" X 12" redwood fastened with aluminum
nails, were lined with polyethylene plastic sheeting, and sunk
i n t o the g r o u n d to a depth of 2 1/2 feet, spaced 4 feet apart.
Examination of the soil in the experimental area did not reveal
the presence of sugar beet nematode.
Sugar beets (Beta vulgaris L. var. U. S. 75) were grown in
the greenhouse in individual a l u m i n u m foil cylinders containing steam-sterilized soil. W h e n the plants were well established,
the a l u m i n u m foil was removed a n d 16 beets transplanted to
each microplot d u r i n g the spring of 1960 (Figure 1). Soil infested with cysts of Heterodera schachtii was added to each of
the microplots at the time beets were transplanted.
1
Cooperative Investigations of the Crops Research Division, Agricultural Research
Service, United States Department of Agriculture and the Beet Sugar Development
Foundation.
.
2
Nematologist and Research Agronomist, respectively, Crops Research Division, Agricultural Research Service, U. S. Department of Agriculture, Salinas, California.
3
Numbers in parentheses refer to literature cited.

VOL.

13, N o . 4, J A N U A R Y

Figure
Heterodera

1.Microplot
schachtii.

1965

containing

315

sugar

beet

plants

infected

with

Soil samples were obtained from each microplot on March


9, 1961, after beets had been removed. The samples were oven
dried, weighed, and processed to recover cysts. Counts of the
number of cysts per 100 grams of soil ranged from 35 to 64 and
averaged 49 per sample.
Legume non-host crops were planted in individual microplots on April 25, 1961, and again on May 25, 1962, each crop
being replicated 5 times in a randomized block design. The crops
used in this test were: Kentucky wonder white-seeded pole
beans (Phaseolus vulgaris L.); California Small White bean;
Alderman peas (Pisum sativum)', White Dutch clover (Trifolium
repens); or Chilean alfalfa (Medicago saliva). Microplots receiving peas or beans contained 4 planting rows spaced 1 foot
apart; seeds of clover or alfalfa were planted broadcast. (Figure
2).
Random samples of soil were taken from all microplots on
October 25, 1961 and October 25, 1962, after legume crops
were removed from the microplots. The samples were oven
dried, weighed, and processed to recover cysts.
Seeds of sugar beets were planted in each of the 25 microplots on February 4, 1963. On March 12, 1963, several plants
were removed from each microplot and taken to the laboratory
where the plants were washed and weighed and the roots stained
in a boiling solution of lactophenol and examined for the
presence of nematode larvae. Counts of larvae are listed by
crops in Table 1.

316

JOURNAL OF THE A. S. S. B. T.

Figure 2.Infested microplot containing legume crops.


Table

1.Effects of l e g u m e crops on p o p u l a t i o n s of Heterodera schachtii.


Replications

Crop
Alfalfa
Clover
Navy bean
Pea
Pole bean
Significance
L S D .05
1

260 1
196
797
463
752

122
397
504
835
320

238
676
554
488
178

4
381
188
646
884
1,077

5
196
205
412
750
1,203

Total

Average

1,197
1,662
2,913
3,420
3,530

239.4
332.4
582.6
684.0
706.0

354.4

F i g u r e s listed a r e a v e r a g e n u m b e r s of l a r v a e p e r g r a m of r o o t s of s u g a r beet.

Beets were t h i n n e d to 20 plants per microplot on March 14,


1963, and harvested and weighed on October 2, 1963. Soil
samples taken on October 10, 1963, were again oven dried,
weighed, and processed to recover cysts.
Results and Conclusions
Soil samples removed from microplots after legumes were
grown for 2 seasons averaged 27.9 cysts p e r 100 grams of soil.
T h i s n u m b e r was a b o u t 43 percent of n u m b e r s of cysts present
at the beginning of the test, b u t significant differences between
legume crops were not apparent.
Data from the larval counts ( T a b l e 1) indicated that alfalfa
or clover reduced populations of sugar beet nematode larvae
significantly more t h a n did beans or peas. However, the nematode-trapping effect of these crops was n o t sufficient to affect
subsequent population increases appreciably when sugar beets
were grown. Samples taken after beets were harvested contained
an average of 128.6 cysts p e r 100 grams of soil with no significant
differences between treatments. U n d e r the conditions of this

VOL.

13,

No.

4,

JANUARY

1965

317

test, enough larvae remained to increase the population more


than 2.6 times when beets followed legumes.
Since alfalfa gave the greatest reductions of nematode populations, this crop may be particularly suited to use in rotations on
nematode-infested soil. T h e data indicate that further experiments with alfalfa and beans in rotation studies are warranted.
Literature Cited
(1)

GOLDEN,

A.

M.

and

THELMA

SHAFER.

1959.

Host-parasite

relation-

ships of various plants a n d the sugar beet nematode, Heterodera


schachtii. U. S. Dept. of Agri. Plant Disease Reptr. 43 (12) : 12581262.
(2) JOHNSON, R. X. a n d G. W. W H E A T L E Y . 1959. T h e effects of different
rotations on sugar beet production in land infested with the sugar
beet n e m a t o d e , Heterodera schachtii, in the Salinas Valley of California. J. A m . Soc. Sugar Beet Technol. 10(4): 286-289.
(3) JONES, F. G. W. 1955. A microplot technique for the study of soil
p o p u l a t i o n s of cyst-forming root eelworms of the genus Heterodera.
Soil Zoology, p p . 390-393.'
(4) JONES, F. G. W. 1956. Soil population studies using microplots. Nematologica 1 (2) : 109-110.
(5) JONES, F. G. W. 1956. Soil population of beet eelworm (Heterodera
schachtii Schm.) in relation to cropping. II. Microplot and field
plot results. A n n . A p p l . Biol. 4 4 ( 1 ) : 25-26.
(6) M A I , W. F. 1953. Small field plots for experiments involving plant
p a t h o g e n i c nematodes. Phytopath. 4 8 ( 5 ) : 263.

Historical Highlights in Sugar Beet Harvest


Mechanization
AUSTIN

A.

ARMER1

Received for publication June 8, 1964

T h e word "Historical" in the title refers to rather recent


chronologylimited mainly to personal experiences of the author
over the past 25 years, together with some study of the previous
art as far back as 1913.
T h e mechanical harvest of sugar beets is, for the purpose
of this account, divided into three operations; topping, digging
a n d cleaning. In machines currently in use, these operations are
more or less combined. Hence a presentation of the genesis of a
few of the leading U. S. makes of commercial harvesting machines
will conclude the account.
Topping
In 1913, T h e Great W e s t e r n Sugar C o m p a n y of Denver,
Colorado, offered a substantial cash prize for a successful beet
harvester. In response to this offer, over 50 machines were submitted, a n d 15 of these were fieldworthy enough for testing and
appraisal. These machines demonstrated a b o u t every combination of elements which has subsequently come i n t o use. Following is a tabular outline of inventors, topping systems, and subsequent applications as of 1913.
Inventor, 1913
1. Arthur
2. Atwood
3. Blevins &
Lewis
4. Crume
5. Dawson

6. Geibig
7. Leyner
8. Murphy

System

Subsequent application

Multiple disk roll finder


narrow knife
Roll finder-fixed narrow
knife
Multiple finger finder
fixed narrow knife
Multiple disk roll finder
Flat Shoe finder, variable
ratio. Knife splits crown

Several contemporary
English harvesters
Lockwood Topper-Windrower
(Present)
Armer experiments
1940-43
See Item 1
Most contemporary U.S.
disk toppers & scalpers
Roscoe Zuckerman 1943
See Item 5

Flat shoe finder


Rotary blade cutter
Multiple disk finder
2 flat cutting disks
Driven roll finder
Oscillating knife

Catchpole (England)
Devey, 1939, Armer
(U.C.) 1940
Powers (U.C.) 1939

Agricultural Engineer, Spreckels Sugar Company, Woodland, California.

VOL.

13, N o . 4, J A N U A R Y

Inventor, 1913
Pruvot,
France
Siedersleben,
Germany
11. Smith

1965

319

System

Subsequent application

Driven roll finder


fixed narrow knife
Multiple finger finder
Concave topping disk

Lockwood, Wescon and Speedy


Topper-Windrowers
Armer (U.C.) 1941
Present Farmhand,
J o h n Deere
Gemco, International
Several contemporary
English harvesters

M u l t i p l e disk finder
Stationary narrow knife

T o p p i n g in the Machine
W h i l e g r o u n d t o p p i n g offers some self-evident advantages,
t o p p i n g in t h e m a c h i n e has certain u n i q u e virtues. These include:
1) All o p e r a t i o n s (digging, topping, loading) can be done
in a single pass d o w n t h e row.
2) H i g h e r speeds a r e possible t h a n with g r o u n d toppers.
3) O p e r a t i o n is possible in m u d or peat, where beets are
i n s e c u r e i n t h e soil.
Some c o n t e m p o r a r y harvesters which top in the machine are
A r m e r ( I r e l a n d ) , M a r b e e t a n d Scott-Viner. Each of these makes
use of a p a i r of c o u n t e r - r o t a t i n g disks, with slightly overlapping
c u t t i n g edges.
Ground Topping
G r o u n d , or " I n P l a c e " t o p p i n g has long appealed to harvester designers, m a i n l y for these reasons:
1) T h e beets a r e rigidly fixed in the ground.
2) T h e beets a r e a p p r o x i m a t e l y uniform in lateral positioning.
3) T h e beets, w h i l e n o t u n i f o r m in height at the crown,
c a n be gaged by a " F i n d e r " which adjusts the vertical
p o s i t i o n of c u t .
4) T o p p i n g a n d t o p saving may be a separate operation
from r o o t lifting a n d loading.
F r o m 1938 to 1945, a comprehensive program of research
a n d d e v e l o p m e n t in sugar beet field machinery was pursued at
the U n i v e r s i t y of California at Davis. T h e d e p a r t m e n t of Agric u l t u r a l E n g i n e e r i n g s u p p l i e d shop, field and laboratory facilities
in a d d i t i o n to a h i g h l y t r a i n e d staff. T h e U. S. Department of
A g r i c u l t u r e p a r t i c i p a t e d , a n d t h e project was financed through
a g r a n t f r o m t h e U. S. Beet Sugar Association. T h e late Prof.
H. B. W a l k e r h e a d e d t h e project.
.
Several " I n P l a c e " t o p p e r s were developed by the project,
a n d d e m o n s t r a t e d a r e m a r k a b l e facility for accurately severing

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JOURNAL OF THE A. S. S. B. T.

Figure 1. In 1939, John Powers, University of California, Davis, designed this oscillating knife, variable-cut topper.

Figure 2.In 1940, Austin Araier,


Armer, University of California redesigned
teed.
the Devey disc topper for greater simplicity and higher operating speed.

the foliage while the roots remained intact in the soil (Figures
1 and 2).
An entirely different topping principle involves the attrition
of the foliagechopping the leaves a n d petioles into bits with
a beating or scrubbing device which is n o t q u i t e severe enough
to damage the root crown. T h e first of the "Scrubbing Brush"
toppers known to this a u t h o r was the rotary brush preceding
the lifter blades of the M u r p h y Digger, o n e of the entrants in

VOL.

13, N o . 4, JANUARY

The Great Western


1913 ( F i g u r e 3).

1965

Sugar

321

C o m p a n y harvest competition

in

Figure 3.The Murphy Digger (1939) was a modified potato digger


with a rotary brush topper.
A m u c h m o r e effective rotary b r u s h was used in an experimental h a r v e s t e r b u i l t by Roscoe Zuckerman of Stockton, California in 1943 ( F i g u r e 4). It w o r k e d satisfactorily in flat-planted
beets is l i g h t soils, b u t t h e m a c h i n e never became operational
on a c o m m e r c i a l scale.

Figure 4.Roscoe Zuckerman (1943) built


rotary brush topper.
P e r h a p s t h e m o s t i m p o r t a n t c o n t r i b u t i o n t o "Attrition T o p p i n g " was t h e e x p e r i m e n t a l work performed in the imd-30 s by

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JOURNAL OF THE A. S. S. B. T,

Mr. J o n a t h a n Garst. T h i s led to his granting an exclusive license


to Olsen Manufacturing Company of Boise, Idaho. T h e i r rubber
flail beater became well known by its trade n a m e "Rotobeater"
(Figure 5).

Figure 5.The Olson Rotobeater was built u n d e r license from Jonathan


Garst, inventor and patentee, in 1944.

Saving T o p s for F e e d
T h e r e have been numberless well-planned experiments performed, all of which demonstrated the feed value of beet tops,
whether fed green-chopped, ensiled, dehydrated, baled or (most
recently) watered.
In 1948, Spreckels Sugar Company cooperated in a largescale demonstration of the value of beet foliage when steamblanched, pressed to remove juices containing soluble salts, and
dried into a meal closely resembling alfalfa meal. A BlackwelderLocke green crop harvester was employed to cut and deliver
clean foliage prior to harvesting the roots. D u r i n g the same
series of trials, Blackwelder M a n u f a c t u r i n g C o m p a n y built an
experimental rotary chopper which blew the chopped leaves into
a truck.
In 1952, Spreckels Sugar Company conducted trials which
involved kiln-drying of chopped beet tops. An excellent, nutritious meal was produced, b u t fuel costs proved to be prohibitive.
In 1956, the Company conducted e x p e r i m e n t s to show how, with
p r o p e r precautions, palatable silage could be m a d e from beet
tops. In both of these trials, chopped beet foliage was delivered
by an experimental Rotobeater supplied by Olson Manufacturing Company.
T h e most recent efforts to harvest a n d utilize beet tops have
m a d e use of the Lockwood, Speedy a n d Wescon toppers (Figure
6).

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13, N o . 4, JANUARY

1965

323

Figure 6 . T h e Wescon topper, like the Lockwood and Speedy, delivers


multiple windrows of beet tops (1964).

Digging
T h e p r e s e n c e of soil with t h e beets as they are d u g has long
been a source of c o n c e r n to inventors of beet harvesting machinery. T h e e a r l i e r a t t e m p t s at mechanical beet harvesting were
mainly a t t e m p t s to a d a p t p o t a t o harvesters to the beet field
(Figure 3).
H o r s e - d r a w n p o t a t o diggers appeared in great variety during
the second half of t h e 19th century. Most of these devices cont e m p l a t e d s o m e m e a n s of sifting loose soil from the potatoes
("Potato C h a i n " is a heritage of this era). Such a procedure
worked w i t h potatoes, w h i c h are rarely grown in heavy cloddy
soil, a n d w h i c h h a v e no h a i r roots by which soil adheres.
A p o t a t o d i g g e r p a t e n t e d in 1872 in the U n i t e d States by a
C a n a d i a n i n v e n t o r h e l d t h e g e r m of a successful beet digger,
b u t t h e p a t e n t e v i d e n t l y escaped notice u n t i l a search was made
in 1949 to d e t e r m i n e t h e patentability of a digger invented by
Carl O p p e l , t h e n r e s i d i n g in Alberta, Canada (Figure 7).

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JOURNAL OF THE A. S. S. B. T.

Meanwhile, the same principle had b e e n invented a n d reduced to practice by Mr. R o b e r t Maynard, an i m p l e m e n t maker
at Whittlesford, near Cambridge, England in 1923 or 1924, and
sold in considerable n u m b e r s for digging sugar beets, particularly
in England's fen soils (Figure 8). Quite independently, Hammer
Brothers of O h i o developed lifter wheels of the finger type
(Figure 9).

Figure 8.Mr. Robert Maynard (1923) built this horse-drawn wheel


type lifter (Photo courtesy British Sugar Corporation, Ltd.).

Figure 9. Hammer Brothers of Ohio


built beet harvesters using the finger-wheel
digger (Photographed in 1932 by E. M.
Mervine).

T h e r e is no evidence that recent U. S. inventors of beet


harvesters were aware of either " M a y n a r d Wheels"-or the 1872
potato digger wheels. Necessity is the m o t h e r of invention, and

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No.

4,

JANUARY

1965

325

the necessity to free sugar beets from soil existed in full measure
as an obstacle to d e v e l o p i n g a successful sugar beet harvester.
T h e " C o l o r a d o Lifter," or d o u b l e blade plow was an effective
beet digger in friable soils. Most of the earlier sugar beet harvesting m a c h i n e s e m p l o y e d double-blade plows. ( T h e origin of
the d o u b l e b l a d e is o b s c u r e i t probably came to this country
from G e r m a n y as t h e horse d r a w n "Bow Plow").
Beet harvesters b u i l t commercially in the U n i t e d States at
this t i m e favor t h e wheel type lifter. T h i s preference is probably based on t h e lower draft of the wheel lifter as compared
to t h e d o u b l e p o i n t lifter. B u t the wheel lifter comes to grief
in heavy, d r y soils. Its s h a r p r i m s slice through the crust, lifting
out a r i b b o n of soil which contains the beets. T h i s ribbon then
cracks a p a r t at each beet, forming large solid chunks, almost
impossible to b r e a k up or separate in any subsequent screening
operation.
Cleaning
In t h e days of h o r s e d r a w n farm implements, the potato digger
r e p r e s e n t e d t h e last w o r d in root harvesting devices. It is no
wonder, t h e r e f o r e , t h a t w h e n inventors t u r n e d their attention
to h a r v e s t i n g sugar beets, they first attempted m i n o r modifications to t h e p o t a t o digger.
T h e s e modified p o t a t o diggers worked almost as well with
sugar beets as they d i d with potatoes; what most people failed
to realize was t h a t t h e potatoes, after being dug, were dropped
back o n t o t h e g r o u n d a n d later picked u p o n e at a timeby
hand.

Figure 10.The Pruvot digger (1913) used a transverse multiple


cleaning screen
the first known application of "Star Wheels."

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JOURNAL OF THE A. S. S. B. T,

T h u s the need became a p p a r e n t for a mechanism more effective in breaking clods by impact than was possible with potato
diggers. Such a mechanism was applied to the Pruvot digger
built by Emile Degremont at Le Cateau, France, in 1913 (Figure
10). T h i s digger replaced p o t a t o chain with a series of four
transverse rotating shafts to which were fastened a series of
"Star Wheels." These wheels behaved like 6-lobed cams, so that
beets and clods were "kicked" from wheel to wheel. T h u s beets
and clods were subjected to both propulsion and violent agitation, with the result that many clods were fractured a n d sifted
out, while beets survived with only m i n o r bruises.
T h e Pruvot digger was entered in T h e Great Western Sugar
Company harvester competition in 1913, and (along with the
Provot topper) worked so well that T h e Great Western Sugar
Company continued to improve a n d perfect it. In 1924, their
chief engineer, Mr. George Rienks, Sr., revived a n d improved
the Pruvot "Star Wheels," a n d developed the famous "Rienks
Screen"a beet cleaning device used in most receiving stations
until recently, and still used in most makes of sugar beet harvester (Figure 11).

Figure 11.The now famous Rienks Screen was developed by Mr.


George Rienks, Sr., based on the star wheel principle.

H a n d Sorting
While beet diggers e q u i p p e d with Rienks screens delivered
clean beets grown in light soils, the beets from California's fertile
but fractious adobe were generally accompanied by numerous
large, hard clods. To separate these clods from the beets. Prof
Roy Bainer (Agricultural Engineering D e p a r t m e n t Head, University of California, Davis) suggested passing beets and clods
over a "picking table," where clods could be manually picked
out of the beets. Acting on this suggestion, Austin Armer designed
two successive models of harvesters employing sorting belts

VOL. 13, N O . 4, JANUARY

1965

327

Figure 12.In 1941 Austin Armer, University of California, developed


this two row beet harvester, using the hand-sorting principle suggested by
Prof. Roy Bainer.

which t r a v e l l e d at half g r o u n d speed. T h i s slow belt speed gave


e n o u g h t i m e t o r unskilled labor to make the separation (Figure
12). E x p e r i e n c e p r o v e d that even w h e n newly dug beets travelled
over a R i e n k s screen, they were accompanied by so many clods
that it was easier to pick beets from clods than clods from beets.
T h e h a n d s o r t i n g p r i n c i p l e was pursued further; Armer designed a h a n d - s o r t i n g revolving t u r n t a b l e as an attachment to
a J o h n D e e r e N o . 53 harvester at Fort Collins, Colorado, in 1942
(Figure 13).. W h e n t h e J o h n Deere N o . 100 beet harvester was
i n t r o d u c e d , it was furnished with a similar turntable adaptable
e i t h e r to r e m o v i n g clods from beets or vice versa (Figure 14).

Figure 1 3 . - I n 1942 Armer applied the hand-sorting principle (turntable) to the John Deere N o . 53 beet harvester.

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JOURNAL OF THE A. S. S. B. T.

Figure 14. The John Deere No. 100 beet harvester (1943) provided
an optional hand-sorting turntable.

T h e John Deere
sorting belt (potato
1941 harvester at
H M I harvester had

No. 200 beet harvester (Figure 15) used a


chain) arranged similarly to that on Armer's
Davis. Likewise, the International Model
an optional sorting belt.

Figure 15.The John Deere No. 200 beet harvester (1951) used sorting
belts, similar to Armer's 1941 experimental machine.

W h e n Marbeet harvesters came i n t o extensive use in 1945,


the clod problem was reduced, a n d h a n d sorting was eliminated
However, the clod a n d trash p r o b l e m was then passed from tn
field to the receiving station, a n d m o r e effective means for separation at the receiving stations became a major development
project.

VOL.

13, N o . 4, JANUARY

1965

329

C o m m e r c i a l Machines in Current Use


In t h e U n i t e d States (1964) sugar beet harvesting machinery
is offered by an impressive list of manufacturers. T h e names
presented h e r e w i t h a r e k n o w n to the author; others may be
available.
B l a c k w e l d e r M a n u f a c t u r i n g Company ("Marbeet")
Deere a n d Company
F a r m h a n d D i v i s i o n of Daffin Corporation
I n t e r n a t i o n a l H a r v e s t e r C o m p a n y ("McCormick")
I n t e r s t a t e M a n u f a c t u r i n g C o m p a n y ("Imco")
K r i e r E n g i n e e r i n g a n d Sales
L o c k w o o d G r a d e r C o r p o r a t i o n (Toppers)
McCallum Manufacturing Company
Parma Incorporated
Speedy M a n u f a c t u r i n g C o m p a n y (Toppers)
W e s t e r n C o n v e y o r C o m p a n y ("Wescon")
T h i s list
that n u m b e r
ing choice is
However,

of a dozen manufacturers embraces at least twice


of m a c h i n e models, so that a somewhat bewilderp r e s e n t e d to a prospective purchaser.
t h e ancestors of this big family were few in n u m b e r .

O n l y o n e m a k e ( M a r b e e t ) represents an u n b r o k e n line of
descent from its p r o t o t y p e t h e experimental spike-wheel harvester b u i l t by L l o y d a n d Lewis Schmidt in 1941 (Figure 16);
financed by A. L. J o n g e n e e l ; christened " M a r i o n Beet W h e e l "
in h o n o r of M r s . M a r i o n Jongeneel, a n d finally contracted to
" M a r b e e t " ( F i g u r e s 17, 18, 19, 20).

Figure 16.The original model of the present Marbeet harvesters was


built by Lewis and Lloyd Schmidt in 1942.

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JOURNAL OF THE A. S. S. B. T

Figure 17.The first commerciaily produced Marbeet harvester (1943).

Figure 18.The first two-row Marbeet (1944).

Figure 19.The Marbeet "Midget" (1950).

VOL. 13, No. 4, JANUARY l965

331

Figure 20.The Marbeet "Twin Row" (1962).

All o t h e r s in t h i s list, except those m a k i n g toppers only, offer


harvesters w h i c h a r e related by blood or marriage to the Keist
harvester (1942-48, F i g u r e 21) a n d the lifter wheels of Carl
Oppel ( F i g u r e s 7 a n d 22).

Figure 21.The Keist Harvester (1944).

Figure 22.

T h e Oppel Harvester (1949).

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JOURNAL OF THE A. S. S. B. T.

All of the rotary flail toppers have a c o m m o n ancestorJ o n a t h a n Garst's patented beater of the late 1930's. T h o s e with
driven-wheel finders actuating fixed slanting knives are reminiscent of the Pruvot topper of 1913 (Figure 23), although this
machine was in all probability u n k n o w n to the present generation of topper designers.

Figure 23.The Pruvot Topper (1913).


T h e many rotating disk toppers bear a close resemblance
to the experimental toppers of Devey a n d A r m e r (Figures 24
a n d 2). H e r e again is, in all probability, an example of indep e n d e n t development which led to devices closely resembling
earlier models, yet n o t directly descended from them.

Figure 24.The Devey Topper (1939).


T h e s e examples of r e c u r r i n g ideas, closely similar yet independently arrived at, suggest t h a t evolution in sugar beet harvesting machinery parallels evolution in n a t u r e . Environment
(in this case the character of t h e sugar beet field in relation to
its harvest) is the basic d e t e r m i n a n t of an organism capable ot
survival (in this case, a beet harvester).

Some Effects of Ionizing Radiation on the Metabolism


a n d Growth of Sugar Beets
M Y R O N S T O U T 1 AND J O H N D. SPIKES 2

Received for publication May 14, 1964

Introduction
Since t h e discovery of X rays in 1895, plants have been used
extensively in studies of the effects of ionizing radiation on living
material. A l m o s t every conceivable type of study has been made
using seeds, p o l l e n , embryos, isolated tissues of various kinds,
whole p l a n t s , a n d isolated subcellular structures such as chloroplasts a n d m i t o c h o n d r i a . Studies have been made using all
kinds of i o n i z i n g r a d i a t i o n such as X rays; alpha, beta and gamma
radiation; n e u t r o n s ; etc. Studies have been made from almost
every p o i n t of view; i.e., in terms of radiation effects on the
physiology, b i o c h e m i s t r y , cytology, genetics, histology, morphology a n d e m b r y o l o g y of plants. T h e general field has been reviewed r e c e n t l y by S p a r r o w (9) 4 , G u n c k e l a n d Sparrow (3), and
Spikes (11). T h e p r e s e n t studies were undertaken primarily to
d e t e r m i n e w h e t h e r t r e a t m e n t with ionizing radiation would improve storage life of harvested sugar beets. At the same time,
several o t h e r aspects of t h e r a d i a t i o n biology of sugar beets were
surveyed.
R a d i a t i o n Effects on Root Respiration
T h e effect of i o n i z i n g r a d i a t i o n on plant storage tissue depends on p l a n t species, r a d i a t i o n dosage and other factors.
Sussman (14) f o u n d t h a t dosages of only a few thousand roentgens (r) significantly e x t e n d e d the storage life of potatoes by
p r e v e n t i n g s p r o u t i n g ; r a d i a t i o n increased the respiratory rate,
however. Gustafson, et al. (4) also found that gamma radiation
increased t h e r e s p i r a t o r y r a t e of potatoes for several weeks; after
this t h e r e s p i r a t o r y r a t e decreased a n d reached a low point in
a p p r o x i m a t e l y seven weeks. Dallyn, et al. (1) showed that the
s p r o u t i n g of o n i o n s c o u l d also be prevented by gamma radiation
with a c o r r e s p o n d i n g increase in storage life. Smock and Sparrow
(7) showed t h a t 2,500 r of g a m m a radiation decreased the respiratory r a t e of C o r t l a n d apples.
1

Physiologist, Agricultural Research Service, Crops Research Division, U. S. DePartment of Agriculture, Logan, Utah.
.
Professor,
Department
of
Experimental Biology, University of Utah, Salt Lake City
3
This
work was supported in part by U. S. Atomic Energy Commission Contract No.
AT(11-1).875
4
Numbers in parentheses refer to literature cited.
2

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JOURNAL OF THE A. S. S. B. T.

Glegg, et al. (2) examined the softening effects of gamma


radiation on red beets (Detroit dark red). T h e r e was little change
in tissue properties up to a "threshold" dosage of a b o u t 300,000
r. Above this, the resistance of tissue to crushing declined rapidly.
T h e y d i d n o t make any measurements on beet respiration.
Snyder a n d W i a n t (8) found that small dosages of high energy
electron radiation stimulated respiratory processes without any
visible injury to sugar beet roots. H i g h e r dosages injured surface
tissues. Since sugar beets often show large respiratory sugar
losses d u r i n g the storage period between harvesting and processing (12), an examination of the effect of ionizing radiation
on sugar beet root respiratory rates was made to determine if
such treatment might offer a useful t e c h n i q u e for reducing
respiratory losses.
Large sugar beet roots were split lengthwise with a saw and
each half was wrapped in moist paper toweling. One-half of
each beet was exposed to gamma radiation from a cobalt 60 source5,
while the other halves were stored in the same r o o m but protected from irradiation by appropriate shielding. Dosages of 5,000
to 20,000 r were used. Following irradiation, all pieces were
stored moist in a refrigerated r o o m at 1 to 2 C. Periodically
thereafter, matched pieces were sawed from each beet-half and
sliced to 1 mm thickness; disks were t h e n cut with a cork borer,
washed, a n d tissue respiratory rates d e t e r m i n e d by the Warburg
technique, as previously reported ( 1 3 ) . Results, as shown in
T a b l e 1, indicate an appreciable increase in respiratory rate
following irradiation. T h i s increased rate d i d n o t subside for
a b o u t 70 days. It would appear, then, that treatment of sugar
beets with gamma radiation in the dosage range indicated would
not represent a useful technique for decreasing respiratory sugar
losses in stored sugar beets.

Table 1.Effect of gamma irradiation on the respiration of sugar beet root tissue.
Values in percentage of matched nonirradiated check (CO60 source).

VOL. 13, N O . 4, J A N U A R Y 1965

335

R a d i a t i o n Effects on Leaf Metabolism


T h e r e a p p e a r to be very few reports in the literature concerning t h e effects of i o n i z i n g radiation on leaf respiration (3). In
the p r e s e n t e x p e r i m e n t s , disks were cut from m a t u r e sugar beet
leaves a n d s a m p l e s i r r a d i a t e d with a cobalt 60 source 5 for varying
dosages. T h e disks w e r e t h e n stored in plastic bags in a cold
room at 2 C. At intervals, samples were removed a n d respiratory
rates d e t e r m i n e d m a n o m e t r i c a l l y . Irradiated leaf disks, as shown
in T a b l e 2, r e s p i r e d m o r e rapidly t h a n u n i r r a d i a t e d checks for
several days. At h i g h e r dosages, respiratory rates then decreased
with t i m e , p r e s u m a b l y as the cells gradually died. Dosages of
over a m i l l i o n r w e r e r e q u i r e d to kill sugar beet leaf tissue immediately.
Table 2. Effect of X-irradiation on the respiration of sugar beet leaf tissue.

Some i m m e d i a t e c h l o r o p h y l l destruction occurs in irradiated


sugar b e e t leaves; however, this is not large in the dosage range
up to several h u n d r e d t h o u s a n d r. In contrast, chlorophyll dissolved in o r g a n i c solvents is relatively sensitive to ionizing radiation. T h e effects of i o n i z i n g r a d i a t i o n on leaf photosynthesis are
complex (3). F o r e x a m p l e , a dosage of 30,000 r will decrease
the r a t e of c a r b o n d i o x i d e fixation in wheat leaves by 50 percent
(15). T h i s s a m e dosage, however, had no detectable effects on
the o x y g e n e v o l v i n g ability ( H i l l reaction) of chloroplasts isolated from t h e i r r a d i a t e d leaves. T h i s probably means that some
enzyme system i n v o l v e d in the carbon dioxide-fixing steps is
relatively sensitive, w h i l e those concerned with water-splitting
and oxygen e v o l u t i o n a r e r a t h e r insensitive. In the present work
no m e a s u r e m e n t s w e r e m a d e on whole-leaf photosynthesis. However, studies w e r e m a d e of t h e effects of radiation on the photochemical o x y g e n e v o l u t i o n (Hill reaction) of isolated sugar beet
chloroplasts H i l l r e a c t i o n activity was relatively sensitive to
6
These irradiations were carried out with a 5,000 curie colbolt60 source and with a
gamma source containing appropriately aged spent reactor fuel rods. The irradiation was
performed by Dugway Proving Grounds, Utah, througn the cooperationi of Dr. C. J.
Christensen of the University of Utah.

336

JOURNAL OF THE A. S. S. B. T.

gamma radiation. T h e activity of irradiated chloroplasts was


measured potentiometrically (10) at a series of different light
intensities. T h i s permitted d e t e r m i n i n g the effects of radiation
separately on the rate-limiting photochemical process and the
rate-limiting dark process of the Hill reaction (6). Dosages up
to 750,000 r actually increased the rate of the photochemical
process of the Hill reaction; above this inhibition occurred, but
loss was very slight, even at 1,000,000 r. T h e dark process was
more sensitive, being decreased about 5 0 % at dosages of 1,000,000
r. Radiation damage to chloroplasts was decreased somewhat by
the usual protective agents such as beta-aminoethylisothiuronium
salts (AET). T h i s work will be published in detail elsewhere.
Radiation Effects on Seed Germination and Respiration
T h e r e is a tremendous literature concerning effects of many
kinds of ionizing radiation on seed germination and the early
growth of seedlings (3). Generally speaking, with increased dosages, seed germination is delayed a n d seedling growth is decreased. N u m e r o u s types of morphological abnormalities are also
produced. In a few species, however, low dosages appear to be
somewhat stimulating. T h e r e are only a few reports concerning
radiation effects on the germination of sugar beet seeds.
In the present work, sugar beet seeds were subjected to X rays
from a 250 K V A machine with dosages r a n g i n g from 3,000 to
1,000,000 r. O n e g r o u p of samples was irradiated in an air-dry
condition, while the other g r o u p was irradiated while moist,
Table 3.Effect of X-irradiation of dry sugar beet seeds on percent germination and
mortality of seedlings. (Decline in percent germination indicates some seedlings died.)

VOL.

13,

No.

4,

JANUARY

1965

337

following 20 h o u r s of w a s h i n g in r u n n i n g tap water. T h e data


on seeds i r r a d i a t e d in t h e dry condition are presented in T a b l e
3. As m a y be seen, g e r m i n a t i o n is delayed, a n d the percentage
of seeds g e r m i n a t i n g is generally decreased with increasing dosage. S e e d l i n g g r o w t h was reduced, a n d a variety of leaf abn o r m a l i t i e s w e r e observed. Seeds irradiated in a moist condition
were c o n s i d e r a b l y m o r e sensitive to ionizing radiation, as shown
in T a b l e 4. T h i s p h e n o m e n o n is well k n o w n (3).
Table 4.Effect of X-irradiation of moist sugar beet seeds on percent germination
and mortality of seedlings. (Seeds washed 20 hours in running tap water and left moist
during irradiation.) (Decline in percent germination indicates some seedlings died.)

Mikaelsen, et al. (5) f o u n d t h a t n e u t r o n dosages of 30-3600


reps h a d little effect on t h e respiration of germinating barley
seeds d u r i n g t h e first 2 days of germination. After this, irradiated
plants s h o w e d a r e d u c e d respiratory rate which was correlated
with r a d i a t i o n - p r o d u c e d r e d u c t i o n in seedling length. In the
present w o r k , s u g a r b e e t seedballs were X-rayed with dosages of
5,000 to 50,000 r. T h e seeds were then soaked, allowed to germinate, a n d t h e p e r c e n t g e r m i n a t i o n a n d respiratory rates determ i n e d at intervals, as s h o w n in T a b l e 5. Germination was progressively i n h i b i t e d by increasing X-ray dosages. Irradiation
s t i m u l a t e d r e s p i r a t i o n for t h e first 48 hours after treatment, even
t h o u g h g e r m i n a t i o n was i n h i b i t e d . After 73 hours, respiration
was lower in t h e i r r a d i a t e d samples, b u t after 138 hours respiration was a b o u t t h e same in the treated and untreated samples.
R a d i a t i o n Effect on Steckling Growth
Steckling s u g a r beets were t r i m m e d , and 16 lots of 10 beets
each w e r e i r r a d i a t e d at dosages from 200 to 1,000,000 r. A cobalt 6 0
source was u s e d for dosages up to 5,000 r; for higher dosages a

338

JOURNAL OF THE A. S. S. B. T.

Table 5.Effect of X-irradiation on the germination and respiration of sugar beet


seedballs. Irradiated samples are reported in percentage of the values of controls.

F i g u r e 1.Effect of g a m m a
23 days after r e p l a n t i n g .

radiation

on

regrowth

of sugar beets

VOL.

13, N o . 4, J A N U A R Y

1965

339

radiation s o u r c e c o n t a i n i n g appropriately aged, reactor fuel-rods


was used. T h e b e e t s w e r e t h e n planted in soil in a l u m i n u m
cylinders. T y p i c a l effects of radiation on beet regrowth after 23
days are s h o w n in F i g u r e 1. N o n e of the beets survived dosages
of 200,000 r or g r e a t e r ; no regrowth was observed at dosages of
100,000 r, a n d a l m o s t all plants were killed at this dosage. T h e
leaves of i r r a d i a t e d beets showed a marked increase in pubescence;
in general, n e w leaves were finely mottled, and a variety of leaf
and p e t i o l e a b n o r m a l i t i e s were observed, including leaf-blade
r e d u c t i o n a n d fusion of adjacent petioles. Some of the irradiated
beets s h o w e d a loss of apical dominance. In other experiments,
groups of s t e c k l i n g beets of three varieties were irradiated with
dosages of 2,000 to 5,000 r a n d t h e n replanted in a field plot.
T h e observed effects were similar to those described above. Radiation effects of these general types have been shown with a variety
of o t h e r p l a n t s (3).
Literature Cited
(1)

D A L L Y N , S. L., R . L. SAWYER a n d A. H . SPARROW.

1955.

Extending

o n i o n storage life by gamma irradiation. Nucleonics 13 (4) : 48-49.


(2)

GLEGG,

R.

E.,

F.

P.

BOYLE,

L.

W.

TUTTLE,

D.

E.

WILSON

and Z.

I.

KERTESZ. 1956. Effects of ionizing radiation on plant tissues. I.


Q u a n t i t a t i v e measurements of the softening of apples, beets and
carrots. R a d i a t i o n Research 5: 127-133.
(3) GUNCKEL, J. E. a n d A. H. SPARROW. 1961. Ionizing radiations- biochemical, physiological and morphological aspects of their efiects on
plants. Encyclopedia of Plant Physiology, Vol. XVI, edited by
W. R u h l a n d , Springer-Verlag, Berlin, pp. 555-611.
(4)

GUSTAFSON, F . G., L. E. BROWNELL a n d R. E. MARTENS.

1957. Influence

of gamma-irradiation on potatoe tubers on the rate of respiration.


American Potato J. 34: 177-182.
(5)

MIKAELSEN,

K.,

I.

P.

BJORNSETH

and

H.

HALVORSEN.

1956.

Experi-

ments on the effects of neutron-irradiation on the respiration of


barley seeds. I. Effects on growth and oxygen uptake in barley
seedlings of different varieties. Physical Plantarum 9: 697-711.
(6)

R I E S K E , J. S., R . L U M R V a n d J. D. SPIKES.

1959.

T h e mechanism of

the photochemical activity of isolated chlcroplasts. III. Dependence


of velocity on light intensity. Plant Physiol. 34: 293-300.
(7) SMOCK, R. M. a n d A. H. SPARROW. 1957. A study of the effect of gamma radiation on apples. Proc. Am. Soc. Hort. Sci. 70: 67-69.
(8) SNYDER, F. W. and D. E. W I A N T . 1959. Effect of high energy electron
irradiation on respiration of whole sugar beet roots. Proc. Am. Soc.
Sugar Beet T e c h n o l . X: 356-358.
(9) SPARROW, A. H. 1960. Uses of large sources of ionizing radiation in
botanical research and some possible practical applications, in:
Large R a d i a t i o n Sources in Industry. International Atomic Energy
Agency, Vienna, p p . 195-218.

340

JOURNAL OF THE A. S. S. B. T.

(10) SPIKES, J. D., R. LUMRY, J. S. RIESKE a n d R. J. MARCUS.

ing oxidation-reduction potentials in plant


Physiol. 29: 161-164.
(11)

1954. Record

preparations.

SPIKES, J. D., R. LUMRY, J. S. RIESKE a n d R. J. MARCUS.

Plant

1963. Radia-

tion effects and peaceful uses of atomic energy in the plant and soil
sciences, in: Radioecology (edited by V. Schultz and A. W. Klement,
Jr.), Reinhold Publishing Corp., New York. pp. 5-11.
(12) STOUT, MYRON. 1957. Respiratory losses from sugar beets soon after
harvest. J. Am. Soc. Sugar Beet Technol. IX (4) : 350-353.
(13)

STOUT, MYRON, a n d J . D. SPIKES,

1957.

Respiratory metabolism of

sugar beets. J. Am. Soc. Sugar Beet Technol. IX (6) : 469-475.


(14) SUSMAN, A. S. 1953. T h e effect of ionizing radiations upon the respiration and oxidases of the potato tuber. J. Cell. Comp. Physiol. 42:
273-283.
(15) ZILL, L. P. and N. E. TOLBERT. 1958. T h e effect of ionizing and ultraviolet radiations on photosynthesis. Arch. Biochem. Biophsy. 76:
196-203.

I m p a c t Effects on Germination and Seedling


Vigor of Sugar Beets1
O. R.

K U N Z E , F . W . SNYDER AND C . W . H A L L 2
Received for publication June 1, 1064

T h e g e r m i n a t i o n percentage of m o n o g e r m sugar beets is often


lower t h a n desired. T h i s is of concern to farmers who wish to
space-plant t h e seed to get a uniform stand.
T h i s research was u n d e r t a k e n to d e t e r m i n e if certain phases
of t h e m e c h a n i c a l h a n d l i n g a n d processing operations of the seed
may affect its g e r m i n a t i n g ability or seedling vigor. Sugar beet
fruits a r e processed before planting. T h i s processing operation,
which involves decortication, removal of fruits containing no
seeds a n d sizing, usually does n o t visibly h a r m the seed. W o r k
by P e r r y a n d H a l l 3 at Michigan State University has indicated
that visible d a m a g e is an unsatisfactory measure for determining
g e r m i n a t i o n decreases of pea beans resulting from processing
operations.
T w o factors, i n h e r e n t in the processing operations, which
may cause g e r m i n a t i o n decreases or loss of seedling vigor are
m o i s t u r e c o n t e n t of t h e seed a n d impact-loadings experienced
d u r i n g t h e processing o p e r a t i o n . H i g h e r or lower moisture contents m a y m a k e t h e seed m o r e or less subject to damage by the
processing o p e r a t i o n . P e r r y a n d H a l l found that pea beans
h a v i n g relatively h i g h m o i s t u r e content are less subject to p r o cessing d a m a g e t h a n a r e b e a n s h a v i n g a lower moisture content.
I m p a c t - l o a d s m a y result e i t h e r from processing e q u i p m e n t
striking t h e n o n - m o v i n g sugar beet fruit or from moving: fruits
striking an object. In e i t h e r case the impact-load may be conc e n t r a t e d at a p o i n t or it may be distributed over an area dep e n d i n g u p o n t h e position of the fruit. Hence there appear to
be at least t h r e e factors related to impact conditions which may
lower seed g e r m i n a t i o n or seedling vigor. T h e s e are m a g n i t u d e
of the i m p a c t energy; the time rate, or period, d u r i n g which
this i m p a c t energy is absorbed by the seed: a n d the p o i n t of
1
Cooperative investigation of the Michigan Agricultural Experiment Station and the
Crops Research Divisionf Agricultural Research Service. U. S. Department of Agriculture.
Approved for publication as Journal Article # 3333, Michigan Agricultural Experiment
Station.
2
Research Assistant. Department of Agricultural Engineering; Plant physiologist Crops
Research Division, Agricultural Research Service, U. S. Department of Agriculture and
Professor, Department of Agricultural Engineering, respectively, Michigan State University,
East Lansing, Michigan.
3
Perry, John S. and C. W. Hall, 1960. Germination of pea beans as affected by
moisture and temperature at imnact-loadings. Michigan Agricultural Experiment Station,
Quarterly Bulletin 43(1): 33-39, Michigan State University, East Lansing, Michigan.

342

JOURNAL OF THE A. S. S. B. T

impact-loading the seed, e.g., seed position as related to impactloads.


Although these factors may be obvious, precise measurement
of them presents problems of considerable magnitude. Theoretically the most severe impact condition would be that in
which the total impact energy is dissipated in a single contact
on the smallest seed surface in the shortest period of time. The
rate of energy absorption would be the greatest u n d e r this condition. Relative to the seed, the most severe impact-loading may
actually be a small impact on the fruit which is transmitted to
a point of the seed that is particularly sensitive.
Materials a n d M e t h o d s
Varieties of sugar beet seeds used were hybrids 62B24-20,
63B11-6 and (SL 126 X 128) ms X 5822-0. All three varieties
were selected because of their demonstrated high germination
capability. T h e y were hand-processed as necessary for each experiment. Fruits were m a i n t a i n e d at 68 F and at 4 0 % relative
humidity (approximately 1 0 % moisture content, dry basis) or
higher prior to treatment a n d were placed on a blotter to terminate the seed within 24 hours after treatment. Only those fruits
which experienced no visual physical damage from the impact
treatments were used in the tests. Daily counts were made of
germinated seed:. Following a 7- or 10-day period, root and shoot
lengths of all seedlings were measured in centimeters. Fruits
with u n g e r m i n a t e d seeds were bisected to d e t e r m i n e if the fruit
contained a developed seed.
Steel-sphere
impact-loads
Sugar beet fruits, positioned on a 1/2-inch steel base-plate,
were impact-loaded by d r o p p i n g steel spheres through glass tubes
(Figure 1). A p p r o p r i a t e cylindrical indentations were made in
the base-plate to restrict or limit seed m o v e m e n t d u r i n g the impacting process. A stand was used to hold the glass tubes in a
vertical position. Clearance between the steel plate and the
end of the glass tubes was a b o u t one-half the diameter of the
sphere used. Glass tubes of different diameters were used to
accommodate the steel spheres of different sizes. Combinations
of tube-lengths a n d sphere-weights were used to get the desired
impact-energies.
Differently shaped indentations were m a d e in the base-plate
to accommodate t h e fruits in various positions. Cylindrical indentations with flat bottoms were utilized while impacting fruits
placed cap down in the base-plate. Cylindrical indentations with
a conical base were used for impacting fruits on the cap. Two

VOL.

13, N o . 4, JANUARY

1965

343

F i g u r e 1.Stand with glass tube, steel sphere and impact base-plate.

1/2-inch steel plates separated by shims were used to hold fruits


which w e r e i m p a c t e d on t h e i r edge. Additional impressions were
m a c h i n e d in t h e steel plates to hold the fruits so that their edges
w o u l d be in a vertical plane.
T h e following imperfections in the above treatments are
recognized: 1) generally t h e fruit was n o t in a completely unstrained c o n d i t i o n w h e n impact-loading occurred, 2) in nearly
every case of impact-loading t h e r e was some a b o u n d of the steel
sphere. T h i s i n d i c a t e d that the total energy had n o t been ret a i n e d b y t h e fruit d u r i n g the initial impact. T h e m a g n i t u d e
of t h e r e b o u n d varied considerably from fruit to fruit.

544

JOURNAL OF THE A. S. S. B. f

Pneumatic
impact-loads
T h e second m e t h o d of impact-loading the individual sugar
beet fruits was developed by F. H. Peto 4 of the British Columbia
Sugar Refining Company in Vancouver, Canada. T h i s method
utilized compressed air, a venturi tube, a n d a steel impact-plate
(Figure 2). T h e compressed air was connected to the venturi
tube. Slight modifications were m a d e in the apparatus to permit
reasonably accurate reproduction of the tests. A gage, which
measured static pressure only, was installed in the air line immediately before the venturi tube. T h e distance between the
end of the venturi tube and the 3/8-inch steel impact-plate could
be varied to whatever distance was desired. A screen cylinder
approximately 8 inches in diameter was b u i l t using the steel
impact-plate as one end. A circular screen section with a 1/2-inch
diameter hole at its center was used to enclose the other end.
Fruits could then be projected against the base-plate through
the 1/2-inch opening. T h e screen cylinder retained all fruits
treated in this m a n n e r .

Figure 2.Screen cylnder with 3/8" steel impact-plate on the left and
sliding circular screen section inserted on the right. Seed was dropped
through small funnel into high velocity air stream. Gage recorded static
pressure.

Fruits were hand-processed and sized into 1/64-inch diameter


classes (7/64, 8/64, 9/64 a n d 10/64). T h e v e n t u r i tube could
n o t accommodate fruits larger t h a n 10/64 inches in size. The air
pressure could be regulated by m a n i p u l a t i o n of a valve. Fruits
were d r o p p e d individually i n t o t h e v e n t u r i t u b e a n d impacted
against the steel plate. All t h e p n e u m a t i c tests were conducted
with a pressure differential of 2 psi a n d a distance of 6 inches
between the v e n t u r i exhaust a n d the steel impact-plate.
4
F. H. Peto described this method in correspondence with F. W Snyder and loaned
the venturi tube to conduct the pneumatic impact experiments.

VOL. 13, N o . 4 , JANUARY 1965

3 4 5

Results
Impact-energies of 63 gram-centimeters applied with a steel
sphere were found to be detrimental to the fruit in any of the
positions listed in Table 1. Each treatment consisted of 25 to
28 seeds and germination tests were over a 10-day period. Impact
on the seedcap caused approximately 50% reduction in seedling
vigor. Treatments on the butt of the fruit or perpendicular to
the radicle of the seed caused about a 30% reduction in growth.
Impact-loads on the micropyle were the least detrimental but
still caused about a 14% decrease in seedling vigor. Seeds impacted by the steel sphere generally germinated. This was true
even if the seedcap had been cracked or broken by the impactload. However, vigor was impaired.
Table 1.Effect of a single 63 gram-centimeter impact on different parts of processed
fruits of sugar beet variety 62B24-20 with 25 to 28 seeds in each treatment.
Average length
Treatment

Root

Shoot

Cm.

Cm.

6.1

2.5

2.2
1.8
1.4

No impact
Impact on butt
Impact on micropyle
Impact perpendicular to radicle
Impact on cap

5.2
4.2
2.5

Percent of control
Root

Shoot

67*
85
69
41

73*
88
72
56

* A\erages for 2 tests.

In another experiment, two steel spheres (0.44 and 1.03


grams) were used to apply impact-loads of various magnitudes
(Table 2). These data represent three sub-experiments. Each
treatment in a sub-experiment consisted of 25 to 28 seeds. Germination tests were over a 7-day period. Impact-energies as small
as 25 gram-centimeters caused some reduction in root and shoot
growth. Treatments of 43 and 65 gram-centimeters with the
heavier sphere had nearly equal effects. The average reduction
in growth was approximately 40% for the root and shoot. The
Table 2.Effect upon seedling vigor of a single impact on seedcap when varieties
63B11-6 were used in sub-experiments A and B and (SL 126 X 128)ms X 5822-0 was used
for sub-experiment C.
Percent of control
Root
Impact energy

Sub-experiment

25 gram-centimeters,
43 gram-centimeters,
51 gram-centimeters,
65 gram-centimeters,

light sphere
heavy sphere
light sphere
heavy sphere

A
98
57
104
55

B
80
49
95
51
81

Shoot
C
98
73
92
36
86

A
79
39
93
65

B
79
64
107

C
83
61
78

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JOURNAL OF THE A. S. S. B, T

variation between experiments suggests that m o r e research may


be necessary to accurately evaluate impact-loads of this magnitude. T h e trend, however, is apparent.
Results of the 51 gram-centimeter impacts with the light
sphere do not agree with the trends produced by the other
impact-loadings. T h e diameter of this sphere was 0.475 centimeters while that of the glass t u b e was 0.500 centimeters. Fired
ends of the glass tubes as received from the factory were generally
contracted enough to p r o h i b i t passage of the sphere. These were
removed before the tubes were used. Otherwise there were no
indications of friction between the sphere a n d the tube. Results
from the treatments ( T a b l e 2) implied that energy was consumed in the drop. Therefore in a n o t h e r experiment, a glass
t u b e with a diameter of 0.600 centimeters was used (Table 3).
Each treatment consisted of 50 to 52 seeds a n d the growth period
was 7 days. A greater reduction in growth was experienced for
both root and shoot with the larger t u b e when 25 gram-centimeters of impact-energy were used. For the 51 gram-centimeter
impact-energies, the data from the larger d i a m e t e r t u b e showed
an 8% reduction in root growth b u t a 2% increase in shoot
growth. T h i s inconsistency is a t t r i b u t e d to the biological variation in the seed. T h e same seed variety was used for sub-experiment C in T a b l e 2 as was used for d e t e r m i n i n g the data in
T a b l e 3.
Table 3.Percent of control growth resulting after a single impact on the seedcap
when the light steel sphere was dropped on (SL 126 x 128)ms x 5822-0 fruits*.
Tube diam. centimeters
Impact-energies gram-centimeters

25

0.500
51

25

0.600
51

Root
Shoot

99
98

94
82

95
97

86
84

* Control root length 6.7, shoot length 3.1 cm.

Pneumatic

impact-loads

Sugar beet fruits may be impact-loaded w h e n conveyed by


air or w h e n striking a surface or object after a free fall through
the air. Fruits were hand-processed a n d the 7-10/64-inch diameter
fruits were used in the p n e u m a t i c system. A i r pressure differential was reduced to a level (2 psi) where virtually no observable physical damage occurred to the fruit from a single impact. Later calculations indicate that this impact-energy was of
the order of 29 gram-centimeters for a fruit having a weight of
0.0125 grams. Repeated application of the impact-load caused
some fruits to shed their caps.

VOL. 13, N o . 4, JANUARY 1965

347

Table 4.Effect of pneumatic impact on germination and vigor of seedlines


of
8
processed sugar beet fruits, (SL 126 X 128)ms X 5822-0.
Percent seeds
germinated

No. impacts

94
95
92
77

0
1
3
5

Average length
Root

Shoot

Cm.

Cm.

4.9

2.0

3.5
3.2

1.5
1.3

Percent of control
Root

Shoot

71
65

75
65

Table 5.Comparison of type* and number of impacts on seedling vigor of handprocessed (SL 126 X 128)ms X 5822-0 fruits.
Growth as percent of con trol**
Treatment
Sphere

Pneumatic

Number of impacts

Fruit diam.
inches

9/64

Root
Shoot

96
97

93
92

92
93

80
80

81
84

8/64

Root
Shoot

79
74

77
74

Average

Root
Shoot

86
84

79
77

72
73
77
79

9/64

Root
Shoot

85
78

78
70

70
64

66
64

8/64

Root
Shoot

60
60

53
53

60
73
64
64

Average

Root
Shoot

__

65
62

60
59

62
68

* Steel-sphere impacts of 32 gram-centimeters or pneumatic impacts using 2 psi pressure


differential.
** 9/64-inch fruitscontrol, root length 6.3, shoot length 2.4 cm.
8/64-inch fruitscontrol, root length 6.8, shoot length 3.2 cm.

T h e p n e u m a t i c impact-load applications reduced vigor similar


to t h a t c a u s e d by t h e steel spheres ( T a b l e 4). Each treatment
consisted of 110 to 120 seeds with a 7-day growth period. T h r e e
impacts r e d u c e d r o o t g r o w t h 2 9 % a n d shoot growth 2 5 % , while
f i v e i m p a c t s r e d u c e d b o t h r o o t a n d shoot growth 3 5 % . T h e data
indicate t h a t t h e seeds w e r e capable of withstanding three imp a c t s of this m a g n i t u d e w i t h o u t a serious decrease in germinat i o n , h o w e v e r , f i v e i m p a c t s r e d u c e d their germination capability
by

17%.

A n o t h e r e x p e r i m e n t u s i n g t h e steel sphere and the p n e u m a t i c


systems of i m p a c t - l o a d i n g confirmed that both methods were
c a p a b l e of r e d u c i n g seedling vigor ( T a b l e 5). Each t r e a t m e n t
consisted of 50 to 56 seeds a n d a 7-day growth period. T h e re-

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JOURNAL OF THE A. S. S. B. T

Table 6.Comparison of type* and number of impacts on percentage germinatior


of seeds in hand-processed (SI. 126 X 128)ms x 5822-0 fruits.
Number of impacts
Treatment

Sphere

Exp. 1
Exp. 2
Average

92

95

98
96
97

93
98
96

95
98
97

Pneumatic

Exp. 1
Exp. 2
Average

96

96

93
94
94

79
81
80

79
84
82

* Steel-sphere impacts of 32 gram-centimeters or pneumatic impacts utilizing 2 psi pressure


differential.

lationship between fruit size a n d a given impact-load is illustrated


in the steel-sphere treatments. T h e seeds in the larger fruits suffered less growth reduction from repeated impacts of a given
energy level than d i d the seeds in the smaller fruits. For the
given conditions, these results imply that fruits with larger than
9/64-inch diameters would have suffered even less reduction
while fruits with smaller t h a n 8/64-inch diameters would have
suffered an even greater reduction in seedling vigor. Similar
reductions were obtained from the p n e u m a t i c treatments. No
a t t e m p t was made to distinguish between energy levels when
8/64 or 9/64-inch d i a m e t e r fruits were used in the pneumatic
system.
Results for the five p n e u m a t i c impacts in T a b l e s 4 and 5 are
in good agreement, each showing an average reduction in root
a n d shoot growth of 3 5 % . Results from the three pneumatic
impacts are not in such close agreement. T h e percentage reductions of root a n d shoot growth usually followed each other
very closely for either m e t h o d of impact-loading. Therefore,
when a sufficiently large n u m b e r of seeds are used in an experiment, a m e a s u r e m e n t of root growth should generally be adequate
to d e t e r m i n e reductions resulting from impact treatments.
G e r m i n a t i o n percentages in this e x p e r i m e n t were not decreased by the steel sphere impact-loading b u t were decreased
by repeated p n e u m a t i c impact-loads ( T a b l e 6). T h e first three
p n e u m a t i c impacts caused virtually no decrease in percentage
germination of the seed. After the fourth impact, however, there
was a definite decrease. T h i s observation is in agreement with
results in T a b l e 4 where the percentage germination decreased
significantly between t h e t h i r d a n d fifth pneumatic impacts
Therefore the fourth p n e u m a t i c i m p a c t seemed to be of critical
significance.

VOL.

13, N o . 4, J A N U A R Y

1965

349

Discussion
Energy
of steel-sphere
impacts
T h e t w o m e t h o d s of impact-loading can be subjected to a
more critical analysis. T h e potential energy of the steel sphere
is e q u i v a l e n t to t h e w e i g h t of t h e sphere multiplied by the height
that it is s u s p e n d e d a b o v e the sugar beet fruit. W h e n t h e sphere
is d r o p p e d , t h e p o t e n t i a l energy is converted to kinetic energy.
At i m p a c t t h e k i n e t i c energy is absorbed by the fruit if no reb o u n d occurs. A s a m p l e energy calculation is shown below.
1.03 g r a m X 31 centimeters = 32 gram-centimeters
If r e b o u n d occurs, t h e fruit returns part of the energy to
the steel s p h e r e . T h i s r e b o u n d may be of a variable nature dep e n d i n g on t h e c o n d i t i o n s of impact. If the fruit shifts its position w h i l e i m p a c t - l o a d i n g occurs, some energy is utilized in
p r o d u c i n g t h i s m o v e m e n t a n d the sphere is deflected against
the glass t u b e . T h e r e s u l t i n g r e b o u n d is of a smaller magnitude
a n d t h e fruit itself often r e b o u n d s . Consequently the second
impact r e s u l t i n g from t h e r e b o u n d may strike the fruit at a
different p o i n t . W i t h a 31-centimeter d r o p for the heavier sphere,
m a x i m u m r e b o u n d s of slightly over 10 centimeters were observed. H e n c e t h e energy r e t a i n e d by the fruit d u r i n g the initial
impact was o n l y a b o u t 20 gram-centimeters. T h e importance
of t h e r e b o u n d s e e m e d less significant, however, than the overall effect of i m p a c t - l o a d i n g t h e fruit. Neglecting friction and
impact losses b e t w e e n t h e sphere a n d glass tube, the total kinetic
energy was finally a b s o r b e d by the fruit.
If t h e s p h e r e is d r o p p e d several times from a given height
on a fruit, t h e n e t effect is n o t additive and does not compare
to a single d r o p i n v o l v i n g t h e same net energy. For example,
a fruit m a y be a b l e to absorb two impact-loads of 25 gramc e n t i m e t e r s each b u t m a y n o t be able to withstand a single
impact-load of 50 gram-centimeters. T h e difference is caused by
the t i m e d u r i n g w h i c h t h e fruit absorbs the energy.
Energy
of pneumatic
impacts
T h e p n e u m a t i c m e t h o d of impact-loading proved to be very
capable of i n j u r i n g t h e seed. Preliminary tests were r u n using
10, 15 a n d 20 psi pressure differentials across the venturi tube.
Impacts p r o d u c e d w i t h these pressure differentials mutilated
nearly every fruit. C o n s e q u e n t l y the pressure difference was
r e d u c e d to a final v a l u e of 2 psi. Calculations were made to
estimate t h e k i n e t i c energy of a sugar beet fruit when accelerated
by this p r e s s u r e difference.
T o m a k e these calculations, the pressure gage and venturi
t u b e w e r e c o n n e c t e d to a pressure regulating valve installed in

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JOURNAL OF THE A. S. S. B. T

a steel pressure cylinder. T h i s container was filled with compressed air a n d weighed. T h e air was then exhausted at a rate
which gave a 2 psi pressure differential across the v e n t u r i tube.
Diameters of the v e n t u r i throat a n d exhaust were 0.188 and
0.290 inches, respectively. T h e time r e q u i r e d to exhaust about
8 0 % of the air was d e t e r m i n e d a n d then the cylinder was weighed
again. By using the time interval r e q u i r e d to release a definite
weight of air, we calculated t h e velocities at the venturi throat
a n d exhaust to be 430 a n d 165 ft/sec, respectively. If lo<v
additional air was d r a w n in t h r o u g h the seed injection tube,
the final velocity at the v e n t u r i exhaust was near 180 ft/sec.
W h e n a fruit was d r o p p e d i n t o t h e v e n t u r i tube, the flow
of air was restricted thus causing an instantaneous pressure
build-up. T h i s caused the air velocity to increase around the
perimeter of the fruit which was b e i n g accelerated. Flotation
velocities for 8/64 a n d 9/64-inch d i a m e t e r hand-processed sugar
beet fruits were found to be approximately 17 ft/sec. With the
assumption of a 5 0 % velocity increase a r o u n d the fruit perimeter, calculations of particle acceleration in an air stream
indicated an impact velocity of 70 ft/sec.
T h e kinetic energy of a fruit weighing 0.0125 grams and
moving with a velocity of 70 ft/sec may be calculated as shown
below.

T h e observable physical damage, if any, was about equal


when the 32 gram-centimeters of impact-energy applied with
the steel sphere were c o m p a r e d with the 29 gram-centimeters
applied with the p n e u m a t i c system. T h e exact nature of the
seed damage resulting from either t h e steel sphere or the pneumatic impact-loads is n o t known. D u r i n g t h e germination tests
several cases of seed fracture were observed. T h e reduction in
seedling vigor as well as some deformed a n d abnormal growths
were the most p r o m i n e n t characteristics of the treated seeds.
T h e a b n o r m a l seedlings possibly developed from injured embryos
which could not support n o r m a l growth. F o r the given magni-

VOL. 13, No. 4, JANUARY 1965

351

tudes of impact-energy, t h e data ( T a b l e 5) indicate that the


physical d a m a g e to t h e t r u e seed was greater from pneumatic
impact-loading.
F r u i t s w h i c h w e r e subjected to repeated treatments with the
steel s p h e r e e v e n t u a l l y failed by chipping or cracking of the
seedcap. T h e n a t u r e of the loading, however, tended to keep the
cap in p o s i t i o n e v e n t h o u g h it may have been cracked or loosened.
H e n c e , if t h e seed h a d the ability to begin germination, it may
have h a d l i t t l e difficulty in emerging from the fruit. T h i s may
have b e e n possible even if t h e seedling was damaged and never
developed n o r m a l g r o w t h .
S h e d d i n g of t h e seedcap was t h e most common physical damage o b s e r v e d w h e n fruits were treated repeatedly with the pneumatic system. T h e i m p l i c a t i o n was, however, that the seed inside
the fruit c o u l d be d a m a g e d w i t h o u t any apparent damage to the
fruit. If this was t h e case, the damaged seed may have never
developed e n o u g h energy to force t h e seedcap from its base, and
hence an u n g e r m i n a t e d seed resulted.
Impact-energy

from

free

fall

H a n d - p r o c e s s e d fruits impelled vertically into the air with


the p n e u m a t i c system traveled to various heights. Possible reasons for t h e h e i g h t variations are: 1) configuration of the fruit,
2) fruit trajectory, 3) density of the fruit and 4) extent to
which fruit was processed.
C o n f i g u r a t i o n of t h e fruit obviously had an influence on its
trajectory. F r u i t s w h i c h stayed in the air stream after leaving
the v e n t u r i w e r e i m p e l l e d h i g h e r t h a n those whose trajectory
carried t h e m o u t of this stream. W e i g h t or density of the fruit
also was an i m p o r t a n t factor. Weights of individual (SL 126 X
128) ms X 5822-0 fruits w i t h diameters of 7/64-inch or larger
varied as m u c h as six-fold. E x t e n t of processing affected the
density of t h e fruits. In this research only hand-processed fruits
were used. O m i t t i n g e x t r e m e values, most fruits when projected
t h r o u g h t h e v e n t u r i at 2 psi pressure differential were impelled
to h e i g h t s of 15 to 20 feet. Since the density of the fruit was
relatively low, t h e a i r h a d a considerable impedance effect. T h i s
may be visualized from a study of the flotation or terminal
velocities of hand-processed sugar beet fruits. Preliminary work
indicated this m a x i m u m velocity to be approximately 25 rt/sec.
T h e r e f o r e , in a v a c u u m , the atmospheric flotation velocity would
be achieved in a p p r o x i m a t e l y eight-tenths of a second, as calculated below.

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JOURNAL OF THE A. S. S. B. 7-

T h e distance w i t h i n which this velocity would be achieved


in a vacuum is
s = (16.1) (.64) = 10.3 ft
In the air, the fruit would n o t accelerate as rapidly and theoretically would never reach its flotation velocity. T h e implication is, however, that near flotation velocity is reached within
a relatively short distance.
If flotation velocity in the air is 25 ft/sec a n d this velocity
is achieved in a vacuum after a 10.3-foot free fall, then the maxim u m kinetic energy of the fruit can be calculated. Assume the
individual fruit has a weight of 0.0125 gram.
0.0125 gram X 10.3 feet = energy, gram-feet
0.0125 X 10.3 X 12 X 2.54 = 3.9 gram-centimeters
T h e kinetic energy of a hand-processed fruit (0.0125 gram)
experiencing a free fall should never be very m u c h in excess of
3.9 gram-centimeters. T h e heaviest b u t n o t necessarily the most
dense hand-processed fruit encountered in this research weighed
0.0312 grams. Its flotation velocity was less than 25 ft/sec. The
impact energy resulting from an u n l i m i t e d free fall of this fruit
would approach 9.8 gram-centimeters. T h e s e energy levels are
q u i t e small when compared to the impact-loads of 25 gramcentimeters or greater per impact which were applied with the
steel spheres. T h e r e f o r e the conclusion may be d r a w n that handprocessed sugar beet fruits normally should n o t experience reduction in vigor or g e r m i n a t i o n capability from impacts received after free falls of virtually any length.
Conveying velocities are often as m u c h as 5 0 % higher than
flotation velocities. If physical damage to the fruit is disregarded,
m a x i m u m velocities are limited only by the equipment used.
T h i s research has indicated that r e d u c t i o n in germination and
seedling vigor is possible before observable physical damage
occurs to the fruit. Conveying velocities equivalent to two. or
three times the flotation velocities may cause fruits to reach energy
levels which are detrimental to g e r m i n a t i o n a n d seedling growth.
N u m e r o u s impacts of a smaller m a g n i t u d e may produce a similar

VOL. 13, N O . 4, JANUARY

1965

353

r e d u c t i o n . T h e r e f o r e c a u t i o n is suggested w h e n pneumatic
systems a r e u s e d to convey sugar beet fruits. T h e m i n i m u m air
velocity w h i c h will successfully convey the fruits is recommended.
Summary and Conclusions
H a n d - p r o c e s s e d sugar beet fruits were impact-loaded by d r o p ping steel s p h e r e s t h r o u g h glass tubes o n t o the fruits a n d also
by a c c e l e r a t i n g t h e fruits w i t h a p n e u m a t i c system a n d projecting
them against a steel impact-plate. Fruits were impact-loaded in
different p o s i t i o n s w i t h a steel sphere. Reduction in seedling
vigor was m o s t a p p a r e n t w h e n fruits were treated on the seedcap. Single impact-loads as small as 25 gram-centimeters applied
by a steel s p h e r e r e d u c e d r o o t growth approximately 6% and
shoot g r o w t h 1 3 % . Single p n e u m a t i c impacts having an estimated
impact-energy of 29 gram-centimeters reduced the root growth
1 5 % a n d t h e s h o o t g r o w t h 2 2 % . Five pneumatic impact-loads
of this s a m e m a g n i t u d e r e d u c e d r o o t a n d shoot growth approximately 3 5 % . S m a l l e r fruits suffered more reduction in germination a n d s e e d l i n g vigor t h a n d i d larger fruits for a given level
of i m p a c t - e n e r g y a p p l i e d repeatedly with a steel sphere. Generally, t h e p e r c e n t a g e r e d u c t i o n s in root and shoot growth were
so similar t h a t r o o t m e a s u r e m e n t s would have been sufficient for
d e t e r m i n i n g t h e r e d u c t i o n in seedling vigor.
T h e o r e t i c a l considerations c o m b i n e d with experimental results i n d i c a t e t h a t n e i t h e r seedling vigor n o r germination should
be r e d u c e d by i m p a c t s r e s u l t i n g from free falls of any distance
by a sugar b e e t fruit. Impact-energies are dependent upon the
velocity a n d w e i g h t of t h e fruit. In practical applications the
impact e n e r g y of 29 gram-centimeters is approximated by an
impact-velocity of 70 ft/sec w h e n the fruit weight is 0.0125 grams.
S e e d l i n g v i g o r a n d percentage germination can be reduced
t h r o u g h t h e a p p l i c a t i o n of impact-loads without visibly harming
the sugar b e e t fruit. Seeds which were impact-loaded three times
with t h e p n e u m a t i c system showed a small reduction in percentage g e r m i n a t i o n . Seeds which were impact-loaded four times
e x p e r i e n c e d a considerably greater reduction in percentage germination. T h e f o u r t h p n e u m a t i c impact seemed to be of critical
significance.

On the Nature of Hatching of Heterodera schachtii.


II. Natural Sources of Hatching Stimulants1
D . R . VlGLIERCHIO AND P . K . Y u 2
Received for publication June 10, 1964

Introduction
T h e hatching response of cysts of Heterodera schachtii has
received m u c h attention since the r e p o r t by Baunacke (l) 3 of
the stimulatory effects of the leachings of sugar beet roots on
larval emergence. If, as Shepherd (4) suggests, hatching is considered a starting point for the process of host infection and
the life cycle of the nematode, the effort a n d attention devoted to
this aspect of nematology in the past was almost inevitable. This
line of work subsequently became extended to the effects of
extracts of other plant organs a n d synthetic chemical agents. As
a result the evidence is overwhelming that leachings of the
appropriate plants contain active agents that stimulate significantly the emergence of larvae from cysts over the normal expected with water.
A great proportion of the research devoted to the phenomenon
of hatching has been concerned with the description of the effect
of physical or environmental factors on cysts or the bioassay
systems (3,7,8,9). T h e research into the chemistry of the hatching process has resolved itself mainly into the testing of a number
of diverse substances from natural a n d synthetic sources for
hatching factor activity (10,12) a n d the identification of a
naturally-occurring hatching factor as yet n o t achieved (2).
Despite the research effort to date little is k n o w n about the
n a t u r e or mechanism of hatching or the role of the naturallyoccurring hatching stimulants in eclosion.
Materials and Methods
Plants used in these studies included dame's violet (Hesperts
matronalis), tomato
(Lycopersicon esculentum)
rape
(Brassica
napus), sugar beet (Beta vulgaris), yellow m i l o (Sorghum vulgare), broccoli (Brassica oleracea, var. botrytti), alfalfa (Medical
sativa) barley (Hordenm
vulgare)
a n d onion (Allium cepa).
Plants were grown u n d e r greenhouse conditions in 6-inch pots
filled with sand a n d watered with modified Hoagland's nutrient
solution (11). W h e n the root systems h a d developed sufficiently
1
Research funds for this study were contributed in part by the Beet Sugar Development2 Foundation.
Associate Nemaiolojrist and Assistant Research Nematologist, respectively Department of Nematology, University of California, Davis, California.
3
Numbers in parentheses refer to literature cited.

VOL.

13, N o . 4, JANUARY

1965

355

they w e r e l e a c h e d profusely with distilled water to remove


soluble c o n s t i t u e n t s from the sand culture. At 24-hour intervals
thereafter each p l a n t was leached with sufficient distilled water
to m a k e up 500 ml of leaching solution. T h e leachings from
each p l a n t w e r e c o m b i n e d every 2 days a n d designated as a batch
so that in a leach p e r i o d there were obtained successively batch
1, 2, 3, etc. T h e leach p e r i o d was arbitrary, b u t less than 3 weeks.
To test t h e effect of lyophilization half of each batch was lyophilized a n d t h e r e m a i n d e r stored in a cold room as a solution.
For assay p u r p o s e s t h e d r i e d material was resuspended in distilled
water a d j u s t e d t o p H 5 w i t h H 2 S 0 4 o r N a O H . T h e redissolved
m a t e r i a l was tested at 5 concentrations overlapping the concentration of t h e o r i g i n a l leachate. T h e refrigerated solution was
tested at n a t u r a l s t r e n g t h as well as 2 dilutions.
L e a c h i n g s from g e r m i n a t i n g seedlings were obtained by placing seeds on a stainless steel screen suspended over a funnel in
a m i s t i n g a p p a r a t u s . T h e q u a n t i t y of seed used for funnel leaching was d e t e r m i n e d p r i m a r i l y by seed size, i.e., previous experim e n t s w i t h s u g a r b e e t g e r m indicated that seed depths greater
t h a n a b o u t 4 to 5 seed diameters were unsatisfactory. T h e collection of t h e leachings was b e g u n w h e n stem or root growth
became e v i d e n t . T h e daily collection from each funnel was
lyophilized a n d stored at 15C.
T h e m i s t i n g a p p a r a t u s consisted of a box 3 feet on edge
c o n s t r u c t e d of a n g l e a l u m i n u m a n d a l u m i n u m sheet and a plexiglass d o o r w i t h a m i s t i n g nozzle at the top center of the chamber.
H o t distilled w a t e r was supplied to the nozzle intermittently
from a h o t w a t e r h e a t e r (with variable temperature controls)
t h r o u g h an a p p r o p r i a t e c o m b i n a t i o n of solenoid valves controlled by a v a r i a b l e on-off interval timer. Fluorescent lights
(6-40 w a t t tubes) w e r e suspended just above the mist chamber
to supply i l l u m i n a t i o n to t h e germinating seedlings.
Cysts of Heterodera schachtii from sugar beets grown in sand
u n d e r g r e e n h o u s e c o n d i t i o n s were selected, separated and stored
by t h e m e t h o d d e v e l o p e d by Viglierchio (5). T h e hatching bioassay u t i l i z i n g t w o sources of cysts was conducted as described
by V i g l i e r c h i o (6). T h e cysts were suspended at the surface ot
the test s o l u t i o n by stainless steel screens in plastic wells cont a i n i n g 0.5 ml of solution sample. T h e cysts a n d solutions (8
replicates/test s o l u t i o n of a concentration series) were incubated
in a h u m i d i t y c h a m b e r at 25C. At 4-day intervals the emerged
larvae t o g e t h e r w i t h test solution were withdrawn for counting
a n d r e p l a c e d w i t h freshly p r e p a r e d test solution. The bioassay
was d i s c o n t i n u e d after t h r e e collections.

JOURNAL OF THE A. S. S. B. T.

356

Results
Leachings from all the plants tested stimulated hatching of
H. schachtii larvae to some degree (Figure 1, 2 a n d 3). It was
evident that the stimulatory activity of the leachate varied as the
leaching progressed. According to expectations the over-all activity of the leachate decreased to the level of water by the end
of the leach period; however, two patterns of larval emergence
in response to leachings were clearly resolved. T h e activity of
milo, alfalfa, and dame's violet leachings decreased to the level
of water hatching in a gradual fashion, whereas the activity of
rape, tomato, broccoli, t u r n i p a n d sugar beet did so erratically
(Figure 1). T h e fluctuations were much too large to be accounted
for by assay variability.
Concentration hatch curves (Figure 2 and 3) for refrigerated
leachate solution indicated that the plant leachings could be
rated in activity from high to low: t u r n i p , rape, dame's violet
and broccoli. T h e leachings of the other plants were very low
in activity, comparable to that of water.
W h e n the plant leachings were lyophilized then resuspended
for assay the hatching activity varied considerably. T h e response
of dame's violet was very high; t u r n i p and rape, high; sugar
beet, onion a n d tomato, moderate; alfalfa, milo, broccoli and
barley, very lowslightly above water. Comparison of the hatching curves of refrigerated solutions and lyophilized material indicated that drying had no effect on the activity of leachings
from t u r n i p , dame's violet, barley and m i l o b u t increased the
activity of leachings from tomato, rape, onion a n d sugar beet.
Drying decreased the activity of leaching of broccoli and possibly
alfalfa.
T h e hatching response to dried leachinors of germinating
seedlings was remarkably similar in all plants tested. Larval
emergence, moderately high, decreased with concentration of
dry solids similarly with all plants tested.
T h e acidity of the leachings of all plants decreased to neutrality as the leach period progressed with t h e greatest change occurring between batches 2 and 3 (Figure 4). Preliminary experiments with sugar beet germ had indicated little change in pH
between leach solution as collected a n d that lyophilized and redissolved. T h e leachings of the other g e r m i n a t i n g seedling increased in pH and most of t h e m became alkaline, pH 8-9. It
became standard practice thereafter before lyophilizing to render
the buffer capacity of the leachings acidic with H 2 S 0 4 until
acidity began to increase rapidly as indicated by a pH meter.

13

N o . 4, JANUARY

1965

357

Figure l . - T h e extraction of hatch factor materials . f r o m various


plant L o t s with progressive leaching as determined by hatching tests of
H. schachtii cysts with successive collections.
Figure 2 and 3 . - T h e cumulative hatch from H. schachtii cysts obtained
with several concentrations of hatch material from several sources prepared as indicated.
Figure 4 . _ T h e change in pH of extracts (original solution and reconstituted solution from dried solids) of several plant roots with
leaching.

JOURNAL OF THE A. S. S. B. T

358

Root weights of the greenhouse leached plants were surprisingly uniform except for the coarse rooted m i l o and fine
rooted dame's violet a n d alfalfa ( T a b l e 1). T h e physical seed
dimensions of milo and tomato a n d dame's violet were not
amenable to the use of uniform weights or n u m b e r s . Percent
germination and g e r m i n a t i o n times also varied as reported. The
dry solid content of the leaching from germinating seedlings
showed little correlation with either the seed n u m b e r or seed
weight. T h e host test for H. schachtii reported in T a b l e 1 was
for the indication of relative susceptibility a n d not necessarily
absolute susceptibility. It was evident that sugar beet was the
best host for H. schachtii b u t that the females were able to mature
more quickly on rape.
Discussion
T h e abnormal environmental regime the plants experienced
d u r i n g the leaching process would effect mineral and nutritional
imbalances resulting in the variable production of stimulatory
agents a n d / o r leaching of competitive inhibitors. It was of
interest to note that milo, alfalfa a n d dame's violet were nonhost plants whereas the others were hosts ( T a b l e 1). Tomato
has been reported as a host of H. schachtii; however, the susceptibility of the Pearson variety was apparently too low to be de
tectable by the test m e t h o d used in this experiment.
It was clear from the hatch curves of refrigerated teachings
(Figure 2 and 3) that the stimulatory potential varied greatly
from one plant to another. It was surprising to observe that
lyophilization, normally considered a gentle procedure, had
altered the hatch characteristics of the leachate of more than
half the plants tested. T h e increase in hatch-factor activity obtained u p o n lyophilization of leachates could be explained by
a loss of inhibitors either through volatilization or chemical
inactivation. T h e change in pH of leachate u p o n drying was
consistent with this notion. T h e loss of volatile organic acids or
bases would explain a change in acidity.
It was unlikely that the e n v i r o n m e n t a l conditions during
lyophilization would have been conducive to synthesis of hatch
factors from precursors. T h e a p p a r e n t inactivation of crude
hatch material u p o n lyophilization as observed in the case or
broccoli and to a lesser degree with alfalfa may have been a
result of active factor volatilization or heat inactivation. In the
process of freeze-drying water was removed from the frozen
solution by sublimation. T h e dry solids r e m a i n i n g behind would
have b e e n at the t e m p e r a t u r e of t h e frozen solution if in contact

VOL.
13, No.. 4, JANUARY

Table 1.Various data for the plants used in this study.

Plants/pot
Root Wt. (g)/pot
Wt. seeds (g)/rep.
No. seeds/rep.
Germination time (hrs.)
Percent germination
Dry solids (g) from seeds
(10 days)
Susceptibility
H. schachtii females
H. schachtii cysts

Turnip

Alfalfa

Rape

Milo

8
70
13
3700
42
96

8
77
10
4400
42
94

15
36
11
5100
42
85

8
73
12
3100
72
93

8
105
22
765
72
96

0.57

0.45

0.45

0.55

0.55

1995
31

1127
15

958
9

0
0

679
556

8
76
9
4700
120
20

Broccoli

Dame's
violet

Tomato
8

Barley

Onion

0
0

0
0

8
71
6
1500
120
85

42
10
5800
120
44

0.74

0.51

0.38

0
0

0
0

0
0

1965

Sugar
beet

JOURNAL OF THE A. S. S. B. T

360

with it. T h e dry solids in contact with their container would


have been at the temperature of the room or the freeze-dryer
warming plates usually 25-30C in this case, while the residual
ice was subliming. Since the lyophilization normally required
some 24 hours, "inactivation" may have occurred. T h i s observation was consistent with literature reports indicating the heat
liability of hatch factor materials from root leachings. The
"inactivation", presumably occurring with all leachings, would
have been masked by a m u c h larger i n h i b i t o r effect. It would
be difficult to determine w h e t h e r a lack of hatch factor activity
was d u e to the absence of one or more hatch factor stimulants
or the presence of an equivalent a m o u n t of i n h i b i t o r material.
Since the bioassays providing the data reported (Figure 2
and 3) were conducted within a few weeks of each other, it was
reasonable to expect that differences in cyst response due to
their increased age would be negligible. In that event one could
conclude that the same hatch stimulatory substances in similar
proportions would be present in hatch stimulating leachings.
T h e diversity of seedlings with stimulatory potential would
preclude specialization and suggest instead that the activity was
d u e to c o m m o n substances, perhaps products or constituents of
metabolism. T h e similarity in response of g e r m i n a t i n g seedlings
with respect to concentration of dry solids appears to be for
tuitous. T h e procedures used in these experiments were not
sufficiently sensitive to detect any correlation between dry weight
of solids a n d weight of seeds or seed n u m b e r .
If hatch stimulation were of value in control applications (4),
these experiments would suggest that a n u m b e r of economically
profitable nonhost plants possess stimulatory activity. It would
not be necessary to consider economically useless plants for such
a purpose. T h e complexity of events occurring in soil, i.e., production, diffusion a n d decomposition of hatch inducing substances would necessitate confirmation of the notion by the
appropriate experimentation.
Literature Cited
(1) BAUNACKE, W. 1922. Untersuchungen zur Biologie und Bekampfung
des Rubennematoden Heterodera schachtii Schmidt. Arb. biol. Abt.
(Anst. Reichsanst.). Berl. 11: 185-288.
(2)

CALAM, C. T., A. R. TODD, and W. S.

WARING.

1949.

T h e potato eel

worm hatching factor. II. Purification of the factor by alkaloid


salt fractionation.Anhydrotetronic acid as an artificial hatching
agent. Biochem. J. 45: 520-525.
(3) FENWICK, D. W. 1952. T h e bio-assay of potato-root diffusate. Ann.
appl. Biol. 39: 457-467.

VOL.

13, N o . 4, JANUARY

1965

361

(4) SHEPHERD, A. M. 1962. T h e emergence of larvae from cysts in the


genus Heterodera. Technical communication No. 32 of the Comm o n w e a l t h B u r e a u of Helminthology.
(5) VIGLIERCHIO, D. R. 1958. Collection and selection of cysts of the
sugar beet nematode, Heterodera schachtii. J. Am. Soc Sugar Beet
T e c h n o l . 10: 318-329.
(6) VIGLIERCHIO, D. R. 1961. A simplified technique for hatching tests
of Heterodera schachtii. Phytopath. 51: 330-332.
(7) W A L L A C E , H. R. 1955. Factors influencing the emergence of larvae
from cysts of the beet eelworm, Heterodera schachtii Schmidt. J.
H e l m i n t h . 29: 3-16.
(8) W A L L A C E , H. R. 1956. Soil aeration and the emergence of larvae from
cysts of the beet eelworm, Heterodera schachtii Schmidt. Ann. appl.
Biol. 44: 57-66.
(9) W A L L A C E , H. R. 1956. T h e effect of soil structure on the emergence
of larvae from cysts of the beet eelworm. Nematologica 1: 145-146.
(10) W A L L A C E , H. R. 1956. T h e emergence of larvae from cysts of the beet
eelworm, Heterodera schachtii Schmidt, in aqueous solutions of
organic a n d inorganic substances. Ann. apl. Biol. 44: 274-282.
(11) W E N T , A. W. 1957. T h e experimental control of plant growth.
Chronica Botanica Co., Waltham, Mass. 388 pp.
(12) W I N S L O W , R. D. 1955. T h e hatching responses of some root eelworms
of genus Heterodera. Ann. appl. Biol. 43: 19-36.

The Performance of Sugar Beet Selections Made


for Purity and Chloride at Three Levels
of Nitrogen Fertility
C.

W.

DOXTATOR,

R.

E.

FINKNER,

R.

AND P. C. HANZAS

H.

HELMERICK

Received for publication June 15, 1964

Purity of beet juice is regarded to be the best criterion for


the d e t e r m i n a t i o n of the a m o u n t of extractable sugar which can
be recovered from the beet. Low purity juice (or juice with
high impurities) means the production of more molasses and
less sugar obtained per ton of processed beets. T h e continued
downward trend of sugar percent a n d low purity in recent years
has caused concern to all beet processors, and attempts to rectify
this situation have been made.
In 1960 J. B. Stark (9) 2 calculated that 1 p o u n d of chloride
carries 6.9 p o u n d s of sugar i n t o molasses. It was stated that
chloride is m u c h m o r e deleterious than a p o u n d of average
impurities, which carries only 1.7 p o u n d s of sugar i n t o molasses.
Since chloride is not removed in carbonation b u t goes into
molasses, the p r o b l e m of high chloride in beets in certain factory
areas may become serious. In 1952 D o x t a t o r a n d Bauserman
(1) reported on 7 chemical constituents (along with percent
sugar a n d percent purity) on 5 varieties grown in replicated
plots in 6 midwestern factory areas of the American Crystal
Sugar Company. Significant differences were found between
varieties for various chemical constituents, one of which was
chloride. Highly significant differences were found in each area
for each of the chemicals studied. O n e factory area was found
to be the lowest of all areas for sucrose percent and purity and
highest in chloride. Sucrose a n d chloride were found to be
negatively correlated (r .97).
T h e r e is ample evidence that the beet responds to simple
breeding methods for chemical constituents. Successful selections have been m a d e for low sodium by D o x t a t o r and Bauserm a n (2) and W o o d (10), for low raffinose by Finkner and
Bauserman (3) a n d W o o d (10), for low aspartic acid and
g l u t a m i n e by F i n k n e r et al. (4);'for p u r i t y as reported by Powers
et al. (8).
1
Plant Breeder, Manager Research Station, Plant Breeder and Research Chemist.
respectively,
American Crystal Sugar Company, Rocky Ford, Colorado.
2
Numbers in parentheses refer to literature cited.

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13, N o . 4, J A N U A R Y

1965

363

T h i s i n v e s t i g a t i o n was u n d e r t a k e n to determine: the effect


of selection for p e r c e n t p u r i t y a n d chloride content on these and
other b e e t c h a r a c t e r s ; t h e effect of nitrogen fertilization on beet
characters; a n d also to d e t e r m i n e if the selections reacted similarly at different n i t r o g e n fertility levels.
Materials and Methods
T h e varieties u s e d in this e x p e r i m e n t were the elite stocks
of 59-401 a n d 59-417, b o t h of which were good sucrose type
m o n o g e r m s . F i e l d selection of roots was made for weight using
a modification of t h e u n i t block m e t h o d of Powers (7). Percent
sucrose was o b t a i n e d polarimetrically, while percent apparent
purity was d e t e r m i n e d from t h e expressed juice of the beet brei.
C h l o r i d e s w e r e d e t e r m i n e d by p a p e r chromatography (5).
Only the p e r c e n t p u r i t y of juice of each beet was used in making
the h i g h a n d low selections in 59-401. Sucrose and weight data
were n o t used. Similarly, in 59-417 only chloride data were
used, w i t h no c o n s i d e r a t i o n given to weight, sucrose or purity
data. R a n d o m selections were m a d e by taking every 20th beet
of t h e 1046 field selected roots of 59-401, and every 13th beet
of the 1023 field selected roots of 59-417. T h e selections, along
with t h e m e a n d a t a a n d s t a n d a r d errors for each selection are
given in T a b l e 1.
Selection for h i g h a n d low purity in 59-401 resulted in a
significantly large difference of 15.22% between the selections;
and selections for h i g h a n d low chlorides in 59-417 resulted in
a five-fold s p r e a d (.91 to . 1 6 % ) . T h e effect of these selections on
sucrose in 59-401 a n d on sucrose a n d purity in 59-417 was large,
as shown in T a b l e 1. H i g h purity beets were found to be of
high sucrose c o n t e n t w h i l e low chloride beets were high in both
sucrose a n d p u r i t y . Differences in all 3 of these characters were
significant. T h e r a n d o m selections ranged between the high and
low selections for t h e 3 characters. Roots of the 6 selections were
p l a n t e d in space-isolated g r o u p s in 1961 and produced adequate
seed for p l o t tests.
In 1962 t h e 6 selections were planted in a split-plot replicated
test at R o c k y F o r d , C o l o r a d o . T h r e e rates of nitrogen fertilization w e r e used: 0 u n i t s , 75 units, a n d 150 units per acre as m a i n
plots, on w h i c h t h e selections were planted as sub-plots. These
sub-plots w e r e single rows 35 feet long in 6 replications with 2
rows of a c o m m e r c i a l variety p l a n t e d on each side of the m a i n
plots to give u n i f o r m c o m p e t i t i o n for each selection and adequate
spacing
b e t w e e n n i t r o g e n treatments. At harvest, each 35 toot
plot was harvested for yield a n d the beets divided into 2 samples
for
sucrose, p u r i t y a n d o t h e r chemical determinations. A p p a r e n t
purity
was d e t e r m i n e d from the expressed juice of the beet brei.

Table 1.Data on mother root selections of 59-401 and 59-417 for high and low chlorides and purity of juice.

Percent on Dry Substance

VOL. 13, No. 4, JANUARY 1965

365

Chlorides, raffinose, a n d n i n e a m i n o acids were determined bv


paper c h r o m a t o g r a p h y . Sodium, potassium, a n d calcium were
d e t e r m i n e d w i t h t h e f l a m e spectrophotometer. T h e microKjeldahl nesslerization m e t h o d (6) was used to determine total
nitrogen.
Experimental Results
T h e results of field a n d laboratory tests of the 6 selections
as an average of t h e 3 n i t r o g e n treatments are given in T a b l e 2.
Data a r e also g i v e n for the 3 nitrogen treatments as an average
of the 6 selections. T h e only selection
nitrogen interaction
that was significant was the o n e for sucrose, which is also presented i n T a b l e 2 .
Selection
Effects
Selection for p u r i t y of juice in 59-401 resulted in the high
selection 61-423 g i v i n g a h i g h e r purity t h a n the low selection
61-415, b u t t h e difference was n o t significant. However, since
the r a n d o m selection 61-409 was intermediate between the two,
some selection effect is indicated. In beet and sugar per acre
yield, a n d p e r c e n t sucrose, the high purity selection 61-423 was
significantly h i g h e r t h a n t h e low purity selection. It was also
significantly low in total n i t r o g e n a n d in 6 a m i n o acidsaspartic,
asparagine, g l u t a m i n e , glycine, gamma a m i n o butyric acid
(G.A.B.A.) a n d a l a n i n e , as well as in total a m i n o acid content.
C o m p a r e d to 61-409 ( r a n d o m selection), 61-423 gave significant
increases in b e e t a n d sugar p e r acre yield a n d significant red u c t i o n s in aspartic acid, glutamic acid and asparagine. In
o t h e r characters differences were not significant at the 5% point.
However, t h e p u r i t y of 61-423 was higher, and lower percentages
were o b t a i n e d for c h l o r i d e , total nitrogen a n d all 9 amino acids.
Selection for c h l o r i d e c o n t e n t in 59-417 resulted in the low
chloride selection 61-421 b e i n g reduced to approximately onehalf t h e h i g h selection, 61-422. Selection 61-421 was lower in
beet yield a n d h i g h e r in sucrose percent than 61-422 by significant m a r g i n s , r e s u l t i n g in a nearly equal sugar per acre yield.
P u r i t y of j u i c e was 1.12% h i g h e r in 61-421, a non-significant b u t
suggestively large increase. Since the r a n d o m selection was interm e d i a t e b e t w e e n t h e p u r i t y values of the low and high chloride
selections, it a p p e a r s t h a t selection for low chlorides did have
the effect of increasing purity. T h e low chloride selection did
not influence total n i t r o g e n c o n t e n t b u t did reduce by significant
percentages, s o d i u m , potassium, aspartic acid, glutamic acid and
valine. T h e G.A.B.A. c o n t e n t of the low chloride selection was
significantly higher, a n d asparagine, glutamine, glycine, alanine,
a n d leucines w e r e all h i g h e r in rank than in the high chloride

Table 2.Yield and chemical results obtained from three selections for purity and three selections for chloride, at three different fertility levels.

VOL. 13, No. 4, JANUARY 1965

367

selection. T o t a l a m i n o acid, although higher for the low chloride


selection was n o t significant. Compared to 61-417 (random selection), 61-421 was significantly higher in beet and sugar per
acre yield a n d sucrose percent. Purity of juice was . 4 5 % higher
a n d a l t h o u g h non-significant, was greater t h a n the high purity
selection 61-423 for p u r i t y , which showed only . 1 7 % higher than
the r a n d o m selection 61-409. Selection 61-421 had significantly
lower a m o u n t s of c h l o r i d e s a n d sodium, b u t was higher in total
nitrogen. It was h i g h e r in all 8 amino acids by significant margins
with t h e e x c e p t i o n of aspartic acid a n d glutamic acid and in
total a m i n o acids.
Nitrogen
Effects
W h e n i n c r e a s i n g a m o u n t s of nitrogen were applied, remarkable differences w e r e o b t a i n e d as an average of the 6 selections.
C o m p a r i n g t h e zero a n d 150 p o u n d rates, sugar per acre was
reduced greatly, a l o n g with sucrose percent and purity. Total
nitrogen, s o d i u m a n d all a m i n o acids were increased significantly
with t h e e x c e p t i o n of g l u t a m i c acid. All other characters showed
m i n o r increases, w i t h t h e exception of beet yield, percent potassium a n d p e r c e n t calcium. In all characters affected, the 75
p o u n d n i t r o g e n r a t e was i n t e r m e d i a t e i n rank.
Selection
Nitrogen
Interaction
T h e o n l y selection
n i t r o g e n interaction was for sucrose.
As shown in T a b l e 2, t h e low purity selection 61-415 had the
highest sucrose c o n t e n t at the 75 p o u n d nitrogen rate, and the
150 p o u n d r a t e d i d n o t r e d u c e the sucrose percent below the
zero r a t e . Lesser variations in 61-421 and 61-422 also contributed
to this i n t e r a c t i o n .
Discussion
In t h i s e x p e r i m e n t t h e selection for high a n d low purity, and
high a n d low c h l o r i d e resulted in progenies which were changed
in the d i r e c t i o n of t h e selection. T h e purity selections, although
differing o n l y slightly, a p p e a r e d to be reliable. Selection for
low a n d h i g h c h l o r i d e gave a highly significant difference. These
differences r e s u l t e d from a selection of approximately 5% of
the beets a v a i l a b l e for each purity selection, and 7.8% of those
available for each c h l o r i d e selection ( T a b l e 1).
It is also of interest to observe the effect of these selections
on sugar p e r acre yield, a n d on the other juice characters. T h e
high p u r i t y selection i m p r o v e d beet a n d sugar per acre yield,
a n d p e r c e n t sucrose; a n d in general reduced the quantity of
a m i n o acids, a n d to a lesser degree nitrogen, chloride a n d sodium.
T h e l o w c h l o r i d e selection m a i n t a i n e d sugar per acre yield,
r e d u c e d b e e t yield p e r acre, increased sucrose percent a n d purity
b u t h a d n o effect o n total nitrogen. Some a m i n o acids were

368

JOURNAL OF THE A. S. S. B. T

increased; others slightly decreased. Sodium along with chloride


was greatly decreased.
It appears therefore, that the purity selection had more effect
on the nitrogen c o m p o u n d s of beet juice t h a n the chloride
selection; the chloride selection h a d m o r e effect in reducing the
mineral elements t h a n the nitrogen compounds.
Although the percent purity data indicate no significant differences, the highest r a n k i n g purity was obtained in the low
chloride selection. It must be pointed o u t however, that the low
chloride selection 61-421 was significantly lower in total nitrogen
a n d in total a m i n o acids t h a n the high selection 61-423. This,
along with the low chloride a n d sodium content of 61-421, indicates that this selection should be high in purity. Since these
two selections came from different parents it is interesting to
note that their r a n d o m selections are significantly different in
nitrogen c o m p o u n d s 61-409 (from 59-401) being high, and
61-417 (from 57-417) b e i n g low.
T h e selection
nitrogen interaction for sucrose is of interest
because some of the selections showed a slightly different reaction to nitrogen applications t h a n the r a n d o m selections 61409 a n d 61-417. T h e low chloride selection 61-421, although
showing a reduction in sucrose percent with increasing rates of
nitrogen, was not significantly different from the zero rate of
nitrogen. T h e r e appears to be a possibility that varieties can
be developed which will p r o d u c e a satisfactory sucrose content
in high nitrogen fertility levels.
T h e data presented indicate that the purity of beet juice
can be increased by b r e e d i n g beets with a low chloride content.
Selecting beets with a low chloride content did not have an
appreciable effect in changing the total n i t r o g e n content of the
beet. In previous investigations F i n k n e r et al. (3) found that
selection for low aspartic acid or low g l u t a m i n e content significantly improved juice purity. It appears that the chemical pathways of chloride a n d a m i n o acid physiology of the sugar beet
plant are not highly associated, a n d that the plant breeder can
select for both low chlorides a n d a m i n o acids simultaneously
T h e results reported in this p a p e r a n d t h e previous paper on
a m i n o acids (3) indicate that t h e b r e e d e r can increase sugar
beet quality significantly a n d still m a i n t a i n yield if selection
pressures are applied for both low chloride a n d low aspartic acio
(or glutamine) content.
Summary
(A) Selections for high a n d low p u r i t y were made in the
elite stock 59-401, which resulted in progenies which differed

VOL.

13, N o . 4, JANUARY

1965

369

in yield, sucrose percent, and in the nitrogen components of


beet juice. T h e high purity selection showed an improvement
over the parent variety in these characters.
(B) Selection for high and low chloride was made in the
elite stock 59-417, which resulted in progenies which differed
in sucrose percent, in chlorides and in sodium and potassium
content. T h e low chloride selection showed an improvement
over the parent variety in these characters as well as in beet and
sugar per acre yield.
(C) Nitrogen applications caused a decrease in percent sucrose and purity, and in sugar per acre yield, and an increase
in the non-sugar compounds of the beet.
(D) A small but significant selection
nitrogen interaction
for sucrose was obtained.
Literature Cited
(1) DOXTATOR, C. W. a n d H. M. BAUSERMAN. 1952. Chemical constituents
of five varieties grown in six midwestern factory areas. Proc. Am.
Soc. Sugar Beet Technol. VII: 307-313.
(2) DOXTATOR, C. W. and H. M. BAUSERMAN. 1952. Parent-progeny tests
for sodium a n d potassium content. Proc. Am. Soc. Sugar Beet
T e c h n o l . V I I : 319-321.
(3) FINKNER, R. E. a n d H. M. BAUSERMAN. 1956. Breeding of sugar beets
with reference to sodium, sucrose and raffinose content. J. Am. Soc.
Sugar Beet T e c h n o l . 9 ( 2 ) : 170-177.
(4)

FINKNER, R. E., C. W. DOXTATOR, P. C. HANZAS and R. H. HELMERICK.

1962. Selection for low and high aspartic acid and glutamine in
sugar beets. J. Am. Soc. Sugar Beet Technol. 12(2): 152-162.
(5) LEDERER, E. a n d M. LEDERER. I n o r g a n i c paper chromatography.
C h r o m a t o g r a p h y 1953 Edition, Chap. 39, p. 338.
(6)

P A Y N E , M E R L E G., L E R O Y POWERS and GRACE W . MAAG.

1959.

Popula-

tion genetic studies on the total nitrogen in sugar beets. J. Am.


Soc. Sugar Beet T e c h n o l . 1 0 ( 7 ) : 631-646.
(7) POWERS, L E R O Y . 1957. Identification of genetically superior individuals a n d the prediction of genetic gains in sugar beet breeding
programs. J. Am. Soc. Sugar Beet Technol. 9 ( 8 ) : 408-432.
(8)

POWERS,

LEROY,

R A L P H E.

FINKNER,

GEORGE E. RUSH, R. R. W O O D

a n d DONALD F. PETERSON. 1959. Genetic improvement of processing


quality in sugar beets. J. Am. Soc. Sugar Beet Technol. 10(7):
578-593.

.
(9) STARK, J. B. 1960. Composition and beet molasses study. Report of
13th M e e t i n g of the Sugar Beet Advisory Committee and the
W e s t e r n Regional Research Laboratory. (Mimeo).
(10) W O O D , R. R. 1954 Breeding for improvement of processing characteristics in sugar beet varieties. Proc. Am, Soc. Sugar Beet Technol.
V I I I ( 2 ) : 125-133.

Resistance to the Sugar Beet Nematode


(Heterodera schachtii) in F 1
Tetraploid Hybrids between Beta vulgaris
and Beta patellaris 1
H E L E N SAVITSKY AND C H A R L E S P R I C E 2
Received for publication June 19, 1964

Introduction
In the genus Beta, only species of the section Patellares Trans.:
B. patellaris Moq., B. procumbens Chr. Sm., a n d B. webbiana
Moq., have proved to be highly resistant to the nematode,
Heterodera schachtii Schmidt
T h e occurrence of a single female n e m a t o d e on a few plants
of B. patellaris as reported by Shepherd (5) a n d by Steele and
Savitsky (6), does not preclude use of hybridization of species of
t h e section Patellares with sugar beet for transmission of nematode resistance to sugar beets. T h e high resistance of species of
the section Patellares to n e m a t o d e infestation distinguishes them
from all other Beta species.
T h e viable semi-fertile polyploid hybrids obtained by H.
Savitsky (4) p e r m i t selection for n e m a t o d e resistance and the
study of manifestation of resistance in a heterozygote, which
is i m p o r t a n t in breeding for resistance a n d for acquiring knowledge of the mechanism of the inheritance of resistance. Studies
of the resistance of F 1 hybrids between sugar beet and B. patellaris
are presented in this report.
T h e hybrids used in this study were produced by the senior
author, infestation of soil a n d the growing of the plants under
test conditions were performed by the j u n i o r author, examination of the plants was performed by b o t h authors.
Materials and Methods
Seed of two parental species, tetraploid B. vulgaris L. (sugar
beet) susceptible to n e m a t o d e a n d tetraploid B. patellaris resistant to nematode, together with F 1 h y b r i d seed obtained from
hybridization of these species, were p l a n t e d in soil in a greenhouse. Viable matings were selected in which seedlings grew on
their own roots. Seedlings which showed good growth and welldeveloped root systems were used for test (Figure 1).
1 Cooperative investigations of the Crops Research Division. Agricultural Research
Service, U. S. Department of Agriculture, and the Beet Sugar Development Foundation
2 Geneticist and Research Agronomist, respectively, Crops Research Division, Agricultural Research Service, U. S. Department of Agriculture. Salinas, California.
3 Numbers in parentheses refer to literature cired.

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13, N o . 4, JANUARY

1965

Figure 1-F1 allotetraploid hybrids (B. vulgaris


amined for n e m a t o d e resistance.

371

B.

patellaris) ex-

T e n seedlings of each p a r e n t species and 10 seedlings of the


F 1 h y b r i d s w e r e t r a n s p l a n t e d in the two-leaf stage into cystinfested soil to test for n e m a t o d e resistance. After 60 days of
growth u n d e r c o m p a r a b l e conditions in a greenhouse, the roots
of all p l a n t s w e r e e x a m i n e d for the presence of female nematodes.
Experimental Results
T h e 10 sugar b e e t plants were heavily infested with female
n e m a t o d e s . N i n e of 10 B. patellaris plants were free of nematodes
and 1 p l a n t h a d 2 females. T h e F 1 hybrids had well developed
root systems ( F i g u r e 2). Of 10 F 1 hybrid plants, 8 plants were
free of n e m a t o d e s , 1 p l a n t had a single comparatively welldeveloped female, a n d on 1 plant, 2 females were found
( T a b l e 1).

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JOURNAL OF THE A. S. S. B. T.

Figure 2.Root system of an allotetraploid F1 hybrid free of females.

T h e plants e x a m i n e d fell i n t o two groups: a highly susceptible g r o u p which contained sugar beets, and a resistant
g r o u p which included B. patellars a n d F 1 hybrids. Resistance in
F 1 hybrids was n o t intermediate between t h e 2 species, B. Vulgaris
a n d B. patellaris, b u t approached closely the resistance ot the
resistant parent, B. patellaris, To d e t e r m i n e whether the grade
of resistance in F 1 hybrids was the same as in B. patellaris an
assumption was m a d e that the r a t i o of the plants with a few
nematodes on the roots to all plants e x a m i n e d was the same in
the population of B. patellaris a n d F 1 hybrids. T h i s assumption
was verified by calculation of the chi-square. T h e value of the
chi-square was 0.3922. T a b u l a t e d value at the 5% level for
d.f. 1 is x 2 0.05 = 3.841 a n d at 1% level x 2 0.01 = 6.63

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T h i s value is m u c h larger t h a n the calculated value of chi-square


which indicates t h a t t h e difference in resistance between B
patellaris a n d F 1 h y b r i d s is n o t significant. T h e data obtained
give no i n d i c a t i o n t h a t F 1 hybrids differ in resistance from the
resistant p a r e n t a l species B. patellaris.
Gaskill (1) r e p o r t e d that F 1 hybrids between Swiss chard
and B. webbiana (4 plants) a n d between Swiss chard and B.
procumbens (2 plants) g r o w n in nematode infested soil, had
no female n e m a t o d e s on the roots. He states, however, that
"most of t h e h y b r i d s were small, and the results cannot be considered as conclusive, b u t they suggest that the high degree of
n e m a t o d e resistance was transmitted to the hybrids."
Conclusion
Resistance to n e m a t o d e (Heterodera schachtii) is a dominant
character. T h e t e t r a p l o i d F 1 hybrids (B. vulgaris X B. patellaris)
did n o t differ in t h e grade of resistance from the resistant parent,
B.
patellaris.
T h e d a t a p r e s e n t e d here, a n d that previously presented by
Gaskill, i n d i c a t e with, a high degree of probability that nematode
resistance t r a n s m i t t e d by all 3 species of the section Patellares,
namely B. patellaris, B. procumbens, and B. webbiana, is domin a n t in t h e F 1 h y b r i d s regardless of whether they are diploids
or tetraploids.
Literature Cited
(1) GASKILL, J. O. 1954. Viable hybrids from matings of chard with Beta
procumbens a n d B. webbiana. Proc. Am. Soc. Sugar Beet Technol.
8 ( 2 ) : 148-152.
(2) GOLDEN, A. M. 1958. Interrelationships of certain Beta species and
Heterodera schachtii, the sugar beet nematode. Plant disease reporter 4 2 ( 1 0 ) : 1157-1162.
(3) H I J N E R , J. A. 1951. De gevoeligheid van wilde bieten voor het bietencystenaaltje
{Heterodera schachtii). Meded. I n s t . V. R a t i o n e l e
Suikerproductie 21: 1-13.
(4) SAVITSKY, H. 1960. Viable diploid, triploid, and tetraploid hybrids
between Beta vulgaris and species of the section Patellares. Am.
Soc. Sugar Beet T e c h n o l . 11 (3) : 215-235.
(5) SHEPHERD, A. M. 1959. Testing p o p u l a t i o n s of beet eelworm,
Heterodera schachtii Schmidt, for resistance-breaking biotypes, using
the wild beet {Beta patellaris Moq.) as indicator. N a t u r e 183
(4668) : 1141-42.
(6) STEELE, A. E. a n d H. SAVITSKY. 1962. Susceptibility of several Beta
species to t h e sugar beet nematode {Heterodera schachtii Schmidt).
Nematologica 8 (3) : 242-243.
(7) W I N S L O W , R. D. 1954. Provisional lists of host plants of some root
eelworms {Heterodera ssp.) Ann. Appl. Biol. 41: 591-605.

Occurrence of the Alfalfa Mosaic Virus


in Sugar Beet in California
R. J. SHEPHERD, D.

H.

H A L L , AND D.

E.

PURCIFULL1

Received for publication June 26, 1964

A n u m b e r of different aphid-borne viruses have been found


in naturally infected sugar beet. Several of these, including the
malva yellows, western yellows a n d yellow n e t viruses are retained by the aphid vector for an indefinite period, whereas
others are stylet-borne with shorter periods of retention by the
insect. In the latter g r o u p are the viruses of beet yellows, beet
mosaic, c u c u m b e r mosaic a n d t h e recently described beet ring
mottle (2) 2 . H e r e the occurrence of a fourth stylet-borne virus,
alfalfa mosaic, is reported in naturally infected beets.
Infected plants were found in 3 different fields in the Sacr a m e n t o Valley in the spring of 1962. All fields were located near
alfalfa plantings in the Colusa C o u n t y area. In the first field in
which the disease was observed, only 2 infected plants were
found. In this case an alfalfa field was located immediately
across a county road from the sugar beet planting. In the second
instance in which the disease was e n c o u n t e r e d a relatively small
localized area in a field was found with a b o u t 25 diseased plants
as if secondary spread h a d occurred w i t h i n the field following
introduction of the disease. An alfalfa field was located less than
one-fourth mile from the infection center in beets. In the third
case, many infected plants were found more-or-less randomly
scattered t h r o u g h o u t o n e edge of a beet field b o r d e r i n g an alfalfa
seed field. Approximately 1 0 % of t h e beet plants were showing
symptoms in that area of t h e field n e a r alfalfa. T h e percentage
of diseased plants, however, decreased markedly at distances away
from the alfalfa with only an occasional p l a n t showing; symptoms
at the opposite edge of the field. T h e proximity of the diseased
plants to alfalfa a n d t h e n a t u r e of t h e symptoms on beets suggested that the disease might be caused by the alfalfa mosaic
virus.
T h e most characteristic symptoms of t h e disease consisted o
p r o m i n e n t yellow blotches a n d ringspots (Figure 1). The most
conspicuous symptoms were e x h i b i t e d on the older leaves of
affected plants with few or no symptoms being manifest on the
u p p e r leaves, t h u s suggesting t h a t plants probably recover rapidly
1
Assistant Professor. Extension Plant Pathologist and Research Assistant, respectively,
Department
of Plant Pathology, University of California, Davis.
3
Numbers in parentheses refer to literature cited.

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375

Figure 1.Symptoms of the alfalfa mosaic virus on naturally infected


sugar beet.

from the disease. T h e irregular blotches and ringspots were


usually sharply delimited from the surrounding green-colored
leaf tissue and were usually not associated with the veins or other
portions of the leaves in any particular way. They were usually
accompanied by chlorotic lines and oak-leaf patterns on affected
laminae.
On younger plants in a 6 to 8 leaf stage, the symptoms were
not generally as conspicuous and consisted chiefly of irregular
chlorotic spots on the older leaves. Little, if any distortion or
blistering was present on any of the diseased plants.
Diseased plants were removed and transplanted in the greenhouse. T h e symptoms persisted on these plants for a period but
the new foliage developed no further signs of the disease. Leaf
tissue showing symptoms was removed and homeginized in 0.05
M phosphate, pH 7.5, and used to innoculate the seedlings of other
species known to be susceptible to the alfalfa mosiac virus. Inoculated primary leaves of beans, Phaseolus vulgaris L. var.
Bountiful, and cowpea, Vigna sinensis, var. Early Wilt Resistant
Ramshorn, developed a few necrotic local lesions following inoculation. These developed as well-defined circular lesions of a
reddish-brown color unlike the more-or-less irregular necrotic
flecks associated with bacteria frequently present in beet tissue
(3). Several of the lesions on bean and cowpea were removed
with a small cork-borer and homogenized in phosphate for further transfer to other plants.

376

JOURNAL OF THE A. S. S. B. T

Tobacco, Nicotiana tabacum L. var. Wisconsin Havana 425


a n d Nicotiana glutinosa L., showed a transient chlorotic etching
followed by complete recovery, a n d cucumber, Cucumis sativus
L. var. National Pickling, developed chlorotic local lesions following inoculation with material from bean a n d cowpea. The
virus was readily recovered from beans, cowpeas, tobacco, Nicotiana glutinosa a n d c u c u m b e r u p o n subsequent inoculation of
cowpea thus demonstrating that a mechanically transmissible
virus was present in diseased beets.
T h e identity of the virus was confirmed by serological tests
using antiserum to a strain of the alfalfa mosaic virus isolated
from alfalfa.
Agar gel diffusion tests were used as these have proved very
convenient for the r o u t i n e identification of the spherical or
shorter rod-shaped p l a n t viruses which diffuse well in agar-gels.
T h i s technique consists of p o u r i n g a shallow layer of melted
agar i n t o a plastic Petri dish a n d allowing it to solidify. Cylindrical wells to accommodate the test reagents are then cut in
the gel with a suitably sized cork-borer, followed by removal of
the agar pieces. T h e a n t i s e r u m is usually added to a larger
center well a n d an extract of the virus infected material under
study to smaller wells radially placed at an a p p r o p r i a t e distance
from the antiserum depot. T h e plates are then incubated for
a day or two at r o o m t e m p e r a t u r e before their evaluation. During this period the virus a n d antiserum diffuse outward toward
one another, m e e t a n d c o m b i n e in o p t i m u m proportions to form
lines of precipitation.
For the tests used here, 11 ml of a solution of purified agar
at a concentration of 0 . 8 % , c o n t a i n i n g 0 . 8 5 % sodium chloride
a n d 0.04% sodium azide was p o u r e d i n t o plastic Petri plates 9
cm in d i a m e t e r a n d allowed to solidify. T h i s gave a layer of
semi-solid gel a b o u t 3 mm in d e p t h . A c e n t e r antiserum depot
8 mm in d i a m e t e r a n d 8 evenly spaced peripheral wells each
4 mm in diameter at a distance of 3 mm from the edge of the
center well were t h e n removed. Sap expressed from naturally
infected sugar beet leaves using a juice extractor was pipetted
i n t o some of t h e peripheral wells. Sap from healthy beets was
placed in o t h e r wells to serve as controls. Following addition
of the antiserum the plates were i n c u b a t e d for 24 hr at room
temperature.
Samples from plants collected at each of three different fields
showed positive reactions for t h e alfalfa mosaic virus. Clearly
a p p a r e n t lines of precipitation were present near the edge of

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4,

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a n t i g e n w e l l s f i l l e d w i t h t h e e x t r a c t e d sap from diseased b u t n o t


f r o m h e a l t h y p l a n t s i n d i c a t i n g t h a t t h e plants were infected
w i t h t h e alfalfa m o s a i c v i r u s . T h e s e results i n addition t o the
host r e a c t i o n s m e n t i o n e d previously a p p e a r a d e q u a t e to show
t h e alfalfa m o s a i c v i r u s was t h e cause of the p r o m i n e n t mottling
o n affected p l a n t s .
T h e d i s e a s e d e s c r i b e d h e r e i s p r o b a b l y distinct from the yellow b l o t c h i n g of l e a v e s r e p o r t e d by B e n n e t t (1) in beets in
C a l i f o r n i a a n d e l s e w h e r e . T h e i r r e g u l a r yellow spots associated
w i t h v i r u s a r e superficially similar to those described by Benn e t t b u t a r e m o r e s h a r p l y d e l i m i t e d from t h e s u r r o u n d i n g green
leaf tissue a n d a r e g e n e r a l l y a c c o m p a n i e d by ringspots a n d line
patterns
In t w o cases a disease similar to that described by
B e n n e t t h a s b e e n o b s e r v e d i n u p p e r San J o a q u i n Valley f i e l d s .
In n e i t h e r case c o u l d v i r u s be recovered by inoculation of beans,
cowpeas or t o b a c c o , n o r w e r e positive serological tests obtained.
L I T E R A T U R E CITED
(1) B E N N E T T , C. W. 1963. Noninfectious yellow splotching of leaves of
y o u n g sugar b e e t p l a n t s in western United States. Plant Disease
R e p t r . 47: 63-64.
(2) D U F F U S , J. E., a n d A. S. COSTA. 1963. Beet ring mottle disease. Phytop a t h o l o g y 5 3 : 1422-1425.
(3)

Y A R W O O D , C. E., E. C. R E S C O N I C H , P. A. ARK, D. E. SCHLEGEL and K. M.

S M I T H . 1961. So-called beet latent virus is a bacterium. Plant Disease


R e p t r . 4 5 : 85-89.

JOURNAL
of the
A m e r i c a n Society of Sugar
Beet Technologists
Volume 13
Number 5
April 1965

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado 80521

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Made in the United States of America

TABLE OF CONTENTS

Author

Page

Relation of water soluble substances in fruits


of sugar beet to speed of germination of
sugar beet seeds---------------------------------------------F. W. Snyder
John M. Sebesoh
J. L. Fairley--------------------379
Preplant weed control on sugar beets----------------------E. F. Sullivan
R. R. Wood
R. L. Abrams
S. G. Walter -----------------389
Breeding sugar beets for resistance to the
cyst n e m a t o d e Heterodera schachtii--------------------Charles Price ----------------397
The chemical destruction of sucrose, fructose, a n d glucose in hot alkaline process
juices and liquors------------------------ S.E. Bischel------------------------- 406
Waste water t r e a t m e n t studies at Tracy,
California ----------------------------------------------------Lloyd W. Norman
James E. Laughlin
L. O. Mills--------------------415
A

new a p p r o a c h to the saccharate cake


purity determination---------------------------------------Karl Schoenrock--------------- 425

Sugar beet tops a n d m o d e r n sugar beet


p r o d u c t i o n ----------------------------------------------------Lionel Harris
D. C. Clanton
M. A. Alexander--------------432
Flotation a n d conveying air velocities for
processed monogerra sugar beet fruits

Injection of preemergence herbicides

Uses for sugar beet pulp R.M.McCready


A.E. Goodban---------------467

O. R. Kunze
C. W. Hall
F. W. Snyder-----------------448
C. R. Kaupke--------------------462

Relation of W a t e r Soluble Substances in Fruits of


Sugar Beet to Speed of Germination of
Sugar Beet Seeds1
F . W . S N Y D E R , J O H N M . SEBESON AND J . L . F A I R L E Y 2
Received for publication June 30, 1964

At least 10 s u b s t a n c e s (9 are organic c o m p o u n d s ) which may


b e p o t e n t i a l l y i n h i b i t o r y t o g e r m i n a t i o n have been isolated from
t h e f r u i t s of s u g a r b e e t s (2,5,6,7,8,11) 3 . Some of the c o m p o u n d s
h a v e n o t b e e n i d e n t i f i e d as to chemical structure. Also, only
l i m i t e d d a t a on t h e q u a n t i t y of i n h i b i t o r s in sugar beet fruits
a r e p r e s e n t l y a v a i l a b l e . T h e c o n c e n t r a t i o n of each of these substances w h i c h w i l l adversely affect g e r m i n a t i o n of sugar beet
seeds r e m a i n s t o b e d e t e r m i n e d critically. O f greater fundam e n t a l a n d p r a c t i c a l i m p o r t a n c e , the causal relation between
c o n c e n t r a t i o n of specific i n h i b i t o r s in the fruit a n d speed of
g e r m i n a t i o n m u s t b e established.
D u y m et al. (3) h a v e c o n c l u d e d that the osmotic pressure
e x e r t e d by i n o r g a n i c substances in t h e seedball of the sugar beet
is r e s p o n s i b l e for t h e i n h i b i t o r y effect on germination. Specific
c o n d u c t a n c e v a l u e s of different seedball extracts may vary in
t h e a m o u n t of electrolytes a n d these differences are reflected by
t h e w h e a t g r o w t h test (10). Froschel (4) observed that demineralized e x t r a c t s of s u g a r b e e t fruits considerably i n h i b i t e d germi n a t i o n of Lepidium a n d o t h e r seeds a n d that the extracts also
e m i t t e d v o l a t i l e substances which caused i n h i b i t i o n . He ind i c a t e d t h e p r e s e n c e of specific orgranic substance(s) as the cause
o f t h e i n h i b i t i o n . A c c o r d i n g t o M a k i n o a n d Miyamoto (6,8),
w a t e r s o l u b l e o x a l a t e , w h i c h may exceed two percent in the fruits
of s o m e s u g a r b e e t varieties, m a y be causally related to germination performance.
T h i s p a p e r r e p o r t s (a) results of physiological a n d chemical
tests w h i c h i n d i c a t e t h a t at least o n e of the mechanisms suggested
as t h e c a u s e of d e l a y e d g e r m i n a t i o n is u n t e n a b l e , (b) a signific a n t c o r r e l a t i o n b e t w e e n water-soluble oxalate in the fruit a n d
speed of g e r m i n a t i o n of sugar beet seeds, (c) evidence that at
least o n e o t h e r i n h i b i t o r besides oxalate also affects speed of
1
Cooperative investigations of the Crops Research Division Agricultural Research
Service, U. S. Department of Agriculture, and the Michigan Agncultural Experiment
Station. Approved for publication as Journal article # 3406, Michigan Agricultural Experiment Station
2

Research Plant Physiologist, Crops Research Division, Agricultural Research Service,


U. S. Department of Agriculture; Professor, Lansing Community College; and Professor
of
Biochemistry, Michigan State University, East Lansing, Michigan respectively.
3
Numbers in paentheses refer to literature cited.

380

JOURNAL OF THE A. S. S. B. T

germination in variety US 401, and (d) data on the distribution


of water-soluble oxalate in the fruits of sugar beet variety US 401.
M e t h o d s a n d Materials
To ascertain some of the characteristics of the inhibitory substances in the water extract of sugar beet fruits (seedballs) of
lot 4401, variety US 401, exploratory investigations were conducted involving specific conductance measurements, cryoscopic
determinations, ashing, dialysis, vacuum distillation, ether extraction, treatment with lead acetate, calcium chloride, activated
charcoal, a n d osmotic additives; addition of suggested inhibitors:
a n d the use of a bioassay technique. T h e bioassav, designated as
the wheat test, has been described previously (10). Briefly, the
test compares the growth of wheat on aqueous extracts of seedballs with the growth of wheat on distilled water in 96 hours.
Thirty-six samples of seed from the variety US 401, each of
which was harvested from a single plant grown in the field at
East Lansing, Michigan, were used in the study of oxalate con
tent. T h e speed of germination of each sample was determined
by the blotter method.
T h e seedball extracts used for the phvsiological studies and
chemical analysis were prepared by soaking air-dried seedballs
in distilled water (usually 1 gm per 10 ml of water) for 18
hours in a refrigerator at approximately 4 C. T h e extract was
decanted a n d filtered.
Oxalate was isolated from the extract according to a modification of the procedure suggested by Suzuki (6) and was as
follows: A 25 ml sample of extract was adjusted to a pH between
5 a n d 6 with acetic acid. Fifteen ml of hot (80 C) 1 5 % calcium
chloride (calcium acetate may also be used) was added to the
extract. T h e m i x t u r e was heated for an h o u r and a half at
approximately 80 C to p r o m o t e crystal enlargement. After cool
ing to r o o m t e m p e r a t u r e , the solution was filtered on No. 2
W h a t m a n paper a n d the precipitate was washed sparingly with
distilled water. T h e precipitate was dissolved in 2 N hydrochloric
acid heated to 80 C a n d t h e solution m a d e alkaline with am
m o n i u m hydroxide d i l u t e d 1:1.
T h e solution was again heated in a water bath for one and
a half h o u r s at 80 C. After cooling to r o o m temperature. it was
filtered. T h e precipitate was washed twice with hot (80 C) 30%
acetic acid a n d then dissolved in h o t (80 C) 1 N sulfuric acid
T h e solution was increased to a v o l u m e of 50 ml. T h i s procedure
differs from that of Suzuki with respect to the addition of the
acetic acid wash. T h e wash was f o u n d to be necessary because

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d e t e r m i n a t i o n s of t h e q u a n t i t y of oxalate by the potassium perm a n g a n a t e m e t h o d a n d b y a colorimetric m e t h o d involving indole


failed t o a g r e e .
A p p a r e n t l y s u b s t a n c e s o t h e r t h a n oxalate were co-precipitated
w i t h t h e c a l c i u m o x a l a t e o r adsorbed t o it. T h e values for
o x a l a t e by t h e i n d o l e m e t h o d were as m u c h as d o u b l e those for
t h e p e r m a n g a n a t e m e t h o d , w h i c h indicates that the impurities
in t h e p r e c i p i t a t e w e r e intensifying the indole colorimetric test
m u c h m o r e t h a n t h e p e r m a n g a n a t e test. B y washing the precipitate w i t h 3 0 % acetic acid (calcium oxalate is essentially ins o l u b l e in acetic acid), t h e i m p u r i t i e s were removed from the
c a l c i u m o x a l a t e p r e c i p i t a t e t o t h e extent that the determinations
of o x a l a t e by b o t h m e t h o d s agree very closely.
T h e q u a n t i t y o f o x a l a t e was d e t e r m i n e d b y titration with
p o t a s s i u m p e r m a n g a n a t e (9) a n d by means of a colorimetric
m e t h o d u s i n g i n d o l e (1). Twenty-five ml of the final solution
w e r e t i t r a t e d w i t h p o t a s s i u m p e r m a n g a n a t e . For the indole
m e t h o d , t h e r e a g e n t was p r e p a r e d by dissolving 100 mg of very
p u r e i n d o l e in 100 ml of c o n c e n t r a t e d sulfuric acid. A standardization c u r v e was p r e p a r e d from 1 N sulfuric acid containing
from 0.1 to 1.0 mg of oxalic acid per ml. To 5 ml of oxalate
s o l u t i o n in a test t u b e w e r e a d d e d 5 ml of indole reagent by
m e a n s of a p i p e t t e . A f t e r w a i t i n g 60 seconds the contents were
m i x e d t h o r o u g h l y . T h e test t u b e was placed in a water bath
a n d h e a t e d to 80-90 C for 45 m i n u t e s . T h e solution was cooled
to r o o m t e m p e r a t u r e , transferred to a cuvette, and its absorbance
m e a s u r e d in a c o l o r i m e t e r at a wave length of 525 m u . A blank,
c o n s i s t i n g of 5 ml of 1 N sulfuric acid a n d 5 ml of indole reagent,
was p r e p a r e d w i t h each set of samples to d e t e r m i n e the zero
reading of the instrument.
T h e p r o p o r t i o n o f t h e i n h i b i t o r y substances removed from
t h e s u g a r b e e t fruits d u r i n g processing was d e t e r m i n e d for a
s a m p l e f r o m 4 different p l a n t s of variety US 401. T h e s e samples
a p p e a r e d to c o n t a i n o x a l a t e as t h e chief inhibitor. A water extract was p r e p a r e d from (a) w h o l e seedballs, (b) processed seedballs, a n d (c) t h e m a t e r i a l r e m o v e d by the hand-processing
o p e r a t i o n . A p o r t i o n of each extract was analyzed tor oxalate
c o n t e n t a n d a n o t h e r p o r t i o n was used for the wheat test.
Results
T h e d a t a ( T a b l e 1 ) d o n o t s u p p o r t the c o n c l u s i o n o f D u y m
et al. (3) that the i n h i b i t o r y effect of the sugar beet seedball
on g e r m i n a t i o n is largely osmotic. If the effect was osmotic and
largely caused by the i n o r g a n i c constituents, then the growth of

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JOURNAL OF THE A. S. S. B. T.

Table 1.Characteristics of a variously treated seedball extract from bulked seed


of lot 4401. US 400 sugar beet.

Diluted 1 : 1

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* One gram seedballs per 4 ml distilled water.


** Undiluted extract.

r e m o v e d t h e t o x i c m a t e r i a l s . Since dialysis removed only a port i o n of t h e toxicity, some of t h e molecules were so largre that
t h e y failed to pass t h r o u g h the pores of the cellophane tube.
A c o m b i n a t i o n of p r e c i p i t a t i o n with basic lead acetate followed
by filtering:, p r e c i p i t a t i n g : t h e excess lead with monobasic potass i u m p h o s p h a t e , a n d dialvsis effectively removed the toxic substances. In o n e test, p r e c i p i t a t i o n with calcium chloride followed
by dialysis was n o t as effective as t h e lead acetate in removing
t h e t o x i c s u b s t a n c e s . In s u m m a r y , the inhibitory action seems
to be c a u s e d by a c o m b i n a t i o n of dialyzable a n d non-dialyzable
s u b s t a n c e s . T h e m o s t p o t e n t o f t h e substances appear t o b e
o r g a n i c c o m p o u n d s w h i c h are dialyzable. Osmotic stress cannot
a c c o u n t for t h e d e g r e e of i n h i b i t i o n observed in the wheat test.
I n t h e p r o c e d u r e s e m p l o y i n g precipitation, the precipitability
of g i v e n o r g a n i c a n i o n s m u s t be established. T h e differences in
g r o w t h of w h e a t on a seedball extract treated with either calcium
or l e a d suggest t h a t some i n h i b i t o r s may be removed by lead
( b u t n o t b y c a l c i u m ) p r e c i p i t a t i o n . T h e inhibitory substances,
v a n i l l i c , caffeic, a n d ferulic acids, previously isolated from sugar
b e e t fruits (5,7), a r e n o t p r e c i p i t a t e d by calcium b u t are prec i p i t a t e d b y lead. T h e presence o f these inhibitors a n d others
c o u l d a c c o u n t for s o m e of t h e inconsistency observed between
lead a n d c a l c i u m p r e c i p i t a t i o n .
Relation of soluble oxalate to speed of germination
B e c a u s e of t h e large q u a n t i t y of soluble oxalate in the fruits
as c o m p a r e d w i t h a n y of t h e o t h e r inhibitors, the influence of
w a t e r - s o l u b l e o x a l a t e on speed of g e r m i n a t i o n seemed a p p r o p r i a t e
to i n v e s t i g a t e . F o r t h e 36 samples examined, the rapid-germinati n g s a m p l e s g e n e r a l l y c o n t a i n e d lower a m o u n t s of water-soluble
o x a l a t e t h a n t h e s l o w - g e r m i n a t i n g samples ( T a b l e 4). The corr e l a t i o n coefficients for speed of g e r m i n a t i o n versus soluable
o x a l a t e i n t h e seedball e x t r a c t a r e - 0 . 6 6 for the p e r m a n g a n a t e

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Table 4.Relation of speed of germination of sugar beet seeds to water-soluble


oxalate in seedball extract of sugar beet variety US 401.
Milligram-percent of oxalate in extract
as determined by
Percentage
germination
Seed
Permanganate
Indole
after
2
days
number

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5,

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F u r t h e r e v i d e n c e t h a t oxalate may be o n e of the principal


i n h i b i t o r s in t h e m a j o r i t y of samples, b u t n o t the only inhibitor
in t h e v a r i e t y US 4 0 1 , is p r o v i d e d by c o m p a r i n g the growth of
w h e a t on s y n t h e t i c s o l u t i o n s a n d on seedball extracts ( T a b l e 5).
T a b l e 5.Growth of wheat on suger beet seedball extracts and synthetic solutions.

* Averages of 4 replications

T h e s y n t h e t i c s o l u t i o n s c o n t a i n e d the same quantity of oxalate


as f o u n d in t h e seedball extracts a n d the same specific cond u c t a n c e , as well as g r e a t e r or lesser quantities. T h e growth of
w h e a t on a s y n t h e t i c s o l u t i o n of oxalate a n d the osmotic additive
(KC1) w a s s i m i l a r to g r o w t h on the extract of seedballs of sample
469, b u t s y n t h e t i c s o l u t i o n s w h i c h h a d similar oxalate and osmotic
v a l u e s t o t h e o t h e r seedball extracts p e r m i t t e d considerably better
g r o w t h t h a n t h e seedball extracts. Additional inhibitory substances m a y b e p r e s e n t i n these samples.
Distribution of water-soluble oxalate in the fruits
A l t h o u g h t h e d a t a ( T a b l e 5) indicate that 3 of the 4 samples
used in t h i s p a r t of t h e study p r o b a b l y have some i n h i b i t o r in
a d d i t i o n to o x a l a t e , t h e 4 were chosen because of large differences
in s p e e d of t e r m i n a t i o n a n d c o n t e n t of oxalate in the seedball
e x t r a c t . T h e r e c o v e r y of t h e corky material after processing and
t h e w e i g h t s of t h e m a t e r i a l s used to prepare the extracts are
g i v e n in T a b l e 6. E a c h p o r t i o n of the material was soaked in 100
ml of w a t e r . T h e r e f o r e , t h e v a l u e of oxalate for the processed
s e e d b a l l s p l u s t h e v a l u e for t h e material removed i n processing

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JOURNAL OF THE A. S. S. B. X.

T a b l e 6 . Q u a n t i t y o f m a t e r i a l ( g r a m s ) f r o m t h e hand-processed s u g a r b e e t seedballs
u s e d t o p r e p a r e extracts for o x a l a t e analysis a n d g r o w t h o f w h e a t .

should equal the value for the whole seedballs (assuming no


los^ of material removed a n d equal solubility for all samples).
T h e oxalate appears to be concentrated in the corky material
of the seedball ( T a b l e 7). Processing w o u l d be particularly beneficial for seedballs that have a high concentration of inhibitors.
T a b l e 7 . S p e e d of g e r m i n a t i o n ( b l o t t e r m e t h o d ) of s u g a r beet seeds a n d growth of
w h e a t o n t h e s e e d b a l l e x t r a c t s (1:10) i n r e l a t i o n t o the d i s t r i b u t i o n o f water-soluble
o x a l a t e in t h e seedballs of 4 p l a n t s of sugar b e e t variety US 4 0 1 .

Discussion
T h e complex nature of the inhibitory action of the suga
beet seedball extract on germination is indicated. Probably
number of effects interact to give the observed germination
sponse. However, the data indicate that organic solutes
largely responsible for the inhibition. At least 2 or&"1* *
stances seem to be involved. At present, only oxalate n;is

VOL. 13, No. 5, APRIL 1965

387

causally r e l a t e d to s p e e d of g e r m i n a t i o n . If the additional substance(s) c o u l d be identified a n d causally related to the inhibitory


action, f u r t h e r progress c o u l d be m a d e in p r e p a r i n g synthetic
s o l u t i o n s t o d e t e r m i n e t h e response t o k n o w n combinations a n d
c o n c e n t r a t i o n s of t h e s e substances. Identifications of all the substances in t h e seedballs w h i c h cause slower germination also
w o u l d a i d i n f o r m u l a t i n g a n empirical procedure for overcoming
the i n h i b i t o r y action.
T h e a p p a r e n t t e n d e n c y for high concentrations o f oxalate
to localize in t h e c o r k y m a t e r i a l of the fruit may indicate that
o t h e r i n h i b i t o r s a r e c o n c e n t r a t e d there.
T h e r e m o v a l of i n h i b i t o r y substances from the seedball extract by t r e a t i n g it w i t h activated charcoal a n d dialyzing is visualized as a t w o - s t e p process. (See T a b l e 3). T h e activated charcoal
r e m o v e s t h e l a r g e m o l e c u l e s t h a t cannot be removed by dialysis,
w h i l e dialysis r e m o v e s t h e smaller molecules or ions which cannot
be absorbed on the charcoal.
Summary
A v a r i e t y of s i m p l e chemical procedures were employed to
p a r t i a l l y c h a r a c t e r i z e t h e i n h i b i t o r y substances in a water extract
of s u g a r b e e t fruits. T h e i n h i b i t i o n was not caused by inorganic
s u b s t a n c e s in t h e f r u i t b u t largely by the presence of organic
substances.
T h e s p e e d of g e r m i n a t i o n by the blotter m e t h o d of 36 samples
of seed of s u g a r b e e t variety US 401 was significantly correlated
(0.6**) w i t h t h e a m o u n t of oxalate in a water extract of the
fruits. S e v e n of t h e samples failed to correlate well, which suggests t h a t an i n h i b i t o r y substance o t h e r than oxalate was dep r e s s i n g s p e e d of g e r m i n a t i o n .
A s t u d y of t h e d i s t r i b u t i o n of oxalate in the sugar beet fruit
r e v e a l e d t h a t o x a l a t e was c o n c e n t r a t e d in the corky material of
the fruit.
Acknowledgment
G e n e r o u s financial s u p p o r t of t h e Farmers and Manufacturers B e e t S u g a r Association, Saginaw, Michigan, made possible
t h e s t u d y of t h e r e l a t i o n of oxalate to speed of germination.
Literature Cited
(1)

B E R O E R M A N , J. a n d J. S. E I X . O T . 1955. Method for direct colorimetric


d e t e r m i n a t i o n of oxalic acid. Anal. Chem. 27: 1014-1015.

(2)

DSKOCK,

P. C, R. F.

H U N T E R and I . ^ M A C D O N A O ,

n a t i o n i n h i b i t o r from sugar-beet.

IMS. A gernu-

J. Exptl. Bot. 4. 272-282.

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JOURNAL OF THE A. S. S. B.

Preplant Weed Control on Sugar Beets1


E. F. SULLIVAN,

R. R. W O O D ,
Received

R.

L. ABRAMS, AND S.

for publication

G. W A L T E R 2

July 1, 1964

Introduction
T o d a y , w i t h t h e u n c e r t a i n t y of availability of h a n d labor,
r e s e a r c h e r s n e e d to d e v i s e effective systems of c h e m i c a l w e e d i n g
on sugar beets. For, the ultimate, complete spring mechanization, r e m a i n s d e p e n d e n t o n t h e d e v e l o p m e n t o f c h e m i c a l c o n t r o l s
t h a t p e r m i t p l a n t i n g t o s t a n d i n a weed-free e n v i r o n m e n t witho u t h a n d l a b o r a n d c u l t i v a t i o n . T h e r e f o r e , t h e systematic screening of available herbicides is an important contribution toward
the r e a l i z a t i o n of t h i s g o a l .
T h e p r e e m e r g e n c e herbicides disodium 3,6-endoxohexahydrop h t h a l i c a c i d ( e n d o t h a l l ) a n d t r i c h l o r o a c e t i c acid ( T C A ) h a v e
given v a r i a b l e r e s u l t s i n surface i r r i g a t e d r e g i o n s a l t h o u g h satisfactory w e e d c o n t r o l s a r e o b t a i n e d u n d e r h u m i d c l i m a t e s a n d
n a t u r a l r a i n f a l l c o n d i t i o n s (2,3,6,8,9) 3 . Lately, p r e p l a n t applications of e t h y l iV, i V - d i - n - p r o p y l t h i o l c a r b a m a t e ( E P T C ) , n-propyl
e t h y l - n - b u t y l t h i o l c a r b a m a t e ( P E B C ) a n d 2,3-dichloroallyl diisop r o p y l t h i o l c a r b a m a t e ( D A T C ) h a v e s h o w n eflEective c o n t r o l of
c e r t a i n w e e d species b u t ineffective c o n t r o l o f k o c h i a i n i r r i g a t e d
r e g i o n s (1,3,7,8,9). R e s e a r c h i n C o l o r a d o a n d M o n t a n a has s h o w n
that t h e c o m b i n a t i o n , P E B C + D A T C , i s m o r e effective i n t h e
c o n t r o l of w i l d o a t , Avena fatua, a n d l a m b s q u a r t e r s , Chenopodium album, w h i l e c o n t r o l l i n g p i g w e e d , Amaranthus retroflexus,
a n d foxtail, Setaria s p p . , t h a n P E B C a p p l i e d a l o n e (8).
T h e o b j e c t i v e s o f t h i s s t u d y w e r e t o d e t e r m i n e f u r t h e r (A)
t h e w e e d c o n t r o l effectiveness o f n e w h e r b i c i d e s a n d (B) t o evaluate t h e r e l a t i v e effectiveness o f h e r b i c i d e c o m b i n a t i o n s i n a n
a t t e m p t t o i n c r e a s e t h e s p e c t r u m o f w e e d c o n t r o l o n s u g a r beets.
Materials and Methods
T h e s p r i n g e x p e r i m e n t s w e r e c o n d u c t e d a t 6 locations, n a m e ly: W i n d s o r a n d S t e r l i n g , C o l o r a d o ; M i t c h e l l a n d Bayard,
N e b r a s k a ; L o v e l l , W y o m i n g ; a n d Billings, M o n t a n a . T h e s e trials
were i n i t i a t e d on M a r c h 27 t h r o u g h A p r i l 20 in the spring a n d
o n J u n e 1 3 a n d J u l y 1 7 i n t h e s u m m e r . T h e n u m b e r o f treatm e n t s p e r t r i a l r a n g e d f r o m 1 2 t o 2 4 a n d t h e n u m b e r o f treatm e n t s a t e a c h l o c a t i o n r a n g e d f r o m 2 4 t o 6 6 a m o n g locations
/ Contribution from T h e Great Western Sugar Company, Agricultural Experiment
^ n ' L o n S m o n t , Colorado.
.
Agronomist, Manager, and Assistant Agronomists, respectively.
Numbers in parentheses refer to literature cited.

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JOURNAL OF THE A. S. S. B. T.

and dates. These treatments were arranged in randomized complete blocks with 2 to 3 replicates. The single herbicides evaluated
in this report are shown in Table 1.
Table 1.Single herbicides evaluated under preplant conditions in the spring and
summer, 1963.

The herbicides were applied preplant at planting time as


sprays incorporated to a depth of li/ to 2 inches. The powerdriven incorporator which was used tilled an area 6 inches in
width, and the tilled soil was compacted with a press wheel. A
variable dosage sprayer, operating at 40 psi, 2.2 mph ground
speed, and equipped with Delavan ES-4 nozzle tips, was used.
The sprayer gave a half-dosage distance of 25 feet. Spray output under constant rate conditions measured 14.3 gpa. The initial
rates of active ingredient varied, depending on the chemical, and
these rates ranged from 4 to 16 pounds per acre. The plot size
was 44 inches by 125 feet, with the herbicides applied in 7-inch
bands on two rows spaced 22 inches apart. The commercial monogerm sugar beet seed used, with variety location dependent,
germinated from 80 to 85%. The seed was planted at the rate
of 6 seeds per foot at the 1-inch soil depth. Soil textures among
locations included sandy loam, silty clay loam, and clay loam,
and these soils were of high fertility.
The experiments were conducted under natural precipitation conditions supplemented with surface irrigation water. Surface irrigation was used to establish beets and weeds at 3 locations
in the spring and on July 17 at Windsor. At other locations,
precipitation was sufficient to maintain the plots without supplemental water until plant counts were made. When the herbicides were applied, soil temperatures at the 2-inch depth averaged 56F in the spring and 79 F in the summer. The spring
soil temperature ranged from 44 to 64F, while the temperature
range at application in the summer was from 72 to 86F. Wind
conditions were calm to moderately windy.
Weed seedings were made, at a shallow depth prior to application of the herbicides, to insure the presence of weeds in ^
test areas. T h e experimental weed seed and synthetic weed stands

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391

contained
pigweed,
Amaranthus
retroflexus;
kochia,
Kochia
scoparia; a n d S e t a r i a m i l l e t , Setaria italica, a m o n g o t h e r species.
At s o m e l o c a t i o n s , v o l u n t e e r l a m b s q u a r t e r s , Chenopodium album;
g r e e n f o x t a i l , Setaria viridis; yellow foxtail, Setaria glauca; a n d
b a r n y a r d g r a s s , Echinochloa crusgalli, w e r e p r e s e n t in m i n o r percentages w h i l e v o l u n t e e r b l a c k n i g h t s h a d e , Solatium nigrum,, p r e sented m a j o r infestations. T h e ratio of broadleaved weeds to
grasses i n t h e u n t r e a t e d c o n t r o l s , a s d e t e r m i n e d b y p l a n t c o u n t s ,
r a n g e d f r o m 7 3 : 2 7 t o 5 1 : 4 9 a m o n g trials. W e e d p o p u l a t i o n
densities i n t h e c h e c k p l o t s r a n g e d from 2 6 t o 100 weeds p e r
s q u a r e foot a n d s u g a r b e e t s h a d e m e r g e d densities r a n g i n g from
1.3 to 3.7 p l a n t s p e r l i n e a r foot of r o w .
W e e d a n d b e e t s e e d l i n g c o u n t s w e r e t a k e n , w i t h i n a w i r e rect a n g l e w h i c h m e a s u r e d 4 by 36 inches, as a m e a s u r e of t r e a t m e n t
effectiveness. T h e c o u n t s w e r e m a d e a t a place i n each r o w
e s t i m a t e d t o h a v e t h e g r e a t e s t w e e d c o n t r o l w i t h t h e least i n j u r y
t o s e e d l i n g b e e t s , a n d t h e p l a c e was r e c o r d e d a s o p t i m u m . B o r d e r
effects w e r e e l i m i n a t e d b y p l a c i n g t h e q u a d r a t e a t e q u i d i s t a n t
i n t e r v a l s t o e a c h side o f t h e b e e t r o w . I n a d d i t i o n , a r e t a r d a t i o n
e s t i m a t e w a s m a d e o n b e e t s . T h e s e o b s e r v a t i o n s w e r e m a d e from
May 16 to M a y 2 8 , on J u l y 1 a n d 2 a n d on A u g u s t 13 for t h e
s p r i n g , J u n e 13, a n d J u l y 1 7 e x p e r i m e n t s , respectively. T h e
o p t i m u m data w e r e recorded a n d calculated as the percentages
of t h e u n t r e a t e d c o n t r o l of s i n g l e or total species. In this study,
the a v e r a g e w e e d c o n t r o l p e r c e n t a g e s of h e r b i c i d e s s h o w i n g less
t h a n 6 0 % c o n t r o l r e m a i n u n r e p o r t e d e x c e p t s t a n d a r d chemicals.
W h i l e , e m e r g e d w r eed s e e d l i n g d e n s i t i e s of less t h a n 2 p e r s q u a r e
foot i n t h e u n t r e a t e d c h e c k s w e r e c o m p o s i t e d a n d r e p o r t e d a s t h e
p e r c e n t a g e c o n t r o l o f o t h e r b r o a d l e a v e d weeds.
N o a t t e m p t w a s m a d e t o a d a p t statistics t o t h e analysis o f
variable dosage results.
R e s u l t s a n d Discussion
Spring

results

T h e a v e r a g e w e e d c o n t r o l p e r c e n t a g e s r a n g e d from 5 0 t o
7 5 w h e n p r e p l a n t h e r b i c i d e s w e r e a p p l i e d i n t h e s p r i n g ( T a b l e 2).
Specific c o m p a r i s o n s s h o w e d t h a t t h e t h i o l c a r b a m a t e s , R 1 9 1 0
a n d E P T C , g a v e p r o m i s i n g c o n t r o l o f p i g w e e d a n d foxtail b u t
insignificant c o n t r o l o f k o c h i a w h i c h h a d b e e n s h o w n e a r l i e r
(8). L i k e w i s e (8), i n t h e a b s e n c e o f w i l d oats, t h e s t a n d a r d h e r b icide, P E B C + D A T C , g a v e 1 4 p o i n t s m o r e b r o a d l e a v e d w e e d
control t h a n t h e percentage control produced by P E B C applied
alone ( T a b l e 2).

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Table 2.Average effects of various preplant herbicides at the optimum response


spring experiments at 6 locations. Treated March 27 through April 20.

3
Number of observations shown in parenthesis followed by combination ratio of acti\_
* Weed densities in the untreated controls averaged: pigweed. 14.5; kochia. 10.5; other
broadleaved weeds 5.9; and foxtail, 18.4 per sq ft. The emerged beet seedlings averaged
3.7 per linear ft of row. Data for foxtail from 4 locations only, all broadleaves omitted
from total species control or average.

T h e data gave evidence that the herbicides evaluated had


three average intensities of: effectiveness, namely, 74, 67 and 52%
control ( T a b l e 2). T h e r e f o r e , c o m p u t a t i o n s showed that the
average difference between the standard t r e a t m e n t , PEBC -f
D A T C , a n d t h e 7 4 % control g r o u p , was 6 percentage points
a m o n g 6 locations ( T a b l e 2). W h i l e , t h e difference between
the m e d i u m g r o u p of 6 7 % control a n d the higher g r o u p averaged
7 percentage points. T h e lowest control g r o u p averaged 52%
which was 21 a n d 14 percentage points less t h a n the average
of the high a n d m e d i u m groups, respectively ( T a b l e 2).
T h e experimental herbicides gave variable control responses
a m o n g broadleaved weeds a n d between broadleaved and grassy
weeds. For example, Pyrazon p r o d u c e d 7 5 % control of all broadleaved weeds b u t ineffective control of foxtail which averaged
4 6 % . T h e Europeans, Fisher (4) a n d L'hoste et al. (5), showed
similar results from preemergence applications of Pyrazon. Conversely, control of foxtail from p r e p l a n t applications of CP32179
a n d T D 2 8 2 ranged from 86 to 81 percentage points while the
broadleaved weed control from the two chemicals averaged 65%
( T a b l e 2). It is significant to n o t e t h a t T D 2 8 2 gave the highest
control of kochia which averaged 7 7 % , while CP32179 gave
4 0 % control of kochia a n d Pyrazon, 6 9 % , at the time the observations were m a d e ( T a b l e 2).
A l t h o u g h o t h e r factors may be responsible, the results showed
t h a t Pyrazon, T D 2 8 2 a n d CP32179 gave 16 percentage points

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m o r e c o n t r o l o f p i g w e e d a n d k o c h i a i n coarse t h a n i n finet e x t u r e d soils w h e n 2 l o c a t i o n s for e a c h soil class w e r e c o m p a r e d


( T a b l e 3). F o x t a i l c o n t r o l f r o m t h e a p p l i c a t i o n o f c h e m i c a l s
o t h e r t h a n C P 3 2 1 7 9 a v e r a g e d 1 0 p e r c e n t a g e p o i n t s less o n sandy
loam t h a n o n clay l o a m soils. I n p a r t i c u l a r , T D 2 8 2 was effective
i n w e e d c o n t r o l w h e n a p p l i e d p r e p l a n t o n sandy l o a m soils.
Table 3.Average effects of Pyrazon, TD282 and CP32179 at the optimum response,
spring experiments at 2 locations each on coarse- and fine-textured soils.
Percent of control 6
Pigweed
Herbicide 5
TD282 (4)
Pyrazon ( 4 )
CP32179 ( 8 )
Pyrazon -f- CP32179 ( 4 ) 3:1

Kochia
Sandy

95
92
94
99

92
72
46
59

Foxtail
loam

Average

soils
75
41
87
70

87~
68
76
76

5
N u m b e r of observations in parenthesis followed by combination ratio of active.
6
Weed densities in the untreated controls averaged for coarse- and fine-textured soil;
respectively: Pigweed, 14.3, 12.8; Kochia. 10.5, 10.6; and foxtail. 14.3 and 22.4 per sq ft.

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T a b l e 4 . A v e r a g e effects o f v a r i o u s p r e p l a n t h e r b i c i d e s a t t h e o p t i m u m response
s u m m e r e x p e r i m e n t s , W i n d s o r , C o l o r a d o . T r e a t e d J u n e 1 3 a n d July 17.

7
N u m b e r of observations s h o w n in p a r e n t h e s i s f o l l o w e d by c o m b i n a t i o n ratio of active
Each e x p e r i m e n t c o n t a i n e d 3 replicates.
8
W e e d d e n s i t i e s in t h e u n t r e a t e d controls a v e r a g e d : P i g w e e d , 38.5; a n d foxtail, 27.2
per sq ft.

the spring applied herbicides ( T a b l e s 2 a n d 4). However, the


effectiveness of the s u m m e r treatments, as measured by percentage control, was 15-20 percentage p o i n t s higher than in the
spring. Similarly, this increased effectiveness was reported for
herbicides applied in the s u m m e r in 1962 (8). Apparently, these
higher control percentages were affected by the summer soil
temperatures which averaged 23F higher t h a n in spring, although t h e presence of kochia in t h e weed populations of spring
a n d absence in the s u m m e r may have been a factor affecting the
results.
A m o n g herbicides a n d species, Pyrazon produced the least
control of foxtail which averaged 7 5 % ( T a b l e 4). T h e control
of foxtail by the other herbicides was effective a n d ranged from
86 to 9 6 % . Nevertheless, Pyrazon gave 29 percentage points
m o r e control of foxtail in t h e s u m m e r t h a n in the spring although
the control difference b e t w e e n seasons for pigweed was 16 percentage points ( T a b l e 2 a n d 4). T h e control of pigweed was
effective for most herbicides, a n d this control ranged from 47%
for R4572 to 9 9 % for Pyrazon. T h e combinations, CPS2179 +
T D 2 8 2 a n d Pyrazon + T D 2 8 2 showed promise, these combinations averaged 93 a n d 9 0 % c o n t r o l of pigweed plus foxtail respectively.
Beet seedling injuries from the s u m m e r application of herbicides were slight with the exception of R4572 which averaged
1 8 % stunting a n d 8% r e d u c t i o n in stand as it had in the spring
(Tables 2 a n d 4). C o m p u t a t i o n s showed that the average beet
p o p u l a t i o n of t h e e x p e r i m e n t a l a p p l i e d in summer were

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p e r c e n t a g e p o i n t s h i g h e r t h a n those o f P E B C a n d P E B G +
D A T C w h i c h a v e r a g e d 1 1 1 % . S t a n d differences a m o n g c h e m icals w e r e u n d e t e c t e d i n t h e s p r i n g ( T a b l e s 2 a n d 4).
Summary
Several s i n g l e h e r b i c i d e s a n d h e r b i c i d e c o m b i n a t i o n s w e r e
e v a l u a t e d a t 6 f i e l d l o c a t i o n s t o d e t e r m i n e t h e i r p r e p l a n t pot e n t i a l for c h e m i c a l w e e d i n g o n s u g a r beets.
T h e results showed that Pyrazon, T D 2 8 2 and CP32179 were
m o r e effective i n w e e d c o n t r o l t h a n P E B C w h i l e P E B C + D A T C
was m o r e effective t h a n P E B C a p p l i e d a l o n e . T h e h e r b i c i d e
combinations Pyrazon + CP32179, P y r a z o n + T D 2 8 2 a n d
C P 3 2 1 7 9 + T E 2 8 2 a m o n g o t h e r s gave effective c o n t r o l o f c e r t a i n
b r o a d l e a v e d a n d grassy w e e d s w i t h o u t u n d u e i n j u r y t o s u g a r
beet s e e d l i n g s .
Specific c o m p a r i s o n s s h o w e d t h a t P y r a z o n p r o d u c e d effective
b r o a d - l e a v e d w e e d c o n t r o l , b u t t h e c h e m i c a l was relatively ineffective i n t h e c o n t r o l o f grass, w h i l e , C P 3 2 1 7 9 a n d T D 2 8 2
gave a d d i t i o n a l c o n t r o l o f grass. T D 2 8 2 effected s u p e r i o r a n d
E P T C a n d P E B C g a v e i n f e r i o r c o n t r o l o f kochia.
T h e results indicated that CP32179, T D 2 8 2 and Pyrazon
w e r e m o r e effective o n l i g h t t h a n h e a v y t e x t u r e d soils.
H e r b i c i d e s a p p l i e d u n d e r s u m m e r c o n d i t i o n s gave h i g h e r
weed c o n t r o l p e r c e n t a g e s t h a n t h e same h e r b i c i d e s a p p l i e d i n
the spring.
Acknowledgement
T h e a u t h o r s wish to express their appreciation on behalf of
T h e G r e a t W e s t e r n S u g a r C o m p a n y t o t h e following c o m p a n i e s
for f u r n i s h i n g t h e h e r b i c i d e s r e p o r t e d i n t h i s s t u d y : A m c h e m
Products, Inc.; M o n s a n t o Chemical Company; Pennsalt Chemicals C o r p o r a t i o n ; a n d Stauffer C h e m i c a l C o m p a n y .
L i t e r a t u r e Cited
(1)

ALLEY,

H.

P.,

E.

W.

CHAMBERLAIN,

1961. W e e d c o n t r o l
M i m e o Circ. 159.
(2)

in

C.

F.

sugar beets.

BECKER,

and

G.

W y o m i n g Agr.

A S C H E M A N , R . E., R . C . Z I E L K E , a n d E. W . STROUBE.

1961.

L.

COSTEL.

Expt.

Sta.,

Herbicides

i n sugar beets. R e s . R p t . N C W C C 18: 69.


(3)

ECKROTH, E.

G.,

E.

M.

H O L S T , a n d C . E. C O R M A N Y .

1958.

Chemical

c o n t r o l of a n n u a l grasses in sugar beets in M o n t a n a , W y o m i n g , a n d


C o l o r a d o . J. A m . Soc. Sugar Beet T e c h n o l . 10: 156-164.
(4) FISCHER, A.
1962.
l-pheiiyl-4-amino-5-chlor-pyridazon-6 (PCA) als ein
n e u e s R u b e n h e r b i z i d . W e e d R e s . 2 : 177-184.

396

JOURNAL OF THE A. S. S. B. f.

(5) L/HOSTE, J., H . D ' I L L E , A. CASANOVA, a n d L. A. DURGEOT.

1963.

Essais

en plein champ de desherbage selectif des betteraves por le chloro


amino-phenyl-pyridazone (PCA). Weed Res. 3: 52-65.
(6) NELSON, R. T. 1954. Grass control in sugar beet with the herbicides
IPC, T C A and DCU. Proc. Am. Soc. Sugar Beet Technol. 8: 130.
134.
(7) SEXSMITH, J. J. 1960. Chemical control of wild oats (Avena fatua L.)
in sugar beets. J. Am. Soc. Sugar Beet Technol. 11: 268-278.
(8)

SULILVAN, E. F., R. L. ABRAMS, a n d R. R. W O O D .

1963.

Weed control

in sugar beets by combinations of thiolcarbamate herbicides. Weeds


11: 258-260.
(9) WICKS, G. A. 1962. Herbicides for control of broadleaf weeds in sugar
beets. Res. Rpt. NCWCC 19: 93-94.

Breeding Sugar Beets for Resistance to the


Cyst N e m a t o d e Heterodera Schachtii
CHARLES

PRICE1

Received for publication July 27, 1964

Introduction
T h e cyst n e m a t o d e Heterodera schachtii S c h m i d t causes a
m a j o r disease o f s u g a r b e e t s i n E u r o p e a n d i n t h e U n i t e d States.
I n t h e U n i t e d States, t h e cyst n e m a t o d e has b e e n severe chiefly
i n areas w h e r e s u g a r b e e t s h a v e b e e n g r o w n for m a n y years. I n
some a r e a s , h o w e v e r , n o t a b l y i n t h e I m p e r i a l Valley o f California, s u g a r b e e t s h a v e b e e n g r o w n a relatively s h o r t t i m e , yet
t h e n e m a t o d e disease is a l r e a d y serious a n d soil infestation is
s p r e a d i n g . P r o b a b l y i n i t i a l n e m a t o d e infestation r e s u l t e d from
cysts c a r r i e d i n p a r t i c l e s o f soil a d h e r i n g t o f a r m e q u i p m e n t
w h i c h h a d b e e n u s e d i n o t h e r areas w h e r e t h e soil was infested
with s u g a r b e e t n e m a t o d e . T h e pest was b r o u g h t t o this c o u n t r y
from E u r o p e , w h e r e i t h a s b e e n k n o w n for over 100 years, t h r o u g h
t h e i m p o r t a t i o n o f s u g a r b e e t seed. I n t h e early 1920's, t h e
a u t h o r f o u n d l i v e n e m a t o d e s i n soil screened from i m p o r t e d
sugar b e e t s e e d ( l ) 3 .
C r o p r o t a t i o n h a s b e e n effective i n keeping: t h e p o p u l a t i o n
of n e m a t o d e s d o w n to a level t h a t will p e r m i t o n e c r o p of beets
to be g r o w n successfully in infested soil o n c e in 4 to 6 years.
T h i s is possible only if nonhost plants are grown in rotation
w i t h s u g a r b e e t s . H o w e v e r , i n t e r v a l s of 4 to 6 years b e t w e e n
sugar b e e t c r o p s s o m e t i m e s a r e n o t c o n d u c i v e t o favorable
economy of small farmers w h o d e p e n d u p o n sugar beets as a
profitable cash c r o p .
C r o p r o t a t i o n , soil f u m i g a t i o n , a n d o t h e r m e a s u r e s h a v e
b e e n r e c o m m e n d e d for c o n t r o l o f t h e n e m a t o d e ; however, these
are n o t fully satisfactory, a n d t h e r e is still serious n e e d for
varieties t h a t a r e t o l e r a n t o f t h e p a t h o g e n . B r e e d i n g research
for t h e d e v e l o p m e n t o f v a r i e t i e s t h a t a r e resistant t o t h e n e m a t o d e
has b e e n a m a j o r a c t i v i t y at t h e U. S. A g r i c u l t u r a l R e s e a r c h
Station, S a l i n a s , C a l i f o r n i a . T h e use o f resistant varieties w o u l d
* Research Agronomist, Croos Research Division, Agricultural Research Service, U. S.
department of Agriculture, Salinas, California.
3
T h e author is indebted to Phvllis Emparan. formerlv a member of the Crops Research
division, Agricultural Research Service. U. S. Department of Agriculture, for help with
statistical analyses, and to C. L. Schneider. Pathologist. Crops Research Division, Agricultural Research Service, l.ogan, Utah, for isolation of the pathogenic fungi from the field
soil used for screening tests.
8
Numbers in parentheses refer to literature cited.

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JOURNAL OF THE A. S. S. B.

r/.

reduce the damage caused by the nematode as well as permit


growers to shorten the interval between crops of sugar beets in
the rotation.
M a t e r i a l a n d Methods
H u n d r e d s of thousands of plants have been examined in
screening tests in an a t t e m p t to find resistance to the cyst nematode Heterodera schachtii. Material examined has consisted of
commercial varieties such as the curly top-resistant varieties
US 22, US 5 6 / 2 , a n d US 33, and some of the Cercospora leafspot-resistant varieties. Also used were promising monogerm
lines, plants from irradiated seeds, and crosses between sucrar
beets and Beta maritima L., as well as crosses between sugar
beets a n d Beta webbiana Moq., which is i m m u n e to the attack
of H. schachtii.
Screening tests in the greenhouse are accomplished by means
of a t e c h n i q u e developed at Salinas, in which seedling sugar beets
are exposed to severe n e m a t o d e attack a n d the most vigorous
plants with fewest cysts are selected for future studies and further
selections. T h e n u m b e r of plants showing any degree of re
sistance is so small that it is necessary to e x a m i n e large populations of sugar beets in o r d e r to obtain a few which show promise
as b r e e d i n g material. Many of these are eliminated later when
tested u n d e r m o r e extensive exposure to nematodes and other
soil borne pathogens.
Viable cysts for use in screening b r e e d i n g material for nematode resistance are collected from fields in which sugar beets have
suffered severe damage from the pathogen. T h e soil is collected
immediately after harvest of the infested crop, and by this
means, freshly formed and highly viable cysts are obtained. The
field soil is screened by means of a m a c h i n e especially designed
for the purpose. T h r e e screens of different size mesh are mounted
in a frame which is agitated by an eccentric. T h e clods and debris
are carried to the e n d of the largest screen a n d returned to the
field; the small soil particles a n d n e m a t o d e cysts pass through
the smallest screen and collect in a container. T h e mixture of
nematode cysts a n d small particles of soil are placed in 50-gallon
d r u m s for use as i n o c u l u m in t h e evaluation of sugar beets for
nematode resistance.
Cyst p o p u l a t i o n in t h e screened field soil is determined by
weighing 100-gram samples of i n o c u l u m from each drum. 1
each of several 100-gram samples contains 200 cysts filled witn
eggs the i n o c u l u m is considered a d e q u a t e . Cysts in the inoculum,
however, frequently exceed 200 p e r 100 grams of soil. Field soil
frequently contains, in a d d i t i o n to n e m a t o d e cysts, root-rotting

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fungi. Rhizoctonia solani Kuehn is the principal pathogenic


fungus isolated from diseased seedlings grown in soil infested
by the inoculum.
Greenhouse
Test
Screening sugar beets for nematode resistance involves planting seeds in sterile sand, and when the seedlings reach the 2-leaf
stage they are transplanted in greenhouse flats (Figure 1) containing a special mix of sterilized light sandy soil to which the
nematode inoculum is added. T h e soil and inoculum are thoroughly mixed so that the cysts are uniformly distributed. After
6 or 8 weeks the plants are immersed in water and the adhering
soil is carefully removed from the roots without detaching the
female nematodes. T h e sugar beet rootlets are tneh examined

Figure 1.Xesting for resistance to Heterodera schachtH. Sugar beet


seedlings are transplanted from sterile sand to flats of nematode infested
soil. US 41 in 2 center rows; new breeding lines on left and right.

for the presence of female nematodes. T h e plants are classified


into 6 categories (0 to 5 inclusive) according to the number of
cysts present on the roots. T h e plants graded 0, 1, and 2, which
have from n o n e to 5 nematodes on the roots, are transferred to
aluminum cylinders where they are exposed further to nematode
attack before final selections for resistance are made. Plants
carded. Frequently, the selected plants which were graded 0, 1,
graded 4 or 5 are considered susceptible to nematode and disand 2 and further tested in aluminum cylinders are graded 4 or

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JOURNAL OF THE A. S. S. R.

5 w h e n removed from the cylinders (Figure 2). Final selections


for resistance to n e m a t o d e are characterized by: a) n o t more
t h a n 3 n e m a t o d e cysts on the roots, b) high vigor, a n d c) comparatively large root size.

Figure 2.Females of Heterodera schetchtii on roots of susceptible


sugar beet grown in infested soil in a l u m i n u m cylinder.

After approximately 8 weeks, sugar beets in aluminum


cylinders are examined, a n d t h e final selections are planted in
8-inch greenhouse pots a n d placed in a favorable location for
growth. After the roots in greenhouse pots have attained approximately 1 inch in diameter, t h e plants are removed to a cold
room, which is e q u i p p e d with lights, a n d t h e temperature maintained at 42 F. T h e plants r e m a i n in t h e 42 temperature 90
to 120 days, d e p e n d i n g u p o n the t h e r m a l i n d u c t i o n requirement
of t h e selection. T h e y are t h e n r e m o v e d to t h e field or green-

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house for s e e d i n c r e a s e o r h y b r i d i z a t i o n . Selections for n e m a t o d e


resistance a r e a g a i n t e s t e d i n f i e l d a n d g r e e n h o u s e a n d a second
cycle of s e l e c t i o n a n d t e s t i n g is c o m p l e t e d . Selections for n e m a t o d e r e s i s t a n c e h a v e also b e e n m a d e from c o m m e r c i a l f i e l d s i n
t h e Salinas, C a l i f o r n i a , a r e a .
Crock

Test

C r o c k s for t h e test ( F i g u r e 3 ) a r e p a r t i a l l y f i l l e d w i t h u n i formly m i x e d f e r t i l e soil. N e m a t o d e cyst i n o c u l u m i s a d d e d t o


t h e soil in half of t h e crocks. E a c h l i n e of s u g a r b e e t to be
e v a l u a t e d is p l a n t e d in a set of 10 crocks each of infested a n d
n o n i n f e s t e d soil. T h e r e a r e 3 such r e p l i c a t i o n s o r 30 crocks
w i t h n e m a t o d e i n o c u l u m a d d e d a n d 3 0 crocks w i t h n o i n o c u l u m .
T h e s u g a r b e e t s a r e t h i n n e d t o a single p l a n t i n each crock. A t
harvest, t o p s a r e r e m o v e d a n d r o o t s a r e w e i g h e d . Yields o f r o o t s
are c a l c u l a t e d o n t h e i n d i v i d u a l r o o t w e i g h t basis a n d each p l a n t
is c o n s i d e r e d a r e p l i c a t i o n .

Figure 3.Crock test of nematode-resistant lines of sugar beet developed


in greenhouse screening program. One plant in each 3-galIon crock of
nematode-infested soil or nematode-free soil.

Field

Test

In t h e field test p l o t ( F i g u r e 4) at t h e U. S. A g r i c u l t u r a l
R e s e a r c h S t a t i o n at Salinas, C a l i f o r n i a , t h e soil is infested w i t h
sugar b e e t n e m a t o d e a n d i t also c o n t a i n s r o o t - r o t t i n g f u n g i ,
p r i n c i p a l l y Rhizoctonia solani. A h i g h p o p u l a t i o n of n e m a t o d e s
i s m a i n t a i n e d i n t h e soil b y m e a n s o f f r e q u e n t a p p l i c a t i o n s , w i t h
a fertilizer drill, o f soil i n f e s t e d w i t h s u g a r beet n e m a t o d e . T h e

400

JOURNAL OF THE A. S. S. B. T.

5 w h e n removed from the cylinders (Figure 2). Final selections


for resistance to nematode are characterized by: a) not more
than 3 nematode cysts on the roots, b) high vigor, a n d c) comparatively large root size.

Figure 2.Females of Heterodera schachtii on roots of susceptible


sugar beet grown in infested soil in aluminum cylinder.

After approximately 8 weeks, sugar beets in aluminum


cylinders are examined, and the final selections are planted in
8-inch greenhouse pots a n d placed in a favorable location tor
growth. After the roots in greenhouse pots have attained approximately 1 inch in diameter, the plants are removed to a col
room, which is e q u i p p e d with lights, a n d the temperature main
tained at 42 F. T h e plants r e m a i n in t h e 42 temperature 90
to 120 days, d e p e n d i n g u p o n the t h e r m a l i n d u c t i o n requirement
of t h e selection. T h e y are then removed to t h e field or green

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house for seed increase or hybridization. Selections for nematode


resistance are again tested in field and greenhouse and a second
cycle of selection and testing is completed. Selections for nematode resistance have also been made from commercial fields in
the Salinas, California, area.
Crock Test
Crocks for the test (Figure 3) are partially filled with uniformly mixed fertile soil. Nematode cyst inoculum is added to
the soil in half of the crocks. Each line of sugar beet to be
evaluated is planted in a set of 10 crocks each of infested and
noninfested soil. T h e r e are 3 such replicationsor 30 crocks
with nematode inoculum added and 30 crocks with no inoculum.
T h e sugar beets are thinned to a single plant in each crock. At
harvest, tops are removed and roots are weighed. Yields of roots
are calculated on the individual root weight basis and each plant
is considered a replication.

Figure 3.Crock test of nematode-resistant lines of sugar beet developed


in greenhouse screening program. O n e plant in each 3-gallon crock of
nematode-infested soil or nematode-free soil.

Field

Test

In the field test plot (Figure 4) at the U. S. Agricultural


Research Station at Salinas, California, the soil is infested with
Migar beet nematode and it also contains root-rotting fungi,
principally Rhizoctonia solani. A high population of nematodes
is maintained in the soil by means of frequent applications, with
a fertilizer drill, of soil infested with sugar beet nematode. T h e

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JOURNAL OF THE A. S. S. B. T.

Figure 4.General view of nematode resistance test, U. S. Agricultural


Research Station, Salinas, California. T h e soil in this field is heavily
infested with the cyst nematode Heterodera schachtii and root-rotting
fungi-

contaminated soil is placed approximately 3 inches deep and


close to the planted row of sugar beets. Selections to be tested
for resistance to nematodes are planted in May, when soil temperatures are favorable for nematode activity and maximum
damage occurs. T h e sugar beets are exposed to nematode attack
at a very earlv stage of growth when they are highly susceptible
to damage. T h e sugar beets are planted on beds and furrow
irrigation is used. Plots of each selected line of sugar beet and
of the commercial varieties used as checks are single row, 20
inches apart and 25 feet long, with 2 to 6 replications of each
line in the test.
Root-rotting fungi play an important part in the damage
attributed to nematodes. T h e punctures made by Heterodera
schachtii undoubtedly facilitate penetration of pathogenic organisms into root tissue. Therefore, a variety of sugar beet that
is resistant to a combination of pathogenic soilborne organisms
and nematode is highly desirable. T h i s phase of the work has
received much attention in the breeding program.
Results
Greenhouse
Test
Screening tests were made in the greenhouse u n d e r conditions of exposure to both nematodes and root-rotting fungi. By
repeated selections and crosses, some lines have been developed
which show remarkable resistance to a combination of root rot
and cyst nematode (Figure 5); b u t much further work is necessary to develop lines of sugar beet that are fully resistant to the
combination of nematodes and other soilborne pathogens under
field conditions. Studies have indicated strongly that when some

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nematode-resistant lines are exposed to the c o m b i n a t i o n they


suffer less d a m a g e t h a n t h e c o m m e r c i a l v a r i e t y U S 4 1 used a s
check. I t i s necessary, h o w e v e r , t o c o n d u c t f u r t h e r tests t o d e t e r m i n e w h e t h e r these l i n e s a r e r e s i s t a n t t o t h e p a t h o g e n i c f u n g i
o r w h e t h e r n e m a t o d e t o l e r a n c e o f t h e s u g a r b e e t gives m o r e
protection against the root-rotting organisms.

F i g u r e 5.Sugar beet seeds p l a n t e d in g r e e n h o u s e flats in soil heavily


infested w i t h t h e sugar beet n e m a t o d e a n d c o n t a i n i n g r o o t - r o t t i n g fungi.
C o m m e r c i a l variety US 41 a n d a m o n o g e r m line, SL 9229, are highly susceptible. A selected l i n e , 101-7, shows r e m a r k a b l e resistance.

Field
Test
T a b l e 1 s h o w s t h e r e s u l t s of tests of 10 l i n e s of s u g a r b e e t
b r e d for r e s i s t a n c e to t h e cyst n e m a t o d e Meterodera schachtii a n d
t w o c o m m e r c i a l v a r i e t i e s , U S 4 1 a n d U S 75, u s e d a s c h e c k s . U S
41 has b e e n u s e d as a c h e c k for several years in n e m a t o d e tests,
Table 1.Root yield of resistant lines of sugar beet grown under severe exposure to
cyst nematode and other soil-inhabiting pathogens. Field test, Salinas, California, 1962.

JOURNAL OF THE A. S. S. B. T.

404

b u t US 75 was used for the first time in the 1962 tests. Sugar
beets were planted in heavily infested soil as already described.
Plots were replicated three times for each line and for each of
the two commercial checks. Of the 10 nematode-resistant lines
included in the test, 7 were significantly superior in yield of
roots per acre to US 41 at the 1% level of significance, and 3
were significantly superior at the 5% level. All nematoderesistant lines tested were superior in yield per acre to US 75
at the 1% level of significance.
Crock Test
T a b l e 2 shows results of 12 entriesUS 41, used as check,
and 11 lines of sugar beet which have been bred for resistance
to the cyst nematode. T h e plants were grown in 3-gallon crocks,
as described u n d e r "Methods". After the tops were removed,
each individual root was weighed and its weight recorded in
grams. T h e r e were 10 crocks in each of three replications, or a
total of 30 beets of each line and 30 beets of US 41. Results
are therefore averages of 30 roots for each entry in the test.
T h r e e 10-beet samples were taken for sucrose determination.
T a b l e 2, column 2, shows weights of roots grown in noninfested
soil, and column 4 shows weight of roots grown in infested soil.
T h e percent loss in weight due to nematodes was calculated on
the basis of difference between weight of roots grown in noninfested soil and those grown in infested soil. T h e loss in weight
Table 2.Root yield and sucrose percentage of resistant lines of sugar beet grown in
crocks containing nematode-infested soil or nematode-free soil. Salinas, California, 1962.
Noninfested

Infested

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varies f r o m 1 % i n l i n e 019 t o 3 0 . 4 % i n U S 4 1 . A l l lines t e s t e d


w e r e h i g h e r i n yield o f r o o t s t h a n U S 4 1 . T h e r e was also a red u c t i o n i n sucrose p e r c e n t a g e d u e t o n e m a t o d e i n all lines tested,
w i t h an a v e r a g e of .81 of a p e r c e n t a g e p o i n t for t h e 11 e n t r i e s .
Summary
S e e d l i n g s u g a r beets w e r e p l a n t e d i n n e m a t o d e i n f e s t e d soil
i n f l a t s i n t h e g r e e n h o u s e a n d l a t e r s c r e e n e d for n e m a t o d e infestation. P l a n t s s h o w i n g t h e least n u m b e r o f cysts w e r e ret a i n e d a n d p l a n t e d singly i n i n f e s t e d soil i n a l u m i n u m c y l i n d e r s
for a s e c o n d e x p o s u r e t o n e m a t o d e s . T h e best o f t h e s e p l a n t s
w e r e r e t a i n e d for seed p r o d u c t i o n . P r o g e n i e s w e r e tested i n
f l a t s , c y l i n d e r s , singly i n 3-gallon crocks, a n d i n r e p l i c a t e d field
tests w h e r e f u r t h e r selections for r e s i s t a n c e w e r e m a d e . L i n e s
w h i c h s h o w p r o m i s e o f a p p r e c i a b l e n e m a t o d e resistance h a v e
b e e n o b t a i n e d f r o m t h e c u r l y t o p - r e s i s t a n t v a r i e t i e s U S 22, U S
5 6 / 2 , a n d U S 33, a n d f r o m s o m e o f t h e C e r c o s p o r a leaf-spotr e s i s t a n t l i n e s . S o m e o f these selections h a v e s h o w n r e s i s t a n c e
to a c o m b i n a t i o n of n e m a t o d e a n d Rhizoctonia root rot.
Literature Cited
(1) F a r m e r s ' B u l l e t i n 1514, p. 6, 1926.

The Chemical Destruction of Sucrose, Fructose,


and Glucose in Hot Alkaline Process Juices
and Liquors
S. E. BlCHSEL 1
Received for publication April 15, 1964

Introduction
Most recently, the destruction of sucrose in hot alkaline
synthetic process juices has been investigated by Pieck (5) 2 and
Carruthers (1).
T h e measurement technique pursued by these investigators
involved the separate measurement of the rate of formation of
invert in a hot alkaline buffered solution containing sucrose and
the rate of invert destruction in a buffered solution under similar
conditions of temperature and p H . In both cases pure sucrose
and invert solutions in conjunction with appropriate buffers
were used. Having obtained the formation rate of invert from
sucrose and the independent rate of destruction of invert, it
was possible to determine the a m o u n t of sucrose destroyed at
any given process condition. All reactions were found to be
of the first order with respect to invert formation and invert
destruction. Carruthers (1) intimated the existence of steady
state with respect to the transient reaction intermediate invert.
Pieck (5) enlarged on the steady state invert concept by measuring invert concentrations u n d e r various pH and temperature
conditions.
It is the intention of this paper to utilize the kinetic concept
of steady state. By doing so, it is possible to solve for the first
order reaction rates describing the destruction of sucrose, glucose
and fructose, and the concentrations of glucose and fructose
within the steady state invert level.
Theory
Steady state as opposed to dynamic state can be explained
in a graphic form. Figure 1 denotes the concentration of sucrose
(A), invert (B), intermediates (C) a n d terminal acidic end
products (D). T h e s e concentrations are plotted with respect to
time. In this model, sucrose concentration is approximately
equivalent to the other reactants. Reaction rates KA, K B and K c
are all high in numerical magnitude. T h i s set of conditions
precludes the existence of steady state. Concentrations of all
reactants involved in the irreversible reaction are subject to
1
8

Research Chemist, The Amalgamated Sugar Company, Twin Falls, Idaho.


Numbers in parentheses refer to literature cited.

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c o n s t a n t c h a n g e . F o r t u n a t e l y , d y n a m i c s t a t e d o e s n o t exist i n
hot alkaline process juices a n d liquors. A n y m e a s u r e m e n t of
r e a c t i o n r a t e s in a d y n a m i c s t a t e r e a c t i o n is e x t r e m e l y difficult.

TIME

>

Figure

1 . D y n a m i c state sucrose destruction.


A > B -+ C > I>
S u c r o s e > I n v e r t > I n t e r m e d i a t e s > A c i d s
J^A
K-R
Kr,

T h e graphic model, F i g u r e 2, stimulates steady state conditions e x i s t e n t i n a c t u a l h o t a l k a l i n e process l i q u o r s a n d j u i c e s .


In t h i s case t h e c o n c e n t r a t i o n of s u c r o s e e x c e e d s t h a t of i n v e r t

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JOURNAL OF THE A. S. S. B. T

by a factor of 300. T h e first order reaction rate describing the


destruction of invert exceeds the reaction rate of sucrose by
a factor of 1000. T h u s , all products of sucrose destruction with
the exception of the acidic e n d products are in steady state with
respect to their individual concentrations in solution. In time,
usually more than five hours, a slight deviation in the steady
state invert concentration may be noted d u e to a small decrease
in the primary reactant sucrose. T h i s may be predicted in accordance with equations developed by Giddings (3).
T h e steady state kinetic principle may be defined as follows:
In any hot alkaline process liquor or juice maintained at
constant pH and temperature, there exists a steady state glucose
and fructose concentration. U n d e r steady state conditions, the
rate of formation of glucose and fructose from sucrose equals
the rate of glucose a n d fructose degradation in solution. Under
these conditions the concentration of both glucose and fructose
remains constant.
Procedure and Equipment
A graphic example of a typical process juice r u n is shown
in Figure 3. U p o n establishing steady state invert concentration,
a weighed dry increment of p u r e fructose is added to the reaction
mixture. T w o or three minutes are allowed for complete mixing
of the fructose in the reactor, before a small sample is withdrawn
and cooled immediately in an ice bath. T h e time of withdrawal
is noted. T h e fructose concentration in excess of the steady
state invert level is determined with triphenyl-tetrazolium chloride reagent. At given time intervals, after the initial sampling,
the fructose concentration is determined. Usually four to five
separate fructose determinations are r e q u i r e d . In approximately
two hours (time for reaction is d e p e n d e n t on solution tempera

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ture and solution p H ) , essentially all fructose will be degraded


and steady state conditions will be restored. T h e same procedure
is duplicated with the addition of a weighed glucose increment
to the reaction solution.
During the course of the reaction the solution pH must be
maintained at some control value. Runs conducted on thin
juice and high brix liquors have displayed typical pH decrease
characteristics. T h e pH of the reaction is maintained with a
few potassium hydroxide pellets mixed in a small volume of
juice from the reaction vessel. Small additions of this solution
throughout the r u n effectively maintain a constant p H .
Reaction system(1) shows the accepted pathway for sucrose
destruction u n d e r hot alkaline conditions at steady state.

(1)

(2)

Reaction system(2) shows the addition of an increment of


fructose to the steady state system (1).

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JOURNAL OF THE A. S. S. B. T.

2. Determine the concentration of (B) and (C) contained within the steady state invert level.
<B> - K2 + K, ' <B> + <C>
<C) " K, + K, * (B> + <C>
When glucose and fructose are produced in equimolar
amounts, as from sucrose, their concentration within the steady
state concentration (B) -j- (C) is indirectly proportional to the
individual rate constants involved.
3. Solve for reaction rate constant Kx describing destruction of
(A) in solution.
Kx (A) Ka (B) + K3 (C)
K2 * (B) - f +_ K3 (C) - f
Ki _
f = factor converting B and C to A equivalents.
4. Solve for percent sucrose destroyed in time (T)
_ K * * ( A > ' T . 100
V
The reaction vessel designed for this project consisted of a
stainless steel vessel of 1.5 liter capacity with pressure fittings
capable of withstanding internal pressures up to 75 psig. This
feature enables the investigator to study destructive reactions
at temperatures far above those obtainable at atmospheric pressure. The reaction vessel was fitted with an internal stirrer,
pressure pH electrodes, electric temperature control, pressure
equalized burret for reagent addition, sample outlets, and standard temperature and pressure gauges.
Results
The concept of steady state invert concentration was investigated
using
first
car
Donation
clarifier
juice.
demonas
describes
carbonation
and
stants
temperatures.
or
strates
enables
the
this
lower
Figure
fructose
juice
K
arange
extrapolation
logarithmic
temperature
the
and
5 temperature
clarifier.
is
indicates
amply
influence
A
in
K
temperature
accordance
ofcovers
increase
The
of
ranges.
the
glucose
of
decreases.
invert
linear
temperature
variation
temperatures
with
in
range
and
concentrations
steady
function
the
The
fructose
of
ofArrhenius
state
the
on
function
70
maintained
found
reaction
C
specific
invert
atto
toFigure
different
for
is
equation
90C
include
concentration
linear,
reaction
velocity
both
in4
was
the
glucose
process
higher
which
whicli
used
first
con
(2).

Figure 5.Reaction rates vs. temperature at a constant pH of 9.70


measured at 2 0 C.
l

r h e diflEerence in reaction constants between glucose and fructose


ndicates that only 36.5% of any steady state invert concentration

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JOURNAL OF THE A. S. S. B. T.

at 80C and 9.70 pH will consist of fructose. Referring to Figure


4, it is noted that the steady state invert concentration at 80C
is 0.055 mg i n v e r t / m l solution. T h u s within this level 0.020
mg/ral is fructose and the remaining 0.035 m g / m l is glucose.
Figure 6 shows the Arrhenius temperature vs. reaction rate
relationships. T h e reaction rate of sucrose is not greatly affected
by increases in temperature at a constant pH of 9.70.

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413

F i g u r e 7 i n d i c a t e s t h e a c t u a l p e r c e n t loss of sucrose, w h i c h
w o u l d b e e x p e c t e d , i n a clarifier after 3 0 m i n u t e s j u i c e r e t e n t i o n
t i m e a t a p H o f 9.70. T h i s p H c o r r e s p o n d s t o a n a l k a l i n i t y o f
.070. It is e v i d e n t t h a t sucrose d e s t r u c t i o n in a clarifier a p p r o x i m a t e s o n l y . 0 2 2 % o f t h e sucrose i n t r o d u c e d . T h i s loss m a y b e
considered negligible.
F i g u r e 8 shows t h e g r a p h i c i n t e r p r e t a t i o n of sucrose d e s t r u c tion in the high green storage t a n k u n d e r various conditions.
D a t a w e r e a r r i v e d a t b y a p p l i c a t i o n o f t h e steady s t a t e m e t h o d
a t t h e f o u r d i f f e r e n t p H levels i n d i c a t e d . T h i s i n t e r p r e t a t i o n
indicates that if the recirculation load is high, the t e m p e r a t u r e
h i g h , s t o r a g e l o n g , a n d h i g h g r e e n p H low, significant losses o f
sucrose d u e t o d e s t r u c t i o n m a y o c c u r . I t s h o u l d b e p o i n t e d o u t
t h a t a n y loss o f sucrose i n t h e process c o n t r i b u t e s n o n s u g a r
b r e a k d o w n products which in t u r n are of a melassigenic n a t u r e .

Figure 8.High green sucrose destruction vs. time at a constant


temperature of 100C. All pH measurements were made at 2 0 C .

Summary
A new kinetic m e t h o d has been presented which is d e p e n d e n t
o n t h e s t e a d y state k i n e t i c c o n c e p t . T h e e q u a t i o n s d e v e l o p e d i n
this p a p e r e n a b l e a n i n v e s t i g a t o r t o o b t a i n t h e f o l l o w i n g information.

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JOURNAL OF THE A. S. S. B. T.

1. T h e steady state invert concentration of any hot alkaline


process liquor or juice.
2. T h e concentration of glucose and fructose contained within
the steady state invert level.
3. T h e first order reaction rate constants for sucrose, glucose
and fructose.
4. T h e percent sucrose destroyed per unit time in any hot
alkaline process juice or liquor.
Acknowledgment
T h e author wishes to express his grateful appreciation to Dr.
J. Calvin Giddings of the University of Utah for a helpful discussion of steady state kinetics.
Literature Cited
(1) CARRUTHERS, A. 1957. T h e destruction of sucrose and of invert sugar in
beet juice processing. Presented to Commission Internationale Technique De Sucrerie. London, England.
(2) DANIELS, F. and R. A. ALBERTY. 1958. Physical Chemistry. John Wiley
8c Sons, Inc. New York. 352 p.
(3) GIDDINGS, J. C. and H. K. SHIN., 1961. Departure from the steady state
in complex reactions with a reactive intermediate. Transactions of
the Faraday Society. 57 (459) : 468-482.
(4) CLASSTONE, S. 1956. Textbook of Physical Chemistry. D. Van Nostrand
Company, Inc. Princeton, New Jersey. 1049 p.
(5) PIECK, R., J. HENRY. 1963. Some additional data on the color formation in sugar solutions by heating between 95 and 135 C and on
the kinetics of the sugar degradation. Presented to Commission
Internationale Technique de Sucrerie Paris, France.

Waste W a t e r Treatment Studies at Tracy, California


LLOYD

W.

NORMAN,

JAMES

E.

LAUGHLIN

AND

L.

O.

MILLS

Received for publication April 30, 1964

Introduction
I n v i e w o f i n c r e a s i n g i n t e r e s t i n a n d r e q u i r e m e n t s for w a s t e
t r e a t m e n t a t b e e t s u g a r m a n u f a c t u r i n g facilities, H o l l y S u g a r
Corporation, in cooperation with Forrest a n d Cotton, Inc., Consulting Engineers, conducted treatment experiments at the Tracy,
C a l i f o r n i a , factory. T h e o b j e c t i v e s w e r e :
1. To characterize the waste at this p l a n t as to organic strength
and biodegradability.
2 . T o d e t e r m i n e t h e effectiveness o f a c t i v a t e d s l u d g e t r e a t m e n t a n d t r i c k l i n g filter t r e a t m e n t a s r o u g h i n g u n i t s a h e a d
o f n o r m a l l a g o o n systems.
3 . T o i n v e s t i g a t e o p e r a t i n g c o n d i t i o n s affecting d e s i g n c o n siderations.
4 . T o establish a n a l y t i c a l t e c h n i q u e s s u i t a b l e for c o n t r o l o f
t r e a t m e n t systems.
5 . T o d e t e r m i n e B O D levels w h i c h m i g h t b e e x p e c t e d i n
t r e a t e d effluent.
Equipment

and Procedures

T h e work of the present investigations becomes naturally


d w i d e d into two phases according to t h e e q u i p m e n t used. On
the o n e h a n d , t h e p i l o t p l a n t t r i c k l i n g filter was e m p l o y e d , w h i l e
o n t h e o t h e r t h e l a b o r a t o r y b e n c h - s c a l e a c t i v a t e d slud<?e u n i t
was u s e d . T h e feed m a t e r i a l t o e a c h w a s t h e s a m e , b e i n g T r a c y
plant waste water.
T r a c y w a s t e w a t e r consists essentially o f b e e t f l u m i n g w a t e r
a n d m a y b e , t h e r e f o r e , s o m e w h a t l o w e r i n B O D t h a n t h e effluent
from p l a n t s w h i c h s e w e r o t h e r w a s t e s t r e a m s . A t T r a c y , p u l p
press w a t e r i s r e c i r c u l a t e d t o t h e Silver s l o p e diffuser, a n d Steffen
waste i s r e c o v e r e d a n d c o n c e n t r a t e d .
T h e e q u i p m e n t u s e d i n t r i c k l i n g f i l t e r s t u d i e s was leased
from t h e D o w C h e m i c a l C o m p a n y a n d d e s i g n e d for u s e w i t h
their " S u r f p a c " c o n t a c t m e d i a . T h i s m e d i a i s a h e a v y s h e e t
plastic m a t e r i a l c o r r u g a t e d i n t w o d i r e c t i o n s t o p r o v i d e s t r e n g t h ,
contact a r e a , a n d efficient c o n t a c t p a t h s for l i q u i d a n d a i r pass a g e I t i s p l a c e d i n t h e u n i t i n a specified m a n n e r t o p r o v i d e
- 8 s q u a r e feet o f c o n t a c t a r e a p e r c u b i c foot o f p a c k i n g c o m p a r e d

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JOURNAL OF THE A. S. S. B. T.

with 20 sq ft/cu ft for stone packing, and 23 sq ft/cu ft for


Raschig rings of equal liquid t h r o u g h p u t capacity.
Figure 1 shows pertinent points of design and operation of
the pilot plant trickling filter. T h e u n i t is a vertical cylinder
(rectangular cross sections can also be used) with a catch basin
at the bottom, ports through which air can enter and circulate

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upward through the packing by natural convection, the packing


s e c t i o n itself, a n d feed c o n t r o l a n d d i s t r i b u t i o n e l e m e n t s a t t h e
top. T h e installed u n i t is shown in Figure 2.
X h e p i l o t u n i t was 3 ft in d i a m e t e r w i t h a p a c k i n g d e p t h of
20 ft. F e e d r a t e was c o n t r o l l e d by a v a r i a b l e orifice w e i r b o x .
A r o t a t i n g d i s t r i b u t o r s p r i n k l e d inflow e v e n l y o v e r t h e p a c k i n g .
X h e w a s t e s t r e a m feed w a s first s c r e e n e d a n d t h e n passed
t h r o u g h a simple settling tank to eliminate sand a n d o t h e r heavy
solids, a s s h o w n i n F i g u r e 3 . I t was t h e n p u m p e d t o t h e w e i r
b o x set a t t h e d e s i r e d flow r a t e . F r e s h feed m i x e d w i t h n u t r i e n t
s o l u t i o n was passed a s e v e n l y a s possible t h r o u g h t h e p a c k i n g
section a n d c o l l e c t e d i n t h e c a t c h b a s i n . T r e a t e d effluent overflowed this basin to discharge.

F i g u r e 3.Screen a n d clarifier s e t u p , X r a c y , C a l i f o r n i a .

X h e a c t i v a t e d s l u d g e u n i t u s e d i n t h e s e s t u d i e s was a laboratory bench model developed at Rice University. It is shown


schematically in Figure 4, a n d the u n i t used in Xracy is shown
in F i g u r e 5.
C o m p o s i t e d r a w w a s t e w a s r e f r i g e r a t e d i n t h e feed vessel.
F r o m t h e r e i t was m e t e r e d t h r o u g h a p e r i s t a l t i c p u m p i n t o t h e
glass r e a c t o r . M e a s u r e d a i r f l o w e n t e r e d t h e r e a c t o r t h r o u g h a
c i r c u l a r diffusion s e c t i o n i n a glass p l a t e . X h e a i r - l i q u i d m i x t u r e
percolated u p w a r d s along the o u t e r sidewall a n d t h e n the l i q u i d
c o m p l e t e d a cycle b y m o v i n g b a c k d o w n a n i n n e r c o n e t o t h e
diffuser a r e a . E f f l u e n t clarified as it m o v e d up a 2-inch p l a s t i c
t u b e a n d was t h e n e v a c u a t e d i n t o a v a c u u m j a r system. X h e
effluent e v a c u a t i o n t u b e was set t o k e e p t h e l i q u i d level c o n s t a n t
and to m a i n t a i n six liters of fluid in t h e vessel. A w a s t e s h u n t
with a t i m e d s o l e n o i d valve p r e v e n t e d excessive b u i l d - u p of
kludge i n t h e r e a c t o r . (If t h e w a s t e s h u n t was set t o m a t c h
sludge g r o w t h r a t e , t h e b a c t e r i a l c u l t u r e w o u l d r e m a i n i n
equilibrium.)

JOURNAL OF THE A. S. S. B. T.
CLARIFIER TUBE

Figure 4.Bench-scale reactor.

Figure 5.Reactor in operation, Tracy, California.

T h e reactor maintained a totally mixed fluid culture, exactly


the aim of many modern activated sludge plants, and allowed
analysis of the kinetics involved in substrate removal.
Results and Discussion
As will be shown later, the characteristics of Tracy waste
water included a relatively constant B O D / C O D ratio which
allowed dependence on C O D for analysis. T h u s , although we
are mainly reporting results here in terms of B O D , most data
were gathered as C O D values.
Because the effectiveness of trickling filtration requires micro
biological growth on the surface of the packing, a preliminai)

VOL. 13, No. 5, APRIL 1965

419

" c o n d i t i o n i n g " o t h e u n i t is r e q u i r e d . T h i s o f t e n takes a w e e k


o r t w o . I n o u r case, o n l y a b o u t 2 4 h r s w e r e r e q u i r e d t o establish
an active growth.
S i n c e m i c r o o r g a n i s m s a r e sensitive t o p H c h a n g e s , t h e r e i s
d a n g e r of l o s i n g t h e m if p r o c e s s u p s e t s (e.g., b o i l i n g o u t e v a p o r a t o r s or l i m e spills) give a s u d d e n s l u g of e i t h e r a c i d or a l k a l i n e
m a t e r i a l . S o m e fairly r a p i d c h a n g e s of as m u c h as 3 to 4 pH
u n i t s w e r e e n c o u n t e r e d , b u t t h e s e lasted 3 0 m i n u t e s o r less, n o t
l o n g e n o u g h t o c a u s e s e r i o u s loss o f m i c r o b i o l o g i c a l activity i n
the pilot unit. T e m p o r a r y reductions in organic removal seemed
to occur, however.
Characterization of T r a c y waste indicated the principal comp o n e n t c o n t r i b u t i n g t o B O D s t r e n g t h was sucrose, a s e x p e c t e d .
Analysis for a v a i l a b l e n i t r o g e n s h o w e d i t was p r e s e n t i n o n l y
trace a m o u n t s . P h o s p h o r u s was p r a c t i c a l l y a b s e n t . A t t h e s e
levels t h e w a s t e clearly l a c k e d sufficient n u t r i e n t s to s u s t a i n bacterial synthesis. T h e r e f o r e , b o t h w e r e a d d e d t o t h e w a s t e s t r e a m
i n t h e f o r m o f c o m m e r c i a l fertilizer dissolved i n w a t e r . A m m o n i u m nitrate and tri-sodium phosphate were the compounds
used.
T h e soluble B O D load i n T r a c y waste varied q u i t e widely,
as w o u l d be expected from n o r m a l variation in sugar content.
T h e r a n g e o f c o n c e n t r a t i o n s w a s f r o m 200 t o 6 0 0 m g / l i t e r s w i t h
a m e a n v a l u e of 4 0 0 m g / l i t e r (See F i g u r e 6.)
E f f l u e n t v a l u e s f r o m t h e t r i c k l i n g filter also v a r i e d r a t h e r
widely, d e p e n d i n g u p o n t h e s t r e n g t h o f t h e i n f l u e n t a s well a s
the h y d r a u l i c feed r a t e . T h e m o s t r a t i o n a l analysis o f p e r f o r m a n c e
700
600

_ 5
400

J
Q
O
CO

300
200

'

% EQUAL. OR L E S S T H A N

Figure 6.Probability plot of BOO in settled raw waste.

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20

30
OXYGEN

40

50
DEMAND

60

70

80

REMOVED <%)

Figure 7.Roughing filter performance.

involved the product of the two (absolute organic loading) per


unit volume of media per u n i t time. T h e loadings used versus
percent removal are shown in Figure 7.
From these data it will be noted that effective B O D removal
was realized u n d e r all feed rates. Best results were obtained, as
far as percent removal is concerned, at low loading rates, but
best total removal in pounds per day was realized u n d e r intensive loading. It should also be noted that the effluent BOD
levels are quite satisfactory to allow a lagoon system to handle
any further reduction required.
W i t h the activated sludge u n i t a different loading parameter
is used, that of lbs of B O D influent per lb of mixed liquor
volatile suspended solids per day. T h e mixed liquor solids are,
of course, a measure of the microbiologically active material
present.
In Figure 8 are presented the data relating u n i t removal rate
to u n i t loading rate; about 8 5 % removal occurred over extreme
ranges in loading. Figure 9 shows effluent organic concentration
seemed independent of loading.
Because the influent B O D is relatively fixed, loadings of
greater magnitude than indicated here were impossible because
the hydraulic t h r o u g h p u t became so great as to carry suspended
solids (active material) out of the unit.
It appears at first glance that the activated sludge process is
fully capable of accepting extremely high loadings while maintaining some 8 5 % removal of the organic load. However, some
very dramatic practical problems arose u n d e r extreme conditions.
Overfeeding unbalanced the biota present u n t i l only one bac-

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1965

421

terial g r o u p , t h e Sphaerotilus, c o u l d b e f o u n d .
B O D removal
r e m a i n e d " n o r m a l " after these slimy f i l a m e n t o u s forms h a d t a k e n
over. B u t t h e s l u d g e w o u l d scarcely settle, a n d t h e m i x e d l i q u o r
gave t h e g e n e r a l i m p r e s s i o n o f b e i n g p o p u l a t e d b y g r e a t masses
of m i l k y seaweed.
I t was n e v e r d e t e r m i n e d i f t h e m i s s i n g species h a d b e e n
w a s h e d o u t o f t h e u n i t , h a d lost o u t i n c o m p e t i t i o n w i t h t h e
Sphaerotilus,, o r b o t h . X h e r e was n o d o u b t , h o w e v e r , t h a t b u l k y
s l u d g e was a likely p r o b l e m u n d e r c o n d i t i o n s of i n t e n s e l o a d i n g .

F i g u r e 8 . L o a d i n g vs. r e m o v a l p e r f o r m a n c e i n reactor.

Figure 9.Loading vs. effluent quality in reactor.

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JOURNAL OF THE A. S. S. B. T.

It is concluded that a healthy system required a loading of


no more than 0.5 p o u n d B O D / p o u n d volatile solids/day. Greater
loading rates may be possible, b u t the campaign ended before
this could be substantiated. It is also concluded that the activated sludge process will not serve as a roughing unit in the
normal sense. T h e system removed 8 5 % of all loads imposed,
far more than the 3 3 % or 5 0 % felt to be sufficient. Roughing
results could be obtained another way, however. A diversion of
3 3 % , or 5 0 % of plant waste (the remainder going directly to
lagoons) could receive activated sludge treatment, then discharge
to the lagoons. W i t h skilled operation to maintain proper conditions, such a system should be satisfactory.
Direct comparison of the two types of treatment systems is
difficult, if not impossible, because of their different natures.
Only an economic analysis based u p o n particular conditions
can be presented. T h i s has been done for pretreating a waste
having 660 m g / l i t e r B O D influent so that at least one-third of
the sugar is removed. Cost calculations give the following results:
Equipment Cost per Million Gal./Day
Trickling Filter
$45,000
Activated Sludge
$30,000
These figures are, of course, applicable only to the specific con
ditions stated. Economic analysis of each situation will be required. Also it is pertinent to point out that operation, control,
manpower requirements, and power requirements will generally
be more favorable with the trickling filter system. In this case,
the annual operating cost for trickling filtration would be about
$1500, where activated sludge would cost $4500.
Analytical Procedures
T h e classic criterion for the pollution potential of organic
wastes is the 5-day B O D (Biochemical Oxygen Demand). Yet
anyone who has undertaken this determination must surely have
longed for a method that is easier to run, more reliable, and
produces quicker results. Such a method is the C O D (Chemical
Oxygen Demand) .Because in most instances no constant relationship between the two can be established, C O D is generally
not acceptable.
In this study a B O D / C O D ratio was established that was
satisfactorily constant. Samples on which both BOD and COD
were determined provide the data from which Figure 10 was
plotted.
T h e results show the mean value of B O D / C O D is 0.67 and
scatter is no greater than the reliability of the B O D determina-

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423

1965

15

2b & 4'6 *'6 s'6 Yo <&


% EQUAL. OR L E S S T H A N

9b

9^

Figure 10.Probability plot of B O D / C O D ratio for settled raw waste.

tions. A s a c o n s e q u e n c e o f t h i s , m u c h o f t h e w o r k r e p o r t e d w a s
based u p o n C O D v a l u e s w h i c h a r e easily a n d r e l i a b l y c o n v e r t e d
to equivalent B O D .
P e r h a p s o n e r e a s o n t h i s c o n v e r s i o n was satisfactory i n t h e s e
tests i s t h e fact t h a t t h e waste w a t e r a n a l y z e d d i d n o t i n c l u d e
e i t h e r p u l p p r e s s w a t e r o r Steffen w a s t e . I t m a y w e l l b e t h a t
varying p r o p o r t i o n s of either, or both, of these in the total waste
would disallow the c o n v e n i e n t use of C O D . Nevertheless, it
would seem a p p r o p r i a t e to check o u t the relationship on any
p a r t i c u l a r s u g a r waste u n d e r s t u d y .
F u r t h e r t i m e s a v i n g w i t h t h e C O D p r o c e d u r e was r e a l i z e d
since i t was f o u n d t h a t r e f l u x d i g e s t i o n t i m e c o u l d safely b e c u t
from t w o h o u r s t o o n e h o u r . S e v e r a l t i m e s t u d i e s o n t h i s aspect
i n d i c a t e d o x i d a t i o n b y t h e d i c h r o m a t e was essentially c o m p l e t e
in 30 minutes, and complete in 45 minutes. O n e h o u r allowed
a sufficient m a r g i n of safety.
P r o b a b l y t h e s a m e factors, n a m e l y t h e p r e p o n d e r a n c e o f
simple sugars i n t h e waste, a l l o w e d t h i s t i m e s a v i n g a s p r o v i d e d
for c o n s t a n t B O D / C O D r a t i o . A g a i n , a n y w a s t e u n d e r c o n s i d e r ation m i g h t b e c h e c k e d for v a l i d i t y o f s u c h a d j u s t m e n t i n p r o cedure.
Summary
S t u d i e s o f waste w a t e r effluents f r o m t h e T r a c y p l a n t , free
o f e i t h e r Steffen w a s t e o r p u l p w a t e r s , w e r e e v a l u a t e d a n a l y t i c a l l y
and s h o w n t o r a n g e i n s e t t l e d r a w w a s t e f r o m 200 t o 6 0 0 m g / l i t e r
ROD loading with a m e a n value of 400 m g / l i t e r B O D .
T r e a t m e n t of this water in a laboratory activated sludge u n i t
and a p i l o t p l a n t t r i c k l i n g filter u n i t i n d i c a t e effective o x i d a t i o n
can b e a c c o m p l i s h e d . W i t h t h e a c t i v a t e d s l u d g e u n i t effluent
C O D c o n c e n t r a t i o n s fell b e t w e e n 5 5 a n d 145 m g / l i t e r w i t h a

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JOURNAL OF THE A. S. S. B. T.

mean value of 100 mg/liter. Overall mean elimination was 8 5 % .


In the case of the trickling filter u n i t B O D reductions of 3 3 %
to 5 0 % were attainable on once-through passes involving organic
loadings of 750 to 350 pounds C O D / 1 0 0 0 cu ft/day.
T h e tests indicated that satisfactory roughing treatment could
be obtained by either treatment on a once through basis with
the trickling filter or by-passing 1 / 3 to 1 / 2 of the waste to the
activated sludge unit. Large changes in pH should be avoided
as well as overloading of the activated sludge system.
A thorough comparison of B O D and C O D values of the
material encountered in the present work gave ratios consistent
enough to be useful for inter-conversion. Eighty percent of the
values for B O D / C O D ratio fell between 0.61 and 0.72 with 0.67
the mean value. From these results it is concluded that the
wastes from most sugar factories might be of consistent enough
composition to allow utilization of the C O D analytical method
with reliable conversion to B O D being possible by establishment
of a factor for the individual plant, such as was established here.
Of special interest was the fact that, in order to obtain optim u m bio-oxidation of the Tracy waste, n u t r i e n t feeding was required. Both nitrogen and phosphorus were added in the form
of commercial fertilizer.

A New Approach to the Saccharate Cake Purity


Determination 1
KARL

SCHOENROCK

Received for publication April 15,

1964

Introduction
A b e e t s u g a r m i l l o p e r a t i n g w i t h a molasses d e s u g a r i n g
process via Steffen s h o u l d k n o w t h e a m o u n t o f n o n s u g a r s i n t r o d u c e d t h r o u g h t h i s o p e r a t i o n . I t h a s b e e n s u s p e c t e d for s o m e
time that the saccharate cake purity values derived t h r o u g h the
classical c a r b o n d i o x i d e gassing m e t h o d a r e n o t r e a l l y i n d i c a t i v e
o f t h e a c t u a l n o n s u g a r l o a d recycled i n t o t h e c a r b o n a t i o n process
of the mill. Xhusly these saccharate p u r i t y values c a n n o t be
used to establish a u s a b l e r e n d e m e n t v a l u e for t h e s a c c h a r a t e
cake.
It has been the general opinion in the industry that the
a c t u a l n o n s u g a r l o a d in s a c c h a r a t e c a k e is of l i t t l e v a l u e since
t h e c a r b o n a t i o n process m a y e l i m i n a t e a d d i t i o n a l n o n s u g a r s
w h i c h s h o u l d b e c r e d i t e d t o t h e Steffen process. T h e classical
c a r b o n d i o x i d e gassing m e t h o d i s a s s u m e d t o d u p l i c a t e c o n d i tions a s p r e v a i l i n g i n factory o p e r a t i o n . N o n s u g a r a d s o r p t i o n
on freshly p r e c i p i t a t e d c a l c i u m c a r b o n a t e is a r e c o g n i z e d fact
a n d m a y b e sizable.
However, conditions d u r i n g carbonation in mill operation
can n o t b e d i r e c t l y c o m p a r e d w i t h c o n d i t i o n s o c c u r r i n g d u r i n g
the gassing w i t h c a r b o n d i o x i d e o f a s a c c h a r a t e c a k e s a m p l e . I n
the l a t t e r case w h e r e t h e c a l c i u m o x i d e : n o n s u g a r r a t i o m a y b e
as h i g h as 10 : 1 c o n d i t i o n s a r e i d e a l for n o n s u g a r a d s o r p t i o n
on calcium carbonate. D u r i n g mill operation, however, the
calcium o x i d e : n o n s u g a r r a t i o i s o n l y a b o u t 1 : 1 .
L a b o r a t o r y tests w e r e c o n d u c t e d t o e x p l o r e t h e r e m o v a l o f
saccharate c a k e n o n s u g a r s d u r i n g c a r b o n a t i o n i n t h e p r e s e n c e
of a l a r g e excess of n o n s u g a r s f r o m r a w j u i c e . A m a t e r i a l b a l a n c e
via single a c i d t r u e p u r i t y d i d n o t s h o w a n y i n c r e a s e d n o n s u g a r
removal for t h e Steffen h o u s e o v e r t h e s t r a i g h t h o u s e u n d e r
equal c o n d i t i o n s o f a c t i v e C a O o n t o t a l n o n s u g a r s i n r a w j u i c e .
The Steffen h o u s e h a d c o n s i s t e n t l y a l o w e r effective a l k a l i n i t y ,
a h i g h e r l i m e salt level a n d m o r e c o l o r i n t h i n j u i c e t h a n t h e
c o m p a r a t i v e s t r a i g h t h o u s e r u n . S a c c h a r a t e c a k e p u r i t i e s calculated f r o m t h e m a t e r i a l b a l a n c e w e r e s u b s t a n t i a l l y l o w e r t h a n
those d e r i v e d f r o m t h e c a r b o n d i o x i d e gassing m e t h o d . F u r t h e r more, i t i s e v i d e n t t h a t s a c c h a r a t e c a k e a s p r o d u c e d i n t h e
1

Research Chemist. T h e Amalgamated Sugar Company, Twin Falls, Idaho.

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JOURNAL OF THE A. S. S. B. T.

Steffen house may be dissolved in raw juice. T h e saccharate


cake nonsugars are thusly released and join a common pool of
nonsugars with those nonsugars coming in with the raw juice.
According to observations d u r i n g this study the elimination
of saccharate cake nonsugars proceeds predominantly through
adsorption on freshly precipitated calcium carbonate. It may
therefore be deducted that saccharate cake purity values by the
carbon dioxide gassing method indicate a substantially higher
purity than can be obtained in mill operation. O t h e r factors
such as pH end point, velocity of gassing, length of time, temperature, etc., have a pronounced influence u p o n the final purity
value. Reproducibility is consequently erratic and the values
are not reliable when based u p o n the carbon dioxide gassing
method. T h e purity values by this method, consequently may
be described as a psychological tool to keep the operators alert.
Methods and Materials
In a search for a better method, saccharate decomposition
with phosphoric acid, oxalic acid, acetic acid, zinc nitrate, amm o n i u m nitrate and a m m o n i u m carbonate has been tried. A
new method based upon the saccharate cake decomposition with
a m m o n i u m carbonate evolved out of this work. If R represents
sugars such as sucrose a n d R' is representative of molasses,
nonsugars carried by saccharate cake the reactions u n d e r consideration may be illustrated as shown below. Ca may stand
here for either single or multiple calcium moles.
Steffen
reaction:
(I) R + R' K+ (Na + ) + 2 C a O = R C a + R ' C a + K+ (Na+)
T h e calcium salts of the nonsugars found in saccharate cake are
normally soluble since their disappearance from the solution
does not occur until insoluble calcium saccharate has been
formed. It is assumed at this stage that these calcium salts are
fixed insolubly on precipitated calcium saccharate where the
saccharate functions as substratum.
Gassing of saccharate cake with carbon dioxide:
(II) R C a + R ' C a + C 0 2 = R + R ' C a + C a C 0 3
Because of the weak ionization of carbonic acid it cannot displace from their calcium salts anions which have a relatively
higher ionization. Precipitation of calcium carbonate does consequently not occur through gassing of these calcium salts with
carbon dioxide. Elimination of these calcium salts may, however, occur through adsorption on freshly precipitated calcium
carbonate formed through carbonation of calcium saccharate.
T h e extent of this elimination is mainly a function of available

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C a O before carbonation. If available C a O is i n a d e q u a t e or if


s o m e c a l c i u m salts a r e n o t easily a d s o r b e d , these n o n s u g a r s a p p e a r
t h e n a s s o l u b l e l i m e salts i n t h e filtrate a f t e r c a r b o n a t i o n .
I n t h e p r e s e n c e o f n o n s u g a r s f r o m r a w j u i c e t h e selectivity
for n o n s u g a r e l i m i n a t i o n via c a l c i u m c a r b o n a t e u n d o u b t e d l y
favors n o n s u g a r s f r o m r a w j u i c e . T h i s i s a r e a s o n a b l e a s s u m p t i o n
b e c a u s e t h e molasses n o n s u g a r s r e i n t r o d u c e d w i t h t h e s a c c h a r a t e
cake r e m a i n e d i n s o l u t i o n i n p r e f e r e n c e t o selectively a d s o r b e d
n o n s u g a r s f r o m r a w j u i c e d u r i n g t h e i r first t i m e t h r o u g h c a r b o n ation. It could be reasoned that the saccharate cake nonsugars
m i g h t b e b y p a s s e d a g a i n i n p r e f e r e n c e t o m o r e easily a d s o r b e d
nonsugars from raw juice.
Decomposition

of

saccharate

cake

-via

ammonium

carbonate:

(III) R CA +R' Ca +(NH4)2 CO3 =


R + N H 4 R ' 4- CaCO3 + N H 3
X h e r e l a t i v e l y h i g h i o n i z a t i o n o f a m m o n i u m c a r b o n a t e causes
v i r t u a l l y c o m p l e t e d i s s o c i a t i o n o f t h e c a l c i u m salts w i t h s u b sequent precipitation of calcium carbonate in a d o u b l e reaction.
C a l c i u m c a r b o n a t e has n o selectivity for t h e a m m o n i u m salts.
It is recognized that the residual a m m o n i u m ion may show a
somewhat larger nonsugar load than actually originating with
t h e s a c c h a r a t e cake. X h e a d d i t i o n a l a m m o n i u m l o w e r s t h e sacc h a r a t e c a k e p u r i t y b y less t h a n o n e p u r i t y p o i n t . H o w e v e r ,
these n o n s u g a r s , if n o t e l i m i n a t e d , force t h e a d d i t i o n of soda
ash t o r e d u c e t h e l i m e salt level i n t h i n j u i c e t o a t o l e r a t e d level.
X h e m e t h o d for t h e d e t e r m i n a t i o n o f s a c c h a r a t e c a k e p u r i t i e s
as described below is based on the reaction as illustrated in
e q u a t i o n III. X h e r e a c t i o n i s b e l i e v e d t o b e s t o i c h i o m e t r i c :
300 g o f s a c c h a r a t e c a k e w i t h a b o u t 1 9 % C a O a n d a t o t a l d r y
s u b s t a n c e o f a b o u t 4 0 % r e q u i r e a b o u t 100 g o f a m m o n i u m
carbonate. X h e use of a m m o n i u m bicarbonate is even m o r e
advantageous. Saccharate cake need not be diluted a n d can
n o r m a l l y b e u s e d a s is. A m m o n i u m c a r b o n a t e i s a d d e d i n
powdered form causing the highly thixotropic cake to be completely fluidized i n t o a n easily f i l t e r i n g m e d i u m .
Procedure
A b o u t 300 g of s a c c h a r a t e c a k e is c h a r g e d i n t o a W a r i n g
Blender. A d e q u a t e a g i t a t i o n is m a i n t a i n e d w h i l e 100 g of a m m o n i u m c a r b o n a t e i s slowly a d d e d . X h e m i x t u r e b e c o m e s r a t h e r
soupy w i t h t h e first a d d i t i o n o f a m m o n i u m c a r b o n a t e . W i t h
hirther addition of the carbonate the m i x t u r e moves t h r o u g h
several t h i x o t r o p i c stages t o a n a l m o s t solidified mass. F i n a l
a d d i t i o n o f a l l r e q u i r e d a m m o n i u m c a r b o n a t e together w i t h

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JOURNAL OF THE A. S.

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forceful agitation will break this thixotropic stage to produce


a fluid mixture. T h e reaction is exothermic and the temperature
may reach 65 G.
Complete cake decomposition is assured after the final thinning stage or as soon as all required a m m o n i u m carbonate has
been dissolved. T h i s requires about 3 minutes with a good
working blender and finely powdered a m m o n i u m carbonate.
T h e mixture is filtered over a Biichner funnel and may be
washed with water if desired (in case a material balance is to
be established).
Ammonia is expelled from the filtrate through steam distillation. A simple steam distillation may be assembled from either
one large or two small electric hot plates; one 1-liter, wide mouth
Erlenmeyer flask; one 1-liter flat bottom boiling flask; 20 inches
of 1 / 4 " i.d. glass tubing and a 1-hole stopper for the boiling
flask. T h e boiling flask is used as the steam generator. Steam
is introduced via the glass tubing into the ammonia-bearing:
filtrate previously charged to the wide-mouthed Erlenmeyer and
vented to the atmosphere. Both flasks should be vigorously
heated to reduce boiling time to a m i n i m u m . Displacement
of the ammonia is usually achieved in about 20 minutes. A pH
end point of around 7 indicates complete removal of free ammonia. T h e exact pH end point may vary somewhat depending
u p o n the type of molasses used in the Steffen house. However,
it was found that a variation in the pH end point of the distillation between 5.5 and 8.5 had an insignificant influence upon
the final purity value or sugar recovery. After cooling, the percent sugar a n d R D S may be determined in the usual manner
the ratio of which expresses the percent purity of the cake.
Purity values for saccharate cake determined by this method
should be between 4 and 10 purity points below those derived
via the C 0 2 gassing method.
Tables 1 and 2 show the material balance for the Steffen
process when employing the a m m o n i u m carbonate and the carbon
dioxide method, respectively. It is very evident that the carbon
dioxide gassing method accounts for only half of the molasses
nonsugars carried in the cake.
T a b l e I. Steffen processsugar b a l a n c e .

( A m m o n i u m carbonate purity m e t h o d . )

V O L . 13, N o . 5, A P R I L 1965
T a b l e 2. Steffen process dry substance b a l a n c e .

429

430

JOURNAL OF THE A. S. S. B. T.

determined on it by the carbon dioxide gassing method. T h e


objective of this test is to check filter operation of the Steffen
house. T h i s is based u p o n the assumption that a relatively large
spread between cake purity and perfect washed cake purity
points toward poor filter operation. However, frequently the
perfect washed cake purity is found to be lower than the original
cake purity. W i t h the a m m o n i u m carbonate cake purity method
some light has been shed on this mysterious behavior. T a b l e 4
shows some results of this study.

Table 4. Effect of time and washing on apparent purity of saccharate cake and
sugar loss in the wash.

A large n u m b e r of other tests have substantiated findings as


presented in T a b l e 4. For example, cake decomposition will
proceed with an increase in time spread between the original
cake and the so-called perfect washed cake. A factory obtaining
a credit for having the perfect washed cake purity not higher
or only little higher than the original cake purity may obtain
just that by leaving the grab sample or composite standing in
a bucket for several hours before proceeding with the perfect
washed operation.
Of particular interest is, however, the trend toward lower
cake purity and increased sugar in the wash with a spread in
the time delay prior to the additional washing. T h i s simply
means that the calcium saccharate is solubilized at a much faster
rate than the nonsugars. It could also mean that the saccharate
cake may not only lose large amounts of sugar through excessive
washing b u t could also be washed to a lower purity. Tests were
conducted to ascertain whether this p h e n o m e n a has an effect in
factory operation. T h e results were inconclusive on the factory
level. T h e extent of cake washing on the filter d r u m is usually
limited to the displacement of mother liquor in the cake. Small
purity changes u n d e r these conditions are not easily unveiled.
More extensive laboratory tests have, however, confirmed the
findings as presented in Table 4. T h e magnitude of reduced

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cake purity through extensive washing was directly traced to


t h e s t a b i l i t y o f t h e c a k e . C a k e s t a b i l i t y was f o u n d t o b e a f u n c t i o n
of lime quality a n d cooler operation such as t e m p e r a t u r e d u r i n g
the liming operation.
Summary

and

Conclusion

In conclusion it is pointed out that the a m m o n i u m carbonate


m e t h o d as presented here provides a simple a n d accurate means
of estimating the t r u e nonsugar load recycled i n t o the mill with
t h e s a c c h a r a t e cake. X h i s h a s h e r e t o f o r e n o t b e e n p o s s i b l e w i t h
t h e c a r b o n d i o x i d e gassing m e t h o d . T h e a m m o n i u m c a r b o n a t e
m e t h o d i s r e l a t i v e l y fast, r e q u i r i n g o n l y a b o u t 1 5 t o 2 0 m i n u t e s
of actual labor which may be performed by unskilled personnel
with e q u i p m e n t already available in most laboratories.

Sugar Beet Tops and Modern Sugar Beet


Production1
L I O N E L HARRIS, 2 D. C. CLANTON AND M. A. ALEXANDER 3
Received for publication July 14, 1964

T h e sugar beet top, a by-product of the susar beet industry,


is a valuable livestock feed (1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and l l ) . 4
D u r i n g the 5-year period (1942-47), Harris (12s) reported
an average yield of 8.4 tons of edible silage from the tops of
sugar beets which yielded 16.5 tons of roots per acre. T h e beets
were topped by hand. T h e feeding value of the beet top silage
was comparable to that of corn silage for fattening lambs (13).
D u r i n g recent years considerable progress has been made
in the development of sugar beet top harvesting machines. Data
have been collected at the Scotts Bluff Experiment Station on
the yields of sugar beet roots and green sugar beet tops harvested
mechanically and yields of edible silage obtained from ensiling
the green tops.
Experiments have been conducted d u r i n g the past 12 years
to determine the feasibility of using beet top silage as the only
roughage in l a m b and cattle finishing rations and the effect
method of harvesting and storing beet tops have on their feed
value.
Materials and Methods
T h e sugar beet top silage, corn silage and alfalfa hay used
in these experiments d u r i n g the period 1951 to 1963 were produced at the Scotts Bluff Experiment Station. Previous to 1957
beet top silage was made by chopping and ensiling beet tops
from the windrow, one to seven days after beets were topped
with a two-row John Deere topper. From 1957 to 1959 beet tops
were harvested for silage with an experimental beet top harvester
consisting of a two-row J o h n Deere topper m o u n t e d on a Ferguson
forage harvester. From 1960 to 1963 unwilted beet tops were
harvested for silage with a Lockwood beet top harvester (Figure
1). Beet tops were chopped and ensiled in piles or cribs, on sloping ground to permit drainage of excess liquid.
Range produced feeder lambs averaging between 65 and 76
pounds were used in the l a m b feeding experiments and Hereford yearling steers were used in the cattle feeding experiments.
i Published with the approval of the Director as Paper No. 1579 Journal Series,
Nebraska
Agricultural Experiment Station.
2
Scotts Bluff Experiment Station. Mitchell, N braska.
8
Animal Science Departmtnt, University of Nebraska, Lincoln.
* Numbers in parentheses refer to literature cited.

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433

F i g u r e 1.Harvesting s u g a r b e e t tops for silage, Scotts Bluff E x p e r i m e n t S t a t i o n . G r e e n t o p s from this field yielded 19.1 t o n s p e r a c r e , b e e t
roots, 19.9, a n d e d i b l e silage 14.2 tons.

A n i m a l s w e r e r a n d o m l y assigned t o g r o u p s . T h e g r o u p f e e d i n g
p r o c e d u r e was u s e d i n all e x p e r i m e n t s . T h e y w e r e fed t w i c e
daily. F r e s h w a t e r was a v a i l a b l e a t all t i m e s . Salt, t h e o n l y
m i n e r a l fed, was g i v e n free c h o i c e . I n d i v i d u a l w e i g h t s w e r e
taken periodically.
T h i s e x p e r i m e n t a l m a c h i n e was d e v e l o p e d a n d m a d e available t o t h e s t a t i o n b y M r . L l o y d S m i t h o f G e r i n g , N e b r a s k a .
Results and Discussion
Yield of roots, tops and edible silage
Yields o f s u g a r b e e t r o o t s , t o p s a n d e d i b l e silaee from v a r i o u s
f i e l d s d u r i n g t h e p e r i o d 1957 t o 1962 a r e s h o w n i n T a b l e 1 .
I n 1 9 6 1 , t h r e e m e t h o d s o f s t o r i n g u n w i l t e d b e e t t o p s silage
were u s e d . O v e r 1 4 t o n s o f e d i b l e silage p e r a c r e w e r e o b t a i n e d
from a field, w h e r e b e e t s y i e l d e d 19.9 t o n s , a n d u n w i l t e d t o p s
T a b l e 1.Yields of sugar beet roots, tops a n d e d i b ' e sila-je (1957-1962).

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JOURNAL OF THE A. S. S. B. T.

19.2 tons. T h e pile of unwilted tops was covered immediately


after harvest with black polyethelene plastic, weighted down
with chopped forage to seal out air and prevent wind damage
to the plastic. T h e edible silage in this experiment was harvested
at a cost of $2.51 per ton (14). W h e n unwilted tops from a
second field yielding 14.0 tons per acre from beets that yielded
15.5 tons, were ensiled in an uncovered stack the yield of edible
silage was only 6.49 tons per acre. Unwilted tops from a third
field were ensiled in cribs and yielded 8.94 tons of edible silage
per acre. T h i s is not a valid comparison of methods of ensiling
beet tops because of the variability in yield between fields. T h e
data are presented as a record of experiences in the making and
feeding of beet top silage. T h e plastic used to cover the pile of
unwilted forage immediately after harvest appeared to reduce
shrinkage and spoilage and has been in standard use at the Station
since 1961.
T h e mean yield of edible silage (1957 to 1962) from all
fields, in procedures used in making silage and methods of storing
silage was 9.48 tons or 1107 pounds per ton of beets.
Beet top silage in lamb feeding experiments
Beet top silage vs. corn silage
T h e beet top silage used in seven experiments conducted
between 1951 and 1960 was made by chopping and ensiling
beet tops in cribs or stacks. For the first four experiments the
tops were chopped from one to three days after the beets were
topped. T h e beet top silage used in the last three experiments
was harvested without wilting. T h e beets were topped, chopped
and ensiled in one operation.
T h e corn silage used in the first experiment was made from
frosted immature corn yielding 30 to 40 bushels of grain per
acre. T h e silage was palatable and readily consumed by the
lambs. T h e corn silage used in each of the other experiments
was made from well matured corn that yielded over 100 bushels
of grain per acre. All the corn silage was stored in trench silos.
Good quality ground alfalfa hay was used in all of the experiments except the first in which the alfalfa hay was only fair
in quality. Dehydrated alfalfa was standard product containing
1 7 % protein. In the first four experiments the concentrate mixture was equal parts by weight of corn, barley and dried beet
p u l p pellets. A mixture of equal parts corn and dried beet pulp
pellets was used in the last three experiments.
Experiment 1 (1951-52): T h e relative feeding value of beet
top silage and corn silage, each supplemented with two levels
of dehydrated alfalfa pellets, or two levels of alfalfa hay was
studied (Table 2). Forty-five lambs were used in each treatment.

5rs
Table 2.The relative feed value ot beet top silage and com silage supplemented with dehydrated alfalfa or alfalfa hay at two levels (45 lambs
per treatment (1951-52).

Concentrate mixture was 13% soybean meal and 29% each: com, barley and dried beet plup pellets.

0*

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JOURNAL OF THE A. S. S. B. T.

Lambs fed beet top silage gained faster and required less
feed per p o u n d of gain than those fed corn silage (Table 2).
Lambs fed dehydrated alfalfa gained faster and required less
feed per p o u n d of gain than those fed alfalfa hay with both
beet tops and corn silage. Little benefit was received from using
the higher levels of alfalfa hay and dehydrated alfalfa pellets.
Lambs receiving alfalfa hay and corn silage did not perform as
well as those receiving alfalfa hay and beet top silage.
Experiments 2, 3, and 4 (1953-56): T h e relative feeding value
of beet top silage and corn silage fed at three restricted levels of
intake was studied in three trials replicated over three years
(Table 3). Fifty lambs per treatment were used each year.
Table 3.The relative feed value of beet top silage and corn silage fed at three levels
(Three-year average 1953-56).

1
Concentrate mixture was 8.8 % soybean meal and 30.4% each: corn, barley and dried
beet pulp pellets.

Beet top silage was fed at restricted rates of 1.0, 1.8 and 2.6
pounds per lamb daily and corn silage on the same basis except
the greatest a m o u n t of corn silage that the lambs would consume was 2.4 pounds per head daily (Table 3). T h e lambs fed
the highest rate of beet top silage (2.6 pounds) would have
consumed more had it been offered to them. Alfalfa hay was
fed free choice and as silage consumption increased consumption
of hay decreased.
Lambs fed the high rate of beet top silage gained faster than
those fed the high rate of corn silage. T h e y also consumed less
feed per p o u n d of gain. Otherwise, the gains of lambs fed the
three rates of beet top silage and the light and m e d i u m rates
of corn silage were comparable.
Experiments 5, 6, 7 (1957-60): T h e relative feeding value of
beet top silage and corn silage fed according to appetite was
studied in three trials replicated over three years (Table 4).
Fifty lambs per treatment were used each year.

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437

Table 4.The relative feeding value of beet top silage and corn i ilage fed ad. libitum
(Three Year Average 1957-60).

Table 5.- Chemical composition of beet top silage and corn silage used in the 1957-58
experiment.

Complete beet top concentrate silage rations for finishing


lambs
W h e n beet tops were harvested and ensiled before wilting,
it was necessary to stack them above ground to permit large
amounts of liquid to drain from the pile. Chemical analysis of
the liquid showed it had nutritive value (10.0% dry matter of
which 6 4 % was nitrogen-free extract).

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JOURNAL OF THE A. S. S. B. T.

To reduce seepage loss, maintain nutrients, adapt silage for


storage in trench or upright silos and for easier handling, various
levels of concentrates were mixed with unwilted beet tops in
1960, 1961 and 1962. T h e amount of concentrates necessary to
prevent loss of liquid varied depending on the moisture content
of the sugar beet tops.
T h e mixed rations were compared with near the same a m o u n t
of concentrates consisting of the same ingredients, fed separately
with beet top silage in three experiments (1960-63) and with
corn silage in two experiments (1960-62). One complete ensiled
ration was used in 1960, three in 1961 and four in 1962 (Table
6). Each treatment was fed to 50 lambs in each experiment.
T a b l e 6.Ensiled b e e t t o p c o n c e n t r a t e m i x t u r e s .

T h e chemical composition of the silage used is shown in


T a b l e 7. T h e composition of the beet top silage was similar
each year, b u t the composition of the mixed rations varied greatly
according to rates of concentrates used. On a dry matter basis
beet top silage contained about 3 5 % ash; most of this was sand.
Experiments 1 and 2: In 1960 and 1961, lambs fed beet top
silage gained significantly faster than lambs fed corn silage with
comparable amounts of concentrates (Tables 8 and 9). Dry
matter intake indicated that beet top silage was more palatable
than corn silage. Dry matter intake less ash was determined
because of the high ash content of the beet top silage.
It was difficult to determine the a m o u n t of concentrates consumed by lambs fed the complete rations. O n e of the objectives
was to determine the difference in feeding value of concentrates
ensiled with beet tops and concentrates not ensiled, but fed
separately. In the first experiment (1960-61), there was no dif
ference in gain from the two methods of feeding concentrates.
Lambs fed the ensiled mixture consumed more dry matter, but
gained little more (Table 8).

1961-62

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JOURNAL OF THE A. S. S. B. T.

Table 8.Relative feeding value of corn silage and beet top silage fed with concentrates ensiled and not ensiled (1960-61).

1
Ration was made by mixing 150 pounds each of corn, beet pulp pellets and dehydrated
alfalfa pellets and 30 pounds soybean meal per ton of unwilted beet tops at time of ensiling.
2
An attempt was made to feed the same amount of concentrate as was fed in the ensiled
concentrate-beet top mixed ration.

In the second experiment lambs fed the concentrates not


ensiled at m e d i u m and heavy rates gained significantly faster
than those fed the ensiled mixtures (Table 9). T h e r e was a
highly significant interaction between levels of concentrate feeding and method of preparing rations. Differences in dry matter
intake may account for some of the differences in gains. However, it appeared that the light rate of feeding concentrates
ensiled with tops produced greater gains than heavier rates,
whereas the reverse was true when concentrates were fed separately.
Experiment 3: In 1962 four different rates of concentrates were
ensiled with beet tops and compared with similar rates of concentrates fed separately (Table 10). In this experiment gains
of lambs increased with increasing amounts of concentrates in
the ration, except for the heavy ensiled concentrate-beet top
ration, which produced a lower gain than the m e d i u m rate of
concentrates ensiled with tops. T h e m e d i u m light and medium
rates of concentrates ensiled with beet tops produced about the
same gains as similar rates of concentrates fed separately. Superior
gains were obtained with concentrates fed separately at the light
and heavy rates. Daily consumption of the complete ensiled
rations declined with increased amounts of concentrates in the
ration. T h e light mixture did not contain enough dry matter
to finish the lambs satisfactorily. T h e best results were obtained
with the medium mixture.

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Xable 9.Relative feeding value of corn silage and beet top silage fed -with varying
amounts of concentrates ensiled and not ensiled.

1
Light mix was 100 pounds each of corn, beet p u l p pellets and dehydrated alfalfa per
ton of unwilted beet tops. Medium mix was 150 pounds each of corn, beet p u l p pellets
and dehydrated alfalfa per ton of unwilted beet tops. Heavy mix was 200 pounds each of corn,
beet p u l p pellets and dehydrated alfalfa per ton of unwilted beet tops.
2
An attempt was made to feed the same amount of concentrate as was fed in the
ensiled concentrate-beet top mixed rations.

X h e larger the a m o u n t of concentrate used in the m i x e d


silage t h e g r e a t e r t h e a m o u n t o f loss d u e t o s p o i l a g e . I n t h e
heavy m i x t u r e i t was e s t i m a t e d o n e t h i r d o f t h e silage w a s lost
d u e t o s p o i l a g e e v e n t h o u g h t h e silage was s t o r e d i n c r i b s
covered w i t h plastic. X h e l a m b s w e r e fed t h e m i x t u r e s u n t i l t h e
good silage was g o n e . T h i s a c c o u n t s for t h e v a r i a t i o n i n t i m e
the l a m b s w e r e o n feed i n t h e t h i r d e x p e r i m e n t . Silo s t r u c t u r e s
are necessary t o p r e v e n t t h e s p o i l a g e i n t h e m i x e d silage.
O f t h e 4 0 0 l a m b s fed t h e e n s i l e d b e e t t o p - c o n c e n t r a t e m i x tures d u r i n g t h e t h r e e - y e a r p e r i o d o n l y o n e d i e d . D i g e s t i v e
disturbances or u r i n a r y calculi were n o t e n c o u n t e r e d .
Beet

top

silage

in

cattle

finishing

rations

T w o e x p e r i m e n t s d e s i g n e d t o c o m p a r e b e e t t o p silage w i t h
corn silage a n d w i t h c o m p l e t e b e e t t o p - c o n c e n t r a t e silage m i x tures w e r e c o n d u c t e d i n 1960-61 a n d 1962-63.
I n t h e first e x p e r i m e n t b e e t t o p silage was c o m p a r e d w i t h
corn silage a s t h e m a j o r r o u g h a g e i n a c a t t l e f i n i s h i n g r a t i o n

Table 10.Relative feeding value of beet top silage fed with varying amounts of concentrate ensiled and not ensiled (1962-63).
Concentrates
not ensiled2

1
Light mix was 50 pounds each corn, beet pulp pellets and dehydrated alfalfa per ton of unwilted beet tops. Medium-light mix was 109 pounds
each of corn, beet pulp pellets and dehydrated alfalfa per ton of unwilted tops. Medium mix was 175 pounds each of corn, beet pulp pellets and
dehydrated alfalfa per ton of unwilted beet tops. Heavy mix was 258 pounds each of corn, beet pulp pellets and dehydrated alfalfa per ton of
unwilted beet tops.
2

An attempt was made to feed the same amount of concentrate as was fed in the ensiled concentrate-beet top mixed rations.

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Each
head
feed
than

treatment was replicated over five lots of seven or eight


each. T h e steers fed corn silage gained faster, using less
per pound of gain and yielded a more desirable carcass
steers fed beet top silage (Table 11).

443

Xable 11.Relative feeding value of beet top silage and corn


cattle (Average of five replications 1960-61).

silage for finishing


Corn
silage

718
2.78
17.1
28.5
615
1025
62-8
17.3
1
Started on 50% ground shelled corn and 50<%> dried beet p u l p pellets and changed to
65% corn and 3 5 % beet p u l p d u r i n g the latter half of the experiment. Each steer received
2 pounds of dehydrated alfalfa pellets and 0.5 pounds soybean meal daily.
2
Hot carcass weight divided by slaughter weight x 100.
8
18, 17, 16 = high, average and low choice, respectively.

In the second experiment two levels of concentrates and


green beet tops were mixed and ensiled (Table 12). T h e mixed
rations were compared with near the same amount of concentrates fed separately with beet top silage and with corn
silage. Chemical composition of the silages used is shown in
Table 13. Replicate lots of eight yearling steers randomly assigned to lots were used for each treatment.
Table 12.Ensiled beet top-concentrate mixtures.

Unwilted beet tops


Corn
Dried beet pulp pellets
Dehydrated alfalfa pellets

Light
pound

Heavy
pound

2000
300
300
75

2000
400
400
100

Steers fed corn silage gained significantly faster than steers


*ed beet top silage with comparable amounts of concentrates
(Table 14). T h e r e was a trend toward greater gains when the
ratio of concentrates to beet tops was increased from 600 to
^00 pounds per ton of beet tops and when concentrate was
ensiled with beet tops instead of fed separately. However, these
differences were not highly significant. T h e difference in gain

JOURNAL OF THE A. S. S. B. 1
T a b l e 1 3 . C h e m i c a l c o m p o s i t i o n o f silages u s e d i n t h e steer f e e d i n g t r i a l s .
B e e t top-

trate rnixtur

T a b l e 1 4 . R e l a t i v e f e e d i n g v a l u e o f c o r n silage a n d b e e t t o p s i l a g e fed w i t h coi


c e n t r a t e ensiled a n d n o t ensiled ( A v e r a g e o f t w o r e p l i c a t i o n s 1962-63).

1
2

Hot carcass weight divided by slaughter weight x 100.


Carcass grade
ade score 16. 17. 18 = low, average and high choice, respectively

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445

i s p r o b a b l y r e l a t e d t o d r y m a t t e r i n t a k e . Steers fed c o r n silage


c o n s u m e d m o r e d r y m a t t e r less ash t h a n steers fed b e e t t o p
silage. T h i s was n o t b e c a u s e t h e b e e t t o p silage r a t i o n s w e r e u n p a l a t a b l e , b u t b e c a u s e t h e y w e r e s o h i g h i n m o i s t u r e a n d ash
c o n t e n t . T h e steers c o u l d n o t eat e n o u g h t o get a n e q u i v a l e n t
a m o u n t of dry matter.
F e e d c o n v e r s i o n closely p a r a l l e l e d t h e w e i g h t gains. T h e steers
fed c o r n silagre w e r e t h e m o s t efficient. H o w e v e r , on a d r y m a t t e r
less ash basis steers fed t h e e n s i l e d c o m p l e t e r a t i o n s w e r e t h e
m o s t efficient.
T h e r e was n o significant difference i n carcass q u a l i t y o r g r a d e
of steers fed t h e different silages.
Steers fed b e e t t o p silage a s t h e m a j o r r o u g h a g e u r i n a t e d
excessively a n d w e r e m o r e l a x a t i v e t h a n t h e steers fed c o r n silage.
T h e l a x a t i v e effect o f b e e t t o p silage i n l a m b s was n o t a s g r e a t
as it was in steers. H o w e v e r , l a m b s fed b e e t t o p silage as t h e
m a j o r rousrhage u r i n a t e d m o r e t h a n l a m b s fed c o r n silage. M u c h
m o r e b e d d i n g was r e q u i r e d t o m a i n t a i n lots w h e r e a n i m a l s w e r e
fed b e e t t o p silage t h a n w h e r e t h e y w e r e fed c o r n silage.
A s m a l l a m o u n t of d r y r o u g h a g e w i t h b e e t t o p silage in a
cattle finishing r a t i o n s h o u l d p r e v e n t t h e l a x a t i v e effect o f t h e
silage. A m i x t u r e o f c o r n a n d b e e t t o p silage w o u l d b e d e s i r a b l e
for t h e s a m e p u r p o s e .
Summary
E x p e r i m e n t s h a v e b e e n c o n d u c t e d d u r i n g t h e past 1 2 years
t o d e t e r m i n e t h e feasibility o f u s i n g b e e t t o p silage a s t h e o n l y
rougHage i n l a m b a n d c a t t l e f i n i s h i n g r a t i o n s a n d t h e effect
i n e t h o d o f h a r v e s t i n g a n d s t o r i n g b e e t tops h a v e o n t h e i r feed
value.
T h e a v e r a g e yield o f g r e e n s u g a r b e e t t o p s (1957-62) was
16.17 t o n s p e r a c r e . T h e a v e r a g e y i e l d o f b e e t r o o t s was 17.93
a n d s u g a r c o n t e n t of t h e b e e t s was 16.0 p e r c e n t . Y i e l d s of e d i b l e
silage (1957-62) f r o m v a r i o u s fields r a n g e d f r o m 6.49 t o n s (734
p o u n d s p e r t o n o f beets) t o 14.16 t o n s (1707 p o u n d s p e r t o n
o f beets). T h e a v e r a g e yield f r o m all a r e a s m e a s u r e d was 9.48
tons (1107 p o u n d s p e r t o n o f b e e t s ) . M u c h o f t h e v a r i a t i o n i n
yields o f e d i b l e silage was d u e t o t h e m e t h o d o f s t o r a g e a n d
a m o u n t of spoilage e n c o u n t e r e d .
L a m b s fed b e e t t o p silage g a i n e d faster a n d m o r e efficiently
than t h o s e fed c o r n silage w h e n b o t h silages w e r e s u p p l e m e n t e d
with e i t h e r d e h y d r a t e d alfalfa o r alfalfa h a y . D e h y d r a t e d alfalfa
was s u p e r i o r to alfalfa h a y as a s u p p l e m e n t to e i t h e r b e e t t o p
milage or c o r n silage.

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JOURNAL OF THE A. S. S. B. T.

Lambs fed either beet top silage or corn silage at three restricted rates gained about the same except those fed the high
rate of beet top silage gained faster than those fed the high
rate of corn silage.
Beet top silage was consumed free choice at the rate of 5.91
pounds per lamb daily, compared with corn silage consumption
at 3.23 pounds. T h e dry matter less ash intake was about the
same for both silage rations. T h e lambs fed beet top silage
gained faster than those fed corn silage.
It was possible to mix concentrates at several rates with unwilted beet tops to produce complete ensiled rations. T h e complete ensiled rations produced good gains, with low death loss
when fed free choice to lambs. T h e lambs fed the complete
mixed silage gained comparably with lambs fed the same amount
of concentrates and beet top silage fed separately.
Yearling steers fed corn silage gained faster and consumed
less feed per p o u n d of gain than steers fed beet top silage. T h e
laxative effect of beet top silage in steers is extreme. Beet top
silage used as the only roughage is not as desirable for cattle
feeding as for lamb feeding.
T h e results of these studies show the efficiency of sugar beet
farming can be improved by feeding beet top silage to livestock.
Literature Cited
(1) HOLDEN, JAMES A. 1926. Lamb feeding experiments in the sugar beet
growing district. Nebr. Agr. Expt. Sta. Bui. 216.
(2) INGRAHAM, ALDEN S. 1936. Sugar beet by-products for fattening lambs.
Wyo. Agr. Expt. Sta. Bui. 216.
(3) JOHNSON, R. F. 1941. Highlights in agricultural research in Idaho.
Idaho Agr. Expt. Sta. Bui. 244.
(4)

WILGUS, H. S., H. P. H. JOHNSON, A. L. ESPLIN and P. B. SMITH.

1948.

Dehydrated sugar beet leaves as a feedstuff. Proc. Am. Soc. Sugar


Beet Technol. pp 778-790.
(5) DUNN, L. E., and C. O. ROST. 1946. Yield and nutrient content of
sugar beet tops. Minn. Agr. Expt. Sta. Bui. 391.
(6) GUILBERT, H. R., R. F. MILLER and H. Goss. 1947. Feeding value of
sugar beet by-products. Calif. Agr. Expt. Sta. Bui. 720.
(7) HARRIS, L., H. P. DAVIS and P. SWANSON. 1943. Sugar beet tops as a
feed for dairy cattle. Nebr. Agr. Expt. Sta. Bui. 353.

V O L . 13, N o . 5, A P R I L 1965

447

(8) M A Y N A R D , Z. J. 1944. F e e d v a l u e of sugar beet by-products in terms


of g r a i n a n d alfalfa hay replaced. T h r o u g h the Leaves. 32:20-24.
(9) HEIDEBRECHT, A. A. 1943.
ferent storage m e t h o d s .
Library.

Utilization of beet tops as affected by difMasters Thesis, Colo. A & M College

(10) W A T S O N , S. J. 1939. Science a n d practice of conservation; grass a n d


forage crops. Vol. 2, Ch. 14, P u b . by Fertilizer a n d Feedstuff s
Journal, London.
(11) S M I T H , P. B. 1947. Survey of sugar beet agriculture.
Sugar Beet T e c h n o l . p p 752-763.

Proc. A m . Soc.

(12)

H A R R I S , L I O N E L . 1948. Studies of beet-top silage p r o d u c t i o n in the


irrigated r o t a t i o n e x p e r i m e n t s at t h e Scotts Bluff, Nebraska, Field
Station. A m . Soc. of Sugar Beet T e c h n o l . Proc. 5: 770-777.

(13)

H A R R I S , L I O N E L a n d M. A. ALEXANDER. 1950. Studies of beet top silage


in l a m b f a t t e n i n g rations. A m . Soc. of Sugar Beet T e c h n o l . Proc.
6: 708-715.

Flotation and Conveying Air Velocities for Processed


Monogerm Sugar Beet Fruits1
O. R. KUNZE, C. W. H A L L AND F. W. SNYDER 2
Received for publication August j, 1964

T h e handling and processing of sugar beet fruits is becoming increasingly mechanized. Little basic information is available relative to the physical and mechanical properties of the
sugar beet fruit which can be used in engineering design. With
an increase in mechanization, there is a greater demand for
basic engineering information. T h i s research concerns flotation
(terminal velocity) and conveying velocities as well as some
observations on the orientation of the sugar beet fruit in a
vertical air stream.
Materials and Methods
A 1-hp electric motor with a centrifugal forward-curved blade
fan connected to a 4-inch (outside diameter) horizontal alumin u m irrigation pipe was used as the air-flow system. A stainless
steel orifice plate and a venturi tube were installed in series in
the pipe. Flange, vena contracta and pressure taps were located
in relation to the orifice plate as recommended by Eckman 3 .
These permitted three pressure-differential readings to be taken
with each rate of flow. A separate measurement was taken with
the venturi tube.
A plenum chamber, 24 X 36 X 30 inches outside dimensions,
constructed from 34-inch plywood was installed at the end of
the pipe. Sheet metal baffles were used to diffuse the air
currents in the chamber. Steel sleeves (2i/ inches outside diameter and 6 inches long) with air-sealing flanges were used to
connect the chamber with the observation tubes which consisted
of 2 1 / 4 -inch (outside diameter) clear acrylic plastic pipe polished
on both interior and exterior surfaces. An air-flow control unit
with a seed retainer was used on the exhaust end of the tubes.
Flotation velocities were determined in the vertical tube
(50 inches long). Conveying velocities were determined in the
1
Cooperative investigation of the Michigan Agricultural Fxnerirrent Station and the
Crops Research Division, Agricultural Research Service, U. S. Departrrent of Agriculture.
Approved for publication as Journal Article #3363, Michigan Agricultural Experiment
Station.
4
Research Assistant and Professor, respectively, Department of Agricultural Engineering, Michigan State University; and Research Plant Physiologist, Crops Research Division,
Agricultural Research Service, U. S. Department of Agriculture, East Lansing, Michigan.
3
Eckman, Donald P. 1958. Industrial Instrumentation. John Wiley & Sons, Inc.
New York.

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h o r i z o n t a l t u b e (17.5 feet l o n g ) . T h e r e s p e c t i v e velocities a r e


a v e r a g e velocities o v e r t h e cross-sectional a r e a o f t h e t u b e . A l l
j o i n t s a n d c o n n e c t i o n s i n t h e air-flow system w e r e a i r t i g h t .
S u g a r b e e t fruits u s e d i n this r e s e a r c h w e r e o f t h e ( S L 126ms
X SL 128) ms X SP 5822-0 m o n o g e r m variety 4 . T h i s is a c o m m e r c i a l v a r i e t y w i t h a h i g h g e r m i n a t i o n c a p a b i l i t y . X h e fruits
w e r e h a n d processed, sized i n t o 1/64-inch d i a m e t e r classes,
e q u i l i b r a t e d (68 F a n d 4 3 . 9 % r e l a t i v e h u m i d i t y ) a n d i n d i v i d u a l l y w e i g h e d b e f o r e t h e tests w e r e r u n . Sizing was d o n e
w i t h s h e e t m e t a l sieves h a v i n g r o u n d p u n c h e d h o l e s o f t h e
sizes i n d i c a t e d . L e n g t h a n d t h i c k n e s s d i m e n s i o n s o f t h e fruits
were not determined.
X h e l i m i t s o f t h e t e r m i n a l o r f l o t a t i o n velocities for a c e r t a i n
size-group w e r e d e t e r m i n e d b y p l a c i n g t h e fruits o n a screen a t
t h e b o t t o m o f t h e v e r t i c a l t u b e . Before t h e fan was s t a r t e d , t h e
air-flow c o n t r o l was closed c o m p l e t e l y . A m i c r o m a n o m e t e r was
used t o m e a s u r e a i r p r e s s u r e differentials. X h e c o n t r o l device
was o p e n e d slowly u n t i l t h e first f r u i t was c a r r i e d i n t o t h e seed
r e t a i n e r . A c o m p l e t e set of r e a d i n g s was t a k e n . X h e air-flow
c o n t r o l was t h e n o p e n e d u n t i l t h e last f r u i t was e j e c t e d f r o m
t h e v e r t i c a l o b s e r v a t i o n t u b e a n d a n o t h e r c o m p l e t e set o f readings was t a k e n . X h e f o r e g o i n g p r o c e d u r e was r e p e a t e d five t i m e s .
X h e a v e r a g e of t h e first set of r e a d i n g s was u s e d to d e t e r m i n e
t h e f l o t a t i o n velocity for t h e l i g h t e s t fruit, w h i l e t h e a v e r a g e
of t h e s e c o n d set was u s e d to c a l c u l a t e t h e flotation velocity of
t h e h e a v i e s t f r u i t i n t h e g i v e n size-group.
C o n v e y i n g velocities w e r e d e t e r m i n e d b y i n j e c t i n g t h e fruits
of a g i v e n size-group i n t o t h e h o r i z o n t a l c o n v e y i n g t u b e . A h o l e
was d r i l l e d i n t h e plastic t u b i n g a p p r o x i m a t e l y 4 feet f r o m t h e
p l e n u m c h a m b e r . S u g a r b e e t fruits c o u l d n o t b e d r o p p e d i n t o
t h e c o n v e y i n g t u b e , b e c a u s e of a p o s i t i v e a i r p r e s s u r e i n s i d e
the t u b e . X h e r e f o r e , t h e h o l e was t h r e a d e d t o a c c e p t a c a r t r i d g e
device m a d e f r o m 3/8-inch steel p i p e . F r u i t s w e r e i n j e c t e d i n t o
t h e system by 1) c o n t r o l l e d i n j e c t i o n a l l o w i n g i n d i v i d u a l f r u i t
dispersion in the air stream, 2) sudden injection of the entire
size-group a n d 3 ) f r u i t s p l a c e d o n t h e b o t t o m o f t h e c o n v e y i n g
t u b e w h i l e system was s h u t d o w n . A w i r e p l u n g e r i n t h e cartr i d g e d e v i c e was u s e d t o c o n t r o l t h e i n j e c t i o n r a t e for t h e first
two m e t h o d s . If t h e f r u i t s d i d n o t c o n v e y at a p r e s e t air-flow
rate, t h e y w e r e r e m o v e d a n d t h e f l o w r a t e was i n c r e a s e d b y i n t e r vals of 1 foot p e r s e c o n d u n t i l t h e y w e r e c o n v e y e d .
* Kindly supplied by the West Coast Beet Seed Company, Salem, Oregon.

JOURNAL OF THE A. S. S. B. T.

450

Results
T h e n u m b e r of fruits in each group, the total weight, the
mean weight per fruit and the standard deviation of the fruits
for each size-group are recorded in T a b l e 1. T h e weights of
the individual fruits in any size-group approach a normal distribution. Hence 68.27% of the observations should be within
one standard deviation of the mean, 95.45% within two standard
deviations and 99.73% within three standard deviations. A study
of each g r o u p revealed that only the lightest fruit in the 9/64inch group and the heaviest fruit in both the 11/64- and 12/64inch groups were more than three standard deviations removed
from the mean. W h e n the fruits in each group were divided
by equal weight intervals into approximately 12 subgroups, the
weight interval containing the mean weight had the lamest
n u m b e r of fruits in every case with only one exception (Table
2). In this exceptional case, the weight interval containing the
most fruits was immediately below the one containing the mean
weight, and the mean weight was the lowest value in its weight
interval.
Table 1. Size, mean weight, and standard deviation for hand-processed sugar beet
fruits of (SL 126ms x SL 128)ms x SP 5822-0 hybrid variety.
Fruit diam.
inches
7/64
8/64
9/64
10/64
11/64
12/64 o r larger

No. of
fruits
90
96
128
127
85
96

Total weight
grams

Mean weight
grams

Standard deviation
grams

.7296
.9500
1.6039
1.8711
1.5126
2.0638

.0081
.0099
.0125
.0147
.0178
.0215

.0016
.0018
.0017
.0021
.0023
.0031

M a x i m u m and m i n i m u m fruit weights in each size-group


are plotted against fruit size in Figure 1. Observations which
may be made are 1) for the first three size-groups, the heaviest
fruit in a given group was nearly three times the weight of the
lightest fruit in that same group, 2) for the last three sizegroups, the heaviest fruit in a given g r o u p was about twice the
weight of the lightest fruit in that same g r o u p and 3) considering the fruits in all size-groups, the heaviest fruit had nearly
seven and one-half times the weight of the lightest fruit. For
the groups with the larger size fruits, the span between the
heaviest and the lightest fruits became greater. T h e plotted
points represent extreme values and the weights of all other
sugar beet fruits used in this research are represented by values
in the shaded area. T h e mean value of each size-group is plotted
as a heavy dot. These, in general, are close to the center of the

Table 2- The weight distribution of hand-processed (SL 126ms x SL 128)ms x SP 5822-0 variety sugar beet fruits after subdivision into
size-groups.

Subgroup containing mean weight

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JOURNAL OF THE A. S. S. B. T.

Figure 1.Maximum and minimum individual weights of (SL 126ms


X SL 128)ms X SP 5822-0 hybrid variety hand-processed sugar beet fruits
plotted against fruit size.

weight span for their respective size-groups. For the larger fruits,
the dots are below the mid-point of the weight span, thus indicating that there were only a few unusually heavy fruits in the
group.
T h e span of flotation velocities for each size-group is illustrated
in Figure 2. Only a slight increase in air velocities was necessary
as the size of the fruit diameters increased. T h e m a x i m u m and
m i n i m u m values plotted represent averages of the average values
calculated from the flange, vena contracta and pressure tap
readings, all used with the 2-inch orifice plate.
Velocities required to convey horizontally the fruits for the
different methods of injection are plotted in Figure 3. T h e
least velocity (23 to 24 fps) was required when the fruits were
injected at a rate which permitted each fruit to be individually
dispersed in the air stream as it fell across the diameter of the
conveying tube. Each plotted point (Figure 3) for this injection method represents the air velocity which successfully
conveyed the size-group in ten successive trials. T h e heavier or
more dense fruits tended to lodge in the tube, but the lighter
fruits had a scouring effect and helped to move the heavier fruits
along.

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/64
IO/B4
!2/fc4
Figure 2 . M a x1/64
i m u m 8FRUIT
and
m9/64
inimum
air 11/54
velocities
required for the
DIAMETER,
(INCHES)
flotation of hand-processed sugar beet fruits of (SL 126ms X SL 128)ms
X SP 5822-0 hybrid variety.

W h e n t h e s u g a r b e e t fruits of a g i v e n size w e r e i n j e c t e d as
a g r o u p , a h i g h e r velocity was r e q u i r e d t o c o n v e y t h e m t h r o u g h
t h e t u b e . X h e g r e a t e r t h e t o t a l w e i g h t o f t h e fruits i n j e c t e d ,
t h e g r e a t e r was t h e a i r velocity necessary t o sufficiently d i s p e r s e
a n d a c c e l e r a t e t h e fruits w h i l e t h e y w e r e f a l l i n g across t h e dia m e t e r o f t h e c o n v e y i n g t u b e . E a c h p l o t t e d p o i n t for t h i s m e t h o d
of i n j e c t i o n r e p r e s e n t s t e n successive trials in w h i c h t h e respective size-groups w e r e successfully c o n v e y e d .
F r u i t s p l a c e d i n t o t h e c o n v e y i n g t u b e w i t h n o a i r flowing
r e q u i r e d t h e g r e a t e s t a i r velocity t o d i s l o d g e a n d c o n v e y t h e m
t h r o u g h t h e t u b e . I n t h i s case, t h e s t a t i o n a r y fruits s u p p o r t e d
each o t h e r i n t h e s t a t i c c o n d i t i o n . X h e lowest a i r velocity i n
the c o n v e y i n g t u b e was a r o u n d t h e t u b e p e r i m e t e r , s o t h e f l o w
rate i n t h e t u b e h a d t o b e i n c r e a s e d u n t i l t h e p e r i m e t e r velocity
was sufficiently h i g h t o give t h e s u g a r b e e t fruits t h e i r i n i t i a l
acceleration. E a c h p l o t t e d p o i n t for t h i s m e t h o d r e p r e s e n t s five
successive trials in w h i c h t h e size-groups w e r e successfully conveyed.
The r e c o m m e n d e d c o n v e y i n g velocity of 38 fps is d e v e l o p e d
in t h e d i s c u s s i o n s e c t i o n of t h i s r e p o r t .

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JOURNAL OF THE A. S. S. B. T.

Figure 3.Velocities required to convey horizontally hand-processed


(SL 126ms X SL 128)ms X SP 5822-0 hybrid variety sugar beet fruits of the
size indicated.

Discussion
Air-flow
equipment
Accurate air-flow measurement presents a problem even with
modern instrumentation. Considerable time and effort was spent
in the design and construction of an orifice meter and a venturi
tube. T h e objective was to use these devices in the same air
stream and then from their pressure differential readings to
calculate air volumes which would coincide. T h e orifice was
machined from an 0.08-inch stainless steel plate. T h e venturi
tube was constructed from 26-gage galvanized sheet metal. A
piece of heavy-duty steel pipe was machined to the appropriate
slopes and throat diameter to give a precision throat section.
A 0.50 diameter ratio (orifice diameter/inside pipe diameter)
was used in the calculations. T h e air-flow system developed less
than 2 inches H 2 0 m a x i m u m static pressure. Therefore, the air

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could be considered as an incompressible fluid and the equation


as shown below was used to determine the orifice flow.

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fruits were placed into the system, they tended to strike each
other in the oscillation process and thereby disrupted the aerodynamic equilibrium position of an individual fruit.
Previous research by Kunze, Snyder and Hall 5 has indicated
the sugar beet seed to be most sensitive to damage when the
fruit is impact-loaded on its cap. From the foregoing observations, the individual fruit in a free fall would position itself
with its cap up and hence would impact on its butt, b u t it
would be impacted on its cap by succeeding falling fruits.
Terminal

velocities

T h e terminal velocity of a sugar beet fruit is the highest


velocity that such a fruit will attain while falling through still
air for an infinite distance; or conversely, the terminal velocity
is that air velocity which is necessary to suspend a sugar beet
fruit in a vertical air stream. In this research, terminal velocity
is defined as that velocity which was necessary to extend the
elliptical flotation path of the fruit just beyond the 4-foot length
of the observation tube.
If a single layer of fruits was observed in the base of the tube
as the airflow control was slowly opened, the fruits could be
seen to orient themselves in the position of cap up. T h e y became
airborne as the air flow was increased. T h e air stream beyond
the screen formed a velocity gradient. As the air moved up the
tube, the velocity in the center increased b u t that around the
tube perimeter decreased. T h i s p h e n o m e n o n caused the fruit to
move up in the center of the observation tube and to drop down
when it moved out to the tube perimeter. T h e air speed at the
center was greater than the terminal velocity for the given fruit,
while the air speed at the tube perimeter was less than the
terminal velocity. As the fruit at the tube perimeter dropped,
it encountered an increasingly higher velocity which halted its
downward motion. T h e n , if it moved into the center of the
tube once more, its elliptical flotation path was repeated.
T h e time that a fruit remained in the center stream was
variable. Generally the oscillations of a fruit were less than 2
feet in height. T h u s with a given air flow, a fruit may have
oscillated in the 2-foot section immediately above the screen. A
slight increase in air flow was necessary to make that same fruit
oscillate in the 1- to 3-foot section of the tube since the center
velocity was still increasing b u t the velocity at the perimeter
of the tube at the 1-foot level was then sufficiently high to suspend
5
Kunzo, O. R., F. W. Snyder and C. W. Hall, Impact Effects on Germination and
Seedling Vigor of Sugar Beets. J. Am. Soc. Sugar Beet Technol. 13(4): 341-353. 1965.

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t h e fruit. A n a d d i t i o n a l i n c r e a s e i n a i r f l o w c a u s e d t h e fruit t o
oscillate t o w a r d t h e t o p o f t h e t u b e w i t h t h e possibility o f b e i n g
ejected i n t o t h e s e e d - r e t a i n e r .
Conveying

velocities

Velocities r e q u i r e d to convey a g i v e n size-group of f r u i t s w e r e


s o m e w h a t d e p e n d e n t u p o n t h e m e t h o d b y w h i c h t h e fruits w e r e
i n j e c t e d i n t o t h e system. F r u i t s w h i c h w e r e i n i t i a l l y a t rest i n
t h e h o r i z o n t a l t u b e b e f o r e t h e a i r flow was s t a r t e d r e q u i r e d t h e
h i g h e s t c o n v e y i n g velocity. T h e lowest velocity w a s r e q u i r e d
for fruits w h i c h w e r e i n d i v i d u a l l y d i s p e r s e d i n t h e c o n v e y i n g
air s t r e a m .
C o n v e y i n g velocities w e r e g e n e r a l l y h i g h e r t h a n f l o t a t i o n
velocities. F o r t h i s r e s e a r c h , a c o n v e y i n g velocity was defined
as t h a t velocity w h i c h c o n v e y e d all t h e s u g a r b e e t fruits of a
given size-group t h r o u g h t h e h o r i z o n t a l t u b e . T h e lowest conveying velocities ( F i g u r e 3) w e r e a b o u t 3 fps h i g h e r t h a n t h e
h i g h e s t f l o t a t i o n velocities, o r n e a r l y 5 0 % h i g h e r t h a n t h e
average flotation velocities ( F i g u r e 2) for t h e r e s p e c t i v e sizeg r o u p s . T h e lowest c o n v e y i n g velocity was i n effect a n a v e r a g e
c o n v e y i n g velocity since f r u i t s w h i c h c o n v e y e d r e a d i l y w o u l d
strike a n d a c c e l e r a t e o t h e r fruits w h i c h w e r e t e n d i n g t o l o d g e
in t h e t u b e . A 5 0 % i n c r e a s e a b o v e t h e flotation velocity of a
sugar b e e t fruit was sufficient t o c o n v e y it. F o r e x a m p l e , fruits
which r e q u i r e d flotation velocities of 16 to 18 fps r e q u i r e d m i n i m u m c o n v e y i n g velocities of 24 to 27 fps. F r u i t s a m p l e sizes
i n this r e s e a r c h w e r e necessarily l i m i t e d a n d t h e r e b y c r e a t e d t h e
possibility t h a t e x t r e m e c o n d i t i o n s w e r e n o t e n c o u n t e r e d . M a x i m u m flotation velocities of slightly o v e r 22 fps w e r e m e a s u r e d .
Velocities of 25 fps s h o u l d be sufficient for t h e flotation of a n y
hand-processed s u g a r b e e t fruits. C o n v e y i n g velocities s h o u l d b e
based o n m a x i m u m f l o t a t i o n velocities. T h e n w i t h a 5 0 % increase i n this f l o t a t i o n velocity, t h e r e c o m m e n d e d c o n v e y i n g
velocity w o u l d be 38 fps.
I n w o r k i n g w i t h coffee fruits, E s c h e n w a l d a n d H a l l 6 considered t h e c o n v e y i n g a i r velocity t o b e 3 0 fps g r e a t e r t h a n t h e
f l o t a t i o n velocity. W h e n t h e i r r e s u l t s for g r e e n coffee fruits, r i p e
fruits, fresh p u l p a n d p u l p e d w a s h e d b e a n s w e r e s t u d i e d , t h e
conveying velocities r a n g e d f r o m 4 6 t o 5 8 p e r c e n t a b o v e t h e
f l o t a t i o n velocities. T h e r e s u l t s h e r e a r e i n g o o d a g r e e m e n t w i t h
the w o r k o n coffee b e a n s w h e n c o n v e y i n g velocities a r e considered t o b e a c e r t a i n p e r c e n t a g e g r e a t e r t h a n f l o t a t i o n velocities.
"Eschenwald, Adolfo and Carl W. Hall, 1961, Air-Flotation, Pneumatic Conveying
velocities, and Airflow Relationships for Coffee Fruits and Coffee Beans. T h e J o u r n a l of
Agriculture of the University of Puerto Rico. 4 5 ( 4 ) : 319-331,

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JOURNAL OF THE A. S. S. B. T.

T h e earlier work by Kunze, Snyder and Hall has indicated


that germination and seedling vigor of sugar beet seeds can be
reduced when fruits are impact-loaded (using a steel sphere)
with as little as 25 gram-centimeters of impact-energy. Experimental results and theoretical calculations showed that a sugar
beet fruit weighing 12.5 milligrams and traveling at a velocity
of 70 fps would have an impact energy of about 29 gram-centimeters. A single application of this impact energy reduced root
and shoot growth of the seedling by 15 to 20 percent.
W h e n an air velocity of 38 fps is used to convey sugar beet
fruits, the fruit never attains this velocity because of friction.
However, if a sugar beet fruit weighing 12.5 milligrams attained
this velocity and was impacted against a stationary object, its
impact-energy would be only 8.6 gram-centimeters. This is well
below the energy-level of impact-loads which have been found
to be detrimental to the seed. In contrast fan manufacturers
recommend air velocities in excess of 70 fps for conveying inert
materials of similar densities.
Summary and Conclusions
Sugar beet fruits of the (SL 126ms X SL 128) ms X SP
5822-0 hybrid variety were hand-processed, sized and equilibrated
at 68F and 43.9% relative humidity. T h e fruits were individually
weighed before a given size-group was subjected to either flotation
or conveying tests. Ambient air was used in the air-flow system
to determine flotation (terminal) and conveying velocities.
Fruits of the same size had a two- to three-fold weight variation. Unsized fruits 7/64 inch or larger in diameter had as much
as a seven and one-half fold variation in weight. Larger diameter
fruits generally required a slightly higher flotation velocity than
did smaller diameter fruits.
Experimental results indicated that flotation velocities varied
from a m i n i m u m of 13.2 fps to a m a x i m u m of 22.1 fps. The
span of flotation velocities for any given size-group was only
slightly less. A velocity of 25 fps should, in general, be adequate
to suspend any hand-processed sugar beet fruits in an air stream.
An individual sugar beet fruit of normal configuration tends
to float in an air stream with the cap of the fruit facing up.
Twisting and spinning of the fruit is induced by collisions with
other fruits in the air stream.
Horizontal conveying velocities were generally higher than
flotation velocities. M i n i m u m conveying velocities were depend-

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e n t u p o n t h e m e t h o d o f i n j e c t i n g t h e fruits i n t o t h e a i r s t r e a m .
T h e lowest velocity (23 t o 2 4 fps) was r e q u i r e d w h e n fruits
w e r e i n d i v i d u a l l y d i s p e r s e d i n t h e a i r s t r e a m . T h e h i g h e s t velocity (30 t o 3 5 fps) w a s r e q u i r e d w h e n t h e fruits w e r e i n i t i a l l y
a t rest i n t h e c o n v e y i n g t u b e b e f o r e t h e air-flow was s t a r t e d .
A i r velocities o f 3 8 fps s h o u l d b e a d e q u a t e t o c o n v e y fruits
h o r i z o n t a l l y i n a t u b e . N o tests w e r e r u n w i t h e l b o w s i n t h e
system.
Previous work by the a u t h o r s has indicated that g e r m i n a t i o n
a n d v i g o r o f s u g a r b e e t seeds m a y b e r e d u c e d w h e n fruits a r e
s u b j e c t e d to i m p a c t - l o a d s of 25 g r a m - c e n t i m e t e r s . If a f r u i t (12.5
m i l l i g r a m s ) a t t a i n e d a velocity of 38 fps, it w o u l d h a v e an i m p a c t e n e r g y of 8.6 g r a m - c e n t i m e t e r s . T h i s is well b e l o w t h e 25 g r a m c e n t i m e t e r e n e r g y level w h i c h i s d e t r i m e n t a l .

Injection of Preemergence Herbicides


C.

R.

KAUPKE1

Received for publication October 19, 1964

Certain preemergence herbicides have proved more effective


when incorporated into the soil at the time of application
(1,5,6). 2 Surface application followed by furrow irrigation frequently effects little or no weed control. Volatilization or photodecomposition may reduce the apparent rates of surface applied
materials before rainfall or overhead irrigation moves them into
the soil.
Many methods of mechanical incorporation have been evaluated by researchers. These methods can be grouped as: a) rotary
tillers (power driven), b) rotary hoes (ground driven), c) harrows, d) subsurface applicators, and e) miscellaneous devices
or procedures such as chain drags and listing-shaping operations.
A general criterion for an optimum method of incorporation
is acceptable weed control and crop response with m i n i m u m
overhead, operating costs, and power consumption. Because of
the many influencing factors such as herbicide, soil type and
moisture content, climate, irrigation practice, crop, and weed
species, little progress has been made in establishing the optimum
method for various field conditions.
Rotary tillers have produced the most consistent results, apparently owing to good mixing of herbicide and soil under
a variety of soil conditions. In addition the rotary tiller aids
in seedbed preparation and destroys weed seedlings if present.
However, costs and power consumption of these units are high
compared to harrows and rotary hoes. U n d e r some soil conditions excessive pulverization has occurred, leading to crusts
which impede crop emergence.
Investigations of high-pressure injection of soil fumigants (4)
and anhydrous ammonia (2) suggested this technique for incorporating herbicides into the soil. Since the necessary penetration depths for herbicides are less than those sought for
fumigants or anhydrous ammonia, lower application pressures
were anticipated. Reduced overall power requirements as compared to rotary tillers were foreseen. Furthermore, such a nontillage method would be practical as a post-seeding operation.
T h e common problem of contaminating the treated zrne with
untreated soil d u r i n g seeding would be eliminated. While rotary

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1965

F i g u r e 1.Assemblies
a subsurface b l a d e .

for d i r e c t i n j e c t i o n

(left)

and

injection over

Orifice sizes of 0.023 and 0.039 inches were used to determine


the variation of penetration depth with flow rate, pressure, and
ground speed. Orifice data and application rates are given in
Table 1.
8

Manufactured by Spraying Systems Co., Bell wood, Illinois.

464

JOURNAL OF THE A. S. S. B. T.
Table 1.Orifice data and application rates.

1
2

Average of 4 orifices.
Based on overall coverage and 1 inch effective width per orifice.

Air-dried Yolo sandy loam passing a No. 4 Tyler mesh was


compacted at i/ psi to a depth of approximately 4-7/8 inches
and loose soil added to fill a 5-inch-deep pan. T h e pan was
placed on a floor and the skid height adjusted to i/4 inch above
the soil surface. Pressures of 150, 250, and 400 psi and tractor
speeds of 1, 2, and 3 miles per h o u r (mph) were used. Depth
of penetration was determined by immediately cutting three
vertical profiles and taking the average wetted depth to the
nearest \/% inch. Effective width of treatment varied from 14
inch for the 0.023-inch orifices at 150 psi and 3 m p h to V 4 inch
for the 0.039-inch orifice at 400 psi and 1 mph. Considerable
heaving of the soil surface occurred u n d e r the latter condition,
indicating increased hydraulic pressure in the soil (2). T h e test
results are shown in Figure 2.
Qualitative tests with a fluorescent tracer were made in the
field by observation at night of profiles illuminated with ultraviolet light. Compacted areas severely reduced penetration. Large
surface clods caused spattering while small clods were displaced
or rotated, producing irregular distribution patterns.
Herbicide tests were made using 0.039-inch orifices at 400
psi and speeds of 1, 2, and 3 mph. Pre-irrigated beds in Yolo
sandy loam were spike-tooth harrowed to a depth of about 2
inches and sledded to form a level bed top. Most of the loose
soil was removed by the sledding operation. Proso millet
(Panicum miliabeum) was sown by hand in an 8-inch band to
insure the presence of a weed. T h e seed was covered by raking
to a m a x i m u m depth of about 1 inch. T o m a t o variety FW36
was seeded 1 inch deep with a sled-mounted planter followed
by the application of a commercial formulation of propyl ethyl
n-butylthiolcarbamate (PEBC) at a constant rate of 4 pounds
active per acre in a 6-inch band (6 nozzles). T h e plot was furrow
irrigated the following day.

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Figure 2-Penetration into soil of a compact water jet.

O b s e r v a t i o n t h r e e weeks after t r e a t m e n t s h o w e d significant


m i l l e t c o n t r o l a t o n l y t h e 1 m p h speed. T h e n a t u r a l w e e d p o p u l a t i o n w a s insufficient t o e v a l u a t e . D e p t h s o f p e n e t r a t i o n a t t h e
h i g h e r speeds w e r e i n a d e q u a t e t o p r o d u c e c o n s i s t e n t c o n t r o l .
T h e a p p l i c a t i o n r a t e o f 4,190 g p a a t 1 m p h (overall basis) a n d
low field c a p a c i t y (acres p e r h o u r ) w o u l d b e e c o n o m i c a l l y p r o h i b i t i v e for m o s t o p e r a t i o n s . T h e r e was n o e v i d e n c e o f h e r b i c i d e
injury to the tomato plants.
Injection
Over a
Subsurface Blade
A m a n i f o l d s i m i l a r t o t h a t d e s c r i b e d a b o v e was m o u n t e d
a b o v e t h e d e c k o f a s u b s u r f a c e b l a d e ( F i g u r e 1). T h e d e c k
provided a positive limit of penetration. T h e blade h a d an
i n c l i n a t i o n of 26 a n d a lift of l-1/2 i n c h e s . M a n i f o l d h e i g h t
was a d j u s t e d t o p r o v i d e s m o o t h f l o w o f soil b e t w e e n t h e nozzles
and deck with m i n i m u m clearance.
D i s t r i b u t i o n as d e t e r m i n e d w i t h a fluorescent t r a c e r was
u n i f o r m in t h e l o w e r 2 i n c h e s of a 2-1/2-inch profile t r e a t e d w i t h
the 0.023-inch orifice a t 150 psi a n d 3 m p h . T h e u p p e r 1 / 2 > i n c h
contained little fluorescent material. H i g h e r pressures t e n d e d
t o c o n c e n t r a t e t h e m a t e r i a l l o w e r i n t h e profile.
H e r b i c i d e tests w e r e m a d e u n d e r f i e l d c o n d i t i o n s s i m i l a r t o
those p r e v i o u s l y d e s c r i b e d . A f t e r s o w i n g a n d r a k i n g i n t h e
millet P E B C was a p p l i e d w i t h 0.023-inch orifices a t 4 p o u n d s
per a c r e w i t h p r e s s u r e s of 150 a n d 4 0 0 psi a n d speeds of 1 a n d

466

JOURNAL OF THE A. S. S. B. T.

3 mph. Blade operating depth was 2-1/2 inches. T o m a t o seeding


was done after the herbicide treatment. All tests showed excellent control of millet and natural weeds. No herbicide symptoms
were observed on the tomato plants.
Summary and Conclusions
Direct injection of herbicides appears feasible only u n d e r
very favorable conditions of soil consistence, structure, and tilth.
Compacted soil severely reduces penetration. Clods reduce penetration and cause irregular distribution patterns. Good control
of proso millet with PEBC at 4 pounds per acre was achieved
with a 0.039-inch orifice at 400 psi and a tractor speed of 1 mph.
However, the corresponding application rate of 4,190 gpa and
low field capacity cannot be justified by the advantages of postseeding incorporation and reduced power consumption.
Injection over a subsurface blade produced a uniform tracer
distribution in the lower 2 inches of a 2 1/2-inch profile with a
0.023-inch orifice at 150 psi and 3 mph. T h e same conditions
provided excellent control of proso millet with 4 pounds per
acre of PEBC. T h e corresponding application rate of 277 gpa
is 6 to 10 times higher than presently used in conjunction with
rotary tillers, but is not impractical in view of reduced costs
and power consumption.

Literature Cited
(1) ANTOGNINI, J. 1960. Soil incorporation of herbicides. Proc. California
Weed Conf. 112-114.
(2) ARYA, S. V. and G. E. PICKARD. 1958. Penetration of liquid jets in soil.
Agricultural Engineering 39(1): 16-19, 23.
(3) CARTER, L. M. and J. H. MILLER. 1961. Progress report on experimental
equipment for soil incorporation of herbicides. Proc. California Weed
Conf. 78-82.
(4) CYKLER, J. F. and R. T. TRIBBLE. 1961. Principles of injection soil
fumigation. Transactions of the ASAE 4 ( 1 ) : 199 202.
(5)

JORDAN, L. S., B. E. DAY, and W. A. CLERX.

1963.

Effect of incorpora-

tion and method of irrigation on preemergence herbicides. Weeds.


11 (2) : 157-160.
(6) MENGES, R. H. 1963. Effect of overhead and furrow irrigation on per
formance of preemergence herbicides. Weeds. 11 (2) : 72-76.

NOTES

SECTION

Uses for Sugar B e e t P u l p


P r o s p e c t s a r e g o o d for h a r v e s t i n g 25 m i l l i o n t o n s of s u g a r
beets f r o m o n e a n d one-half m i l l i o n acres i n t h e U n i t e d States
i n 1964. M o r e t h a n o n e m i l l i o n t o n s o f d r i e d b e e t p u l p will
b e a v a i l a b l e for feed o r i n d u s t r i a l use. A t p r e s e n t , p u l p a n d
m o l a s s e s - p u l p enjoy a r e a d y a n d p r o f i t a b l e m a r k e t a s a n i m a l
feed, a l t h o u g h t h i s s i t u a t i o n has n o t always p r e v a i l e d a n d i t
c o u l d c h a n g e a t a n y t i m e . Because a c o m m o d i t y s h o u l d h a v e
several uses, i t seems w o r t h w h i l e t o focus s o m e a t t e n t i o n o n
possible n e w i n d u s t r i a l uses for this v a l u a b l e b e e t b y - p r o d u c t .
F o u r carbohydrates m a k e up a b o u t 7 6 % of the dry weight
o f b e e t p u l p i n essentially t h e f o l l o w i n g p r o p o r t i o n s : g a l a c t a n ,
6 % ; a r a b a n , 2 0 % ; cellulose, 2 5 % ; a n d p e c t i n , 2 5 % . P r o t e i n ,
ash, acetyl, a n d o t h e r c o n s t i t u e n t s m a k e u p t h e r e m a i n d e r . S u g a r
beet g u m ( p r i m a r i l y g a l a c t a n a n d a r a b a n ) i s w a t e r - s o l u b l e b u t
difficult t o e x t r a c t f r o m u n t r e a t e d p u l p . B e e t cellulose f r o m
cossettes is c o m p o s e d of r e l a t i v e l y s h o r t fibers as c o m p a r e d w i t h
w o o d p u l p cellulose. P e c t i n i n b e e t p u l p i s w a t e r - i n s o l u b l e ,
but can be extracted u n d e r mild acidic conditions. T h i s pectin
has p o o r g e l a t i o n p r o p e r t i e s . B e e t c e l l u l o s e a n d g u m a r e relatively i n e r t c h e m i c a l l y b u t p e c t i n i s n o t . B e e t p e c t i n c o n t a i n s
acetyl esters, free a n d p a r t i a l l y n e u t r a l i z e d c a r b o x y l g r o u p s , a n d
a general structure that can be degraded u n d e r certain conditions.
A d d i t i o n o f l i m e t o p u l p r e s u l t s i n several m a j o r c h a n g e s
w h i c h vary i n c h a r a c t e r w i t h t h e t e m p e r a t u r e o f r e a c t i o n . W h e n
l i m e i s a d d e d t o p u l p a t a b o u t 7 0 C , esters a r e h y d r o l y z e d a n d
the p e c t i n m o l e c u l e i s d e g r a d e d . C a l c i u m i o n s r e a c t w i t h t h e
carboxyl groups of the degraded pectin molecules. T h e n e t
effect is s o l u b i l i z a t i o n of t h e b e e t g u m a n d d e s t r u c t i o n of t h e
pulp structure.
A d d i t i o n of lime to p u l p at 3 5 C or below results in an
almost i n s t a n t a n e o u s t o u g h e n i n g of tissues b e c a u s e c a l c i u m ions
react w i t h free c a r b o x y l g r o u p s t o cross-link t h e p e c t i n m o l e c u l e s
i n t o a t h r e e - d i m e n s i o n a l l a t t i c e of g i a n t colecules. Esters a r e
hydrolyzed, b u t t h e r a t e o f d e g r a d a t i o n o f t h e p e c t i n m o l e c u l e
i s low. A f t e r t h e m e t h y l esters h a v e b e e n s u b s t a n t i a l l y r e m o v e d ,
the p e c t i n c h a i n i s s t a b l e t o h o t a n d c o l d a l k a l i b e c a u s e m e t h y l
ester g r o u p s a r e necessary for a l k a l i n e d e g r a d a t i o n o f p e c t i n .

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JOURNAL OF THE A. S. S. B. T.

Water soluble gum


H o t lime solution converts beet p u l p to a fluid mass which
permits soluble gum to pass into solution. T h e solution can
be carbonated and filtered, and the crude gum isolated by simple
drying. Over 200,000 tons of crude gum is available from pulp,
and an industrial use for it should be found. T h i s process does
not utilize the entire pulp; a residue of spent lime, cellulose,
and degraded pectin remains as waste.
Pressed Board
Preliminary experiments have been conducted on the use
of beet pulp and limed beet p u l p as a structural constituent and
bonding agent in pressed boards. Results are promising since
the bonding action of pulp appears to be good. Much practical
research must be conducted on the properties of boards made
from beet pulp, particularly with respect to additives such as
fiber and waterproofing agents, before the industrial potential
can be evaluated.
Thickening
agent
Untreated pulp contains pectin in an insoluble form that
cannot be used as a gelation agent. Alkali-treated pulp, dried
and finely powdered, may be mixed with sodium ethylene diamine tetraacetate, d i a m m o n i u m phosphate, and water to form
a thixotropic gel having physical properties that may make it
suitable for use in forest fire retardants. Tests of this processed
pulp will be conducted in cooperation with agencies having this
primary responsibility.
T h e possibility of manufacturing a valuable gum material
from p u l p is good. However, utilization of p u l p directly as an
ingredient for board manufacture or as an industrial thickening
agent appears to be better. Further research to investigate industrial uses should prove rewarding.
R. M. McCready
A. E. Goodban
Western Regional Research Laboratory
Agricultural Research Service
U. S. Department of Agriculture
Albany, California 94710

JOURNAL
of the
American Society of Sugar
Beet Technologists
Volume 13
Number 6
July 1965

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TABLE OF CONTENTS

Author
A d d i t i o n a l hosts of the beet
Aphanomyces cochlioides Drechs
Xhe Findlay
system

flume

and

water

condenser

mold

of

sugar

469

S. L. Force

478

/. B. Stark

492

H. G. Walker, Jr.
B. A. Ricci
J. C. Goodwin

503

C. S. Baldwin
C. E.
Broadwell
J. F. Davis

509

water

Ion exclusion purification of molasses


Xhin-layer c h r o m a t o g r a p h y
carbohydrates

C. L. Schneider

beet

P r o d u c t i o n practices affecting y i e l d a n d
sugar c o n t e n t of sugar beets g r o w n in O n tario, C a n a d a 1 9 6 1 a n d 1962

Host-parasite relation of Nacobbus batatiformis a n d t h e sugar beet a n d o t h e r hosts M. L. Schuster


Robert
Sandstedt
Larry W. Estes

523

470

JOURNAL OF THE A. S. S. B. T.

described method (13). Concentration of zoospores was determined with a bright-line counting chamber.
In order to reduce the likelihood of seedling infection by
seed-borne fungi, seeds of most of the species included in inoculation tests were soaked in an aqueous suspension of ethyl mercury
phosphate (1000 ppm) for 20 minutes, rinsed in r u n n i n g tap
water for 30 minutes, then dried.
Bioassays to determine the occurrence of A. cochlioides in
plants exposed to the fungus were conducted as follows: Small
pieces of hypocotyls and roots were immersed in a shallow layer
of sterile water in Syracuse dishes and incubated at 20-25 C.
T h e presence of A. cochlioides was indicated by the production
of characteristic zoosporangia protruding from the tissues after
approximately 16 hours, and by characteristic oospores or oogonia
within the tissues. Identification was based on descriptions published by Drechsler (5) and Scott (4).
Host Range Survey
Seedlings of 98 species representing 40 plant families were
exposed to zoospores of A. cochlioides in a series of tests. Included were many of the important crops and weeds of the Great
Lakes region. In each test, seedlings of a highly susceptible
sugar beet variety were included as a check on the infection
potential of the zoospore inoculum.
Fifty-three species were first subjected to an extremely rigorous laboratory test. Seeds were germinated on moist blotting
paper in petri dishes or glass jars. After germination, 20 ml
water containing approximately 25,000 zoospores were poured
into each dish or jar containing the seedlings and were decanted
2 days later. W i t h i n 3 days after exposure to zoospores, susceptible
seedlings began to d a m p off. Although the extremely artificial
environment doubtlessly predisposed plants of susceptible species
to infection, the laboratory test was useful in that it provided
a relatively quick method of isolating nonsusceptible species.
All species susceptible in the laboratory test were included
in subsequent inoculation tests in the greenhouse. Also included
were 48 species that were not tested previously in the laboratory.
Plants were grown in 4-inch pots of steam-sterilized soil. When
seedlings were emerging, 50 ml of water containing approximately 150,000 zoospores were poured into the soil of each pot.
Total n u m b e r of blighted seedlings was determined approximately 30 days after exposure to zoospores. These included plants
with root rot and plants that had damped off.
Seedlings of 30 species in the following families became
blighted when exposed to A. cochlioides zoospores in the greenhouse: Aizoaceae, Amaranthaceae, Caryophyllaceae, Hydrophyl-

VOL.

13,

No.

6,

JULY

1965

471

laceae, L i n a c e a e , P a p a v e r a c e a e , P o r t u l a c a c e a e , a n d S o l a n a c e a e
( T a b l e 1). Species s u s c e p t i b l e i n t h e l a b o r a t o r y test w e r e also
s u s c e p t i b l e i n t h e g r e e n h o u s e test.
Species s u s c e p t i b l e to artificial e x p o s u r e s of t h e p a t h o g e n
w e r e s u b s e q u e n t l y g r o w n i n t h e g r e e n h o u s e o r f i e l d i n soil
n a t u r a l l y infested w i t h A. cochlioides. P l a n t s w i t h s y m p t o m s of
damping-off, w i l t i n g , r o o t r o t , a n d d i s c o l o r a t i o n o f t h e l o w e r
s t e m w e r e bioassayed for t h e p r e s e n c e of A. cochlioides.
A.
cochlioides was i d e n t i f i e d in p l a n t s of 19 species, besides Beta
vulgaris, t h a t w e r e g r o w n in n a t u r a l l y infested soil ( T a b l e 1).
P u r e c u l t u r e s of A. cochlioides w e r e isolated f r o m Chenopodium
album,
Spinacia
oleracea,
Tetragonia
expansa,
Mollugo
verticillata a n d Saponaria ocymoides a n d w e r e p a t h o g e n i c on s u g a r
b e e t seedlings i n g r e e n h o u s e i n o c u l a t i o n tests.
A. cochlioides was n o n p a t h o g e n i c on t h e following; species:
A s c l e p i a d a c e a e , Asclepsis syriaca L . ;
B a l s a m i n a c e a e , Impatiens
balsamina L.; B o r a g i n a c e a e , Anchusa azurea M i l l . ; C o m p o s i t a e ,
Ambrosia trifida L., Aster macrophyllus L.,
Calendula
officinalis
L., Carthamus tinctorius L., Solidago sp., Tagetes sp., Zinnia sp.;
Convolvulaceae,
Ipomoea
purpurea
Lam.;
Cruciferae,
Alyssum
saxatile L., Brassica arvensis (L.)
R a b e n h , B. rapa L., Lepidium
virginicum
L., Raphanus sativus L., Thlaspi
arvense
L.; Cuc u r b i t a c e a e , Cucumis melo L., C.
sativus
L., E u p h o r b i a c e a e ,
Euphorbia serpyllifolia Pers., Ricinus communis L.;
Geraniaceae,
Geranium sp.; G r a m i n e a e , Avena sativa L., Echinochloa crusgalli
(L.)
B e a u v . , Hordeum vulgare L., Secale cereale L.; Setaria
glauca L. (Beauv.), Zea mays L., L a b i a t a e , Monarda fistulosa L . ;
Leouminoseae,
Astragulus
canadensis L., Medicago
sativa
L.,
Melilotus alba Desr., Pisum sativum L., Glycine max
(L.)
Merr.,
Trifolium pratense L.; L i l i a c e a e , Allium cepa L.;
Malvaceae,
Althaea
rosea C a v . ;
Nyctaginaceae,
Abronia
umbellata
Lam.,
Mirabilis jalapa L . ; O n a g r a c e a e , Clarkia elegans D o u g l . ; P e d a l iaceae, Sesamum indicum L . ; P i n a c e a e , Pinus banksiana L a m b . ;
P l a n t a g i n a c e a e , Plantago
major L.;
P l u m b a g i n a c e a e , Armeria
pseud-armeria
Mansfeld;
Polemoniaceae,
Phlox
drummondii
Hook;
P o l y g o n a c e a e , Fagopyrum esculentum M o e n c h , Polygonum
erectum L., P. pensyIvanicum L., P. persicaria L., Rumex crispus
L.,
R.
mexicanus
Meisn.;
Primulaceae, Anagallis
linifolia
L.,
R a n u n c u l a c e a e , Delphinium ajacis L.; R o s a c e a e , Geum chiloense
Balbis,
Potentilla
arguta
Pursh;
Scrophulariaceae,
Verbascum
thapsus L . ; S o l a n a c e a e , Lycopersicon esculentum M i l l . , Nicotiana
rustica
L.
Petunia
hybrida
Vilm.,
Solanum
melongena
var.
esculentum N e e s ;
Tropealaceae,
Tropeolum
majus L . ;
Umbellferae, Anethum graveolens L . ;
Urticaceae,
Urtica gracilis A i t . ;
V e r b e n a c e a e , Verbena hybrida Voss; V i o l a c e a e , Viola tricolor L.

Table 1. Reaction of 30 plant species to artificial and natural exposures of Aphanomyces cochlioides.

1
2

Authorities for scientific and common names are: Bailey (1), Engler and Prantl (7) and Fernald (8).
Symptoms confined to a slight discoloration of the roots.

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JOURNAL OF THE A. S. S. B. T.

Differences in Susceptibility Between Some Host Species


Differences in degree of susceptibility to A. cochlioides between 8 species that had been infected in the host range survey
were experimentally demonstrated in the greenhouse. Seedlings
of the following species, representing 4 plant families, were grown
in 4-inch pots of steam-sterilized soil and exposed to zoospores
(approximately 150,000 per pot) about 3 weeks after planting,
when all seedlings had emerged: Amaranthus blitoides, Lychnis
alba, Saponaria officinalis, Beta vulgaris, Chenopodium album,
Salsola kali, Spinacia oleracea, and Portulacea oleracea. Symptoms
of disease included damping-off and root rot. Approximately
40 days after exposure to zoospores, significant differences between some species in susceptibility were apparent (Table 2)
and ranged from 0% plants diseased, for Lychnis alba and
Portulacea oleracea, to 100% for Beta vulgaris.
Table 2. Relative susceptibility of 8 plant species exposed to Aphanomyces cochlioides zoospores in the greenhouse after emergence of seedlings.

LSD (P = .05)
36.3
Includes plants damped off and those with root rot.
Results expressed as mean of two 4-inch pots. Percentage data were converted to degrees
for statistical analysis.

1
2

Pathogenicity of Isolates from Different Hosts Compared


T h e pathogenicity of 10 cultures of A. cochlioides, isolated
from Beta vulgaris, Spinacia oleracea, and Chenopodium album
grown in soils from several sugar beet-growing areas of the United
States 4 was compared in the greenhouse. Seedlings of moderately
resistant sugar beet variety US 400 were exposed to zoospores
of each culture (approximately 150,000 per pot) 14 days after
4
Samples of Michigan and Montana soils were kindly furnished by H. W. Bockstahler
and M. M. Afanasiev, respectively.

V O L . 13, N o . 6, J U L Y 1965

475

p l a n t i n g , w h e n e m e r g e n c e was c o m p l e t e . T h i r t y days after exp o s u r e t o zoospores, each p l a n t was assigned o n e o f t h e f o l l o w i n g


n u m e r i c a l r a t i n g s a c c o r d i n g to severity of s e e d l i n g b l i g h t : 0 ( n o
s y m p t o m s ; 1 ( l i g h t s y m p t o m s ) ; 2 ( m o d e r a t e s y m p t o m s ) ; 3 (severe
s y m p t o m s ) ; 4 ( d e a d ) . I n c i d e n c e a n d severity of b l i g h t c a u s e d
by each isolate w e r e t h e n c o n v e r t e d to a s i n g l e p e r c e n t a g e v a l u e
b y a m e t h o d s i m i l a r t o t h a t o u t l i n e d b y L e C l e r g (9). T h e average i n f e c t i o n t y p e o f t h e p l a n t s i n each p o t was o b t a i n e d b y
a d d i n g t h e p r o d u c t s o f each i n f e c t i o n t y p e t i m e s t h e n u m b e r o f
plants of that type a n d d i v i d i n g this sum by the n u m b e r of
p l a n t s e x p o s e d t o zoospores. T h i s n u m b e r was c o n v e r t e d t o a
percentage value by dividing it by 4, the value of the highest
i n f e c t i o n type, a n d m u l t i p l y i n g b y 100.
V a r i e t y U S 4 0 0 was s u s c e p t i b l e t o each c u l t u r e ( T a b l e 3).
N o significant differences i n v i r u l e n c e w e r e n o t e d b e t w e e n cult u r e s f r o m t h e 3 h o s t species n o r from t h e 5 l o c a t i o n s .
Table 3. Relative susceptibility of sugar beet variety US 400 to 10 cultures of
Aphanomyces cochlioides isolated from sugar beet, spinach, and lamb's quarters (Chenopodiutn album) and from different areas.

LSD (P = .05)
N.S.
Sub-cultures from single zoospores are designated by a capital letter following the culture number.
2
Results expressed as mean of 4 pots. Incidence and severity of disease expressed as a
single percentage value transformed to degrees for statistical analysis.
1

Discussion
A. cochlioides h a s b e e n r e p o r t e d to o c c u r in soils in w h i c h
crops o f s u g a r b e e t s a p p a r e n t l y h a v e n e v e r b e e n g r o w n (4). S o m e
of t h e c o m m o n w e e d s s h o w n to be hosts of A. cochlioides including
Chenopodium
album,
Amaranthus
blitoides,
A.
retroflexus a n d Mollugo verticillata m a y well a i d in t h e s u r v i v a l of
the f u n g u s i n a g r i c u l t u r a l soils. A r e p o r t e d i n c r e a s e i n i n c i d e n c e

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JOURNAL OF THE A. S. S. B. T.

of black root disease of sugar beets on land previously occupied


by a dense stand of Amaranthus (3) is cited as an example.
Weed hosts that were shown to be highly resistant to infection
by A. cochlioides, such as Portulacea oleracea and Lychnis alba,
may play a less effective role in survival of the fungus than the
more susceptible hosts. Spinach and ornamental plant hosts
such as Saponaria ocymoides, although extremely susceptible,
probably contribute little to the occurrence of the fungus in
soils where the sugar beet crop is grown.
In this study and in studies of other investigators (2, 11),
no major crop grown in rotation with sugar beets in the Great
Lakes region was shown to be a natural host of A. cochlioides.
Nevertheless, care must be exercised in the choice of crops to
precede sugar beets, since it has been shown that black root
disease is usually more severe when sugar beets follow lateplowed plantings of alfalfa or sweet clover than when sugar
beets follow corn or rye (3). Inasmuch as there is no evidence
that the legume crops are hosts of A. cochlioides, one explanation
for the increase in black root disease when beets follow alfalfa
or sweet clover may be that the residues of these crops and the
accompanying microflora are more favorable for increase of the
pathogen than those of corn or rye.
Summary
T h e reaction of 94 plant species to pure culture of Aphanomyces cochlioides was determined. Twenty-eight new experimental hosts from the following families are reported: Aizoaceae,
Amaranthaceae, Caryophllaceae, Chenopodiaceae, Hydrophyllaceae, Linaceae Papaveraceae, Portulacaceae. Nineteen species
in addition to Beta vulgaris were found to be natural hosts of the
fungus. Differences in relative susceptibility to A. cochlioides
between several host species were experimentally demonstrated.
Cultures of A. cochlioides isolated from Spinacia oleracea and
Chenopodium album were as pathogenic on Beta vulgaris as cultures isolated from Beta vulgaris.
Literature Cited
(1) BAILEY, L. H. 1949. Manual of cultivated plants. Revised edition
of the Hamilton Company, New York. pp 1116.
(2) BUCHHOLTZ, W. F. 1944. Crop rotation and soil drainage effects on
sugar beet tip rot and susceptibility of other crops to Aphanomyces
cochlioides. Phytopathology 34: 805-812.
(3)

COONS,

G.

H., J.

E.

KOTILA

and H.

W.

BOCKSTAHLER.

1946.

Black

root of sugar beets and possibilities for its control. Proc. Am. Soc.
Sugar Beet Technol. 4: 364-380.

V O L . 13, N o . 6, J U L Y 1965

477

(4) D O W N I E , A. R. 1942. Damping-off a n d r o o t r o t of sugar beets caused


by Aphanorayces cochlioides Drechs. P h . D . Thesis, University of
Minnesota (unpublished) .
(5) DRECHSLER, C H A R L E S . 1929. T h e beet water mold a n d several related
root parasites. J. Agric. Res. 28: 309-361.
(6) DRECHSLER, C H A R L E S .
1954. Association of Aphanomyces cladogamus
with severe r o o t r o t of pansies. Sydowia 8: 334-342.
(7) ENGLER, a n d P R A N T L .
1934. Die Naturlichen Pflanzenfamilien. Vol.
16 W i l h e l m E n g e l m a n n , Leipzig.
(8) FERNALD, M. L. 1950. Gray's N e w M a n u a l of Botany. 8th edition
A m e r i c a n Book C o m p a n y , N e w York. pp 1632.
(9) L E C L E R G , E. L. 1939. M e t h o d s of d e t e r m i n a t i o n of physiologic races
of Rhizoctonia solani on the basis of parasitism on several crop
p l a n t s . P h y t o p a t h o l o g y 29: 609-616.
(10) M C K E E N , C. D. 1952. Aphanomyces cladogamus Drechs., a cause of
damping-off in p e p p e r s a n d certain o t h e r vegetables. C a n . J. Bot.
30: 701-709.
(11) M C K E E N , W. E. 1949. A study of sugar beet root-rot in Southern
O n t a r i o . C a n . J. Res. C 27: 284-311.
(12) SCHNEIDER, C. L. 1952. H o s t r a n g e of t h e sugar beet root rot fungus,
Aphanomyces cochlioides.
P h y t o p a t h o l o g y 42: 18
(abstract) .
(13) SCHNEIDER, C. L. 1954. M e t h o d s of i n o c u l a t i n g sugar beets with
Aphanomyces cochlioides Drechs. Proc. A m . Soc. Sugar Beet T e c h nol. 8(1) : 247-251.
(14) SCHNEIDER, C. L. 1958. F u r t h e r studies on the host r a n g e of Aphanomyces cochlioides. P h y t o p a t h o l o g y 48: 463-464. (abstract) .
(15) SCOTT, W I L L I A M W A L L A C E . 1961. A m o n o g r a p h of the genus A p h a n o myces. Va. Agr. E x p t . Sta. Tech. B u l . 151: 1-96.

The Findlay Flume and Condenser W a t e r System


S.

L.

FORCE 1

Received for publication August 4, 1964

In the spring of 1955 T h e Great Western Sugar Company


acquired two beet sugar factories in Ohioone in the City of
Fremont, the other in Hancock County immediately adjacent to
Findlay.
Ohio law requires industry to obtain a license to discharge
wastes of any kind into the water resources of the state. Fremont's lagoon system was acceptable. Findlay's was not and a
temporary permit was issued to operate one campaign using the
existing waste system, with the understanding that a satisfactory
system would be developed to direct all out-flows through the
City Sewage T r e a t m e n t Plant.
T h e temporary permit specified wastes generated d u r i n g the
1955-56 campaign would be handled in the same manner as during the 1954 factory operation, i.e., a closed recycle system.
Basically, this system was as follows:
1. P u l p press and separator water returned to the cell type
diffusion battery.
2. Condenser waters recycled in a closed system through spray
ponds.
3. Flume water recycled through a large settling pond.
4. Lime m u d water returned to the flume water.
5. T h e flume pond water to be pumped at a controlled rate
to the Blanchard River after campaign and only during
the high river flows d u r i n g the spring runoff.
T h i s system had three principal problems:
1. T h e flume water pond accumulated all of the excess water
used in the plant. All waters had to be contained and
none could be directed to the Blanchard River until after
campaign. T h u s water build-up had to be carefully controlled.
2. T h e flume water system had no heat in its cycle and made
cold weather fluming difficult.
3. T h e degree of pollution in the system was high, approximately 2500 ppm BOD.
Late in 1955 the Ohio Highway Department appeared with
plans for a bypass highway crossing the factory site. T h e road
and right-of-way eliminated a major part of the land on which
1

District Superintendent, The Great Western Sugar Company, Denver, Colorado.

V O L . 13, N o . 6, J U L Y 1965

479

the flume water ponds were built. T h i s added to the problem


b y severely r e d u c i n g t h e l a n d a r e a s a v a i l a b l e for p o n d s . U n d e r
these c i r c u m s t a n c e s , t h e d e v e l o p m e n t a n d d e s i g n of a waste
t r e a t m e n t system for t h i s factory was i n s t i t u t e d . A f t e r considera b l e t i m e a n d effort, a system w a s d e v e l o p e d c a l l e d " T h e F i n d l a y
F l u m e a n d C o n d e n s e r W a t e r System," w h i c h has w o r k e d very
well, a n d m u c h has b e e n l e a r n e d a b o u t its o p e r a t i o n a n d m a i n tenance.
T h e n e w facility was d e s i g n e d a n d b u i l t a t t h e F i n d l a y , O h i o ,
plant of the N o r t h e r n O h i o Sugar C o m p a n y , a subsidiary of
T h e Great Western Sugar Company, in the spring a n d summer
o f 1956. T h e d e s i g n objectives w e r e :
1. M i n i m u m v o l u m e of w a t e r .
2. W a r m e r flume water.
3. Separate i m p o u n d i n g of lime a n d m u d .
4. Reduced B O D content of the water.
5 . R i g i d c o n t r o l o f i n p l a n t w a t e r use.
6. A closed system w i t h all final wastes d i r e c t e d to t h e city
sewage t r e a t m e n t p l a n t .
7 . M i n i m u m l a n d a r e a s w e r e a v a i l a b l e for p o n d s a n d a n y
p l a n d e v e l o p e d h a d t o b e c o m p a t a b l e w i t h t h i s physical
limitation.
T h e F i n d l a y factory is a non-Steffen o p e r a t i o n . It h a s a d r y e r
a n d t h e p r e s s e d p u l p w a t e r i s r e t u r n e d t o t h e diffuser. T h e basic
features o f t h e F i n d l a y system a r e s h o w n o n t h e d r a w i n g s
of t h e flow s h e e t ( F i g u r e 1), a n d g e n e r a l p l a n ( F i g u r e 2).
R e f e r r i n g t o t h e f l o w sheet, t h e system i s o u t l i n e d i n steps a s
follows:
1 . F l u m e w a t e r c a r r y i n g b e e t s i n t o t h e factory i s s e p a r a t e d
from the beets at the beet wheel a n d flows to the flume
sewer s u m p p u m p .
2 . T w o f l u m e sewer p u m p s , e a c h r a t e d a t 4 0 0 0 g p m a t 4 0
ft T D H are used to p u m p the flume water to the grit
separator.
3. T h e grit separator is a l i q u i d cyclone to r e m o v e heavy
solids. T h e solids a r e p u m p e d t o t h e s l u d g e p o n d .
4 . T h e s e p a r a t o r overflow i s d i v i d e d b e t w e e n 3 L i n k - B e l t
C A # 1 5 1 2 v i b r a t i n g s c r e e n s t o r e m o v e l i g h t solids, t o p s ,
w e e d s a n d o t h e r o r g a n i c d e b r i s . T h e s e solids a r e h a u l e d
a w a y for use a s feed, g r e e n f e r t i l i z e r o r o t h e r d i s p o s a l . T h e
screens a r e 5 ft w i d e a n d 12 ft l o n g a n d a r e e q u i p p e d
with T y l e r ton cap # 7 4 0 M o n e l wire mesh screen.
5. Screened water is then divided equally between two 63
f t 8 i n d i a m b y 1 2 f t h i g h D o r r clarifiers. T h e s e u n i t s

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JOURNAL OF THE A. S. S. B. T.

Figure 1.Flow sheet of flume, condenser and fresh waters, Findlay,


Ohio, plant, Northern Ohio Sugar Company.

work on a cross-flow principle and have a retention time


of 67.5 m i n at 2450 gpm. Settleable solids fall to the bottom of these tanks, and are subjected to a center sludge
drawoff by the sweep mechanism and p u m p e d to the
sludge pond with a 4 in manganese-lined p u m p .
6. T h e clarified flume water is p u m p e d to the spray pond.
T h i s p u m p is rated at 5000 gpm at 48 ft T D H . T h e pipe
line is 18 in OD steel pipe with Dressier couplings and
is 1920 ft long.
7. T h e spray pond has four headers each having 30 spray
nozzles spaced 15 ft apart. T h e nozzles are Marley #2004
one-piece design with a 1-1/16" diam outlet orifice capable
of passing 44.4 gpm at 10 psig pressure. Cooling realized
ranged from 10 to 15 C depending on weather conditions.
8. T h e spray-cooled water is then p u m p e d back to the main
water " s u m p " through the excess water p o n d (Figure 2),
which serves as a b u m p e r tank. T h e spray pond return
p u m p is rated at 5000 gpm at 40 ft T D H .

VOL.

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No.

6,

JULY

1965

481

Figure 2.General plan of water and waste system, Findlay, Ohio,


plant, Northern O h i o Sugar Company.

9 . F r o m t h e m a i n w a t e r s u m p t h e clarified a n d spray-cooled
flume water is p u m p e d to the "main water tank," which
s u p p l i e s all t h e c o n d e n s e r s a n d o t h e r m i s c e l l a n e o u s uses
not r e q u i r i n g clean water.
10. T h e c o n d e n s e r w a t e r i s c o l l e c t e d i n seal t a n k s a n d p i p e d
to the "flume water tank".
11. T h i s c o n d e n s e r w a t e r i s t h e n p u m p e d t o t h e f l u m e s b y t h e
4 0 0 0 g p m f l u m e w a t e r p u m p t o c o m p l e t e t h e closed system
cycle.
N o n e o f t h e r e c y c l e d w a t e r e n t e r s t h e s u g a r - m a k i n g process.
Diffuser s u p p l y m a k e - u p c o m e s f r o m a fresh well w a t e r c i r c u i t
which serves a s c o o l i n g w a t e r for crystallizers, p u m p j a c k e t s a n d
other uses r e q u i r i n g c l e a n , c o l d w a t e r . T h i s i s t h e n d i r e c t e d
i n t o t h e process w a t e r a s diffuser m a k e - u p w a t e r . Excess well
and city w a t e r n o t r e q u i r e d for m a k e - u p i s sent t o t h e B l a n c h a r d
River. T h i s i s p e r m i s s i b l e since t h i s excess well w a t e r d o e s n o t
contain a n y p o l l u t a n t s a n d p r e v e n t s a n u n d e s i r a b l e v o l u m e
b u i l d - u p i n t h e recycle system.
W a t e r accumulating in the sludge a n d lime ponds is reused
in t h e system t h u s e l i m i n a t i n g a b u i l d - u p of w a t e r f r o m t h e s e
sources.

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JOURNAL OF THE A. S. S. B. T.

In the early summer the lagoon water is p u m p e d at a controlled rate of 150 gpm during the hours from 6:00 pm to 6:00
am to the Municipal Sewage Plant. T h e r e is a waste water
disposal contract with the city with a service fee figured on the
basis of pounds of B O D and suspended solids passing through
the city disposal system.
As would be expected, the BOD and suspended solids are
subject to wide variations. In October and early November the
BOD is generally around 700 ppm. By the end of the operating
season, approximately the 1st of January, the B O D approaches
2600 ppm, total solids range from 1500 to 2000 ppm, the water
temperature is 1C and the pH approximately 6.5. By midMarch the water temperature is 8C, pH 7.7 and the B O D 700
ppm. W h e n the final wastes are retained until June 1st, BOD's
as low as 70 ppm have been obtained with water temperature
approximately 16C and a pH in the neighborhood of 7.7.
No serious odor problems have been encountered by holding
the waste waters until late May. For a short period in March
odors are noticeable downwind from the ponds for a distance
of about 200 ft, b u t about two weeks later they become less
noticeable. T h e odors emanating from the lime pond, the mud
pond and the spray pond are always a great deal more pronounced than from water stored in the excess water pond. Longer
retention time d u r i n g warm weather reduces the discharge costs.
T h e total volume of water delivered to the city treatment
plant varies from a low of 1,616,700 gal in 1958 to a high of
6,446,100 gal in 1959. T h e system is estimated to hold 9,500,000
gal.
T h e Findlay system originally went into operation during
the 1956-57 campaign. T h e contract with the city called for
payment of $20 per 100 lb of B O D disposed through the city
system. Immediately prior to the 1961-62 campaign the contract
was revised, and service charges are presently $16 per 100 lb BOD
in excess of 200 ppm and $12.50 per 100 lb of suspended solids
in excess of 240 ppm. Fifty per cent is then added to the bill
because the factory is outside the city limits.
T a b l e 1 shows the volumes treated and the costs for the past
nine years. T h e 1955-56 data was included to show a comparison
between the old and new systems. T h e treatment costs up to
the 1961-62 sugar year were based on the B O D average for the
preceding year. For the first year a B O D of 1500 was assumed.
It is interesting to note the a m o u n t of water p u m p e d to the
treatment plant d u r i n g the operating season. W i t h proper inplant water use and control, it is not necessary to unload the
system d u r i n g campaign. D u r i n g the 1963-64 campaign, the

Table 1.Nine-year costs and volumes treated, Findlay flume and condenser water system.

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water build-up and inplant water usage was so well controlled


that no water was p u m p e d to the city treatment plant. Since
no serious odor problems have developed up to July 1, 1964,
the water is still in the lagoons and it will probably not be
necessary to discharge any wastes to the city sewage plant.
W h e n the pond water was emptied J u n e 18, 1963, the B O D
and suspended solids were so low that treatment costs were
eliminated. T h i s system can be operated so that no water is discharged d u r i n g the processing period and i the ponds are
drained alter the first of June, the B O D and solids content will
be such that u n d e r the existing treatment contract, the cost
will be eliminated.
T a b l e 2 shows some of the information developed during
the first and last part of campaign as well as the campaign
averages at various points in the system. Note the suspended
solids in the lime pond overflow. T h i s reflects the fact that
there was an insufficient volume for the lime m u d and when
freezing weather added its contribution of ice, there were practiTable 2.Findlay waste water system1963-64 campaign

V O L . 13, N o . 6, J U L Y 1965

485

cally n o solids e l i m i n a t e d . T h i s was n o t g o o d for t h e c o n d e n s e r


leg l i n e s a n d as a m a t t e r of fact, c a u s e d a s h u t d o w n to c l e a n t h e
low r a w p a n c o n d e n s e r leg l i n e .
F i g u r e 3 shows B O D b u i l d - u p d u r i n g t h e 1963-64 c a m p a i g n
o n t h e m a i n w a t e r t a n k a n d D o r r overflow w a t e r . A l s o s h o w n
i s t h e b u i l d - u p o f s u s p e n d e d solids i n t h e m a i n w a t e r t a n k a s
w e l l a s t h e effluent f r o m t h e l i m e p o n d a n d t h e B O D d r o p
d u r i n g t h e first p a r t o f c a m p a i g n . A l l p r e v i o u s d a t a g e n e r a t e d
a t b o t h O h i o factories d u r i n g e a c h c a m p a i g n since 1956 shows
this s a m e basic t r e n d . T h i s i s n o d o u b t d u e t o t h e fact t h a t
slicing i s s t a r t e d w i t h t h e p o n d s r e l a t i v e l y e m p t y a n d t h e reduction is accomplished by dilution.
Insufficient l i m e p o n d
settling v o l u m e is a p p a r e n t from D e c e m b e r 9 to the e n d of

Figure 3 . C o n t a m i n a n t buildup, waste water system, Findlay, Ohio,


1963-64 campaign.

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JOURNAL OF THE A. S. S. B. T.

campaign. T h e spectacular rise in the suspended solids in the


lime pond overflow indicates there were very few solids removed. Another interesting observation is that the B O D seems
to level out at the end of campaign at a point lower than
would be expected.
Figure 4 is a water balance. T h e flows were obtained by
p u m p curves, amperage readings and pressure. A BOD balance
can be made based on this table, using the campaign average
BOD's shown on T a b l e 2. A B O D balance has not been included
in this report as it does not balance as well as was hoped. Figure
4 did show there was not a significant d r o p in total pounds of
BOD's across the spray pond which averagely reduces the pond
water temperature 21 F.
Dike areas are as follows:
Acres
1. Excess Water Pond
1.11
2. Dorr Sludge Pond
5.80
3. Lime Settling Pond
5.40
4. Spray Pond
1.36
T h e Fremont water recycle system is based on a closed circuit
flume system with the condenser water returned to the river.

Figure 4.Diagram of the flume and condenser water system, Findlay,


Ohio, 1963.

Table 3.Average figures on Fremont waste water in the lagoons for the campaign and the discharge period.

* B.O.D. samples heavily diluted with water pumped into lagoon from flooded area.

Table 4.BOD, suspended solids, dissolved solids, temperature and pH on the spray pond, Findlay, Ohio, Intercampaign.

*BOD determined actually 42 and 38 respectively. These results were incorrect because the ponds now contain dissolved oxygen which we failed to
consider.

Table 5.Same data as in Table 4 on the excess water pond, 1964.

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JOURNAL OF THE A. S. S. B. T.

Lagoon water is discharged to the Sandusky River only d u r i n g


the period of high river flows in the spring. T a b l e 3 shows
average figures from data obtained on Fremont waste water in
the lagoons both for the campaign and the discharge periods
to the Sandusky River. T h i s is included as a matter of interest
and for comparison with the Findlay system.
Table 6.Coliform counts on samples taken June 10, 1964, from the spray, lime and
excess water ponds.

A word of explanation might be in order regarding the techniques used and the
interpretation of this data. One hundred millileter samples were collected on June 10,
1964 in sterile bottles. Serial dilutions of 100. 1000, and 10,000 were made and plated for
a colony count on nutrient agar. Also. 0.1, 1.0, and 10ml samples were inoculated into
lactose broth. This is a method of approximating the numbers of coliform organisms based
on statistical probability. It also constitutes a presumptive test for the presences of the
coliform organisims. After 24 hours, samples were taken from the positive lactose broth
tubes and streaked on EMB agar. After an additional 24 hours, all plates showed the
presence of Escherichia coli. This test is considered a confirmation test in the standard
analysis of water supplies.
The plate count shows the numbers of bacteria to be verv high in the spray pond with
fewer bacteria per millileter in the lime and excess water ponds.
The most significant aspect of this analvsis is the LARGE NUMBERS of coliform organisms, particularly E. coli. This means that this water is heavily polluted from a Mater
sanitation viewpoint. It does not mean that these impoundments were necessarily contaminated directly with domestic sewage. Although the human colon is the primary source
of the coliform organisms in nature, these organisms are not necessarily parasites but can
establish themselves wherever the proper conditions of nutrients, temperature, etc. prevail.
It does mean however, that these bodies of water are of a potential danger to domestic
sources of drinking water if the water is not treated or disposed of in the proper way.
* Using lactose broth which is a presumptive test for the coliform group of bacteria.
** Using EMB agar which is selective for Escherichia coli.
Data by Dr. John W. McClymont, Professor of Botany and Bacteriology, Findlay College,
Findlay, Ohio.

T a b l e 4 shows B O D , suspended solids, dissolved solids, temperature and pH on the spray pond with comments from the
end of campaign through J u n e 17, 1964. T a b l e 5 shows the same
data on the excess water pond. T h i s is the first time the ponds
have been closely checked after campaign and prior to discharge.
T h e r e were some surprising rapid changes: the marked increase
in filterability of the waste water on May 27, which coincided
with the appearance of the green algae on the spray pond; the
disappearance of the green algae d u r i n g the week ending June

V O L . 13, N o . 6, J U L Y 1965

491

11th i n t h e spray p o n d , a n d t h e take-over o f t h e s m a l l b u g s


followed o n e w e e k l a t e r b y t h e d i s a p p e a r a n c e o f t h e b u g s a n d
r e a p p e a r a n c e o f t h e g r e e n algae. T h e s e t a b l e s g e n e r a t e t h e following questions:
1. C o u l d t h e dissolved solids be c h e m i c a l l y p r e c i p i t a t e d ?
2. Could nutrients be added to lagoon water to aid the desired
growths?
3. C o u l d i n n o c u l a t i o n w i t h a specific o r g a n i s m , after t h e t e m p e r a t u r e s b e c o m e a c c e p t a b l e , speed u p t h e r e d u c t i o n o f
solids o r B O D ?
T a b l e 6 shows c o l i f o r m c o u n t s o n s a m p l e s t a k e n J u n e 10,
1964. f r o m t h e spray p o n d , t h e l i m e p o n d a n d t h e excess w a t e r
p o n d s . T h e s e d e t e r m i n a t i o n s , a s well a s t h e e x p l a n a t i o n s , w e r e
m a d e b v D r . Tohn W . M c C l y m o n t , Professor o f B o t a n y a n d
B a c t e r i o l o g y o f F i n d l a y College, F i n d l a y , O h i o . D r . M c C l y m o n t
identified t h e s m a l l b u g s i n t h e s p r a y p o n d a s Daphnia.
In c o n c l u s i o n , t h i s system fulfilled its d e s i g n objectives. Since
wastes a r e d i s c h a r g e d t h r o u g h t h e city sewage t r e a t m e n t p l a n t ,
a p e r m i t is n o t necessary each y e a r to d i s c h a r g e wastes. T h i s is
most d e s i r a b l e since t h e costs o f w a s t e w a t e r t r e a t m e n t b y t h e
city h a v e n o t b e e n excessive. T h e f o l l o w i n g deficiencies w e r e
noted:
1 . F l u m e w a t e r i s t o o h o t i n O c t o b e r . T h i s c o u l d b e corr e c t e d b y h a v i n g t w o closed w a t e r systems:
a. A closed flume w a t e r system.
b . A closed s p r a y p o n d c o n d e n s e r w a t e r system. W h e n t h e
w e a t h e r gets cold, t h e h e a t i n t h e f l u m e system i s n o t
excessive a n d i s d e s i r a b l e . A t t h i s t i m e t h e s e p a r a t e
systems c o u l d b e c o m b i n e d .
2 . L i m e a n d m u d p o n d effluent i s u n d e s i r a b l e . I f t h e l i m e
m u d transport water could be reduced to the point where
t h e l i m e p o n d overflow c o u l d b e d i s p o s e d o f b y o p e n
f i e l d i r r i g a t i o n , t h i s w o u l d i m p r o v e t h e system. A n o t h e r
possibility w o u l d b e t o u s e t h e l i m e p o n d overflow a s
l i m e m u d t r a n s p o r t w a t e r . T h e same p r o c e d u r e s c o u l d
b e c o n s i d e r e d for t h e m u d p o n d overflow, b u t t h i s i s n o t
a s c r i t i c a l a s t h e l i m e p o n d overflow. T h e l i m e p o n d
overflow s h o u l d b e e l i m i n a t e d from t h e recycle w a t e r
system.
Acknowledgment
M a n y persons helped in c o m p i l i n g this paper: D. G o r d o n ,
Superintendent at Findlay; Lee Coons, Cashier at Findlay; W a y n e
Argabrite a n d Shelby Knepper, Engineers; a n d H a r r y Dougherty,
Chief C h e m i s t , w h o r a n t h e m a n y , m a n y d e t e r m i n a t i o n s t h a t
go into the d e v e l o p m e n t of tables in this paper.

Ion Exclusion Purification of Molasses


J.

B.

STARK 1

Received for publication August 7, 1964

T h e separation of ionic from non-ionic substances using ion


exclusion is only a little over ten years old. Ton exclusion purification arose from the observation that at equilibrium the concentration of a strong electrolyte is lower within the aqueous
portion of the resin phase than in the surrounding solution.
T h e electrolyte is partially excluded from the water in the resin
bead. Non-ionized or weakly ionized compounds generally have
a relatively high concentration within the resin bead compared
to highly ionized compounds. W h e n the differences in relative
concentrations inside and outside the resin bead are sufficiently
large, two substances may be separated readily on a resin column.
Essentially ion exclusion purification is a chromatographic
technique. T h e components to be separated are loaded on the
column and washed through with water as the eluant. Compounds having a lower relative concentration in the resin bead
will be eluted more rapidly and hence separate from components
having a higher relative concentration in the resin bead. Nonionized or weakly ionized compounds may also separate from
each other if there is an adequate difference in their relative
concentrations between the solution inside and outside the resin
beads.
Recovery of sugar from molasses by ion exclusion offers a
n u m b e r of advantages over ion exchange. A cation exchanger is
cheaper and more stable than an anion exchanger. Operation
near pH 7 precludes sucrose inversion. T h e r e is no expense for
costly regenerants as water elutes both the sucrose and impurities
from the column. Fewer resin beds should be required as only
one resin is utilized.
Wheaton and Bauman (1,2) 2 discussed theory of ion exclusion and its potential applications in 1953. Patents have been
issued for the separation of ionic materials having different
degrees of ionization (3) and for the separation of certain organic compounds (4). T h i s latter patent (4), describes separations for glucose-acetone and sucrose-glycerol-triethylene glycol.
A later article by Asher (5) discusses sugar purification by ion
exclusion and gives examples of the separation of salts from
dextrose or sucrose. Color and strong ionic materials present
1
Western Regional Research Laboratory, Western Utilization Research and Develop
ment Division, Agricultural Research Service, U. S. Department of Agriculture, Albany.
California.
2
Numbers in parentheses refer to literature cited.

V O L . 13, N o . 6, J U L Y 1965

493

i n molasses o r c r u d e c a n e s u g a r w e r e s e p a r a t e d from t h e sucrose.


A p a t e n t by W h e a t o n (6) on i o n e x c l u s i o n p u r i f i c a t i o n d e s c r i b e s
s i m i l a r e x p e r i m e n t s i n c l u d i n g t r e a t m e n t o f c a n e molasses t o
r e m o v e i o n i c c o m p o u n d s . S i m p s o n a n d W h e a t o n (7) give a
g o o d r e s u m e of t h e effects of a n u m b e r of v a r i a b l e s t h a t influence t h e efficiency of i o n e x c l u s i o n s e p a r a t i o n p a r t i c l e size,
f l o w r a t e , crosslinkage, a n d feed v o l u m e t o c o l u m n v o l u m e r a t i o
u s i n g e t h y l e n e glycol as a test c o m p o u n d . S i m p s o n a n d B a u m a n (8) d e s c r i b e a p r o c e d u r e for r e c y c l i n g effluents from an
i o n e x c l u s i o n s e p a r a t i o n o f e t h y l e n e glycol f r o m s o d i u m c h l o r i d e
w h i c h c o u l d possibly b e a d a p t e d t o s u g a r l i q u o r p u r i f i c a t i o n .
A m o r e recent publication by N o r m a n , R o r a b a u g h , and Keller
(9) discusses s o m e of t h e a d v a n t a g e s of i o n e x c l u s i o n as a p p l i e d
t o s u g a r j u i c e p u r i f i c a t i o n . T h e y also p o s t u l a t e o p e r a t i n g req u i r e m e n t s for a c o n t i n u o u s H i g g i n ' s c o n t a c t o r (10) w h i c h
m i g h t b e u s e d t o p u r i f y s u g a r b e e t thick j u i c e d i l u t e d t o 4 0 B r i x .
N o n e o f these p a p e r s i n d i c a t e a n y s e p a r a t i o n o f r e d u c i n g
sugars, i.e., glucose or fructose from sucrose a n d o n e (4) specifically states t h a t glucose c a n n o t b e s e p a r a t e d f r o m sucrose using:
i o n e x c l u s i o n . H o w e v e r , p a p e r s by J o n e s et al. (11,12) a n d
C a r r u t h e r s et al. (13) d e m o n s t r a t e t h e feasibility of s e p a r a t i n g
m o n o , d i a n d t r i s a c c h a r i d e s for a n a l y t i c a l p u r p o s e s o n D o w e x
5 0 W , ( 2 % D V B , 200-400 m e s h , L i f o r m ) u s i n g c o l u m n s 100150 cm l o n g a n d 1-2 cm d i a m e t e r . T h e s e a n a l y t i c a l c o n d i t i o n s
would be impractical industrially. Cations in the sugar liquors
would displace l i t h i u m and the load a n d rate of elution are too
low.
T h e r e i s l i t t l e i n f o r m a t i o n i n a n y o f these p a p e r s t o i n d i c a t e
w h e t h e r t h e i m p u r i t i e s o t h e r t h a n salts a n d color p r e s e n t i n
s u g a r l i q u o r s , such as a m i n o acids, salts of w e a k acids, saccarides
a n d o t h e r s , m a y b e s e p a r a t e d f r o m sucrose b y ion e x c l u s i o n .
I t s e e m e d possible t h a t , i n a d d i t i o n t o s e p a r a t i n g s t r o n g salts
a n d c o l o r e l u t e d b e f o r e sucrose, o t h e r i m p u r i t i e s m i g h t b e e l u t e d
after sucrose e n a b l i n g f u r t h e r p u r i f i c a t i o n . T h i s p a p e r d e s c r i b e s
a n u m b e r o f e x p e r i m e n t s u s i n g c a n e a n d b e e t molasses t h a t
show t h e s e p a r a t i o n of s o m e molasses c o m p o n e n t s by ion exclusion.
M a t e r i a l s a n d Procedures
T h e r e s i n was D o w e x 5 0 W , 50-100 m e s h . T h e p o t a s s i u m
form of t h e e x c h a n g e r was u s e d since this f o r m of t h e r e s i n
would most nearly a p p r o x i m a t e the form that w o u l d develop
d u r i n g factory o p e r a t i o n s o n b e e t molasses. S t u d i e s c o m p a r e d
the efficiency o f X-4 a n d X-12 crosslinkages a t r o o m t e m p e r a t u r e
a n d a t 9 0 C . T w o P y r e x glass c o l u m n s w e r e j o i n e d t o c o a r s e

494

JOURNAL OF THE A. S. S. B. T.

sintered glass funnels: one for use at room temperature was


90 mm tubing 135 cm long containing 6500 ml of resin 115 cm
high; the other was 50 mm tubinsr 115 cm long jacketed for
operation at 90C and containing 1800 ml of resin 100 cm high.
Work with columns of this size is generally considered suitable
for scaling up to industrial sized units. T e m p e r a t u r e in the
second column was maintained by p u m p i n g hot water through
the jacket while loading with hot molasses and using hot water
for elution. T h e normal flow rate of 1 m l / c m 2 / m i n was controlled with a capillary outlet; minor adjustments in rate being
made by raising or lowering the effluent reservoir. Load volumes
were 15% of the resin bed volume, 975 ml for the large column
and 270 ml for the smaller column. Contraction of 5-10% in
the resin bed d u r i n g loading and subsequent expansion during
elution do not interfer with column operation. Effluent fractions equal to 1/20 of the bed volume were collected and analyzed.
T h e following general analytical methods were used: Reducing sugars were determined by Munson-Walker copper reduction. Sucrose was determined by invertase inversion followed by copper reduction. Total nitrogen was determined by
the Kjeldahl method and amino nitrogen by the Van Slyke
procedure. Ash was determined by heating at 550C and weighing the residue. Chloride was determined using an AmincoCotlove automatic chloride titrator. Color determinations were
made with a Beckman Model B spectrophotometer measuring
adsorption at 720 and 425 u\fx and reported in arbitrary units.
Experiments and Results
An attempt was made to separate sucrose from glucose at
room temperature using the large column with X-12 crosslinked
resin. A solution containing approximately 190 g each of glucose
and sucrose in 975 ml of water was loaded on the column and
eluted with water. T w o runs were made at flow rates of 1
m l / c m 2 / m i n and one-third that rate. T h e data for these runs
are presented in T a b l e 1 and Figure 1. T h i s and other figures
plot V e /V T (ratio of effluent volume to resin bed volume) against
C e / C T (ratio of effluent concentration to original concentration).
Results obtained using columns of different sizes may be compared directly on this basis. Although the separation of sucrose
from glucose was not complete, nearly 5 0 % of the sucrose was
in the first three fractions at greater than 8 0 % purity. T h e separation of salt from sucrose described by Asher (5) combined
with these results demonstrate the possibility of separating cane
molasses into a salt-rich fraction, a sucrose-rich fraction, and
finally an invert-rich fraction.

VOL.

13,

No.

6, J U L Y

495

1965

F i g u r e 1.Separation o f sucrose a n d glucose u s i n g D o w e x 5 0 W ( 1 2 %


DVB, 50-100 mesh, K form) at 2 5 C . R u n 1. L o a d : 190 g e a c h of sucrose
a n d glucose i n 975 m l s o l u t i o n . E f f l u e n t r a t e : 1 m l / s q c m / m i n . R u n 2 .
L o a d : 200 g e a c h of sucrose a n d glucose in 975 ml solution. E f f l u e n t r a t e :
i/3 m l / s q c m / m i n .
T a b l e 1.Separation of sucrose from glucose at 25C using Dowex 50 W ( K ) X-12Percent of total
Sucrose
Fraction
8
9
10
11
12
13
14
15
16
17

Total g
1

Glucose

Sucrose purity

R u n 21

Run 1

Run 2

Run 1

Run 2

2.0

2.5

19.7
26.3
23.8
13.3
8.1
4.0
2.0
0.7
0.3
188.4

19.0
25.6
26.5
15.3
6.3
3.5
1.0
0.2

0.2
0.9
6.5

0.0
0.3
4.0

16.4
22.9
21.4
16.3
9.5
4.0
1.9
188.8

13.5
27.3
33.5
17.1
3.6
0.5

92.1
95.5
80.1
59.1
36.7
27.3
19.7
17.4
14.5
15.5

99.0
98.6
86.3
65.9
35.6
15.7
17.0
21.1
32.0

Run 1

203.6

201.6

Flow rate of 1/3 m l / c m 2 / m i n

A series of e x p e r i m e n t s w e r e c o n d u c t e d u s i n g X-12 r e s i n at
25C a n d 9 0 C a n d X-4 r e s i n a t 9 0 C . D i l u t e b e e t molasses, 4 0
RDS, was l o a d e d o n t h e l a r g e c o l u m n c o n t a i n i n g X-12 r e s i n
and w a s h e d t h r o u g h w i t h w a t e r a t r o o m t e m p e r a t u r e . T h e r e sults o f t h i s e x p e r i m e n t a r e s h o w n i n T a b l e 2.1 a n d g r a p h e d
i n F i g u r e 2 . T h e c u r v e s for ash a r e o m i t t e d i n t h i s a n d o t h e r

496

JOURNAL OF THE A. S.

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T.

Figure 2.Beet molasses purification using Dowex 50W (12% DVB,


50-100 mesh, K form) at 25C. Load: 480 g 65 purity beet molasses in
975 ml solution.
T a b l e 2.1Purification of beet molasses at 25C using D o w e x 50 W ( K ) X-12.
Percent of total

Fraction

Sucrose

Nonsucrose
solids

8
9
10
11
12
13
14
15
16
Total g

2.6
16.9
24.6
23.0
16.7
9.8
4.0
1.6
0.6
262.0

3.3
15.9
21.8
21.8
18.5
11.4
4.9
1.9
0.5
132.3

Nitrogen

Amino
ni trogen

Ash

Chloride

Purity

2.0
9.7
13.6
17.7
18.8
16.9
11.9
6.9
2.5
1.2

3.3
18.8
26.8
22.8
16.0
9.1
2.6
0.5
0.0
56.4

2.2
15.5
27.2
26.2
18.1
8.3
2.1
0.3
0.0
8.4

61.5
67.8
69.1
67.6
64.2
63.1
61.8
62.6
69.9

4.7
10.6
15.7
20.4
20.2
15.2
8.2
3.7
1.3
7.9

T a b l e 2.2Purification of cane molasses at 25C with D o w e x 5 0 W ( K ) X-12


Percent of total

Fraction
8
9
10
11
12
13
14
15
16
17
Total

Sucrose

Reducing
sugars

2.9
17.2
22.2
19.1
18.1
11.7
5.7
1.7
0.8
0.5
148.7

0.6
3.4
8.5
13.4
17.1
19.0
17.0
11.3
6.3
3.5
98.9

Solids less
sucrose &
red. sugars
6.0
20.6
22.2
20.7
13.5
9.9
4.5
1.3
0.7
0.6
124.7

Nitrogen

Chloride

Purity

6.7
18.7
19 3
16.7
14.0
10.0
5.3
3.3
3.3
27
2.4

2.1
17.3
25.3
23.4
18.7
11.6
1.4
.1

35-1
46.9
47.8
42.1
44.3
35.9
27.2
16.7
IS.8
16.2

9.5

497

VOL. 13, No. 6, JULY 1965

g r a p h s s i n c e t h e ash v a l u e s c o r r e l a t e very closely w i t h t h e c h l o r i d e


values. T h e r e is o n l y a slight s e p a r a t i o n of sucrose a n d n o n sugars. T h e d a t a f r o m a s i m i l a r r u n u s i n g 490 g o f 4 0 p u r i t y
refiners c a n e molasses s h o w s essentially t h e s a m e r e s u l t s e x c e p t
t h e r e i s c o n s i d e r a b l e s e p a r a t i o n o f sucrose f r o m r e d u c i n g sugars
( T a b l e 2.2).
R u n s w e r e m a d e u s i n g X-12 a n d X-4 r e s i n s a t 9 0 C w i t h
b o t h b e e t a n d c a n e molasses. T h e r e s u l t s o f t h e r u n u s i n g b e e t
molasses o n X-12 r e s i n i n t h e s m a l l c o l u m n a r e p r e s e n t e d i n
T a b l e 3.1 a n d F i g u r e 3 . X-12 s e p a r a t e s b e e t molasses c o m p o n e n t s slightly b e t t e r a t 9 0 C t h a n a t r o o m t e m p e r a t u r e b u t
n o t sufficiently b e t t e r t o b e i n d u s t r i a l l y significant. T h e h i g h e r

Figure 3.Beet molasses purification using D o w e x 5 0 W ( 1 2 % DVB,


50-100 mesh K form) at 9 0 C . Load: 133 g 65 purity beet molasses in 270
ml solution.
Xable 3.1Purification of beet molasses at 90C using Dowex 50 W(K) X-12.
Percent of total
Nonsucrose
Amino
Ash
Fraction
Chloride Sucrose solids Nitrogen
nitrogen
Color
Purity
7
8
9
10
11
12
13
14
15
16

0.2
6.0

19.0
27.3
30.2
17.4

3.9

14.8
23.9
28.2
23.3
4.5
1.0
0.5

2.3
Total g
75.4
1
Arbitrary color units at 425 m.

0.2
.05

16.2
23.8
26.3
23.2
7.7
1.7
0.4

37.4

0.2
3.7

10.6
16.7
21.1
26.5
15.3
4.1
1.9

2.3

5.6

16.8
25.9
30.5
20.5
0.7
0.1

3.1
8.9

12.5
15.6
21.1
20.6
12.7
5.5

15.2

.4

0.6

13.0
24.5
30.0
19.3
9.1
2.0
0.9
0.6

50.71

64.8
66.9
68.4
67.0
53.8
55.6

498

JOURNAL OF THE A.

S.

S.

B.

T.

temperature slightly increases the m a x i m u m concentration of


the individual components, b u t there is little other advantage.
T a b l e 3.2 presents a similar r u n using 141 g of 40 purity cane
molasses. T h e results for cane molasses is also poor except for
the separation of reducing sugars from sucrose.
T h e most successful separations were made with X-4 resin
at 90C. R u n s were made with diluted beet or cane molasses
using the small column. T h e analytical results for the r u n
made with beet molasses are presented in T a b l e 4 and Figure 4.
Nearly 5 0 % of the sucrose was eluted at 80 purity or higher.
T a b l e 3 . 2 P u r i f i c a t i o n of c a n e molasses at 90C u s i n g D o w e x 50 W ( K ) X-12.
Percent of

Fraction

Sucrose

Reducing
sugars

Solids less
sucrose &
reducing
sugars

7
8
9
10
11
12
13
14
15
Total g

0.0
4.4
19.1
26.4
27.9
18.0
3.4
0.7
0.1
42.7

0.0
1.0
2.4
7.1
18.5
32.2
29.7
7.9
1.4
31.0

0.2
9.7
22.4
26.3
23.8
12.7
3.2
1.2
0.5
38.5

total

Chloride
7.1
23.5
29.2
28.4
11.7
0.1
2.8

Nitrogen

Color

Purity

0.4
12.1
22.5
25.0
21.4
10.7
4.2
2.3
1.6
0.6

0.2
15.9
27.8
29.3
20.3
5.4
0.5
0.3
0.3
327.6 1

0.0
31.6
46.5
47.8
44.5
34.1
12.3
8.8
7.7

Arbitrary color units at 425 m.

Figure 4.Beet molasses purification using Dowex 50W (4% DVB,


50-100 mesh, K form) at 90C. Load: 133 g 65 purity beet molasses in
270 ml solution.

VOL.

13, N o . 6, J U L Y 1965

499

Xable 4.Purification of beet molasses at 90C using Dowex 50 W(K) X-4.


Percent of total

Fraction

Sucrose

7
8
9
10
11
12
13
14
15
16
17

0.6
3.9
12.1
21.9
29.5
19.6
12-1
0.2

Total g

76.2

Chloride
0.2
1.7
5.9
11.6
16.8
22.1
29.7
12-0

2-3

Nonsucrose
solids

Nitrogen

Amino
nitrogen

Color

Ash

Purity

0.3
3-0
6.9
10.7
14.3
17.0
19-6
11.7
8.4
6.8
1.2

0.4
2.3
4.8
7.4
10.1
11.7
13.2
14.7
16.9
14.5
3.8

0.1
1.6
4.3
7.2
9.8
12-1
12-7
19.0
11.8
12.4
9-2

1.7
10.1
15.0
19.2
21.7
15.5
7.0
6.4
2.4
0.8
0.3

0.3
2.9
7.3
12.6
16.9
21-1
25.8
11.6
1.1
0.3
0.1

9.1
34.3
57.4
68.0
82.7
81.5
77.0
27.6

2-2

0.3

48.7 1

15.0

40.2

Arbitrary color units at 425 nut.

Most i m p u r i t i e s are e l u t e d before t h e m a x i m u m sucrose concentration, b u t some a p p e a r later. I m p u r i t i e s eluted later are
p r i n c i p a l l y a m i n o acids a n d o t h e r n i t r o g e n c o n t a i n i n g c o m pounds.
T h i s e x p e r i m e n t was r e p e a t e d u s i n g c a n e molasses, ( T a b l e
5 a n d F i g u r e 5). M o r e t h a n 6 5 % o f t h e s u c r o s e was e l u t e d a t
68 purity or higher. T h i s purity is a considerable i m p r o v e m e n t
over t h a t o b t a i n e d u s i n g X-12 a t 9 0 C w h e r e t h e best f r a c t i o n
h a d a 4 8 p u r i t y . I t i s likely t h a t t h e s u b s t a n c e s s h o w n i n t h e
Table 5.Purification of cane molasses at 90C using Dowex 50 W(K) X-4.
Percent of total
Solids less
sucrose &
Fraction

Sucrose

sugars

7
8
9
10
11
12
13
14
15
16
17
18

0.1
1.0
6.7
17.8
32.9
34.0
7.5
0.1

0.8
1.1
1.5
1.5
3.8
19.8
36.9
30.1
4.3

Total g

45.6

30.5

Arbitrary color units at 425 m.

sugars

Chloride

Nitrogen

Ash

Color

Purity

1.3
6.8
11.5
16.2
18.9
19.0
16.2
2-9
2.9
2.4
1.1
0.8

0.4
2.4
7.6
15.1
22.1
28.4
23.7
0.1

1.6
8.8
14.5
17.6
17.9
13.8
11.6
6.3
3.5
2.5
1.9

0.7
4.2
10.3
16.7
22.3
23.9
20.6
0.6
0.3
0.2
0.2

3.8
12-3
18.4
21.8
27.2
7.1
6.0
1.0
0.8
0.6
0.5
0.4

1.4
6.6
29.3
52.3
68.1
68.6
21.7
O.3

36.3

2.8

0.6

15.9

323.0 1

500

JOURNAL OF THE A. S. S. B. T.

Figure 5.Cane molasses purification using Dowex 50W (4% DVB,


50-100 mesh, K form) at 90C. Load: 141 g 40 purity cane molasses in
270 ml solution.

early portions of the reducing sugar curve are not reducing


sugars but higher molecular weight compounds or salts that
exhibit reducing properties. Comparison of the color curves
for the beet and cane molasses runs shows that both depart considerably from a smooth curve at fractions 12-14. This anomaly
is due to some colored substance that is excluded less from the
resin than the majority of colored compounds.
T h e difference in the nature of the nitrogen compounds
present in beet molasses from those in cane molasses is shown
by a comparison of the separation of beet and cane components
on Dowex 50 W ( 4 % DVB, 50-100 mesh, K form) at 90C.
Most of the cane molasses nitrogen compounds are eluted in the
first half of the run, b u t most beet molasses nitrogen compounds
are eluted in the later half. T h e late elution of beet molasses
nitrogen compounds is primarily due to the presence of relatively larger amounts of amino acids and betaine. T h e separation of reducing sugars from other material is quite good. More
than 7 0 % of the reducing sugars can be obtained at 78 purity
compared with an original 27 purity. More than half of the
non-reducing fraction is sucrose so that the total sugar purity
(no. 15-18) is 90.

VOL.

13,

No.

6,

JULY

1965

501

Conclusions
E x a m i n a t i o n of t h e r e s u l t s of these e x p e r i m e n t s i n d i c a t e s
t h a t X-4 r e s i n a t 9 0 C gives t h e best p u r i f i c a t i o n . U n d e r these
c o n d i t i o n s 5 0 % o f t h e sucrose i n b e e t molasses was s e p a r a t e d
a t 8 0 p u r i t y o r h i g h e r . O n a c o m m e r c i a l scale these fractions
could be r e t u r n e d to i n t e r m e d i a t e pans using liquors of this
p u r i t y for crystallization a n d sucrose recovery. F r a c t i o n 16, 77
purity, containing 1 2 % of the sugar could be r e t u r n e d to the
low r a w p a n s b u t fractions 12-13 c o n t a i n i n g 3 4 % o f t h e s u g a r
a r e o f s u c h low p u r i t y t h a t u n d e r p r e s e n t p r a c t i c e t h e y s h o u l d
n o t b e r e t u r n e d t o t h e s u g a r e n d o f t h e factory. M o s t o f t h e
s u g a r i n t h e s e fractions m a y b e r e c o v e r e d b y u s i n g t h e m t o d i l u t e
fresh molasses for t h e n e x t c o l u m n l o a d . The i m p u r i t i e s ret u r n e d t o t h e s u g a r e n d w o u l d o f c o u r s e f o r m s o m e molasses
a n d l a t e r b e recycled t h r o u g h t h e c o l u m n . U s i n g i o n e x c l u s i o n
i t w o u l d n o t b e necessary t o o b t a i n o n l y h i g h l y p u r i f i e d fractions.
H i g h p u r i t y fractions w o u l d b e r e t u r n e d t o t h e s u g a r e n d o f
t h e factory for c o n c e n t r a t i o n a n d crystallization, fractions h i g h
i n salts a n d l o w i n sucrose w o u l d b e d i s c a r d e d , a n d fractions
i n t e r m e d i a t e i n p u r i t y w o u l d b e used t o d i l u t e fresh molasses
a n d recycled o n t h e c o l u m n .
X h e r e s u l t s o f these e x p e r i m e n t s show t h a t static i o n e x c l u s i o n
c o l u m n s of D o w e x 50 W ( 4 % D V B , 50-100 m e s h , K) c a n be
used t o r e c o v e r sucrose f r o m b e e t a n d c a n e molasses a n d i n v e r t
from c a n e molasses.
O p e r a t i o n o f t h e c o l u m n s a t 9 0 C gives t h e best overall
s e p a r a t i o n o f molasses c o n s t i t u e n t s , t h e h i g h e s t c o n c e n t r a t i o n
of solids, a n d l o w e r s t h e possibility of f e r m e n t a t i o n t a k i n g p l a c e
d u r i n g purification.
Acknowledgment
X h e a u t h o r wishes t o t h a n k t h e f o l l o w i n g p e o p l e for a n alytical d e t e r m i n a t i o n s : M r . E . F . P o t t e r ( r e d u c i n g s u g a r s a n d
sucrose) a n d M r . H . W r i g h t (solids, ash, t o t a l n i t r o g e n , a n d
a m i n o n i t r o g e n ) . X h e a u t h o r i s i n d e b t e d t o t h e Spreckels S u g a r
Company a n d to California and Hawaiian Sugar Refining C o r p o r a t i o n for s a m p l e s of b e e t a n d c a n e refiners molasses.
Reference to a c o m p a n y or p r o d u c t n a m e does n o t imply
approval or recommendation of the product by the U . S . Department of Agriculture to the exclusion of others that may be
suitable.
(1)

L i t e r a t u r e Cited
W H E A T O N , R. M. a n d W. C. B A U M A N . 1953. Ion exclusion: A u n i t
o p e r a t i o n utilizing ion e x c h a n g e materials. I n d . E n g . C h e m . 4 5 :
228-233.

502

JOURNAL OF THE A. S. S. B. T.

(2) WHEATON, R. M. and W. C. BAUMAN. 1953. Non-ionic separations


with ion exchange resins. Ann. N.Y. Acad. Sci. 57: 159-176.
(3) BAUMAN, W. C, U. S. patent 2,684,331 (1954) [C.A. 48: 11853 (1954) ];
Dow Chemical Company.
(4) WHEATON, R. M., U. S. patent 2,911,362 (1959) [C.A. 54: 1946 (1960) ];
Dow Chemical Company.
(5) ASHER, D. R. 1956. Sugar purification by ion exclusion. Ind. Eng.
Chem. 48: 1465-1466.
(6) WHEATON, R. M., U. S. patent 2,890,972 (1959) [C.A. 53: 15612 (1959) ];
Dow Chemical Company.
(7) SIMPSON, D. W. and R. M. WHEATON.

1954.

Ion exclusioncolumn

analysis. Chem. Eng. Prog. 50: 45-49.


(8) SIMPSON, D. W. and W. C. BAUMAN. 1954. Concentration effects of recycling in ion exclusion. Ind. Eng. Chem. 46: 1958-1962.
(9) NORMAN, L., G. RORABAUGH and H . KELLER.

1963. Ion exclusion p u r i -

fication of sugar juices. J. Am. Soc. Sugar Beet Technol. 12: 363370.
(10) HIGGINS, I. R. U. S. patent 2,815,322 (1957) [C.A. 52: 4076 (1958)].
(11) JONES, J. K. N.; R. A. W A L L and A. O. PITTET.

1959.

sugars on ion exchange resins. Chem. and Ind.


(12) JONES, J. K. N., R. A. WALL and A. O. PITTET.

Separation of

1196.

1960.

T h e separation

of sugars on ion-exchange resins. Can. J. Chem. 38: 2285-2289.


(13)

CARRUTHERS, A., J. V. DUTTON, J. F. T. OLDFIELD, C. W. ELLIOIT, R.

K. HEANEY and H. J. TEAGUE. 1963. Estimation of sugars in beet


molasses. Sixteenth Annual Technical Conference of the British
Sugar Corp. Ltd.

Thin-Layer Chromatography of Sugar Beet


Carbohydrates
H.

G.

WALKER, JR.,
Received

B.

A.

RICCI

for publication

August

AND J .
20,

C.

GOODWIN1

1964

T h e use of thin-layer c h r o m a t o g r a p h y ( T L C ) for analysis of


sugars i n simple m i x t u r e s has b e e n r e p o r t e d b y several a u t h o r s
(3,5,6a,8) 2 . T h i s n e w t e c h n i q u e has b e e n f o u n d t o b e e x t r e m e l y
useful for q u a l i t a t i v e analysis of t h e c a r b o h y d r a t e s in c r u d e
sugar beet juices. A s a n a l t e r n a t i v e t o p a p e r c h r o m a t o g r a p h y ,
T L C offers c o m p a r a b l e i n f o r m a t i o n i n a s h o r t e r t i m e w i t h o u t
sacrificing t h e simplicity o f t h e o l d e r m e t h o d . W e believe t h a t
this n e w t e c h n i q u e m e r i t s t h e c o n s i d e r a t i o n of all in t h e b e e t
i n d u s t r y w h o have occasion t o r u n p a p e r c h r o m a t o g r a m s .
Several books a r e n o w available t h a t d e s c r i b e t h e general
t e c h n i q u e s a n d e q u i p m e n t o f T L C (1,9,11). T h i s p a p e r will
describe t h e T L C analysis of c a r b o h y d r a t e s in sugar b e e t materials t h a t w e have f o u n d simplest. I n o u r e x p e r i e n c e t h e
t e c h n i q u e s described by o t h e r w o r k e r s d i d n o t give a completely
satisfactory q u a l i t a t i v e p i c t u r e of t h e c a r b o h y d r a t e c o n t e n t of
c o m p l e x b e e t m i x t u r e s . T L C analysis o f o t h e r b e e t c o n s t i t u e n t s
such as a m i n o acids or o r g a n i c acids has n o t b e e n investigated
i n this l a b o r a t o r y , b u t w o u l d p r o b a b l y n o t b e difficult.
Materials and Methods
T h e thin-layer a p p l i c a t o r , 8 X 8-inch glass plates, p l a t e h o l d ers, a n d t a n k s used were s t a n d a r d c o m m e r c i a l i t e m s 3 .
A variety of m a t e r i a l s w e r e tested as t h i n layer a b s o r b e n t s .
N e i t h e r c o m m e r c i a l silica gel G n o r K i e s e l g u h r G p e r f o r m e d
as well as a m i x t u r e c o n t a i n i n g e q u a l p a r t s of C e l i t e Analytical
F i l t e r A i d 4 a n d a n h y d r o u s c a l c i u m sulfate (analytical grade)
buffered w i t h s o d i u m acetate. A slurry m a d e from 10 g each of
Celite a n d c a l c i u m sulfate m i x e d w i t h 50 ml of 0.02 M s o d i u m
acetate gave e n o u g h m a t e r i a l to coat five glass plates w i t h a
layer 250 thick. After c o a t i n g , t h e plates w e r e a i r d r i e d a n d
t h e n a c t i v a t e d by h e a t i n g at 125 C for one-half h o u r .
S o l u t i o n s t o b e c h r o m a t o g r a p h e d w e r e adjusted t o 8-10%
solids ( R D S ) w i t h o u t o t h e r p u r i f i c a t i o n o r t r e a t m e n t . O n e o r
1

Western Regional Research Laboratory, Western Utilization Research and Development Division, Agricultural Research Service, U. S. Department of Agriculture, Albany,
California.
2
Numbers in parentheses refer to Literature Cited.
3
Brinkman Instruments, Inc., Westbury, N. Y.; Research Specialties Co., Richmond,
California.
4
Johns Manville, New York, N. Y.

504

JOURNAL OF THE A. S. S. B. T.

two microliters of each sample was placed at the origin line


as in paper chromatography. Less disturbance of the absorbent
layer resulted when a microsyringe with a capillary wire needle
was used to apply the droplet than when a glass-tipped micropipette was used.
Chromatograms were developed by the ascending method in
tightly closed rectangular tanks lined with filter-paper wicks for
improved vapor equilibration. As long as the tank covers fit
tightly, the same solvent could be used for several days. Results
were better when equilibrating wicks were present than when
they were absent. A double development technique gave maxim u m resolution of the components of interest. Plates were
developed first in solvent A (ethyl acetate, 65; 2-propanol, 23;
water, 12) (8) and then, after air drying, in solvent B (ethyl
acetate, 55; 2-propanol, 30; water, 15). Solvent migration in each
case was 15 cm instead of the usual 10 cm. Total time for the
double development with intervening drying was about three
hours.
After development, plates were dried and sprayed with Stahl's
indicator (8) composed of 0.5 ml anisaldehyde, 0.5 ml concentrated sulfuric acid, and 9.0 ml 9 5 % ethanol. Color development was brought about by heating at 125 C for 10-15 minutes.
Results and Discussion
Although other workers have suggested different conditions,
we found that the above procedure gave o p t i m u m separation
of the carbohydrates in beet materials. W i t h the solvents chosen,
only the carbohvdrate materials move to any extent. Spraying
irrigated T L C plates with other indicators (ninhydrin or bromcresol blue) did not reveal migration of any amino acid or
organic acid components to the carbohydrate area. Figures 1
and 2 show typical chromatograms. Figure 1 shows that a mixture of pure beet carbohydrates, namely fructose, glucose, sucrose, raffinose, and galactinol distinctly separated into its component parts. We omitted kestose for lack of a suitable standard;
it would have fallen in the gap between sucrose and raffinose.
T h e separation was especially distinct when the components were
all present in about equal quantities as in D, E, F of Figure 1.
As the concentration of sucrose increased to 200-300 times that
of the other components, the resolution of the reducing sugars
became less distinct, b u t the other sugars remained unaffected.
T h i s trend can be seen in Figure 1, A, B, C.
Figure 2 shows a thin layer chromatogram of actual beet
juices. Colloidal material in raw juice (B), high pH in intermediate juices (C and D), or high ash (E and F) in final

VOL.

13,

No.

6,

JULY

1965

505

F i g u r e 1.TLC s e p a r a t i o n of p u r e sugars n o r m a l l y f o u n d in beet


molasses, (1) fructose, (2) glucose, (3) sucrose, (4) raffinose, (5) galactinol.
A, B, a n d C c o n t a i n 50, 100, 150 g respectively, of sucrose. Fructose,
glucose, raffinose, a n d galactinol a r e all c o n s t a n t at the 0.6 g level. In
D, E, a n d F, 0.6 g of each c a r b o h y d r a t e is present.

F i g u r e 2 . T L C s e p a r a t i o n of carbohydrates in actual beet juices; A,


s t a n d a r d solution c o n t a i n i n g fructose, glucose, sucrose, raffinose, a n d
galactinol; B, diffusion juice; C, first c a r b o n a t i o n juice; D, p h o s p h a t e d
thin j u i c e ; E, straight house molasses; a n d F, b a r i u m process molasses, 1.0
l of each 1 0 % R D S juice a p p l i e d at origin. Sugars a r e (1) fructose, (2)
glucose, (3) sucrose, (4) kestose, (5) raffinose, (6) galactinol, (7) a n d (8)
unknowns.

506

JOURNAL OF THE A. S. S. B. T.

molasses did not materially affect the resolution. Comparison


of factory diffusion juice (B) and first carbonation juice (C)
showed that processing did a good job in decreasing the levels
of reducing sugars and, apparently, of kestose. D was a phosphated thin juice sample (2) prepared from stored beets showing abnormally high reducing sugars and kestose. E and F were
molasses samples showing the contrast between straight house
(E) and barium saccharate process products (F).
We do not list a table of R f values for several reasons. Experience has shown that differences in plate preparation or
changes in relative concentration of components influence the
rate of migration of the components. T h e relative order of the
components remains unchanged, however, and the overall pattern
is recognizable regardless of the actual concentrations (Figures
1 and 2). A slight increase in R f values occurred as plates aged
after activation. T h e adsorbent properties of the calcium sulfate
thin layer changed gradually with prolonged exposure to atmospheric moisture and reheating in the oven did not restore
original activity. T h e changes in crystal size and shape associated
with calcium sulfate aging are complicated (4), so slight changes
in adsorbent properties were not unexpected. Nevertheless, T L C
plates gave fairly consistent R f values for several days after
preparation and, regardless of the plate's age, the characteristic
pattern of the various sugars was easily recognizable. Calcium
sulfate without Celite was not a satisfactory thin layer material
because it did not adhere to the glass very well.
T h e good separations obtained in this study seemed to depend on the use of extremely small quantities of carbohydrates.
O p t i m u m resolution was in the 0.5-3.0 g range for each component and as little as 0.1 g of most materials could be detected
by Stahl's indicator. When amounts of sugars commonly used
in paper chromatography were p u t on o u r T L C plates, it was
impossible to obtain good separations. Because such small
quantities of sugars were used in these T L C experiments, some
of the common paper chromatography sprays (7) were not sensitive enough to locate the individual components. N o r was
spraying with sulfuric acid followed by charring with heat sensitive enough. A silver nitrate (7) spray was as sensitive as
Stahl's anisaldehyde-H 2 S0 4 , b u t produced more background color.
T h e double development technique (10) described in the
experimental section was necessary for good resolution. Single
development with either solvent A or solvent B was unsatisfactory as was reversing the order of development by using solvent

VOL.

13, No. 6, JULY

507

1965

B first. In o u r o p i n i o n , t h e increased r e s o l u t i o n from d o u b l e


d e v e l o p m e n t justified t h e m o r e t i m e - c o n s u m i n g t e c h n i q u e .
S e m i - q u a n t i t a t i v e analysis of b e e t sugar c a r b o h y d r a t e s by
visual c o m p a r i s o n w i t h k n o w n s t a n d a r d s was n o t t o o a c c u r a t e .
I n o n e instance, w e h o p e d t h a t m i n o r q u a n t i t y changes i n a
series of raffinose samples c o u l d be e s t i m a t e d by this m e t h o d .
Samples c o n t a i n i n g 0.25, 0.50, 0.75, 1.00, 1.25, a n d 1.50 g
of raffinose w e r e d e v e l o p e d a n d t r e a t e d in t h e u s u a l m a n n e r . A
difference of 0.5 g/l of raffinose c o u l d be d e t e c t e d , b u t a
difference of 0.25 g/l c o u l d n o t be seen w i t h c e r t a i n t y . We
c o n c l u d e d from this a n d o t h e r e x p e r i m e n t s t h a t gross o r d e r o f
m a g n i t u d e s c o u l d b e established readily for v a r i o u s c o m p o n e n t s
by visual c o m p a r i s o n , b u t t h a t precise analysis was n o t feasible.
U n f o r t u n a t e l y , since we used considerably smaller q u a n t i t i e s of
m a t e r i a l t h a n G e e (3), i t was n o t possible t o e m p l o y h e r techn i q u e s for q u a n t i t a t i v e analysis of raffinose a n d sucrose. We
c o n t e m p l a t e f u r t h e r w o r k o n t h e use o f T L C for q u a n t i t a t i v e
analysis of b e e t c a r b o h y d r a t e s .
Very recently, P r e y a n d coworkers (6b) p u b l i s h e d a m e t h o d
for t h e q u a n t i t a t i v e d e t e r m i n a t i o n of raffinose in b e e t molasses
using T L C plates w i t h silica gel G a d s o r b e n t a n d a n acetonewater d e v e l o p e r ( 9 : 1 , vol/vol). I n t h e b e e t juices w e e x a m i n e d
by his t e c h n i q u e , t h e r e s o l u t i o n of c o m p o n e n t s was faster b u t
n o t as g o o d as we r e p o r t . Q u a n t i t a t i v e results based on visual
c o m p a r i s o n of spots a p p e a r e d to be s i m i l a r to those from o u r
method.
Summary
T h e use o f t h i n layer c h r o m a t o g r a p h y t o separate fructose,
glucose, sucrose, kestose, raffinose, a n d galactinol in b e e t juices
i s described. T h e m e t h o d i s r e c o m m e n d e d a s a n a l t e r n a t i v e t o
p a p e r c h r o m a t o g r a p h y for r a p i d q u a l i t a t i v e analysis of t h e carb o h y d r a t e s i n sugar b e e t m a t e r i a l s .
R e f e r e n c e t o a c o m p a n y o r p r o d u c t n a m e does n o t i m p l y
approval or r e c o m m e n d a t i o n of the product by the U . S . Dep a r t m e n t of A g r i c u l t u r e to t h e exclusion of o t h e r s t h a t m a y
be s u i t a b l e .

(1)

Literature Cited
BOBBITT, J. M. 1963. T h i n Layer Chromatography. Reinhold P u b lishing Corporation, New York, 208 p p .

(2)

CARRUTHERS,

(3)

assessment of beet quality. I n t . Sugar J. 58: 72-74.


G E E , M. 1962. T h i n layer chromatography of sucrose esters a n d
mixtures of raffinose a n d sucrose. J. Chromatog. 9: 278-282.

A.,

and

J.

F.

T.

OLDFIELD.

1961.

Methods

for

the

JOURNAL OF THE A. S. S. B. T.

508

(4)

(5)

(6a)

MELLOR, J. W. 1923. A Comprehensive Treatise on Inorganic and


Theoretical Chemistry, Vol. III, p. 770-773. Longmans Green
and Co., London.
PASTUSKA, G. 1961. Untersuchen uber die qualitative und quantitative
Bestimmung der Zucker mit Hilfe der Kieselgelschicht-Chromatographie. Z. Anal. Chem. 179: 427-429.
PREY,

V.,

H.

BERBALK

and

M.

KRAUSE.

1961.

Die

Dunnschicht-

Chromatographie der Kohlenhydrate. Mikrochemica Acta. 968-978.


(6b)

PREY, V.,

(7)

1964. Dunnschichtchromatographische Bestimmung der Raffinose


in Melassen. Z. Zuckerind. 14: 135-136.
SMITH, I. 1960. Chromatographic and Electrophoretic Techniques,
Vol. I, Ed. 2, 250-252. Interscience Publishers, Inc., New York,
New York.

(8)

STAHL, E., and U. KALTENBACH.

(9)
(10)
(11)

W.

BRAUNSTEINER,

R.

GOLLER and

F.

STRESSLER-BUCHWEIN.

1961. Dunnschicht-Chromatographie,

VI Mitteilung. Spurenanalyse von Zucker-gemischen auf Kieselgur


G-Schichten. J. Chromatog. 5: 351.
STAHL, E. 1962. Dunnschicht-Chromatographie. Springer-Verlag, Berlin-Gdttinger-Heidelberg, 543 pp.
TATE, M. E. and C. T. BISHOP. 1962. Thin layer chromatography of
carbohydrate acetates. Can. J. Chem. 40: 1043-1048.
TRUTER, E. V. 1963. Thin Film Chromatography. Interscience, New
York, New York. 205 pp

Production Practices A f f e c t i n g Yield and Sugar


Content of Sugar Beets Grown in Ontario, C a n a d a
1961 and 1962 1
C.

S.

BALDWIN,

C.

E.

BROADWELL

AND

J.

F.

DAVIS2

Received for publication September 8, 1964

T h i s study is a cooperative project involving p e r s o n n e l of


the F a r m e r s a n d M a n u f a c t u r e r s Beet Sugar Association, Saginaw,
Michigan, M i c h i g a n State University, East L a n s i n g , M i c h i g a n ,
Canada a n d D o m i n i o n Sugar C o m p a n y , Ltd., C h a t h a m , O n t a r i o ,
and t h e W e s t e r n O n t a r i o A g r i c u l t u r a l School, R i d g e t o w n , Ontario.
T h e objective i s t o investigate t h e p r o d u c t i o n practices c u r rently in use by O n t a r i o sugar b e e t growers a n d correlate these
practices w i t h the r e s u l t a n t yield of roots, gross p o u n d s of sugar
per acre a n d t h e p e r c e n t sugar. D a t a o n p e r c e n t sugar w e r e
available because t h e C a n a d a a n d D o m i n i o n Sugar C o m p a n y
sample each load of beets delivered by t h e farmers for sugar
content.
D a t a were collected from a p p r o x i m a t e l y 1700 farmers r e p resenting some 20,000 acres of beets. T h e d a t a were coded a n d
p u n c h e d o n I B M cards a n d t h e n analyzed t h r o u g h t h e facilities
of t h e M i c h i g a n State University a n d u n d e r d i r e c t i o n of p e r sonnel of t h e Soil Science D e p a r t m e n t .
V a r i o u s c o m p a r i s o n s a r e possible w i t h these d a t a a n d p a r ticular i m p o r t can b e placed o n t h e p e r c e n t sugar a n d gross
p o u n d s of sugar p e r acre as affected by t h e various p r o d u c t i o n
practices.
Procedure
S t a n d a r d I B M cards were used a n d t h e following i t e m s c o d e d
on t h e cards:
1.
2.
3.
4.
5.
6.

District. Three districts.


Year. 1961 to 1963.
Fieldman. Nine in total.
Soil texture. Clay and clay loams, sand and sandy loams, combinations.
Tile drained. Yes, no, partly.
Pounds seed per acre, (monogerm:: 0-0.5, 0.6-0.9, 1.0-1.4, 1.5-1.9, 2.0-2.4, 2,5-2,9,
3.0-3.4, 3.5-3.9, over 3.9.
7. Pounds seed per acre (processed). Same breakdown as monogerm.

1
Contribution of the Soil Science Department of Michigan State University, the
Western Ontario Agricultural School, and the Canada and Dominion Sugar Company, Ltd.
Authorized for publication by the Director as Journal Article No. 3390 of the Michigan
Agr. Expt. Sta., E. Lansing, Mich. Partial requirement of the senior author for fulfillment2 of the Ph.D. degree.
Graduate student in Soil Science, Michigan State University and Chief Instructor and
Extension Specialist, Western Ontario Agricultural School; and Research Supervisor, Canada
and Dominion Sugar Company; and Professor of Soil Science, Michigan State University,
respectively.

JOURNAL OF THE A. S. S. B. T.

510

8. Pounds seed per acre (whole). 0-0.9, 1.0-1.9, 2-0-2.9, 3.0-3.9, 4.0-4.9, 5.0-5.9,
6.0-6.9, 7.0-7.9, over 7.9.
9. Pounds seed per acre. Combination (whole-processed-monogerm). 0-0.5, 0.6-0.9,
1.0-1.4, 1.5-1.9, 2.0-2.4, 2.5-2.9, 3.0-3.4, 3.5-3.9, 4.0-4.5, over 4.5.
10. Previous crop (1st year). Corn, vegetables, beans, wheat, spring grain, clover,
alfalfa, sweet clover, grass sod, tobacco, beets, potatoes, others.
11. Legumes preceding years. 1st year, 2nd year, 3rd year, none.
12. Manure application (tons per acre). 1-4, 5-9, 10-14, 15-19, over 19, none.
13. Manure and year of application. 1963, 1962, 1961, 1960, 1959, 1958, 1957, none.
14. Plowing practice. Fall, spring, none.
15. Depth of plowing (inches). Less than 3.9, 4.0-5.9, 6.0-7.9, 8.0-9.9, 10.0-11.9, over
12.0, not plowed.
16. Soil test. Yes, no.
17. Soil test recommendation followed. Yes, no, partly.
18. Times worked between plowing and planting. 1, 2, 3, 4, 5, 6, 7, 8. 9, over 9.
19. Fertilizer application method. Plow down, broadcast, drill, combination.
20. Pounds fertilizer with drill (pounds per acre). None, 1-99, 100-199, 200-299, 300399, 400-499, 500-599, 600-699, 700 and over.
21. Total pounds fertilizer used (pounds per acre). None, 1-199, 200-399, 400-599,
600-799, 800-999, 1000-1199, 1200-1399, 1400 and over.
22. Fertilizer ratio used with drill. O-x-x, 1-1-1, 1-2-3, 1-3-1, 1-4-2, 1-4-4, 16-5,
1-6-3, others.
23. Nitrogen material used. Ammonium nitrate, urea, anhydrous ammonia, nitrateurea, aqua ammonia, cyanamid, others, none, combination.
24. Nitrogen application method. Pre-plant, side-dress, combination, none.
25. Time of side-dressing nitrogen. Before June 1, June 1-14, June 15-30, July 1-14,
July 15-31, August 1-14, August 15 and later, no side-dressing.
26. Total pounds per acre nitrogen used. 0-19, 20-39, 40-49, 50-59. 60-69, 70-79, 8089, 90-99, 100-119, 120 or more.
27. Total pounds per acre phosphate (P 2 O 5 ) used. 0-49, 50-74, 75-99, 100 124, 125149, 150-174, 175-199, 200-224, 225-249, 250 or more.
28. Total pounds per acre potash (K2O) used. 0-24, 25-49, 50-74, 75-99, 100-124, 125149, 150-174, 175-199, 200-224, 225 or more.
29. Date of planting, actual planting date or average planting date, e.g. Mar. 1-31,
Apr. 1-30, May 1-31, June 1-30.
30. Row width (inches). Less than 22, 22, 24, 26, 28, 30, 32. 34, 36, 38 and over.
31- Date of harvest. Before Oct. 1, Oct. 1-7, Oct. 8-14, Oct. 15-21, Oct. 22-28, Oct.
29-Nov. 4, Nov. 5-11, Nov. 12-18, after Nov. 18.
32. Minor elements. No minor elements, boron, manganese, sodium, magnesium, zinc,
others, combination.
33. Acres harvested.
34. Total tons beets.
35. Percent sugar.
36. Total tons sugar.
37. County. Eight counties.
38. Townships. Eighty townships.
39. Tons per acre beets.
40. Date planted. Before Mar. 21, Mar. 21-30, Apr. 1-10, Apr. 11-20, Apr. 21-30, Mav
1-10, May 11-20, May 21-30, May 31-June 9, after June 9.

T h i s coding required 60 spaces on the standard IBM card.


Prior to the card coding the data were recorded on standard
tabulation sheets, then submitted to the Statistical Laboratory
where the card punching and the analyses were made.
Following is a list of comparisons made:
1.
2.
3.
4.
5.
6.

Coarse-textured versus fine-textured soils.


Tile drainage.
The effect of previous crops on yields.
The effect of legumes in preceding years.
Manure applicationamount.
Manure, year of application.

VOL.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.

13, N o .

6, J U L Y

1965

511

Fall plowing versus spring plowing.


Depth of plowing.
Soil test taken.
Number of times field was worked prior to planting.
Method of fertilizer application.
Pounds of fertilizer applied with the drill.
Total pounds of fertilizer applied.
Fertilizer ratio used with the drill.
Nitrogen material used.
Method of applying additional nitrogen.
Time of side-dressing additional nitrogen.
Total pounds of nitrogen applied.
Amounts of phosphate applied.
Amounts of potash applied.
Date of planting.
Row width.
Date of harvest.
Relationship of planting date and harvest date.
Relationship of nitrogen materials and time of side-dressing of nitrogen.
Relationship of tile drainage and total pounds of fertilizer used.
Relationship of plowing practice and number of times field worked.
Relationship of plowing practice and depth of plowing.
Relationship of plowing practice and date of planting.

Results a n d Discussion
D a t a for 1961 a n d 1962 a r e p r e s e n t e d t o g e t h e r w i t h t h e
weighted averages for t h e t w o years. Unless otherwise stated,
the discussion a c c o m p a n y i n g each table p e r t a i n s to t h e w e i g h t e d
averages. A p p r o x i m a t e l y 80 p e r c e n t of t h e acreage of beets is
grown on fine-textured soils. T h e r e f o r e , possibly m o r e confidence can be placed on results from these soils. H o w e v e r , w h e r e
results from coarse-textured soils a r e p r e s e n t e d sufficient acreage
exists so t h a t r e a s o n a b l e confidence is assured.
In O n t a r i o , fall p l o w i n g is the accepted practice on t h e finet e x t u r e d soils. R e s u l t s from this survey ( T a b l e 1) t e n d to s u b stantiate this. H o w e v e r , such a practice is n o t t h e case w i t h
coarse-textured soils w h e r e s p r i n g p l o w i n g is t h e r e c o m m e n d e d
practice. T h e results, however, i n d i c a t e a similar t r e n d as on
the fine-textured soil, t h a t fall p l o w i n g is s u p e r i o r to s p r i n g
plowing insofar as t h e yield of beets a n d gross sugar a r e c o n cerned.
T h e c o m p a r i s o n of t h e d e p t h of p l o w i n g in T a b l e 2 indicates
that on b o t h t h e coarse- a n d t h e fine-textured soils t h e r e is a
tendency for t h e yield of beets a n d gross sugar to increase as
depth of p l o w i n g increases to 8 inches in d e p t h . On t h e finetextured soil this influence c o n t i n u e s to t h e 8 to 10 inch d e p t h .
The d e p t h of p l o w i n g d i d n o t affect t h e p e r c e n t sugar of roots
produced o n t h e coarse-textured soils b u t o n t h e f i n e - t e x t u r e d
soils t h e r e was a t r e n d for t h e p e r c e n t sugar to decrease w i t h
increasing d e p t h s of p l o w i n g .

Table 1.Beet yields and percent sugar as affected by time of plowing, 1961 and 1962.
1961

Plowing
practice

Acres

Sugar

Beets
T/A

Weighted average

1962

Gross
sugar
Lbs/A

Acres

Beets
T/A

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

15.88
15.69
16.34

5771
5405
5265

13,388
454
388

17.7
16.4
16.3

15.44
15.41
15.53

5475
5050
5048

15.39
15.48
15.54

5637
4893
5233

2732
1232
34

18.0
16.4
15.0

14.96
15.04
15.53

5386
4969
4658

Sugar

Gross
sugar
Lbs/A

Fine-textured soils
Fall
Spring
No

5881
182
305

17.1
15.1
16.3

14.88
14.98
15.31

5097
4519
4989

Fall
Spring
No

1148
531
16

17.5
17.1
12.9

14.36
14.45
15.51

5040
4937
4012

7507
272
83

18.2
17.2
16.1

Coarse-textured soils
1584
701
18

18.3
15.8
16.8

Table 2.Beet yields and percent sugar as affected by depth of plowing, 1961 and 1962.
1961
Depth of
plowing
inches

Acres

Beets
T/A

Weighted average

1962

Sugar

Gross
sugar
Lbs/A

Acres

Beets
T/A

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

16.07
15.79
15.62

5596
5889
5516

5205
7826
793

16.8
18.2
18.0

15.58
15.37
15.30

5285
5581
5519

15.70
15.24
16.00

4935
5716
4800

1660
2097
170

16.4
18.4
17.3

15.03
14.97
14.96

4923
5502
5177

Sugar

Gross
sugar
Lbs/A

Fine-textured soils
8.0
8.0-9.9
10.0-11.9

2365
3251
447

16.4
17.4
18.3

14.99
14.79
15.05

4912
5147
5522

8.0
8.0-9.9
10.0-11.9

777
709
156

17.2
17.6
17.5

14.26
14.44
14.87

4909
5082
5211

2840
4575
346

17.1
18.7
17.7

Coarse-textured soils
883
1388
14

15.7
18.8
15.0

VOL.

13,

No.

6, JULY

1965

513

The beneficial effect of m i n i m u m tillage is d e m o n s t r a t e d in


T a b l e 3. T h e t i m e s w o r k e d refer to t h e n u m b e r of times a field
was tilled after p l o w i n g a n d p r i o r t o p l a n t i n g . M i n i m u m tillage
is defined as t h e least a m o u n t of tillage r e q u i r e d to p r o d u c e a
satisfactory seedbed. Yields are m a i n t a i n e d at a h i g h level w h e r e
fields a r e w o r k e d only 2 or 3 times p r i o r to p l a n t i n g . T h e highest
yield of beets a n d gross sugar p e r acre occurred on fields t h a t
were w o r k e d twice. An a d d i t i o n a l a d v a n t a g e of m i n i m u m tillage
is t h e r e d u c t i o n in expense involved in seedbed p r e p a r a t i o n .
W i t h few exceptions, b e e t p l a n t i n g i n O n t a r i o occurs d u r i n g
the m o n t h s of A p r i l a n d May. F o r this survey, t e n day p l a n t i n g
intervals w e r e r e c o r d e d ( T a b l e 4). Beets p l a n t e d on coarset e x t u r e d soils d u r i n g t h e m o n t h of A p r i l p r o d u c e d a h i g h e r
yield of roots a n d gross sugar t h a n w h e r e beets w e r e p l a n t e d
at a later d a t e . T h e A p r i l 11 th-20th p e r i o d h a d t h e highest p e r cent sugar c o n t e n t .
On fine-textured soils, t h e effect of d a t e of p l a n t i n g , T a b l e 4
shows a s o m e w h a t different t r e n d t h a n t h a t for t h e coarset e x t u r e d soils. T h e average yields of beets a n d gross sugar decreased from t h e earliest p l a n t i n g d a t e to t h e latest p l a n t i n g
date. A m a r k e d decline in yield o c c u r r e d at t h e e n d of A p r i l ,
similar t o t h e coarse-textured soils. T h e p e r c e n t sugar increased
up to t h e e n d of A p r i l a n d t h e n decreased sharply after this
date. T h i s u n d o u b t e d l y a c c o u n t s for t h e lower yield of gross
sugar w h i c h o c c u r r e d at this d a t e . In O n t a r i o , t h e results of
this survey t e n d to s u b s t a n t i a t e t h e r e c o m m e n d e d practice of
p l a n t i n g beets a t a n early d a t e .
I n t h e E a s t e r n U n i t e d States a n d O n t a r i o , t h e r e have b e e n
considerable d a t a o b t a i n e d d u r i n g t h e past years o n t h e influence
of r o w w i d t h on t h e yield of beets. In O n t a r i o , t h e r e c o m m e n d a tion is to p l a n t beets in a 24-inch r o w w i d t h . T a b l e 5 illustrates
for fine-textured soils a r a t h e r m a r k e d a d v a n t a g e of n a r r o w r o w
width p l a n t i n g s , i.e. insofar as t h e yield of beets a n d gross sugar
is c o n c e r n e d . B e y o n d t h e 26-inch r o w w i d t h , t h e r e is a r a t h e r
definite r e d u c t i o n o c c u r r i n g in t h e yield of beets a n d gross sugar.
For t h e coarse-textured soils t h e same general t r e n d occurs,
but t h e r e seems to be possibly even a m o r e distinct r e d u c t i o n
i n yield b e y o n d t h e 26-inch r o w w i d t h . T h e influence o n t h e
yield of gross sugar arises from beet t o n n a g e differences r a t h e r
than from an influence on t h e p e r c e n t sugar.
On fine-textured soils ( T a b l e 6), t h e greatest response to
m e t h o d of fertilizer a p p l i c a t i o n o c c u r r e d w h e r e a c o m b i n a t i o n
of drill a n d broadcast m e t h o d s w e r e used. T h e drill m e t h o d of
fertilizer a p p l i c a t i o n refers to fertilizer t h a t was a p p l i e d at p l a n t ing t i m e , in a b a n d to o n e side a n d below the seed or directly

Table 3.Beet yields and percent sugar as affected by the number of times a field was worked prior to planting, 1961 and 1962. (Fine textured
oils)

19*2

1961

Acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

Acres

Beets
T/A

Weighted average

Sugar

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

154

19.4

14.43

5605

89

19.2

16.16

6202

243

19.3

15.06

386

16.6

14.60

4845

732

18.7

16.04

5996

1100

18.0

15.56

5611

1498

16.7

15.02

5007

1472

17.4

16.07

5595

2970

17.1

15.54

5298

1534

16.8

14.79

4976

1734

18.0

15.80

5696

3268

17.4

15.33

5358

1329

17.5

15.01

5243

2255

18.3

15.83

5780

3584

18.0

15.53

5581

864

17.2

14.90

5121

1106

18.6

15.78

5854

1970

18.0

15.39

5533

366

16.3

14.66

4791

265

17.4

15.71

5458

631

16.8

15.10

5071

5824

Table 4.Beet yields and percent sugar as affected by date of planting, 1961 and 1962.
1961

Date of
planting

Acres

Beets
T/A

Weighted average

1962

Sugar

Gross
sugar
Lbs/A

Acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

Coarse-textured soils
Apr. 1-10
April 11-20

133

20.2

13.77

5569

52

19.8

16.64

6584

185

20.1

14.58

5854

56

20.4

14.15

5796

702

18.8

15.67

5903

758

18.9

15.56

5895

56

18.9

15.12

5707

791

19.6

15.03

5905

847

19.6

15.04

5892

May 1-10

361

17.9

14.15

5062

320

13.3

15.59

4146

681

15.7

14.83

4632

May 11-20

758

16.6

14.68

4885

110

15.8

15.76

4965

868

16.5

14.82

4895

May 21-30

174

16.7

14.17

4724

68

12.4

15.86

3938

242

15.5

14.64

4503

After May 30

157

16.3

14.38

4675

260

14.5

15.33

4443

417

15.2

14.97

4530

5984

April 21-30

Fine-textured soils
Apr. 1-10

641

20.0

14.65

5871

666

19.5

15.65

6092

1307

19.8

15.16

Apr. 11-20

218

19.7

13.73

5416

3343

18.6

15.82

5872

3561

18.9

15.69

5844

Apr. 21-30

151

19.1

14.77

5633

2289

18.0

15.99

5765

2440

18.1

15.91

5757
5198

May 1-10

1068

17.5

14.94

5216

769

16.1

16.06

5173

1837

16.9

15.41

May 11-20

2801

16.4

14.90

4892

344

15.9

15.86

5041

3145

16.4

15.00

4908

May 21-30

1076

15.3

15.32

4676

62

13.4

16.27

4367

1138

15.2

15.37

4659

Table 5.Beet yields and percent sugar as affected by row width, 1961 and 1962.
1961
Row
width
inches

Acres

Weighted average

1962

Beets
T/A

Sugar
%

Gross
sugar
Lbs/A

17.1
18.9
15.6
16.1

14.82
14.89
15.38
14.39

5055
5625
4793
4627

Acres

Beets
T/A

Gross
sugar
Lbs/A

15.89
15.66
16.06
16.23

15.32
15.20
15.74

Sugar

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

5765
6018
5503
5316

9886
1899
2155

17.6
19.1
16.4
16.2

15.40
15.36
15.76
15.13

5441
5867
5190
4903

5806
5691
4564

2391

18.3
18.3
15.0

14.87
14.74
15.42

5465
5403
4625

Fine-textured soils
24
26
28
30

451?

731
951
129

5374
1168
1204

86

18.1
19.2
17.1
16.4

215

Coarse-textured soils
24
26
28

1111

230
349

17.7
17.8
16.1

14.35
14.14
14.77

5073
5028
4748

1280

300
712

18.9
18.7
14.5

530
1061

Table 6.Beet yields and percent sugar as affected by fertilizer application method. 1961 and 1962.

Acres

Beets
T/A

Weighted average

1962

Sugar

Gross
sugar
Lbs/A

Acres

Beets
T/A

Total
acres

Beets
T/A

Sugar

15.68
16.08
15.84

5571
5695
5824

2067
4720
7371

17.6
16.7
18.2

15.36
15.50
15.44

5411
5177
5630

15.67
15.89
15.25

5747
5129
5382

594
694
2703

18.0
15.8
17.7

15.39
15.26
14.84

5494
4869
5271

Sugar

Gross
sugar
Lbs/A

Fine-textured soils
923
2486
2920

17.4
15.7
18.0

14.97
14.99
14.82

5213
4711
5335

Broadcast
Drill
Combination

187
338
1163

16.7
15.7
17.9

14.77
14.59
14.30

4944
4595
5123

1144
2234
4451

17.8
17.7
18.4

<

Broadcast
Drill
Combination

Coarse-textured soils
407
356
1540

18.3
16.1
17.6

A.

Gross
sugar
Lbs/A

OURNAL OF THE

1961
Fertilizer
application
method

VOL.

13, N O . 6, J U L Y

1965

517

below the seed. T h e broadcast m e t h o d seemed to h a v e an advantage over t h e drill m e t h o d insofar as t h e yield of beets a n d
gross sugar are c o n c e r n e d . H o w e v e r , t h e p e r c e n t sugar was
n o t influenced by t h e m e t h o d of fertilizer a p p l i c a t i o n . On
coarse-textured soils t h e greatest yield response o c c u r r e d w h e n
the fertilizer was broadcast. Broadcasting the fertilizer showed
a p r o n o u n c e d increase in t h e yield of beets a n d gross sugar
over t h e drill m e t h o d a n d a lesser response over t h e c o m b i n a t i o n
m e t h o d . O n t h e coarse-textured soils the p e r c e n t sugar a p p e a r e d
to be influenced by t h e m e t h o d of fertilizer application, with
the broadcast m e t h o d s h o w i n g t h e highest p e r c e n t sugar.
T h e effect of total p o u n d s of potash p e r acre a p p l i e d on
fine-textured soils on yields of roots a n d gross sugar is i n d i c a t e d
in T a b l e 7. V a r y i n g t h e rates of potash h a d virtually no influence
on t h e p e r c e n t sugar. H o w e v e r , t h e yield of beets a n d gross
sugar increased as t h e a m o u n t of potash increased over t h e
e n t i r e r a n g e of potash a p p l i c a t i o n . In O n t a r i o , it is generally
accepted t h a t most of t h e fine-textured soils in t h e beet area
are fairly well s u p p l i e d w i t h potassium; therefore, little response
to potash fertilization is a n t i c i p a t e d .
W i t h t h e coarse-textured soils, T a b l e 7, as with t h e finet e x t u r e d soils, t h e r e seems to be a p p r o x i m a t e l y the same general
t r e n d o c c u r r i n g w i t h t h e e x c e p t i o n b e i n g t h a t possibly m i n o r
fluctuations exist in t h e p e r c e n t sugar.
A n i n t e r e s t i n g c o m p a r i s o n was o b t a i n e d o n t h e fine-textured
soils c o n c e r n i n g t h e m e t h o d of a p p l y i n g t h e n i t r o g e n fertilizer.
T h e r e s e e m e d to be little, if any, influence on t h e p e r c e n t sugar
d u e to t h e various m e t h o d s of a p p l y i n g t h e n i t r o g e n .
H o w e v e r , such was n o t t h e case w h e n t h e yield of beets a n d
gross sugar a r e considered. T h e pre-plant m e t h o d of a p p l y i n g
n i t r o g e n ( T a b l e 8) h a d a distinct a d v a n t a g e over t h e side-dress
m e t h o d or a c o m b i n a t i o n of m e t h o d s . T h e side-dress m e t h o d
had a c o n s i d e r a b l e a d v a n t a g e over t h e " c o m b i n a t i o n " m e t h o d .
On t h e coarse-textured soils t h e r e a p p e a r e d to be a r a t h e r
definite influence of t h e m e t h o d of fertilizer a p p l i c a t i o n on t h e
percent sugar. T h e same general t r e n d o c c u r r e d o n t h e coarset e x t u r e d soil as w i t h t h e fine-textured soil b u t was less well
defined.
T h e effect of t i m e of side-dressing n i t r o g e n is s h o w n in T a b l e
9. P r e s e n t r e c o m m e n d a t i o n s in O n t a r i o are t h a t n i t r o g e n s h o u l d
be side-dressed as early as possible b u t no later t h a n m i d - J u l y
due to t h e possibility of a r e d u c t i o n in sugar p e r c e n t that may
result. T h e results of this survey illustrate on fine-textured soil
that t h e r e seems to be no effect on the p e r c e n t sugar d u e to
any of t h e a p p l i c a t i o n times recorded. H o w e v e r , t h e yield of

Table 7.Beet yields and percent sugar as affected by total pounds KitO/acre, 1961 and 1962.
1961

Total Ki-0
Lbs/A

Weighted average

1962
Gross
sugar
Lbs/A

Sugar

Acres

0-24

545

15.5

14.84

4615

486

18.6

16.32

25 - 49

1657

15.7

15.11

4740

1571

17.0

16.00

50 - 74

1432

16.8

14.96

5022

1900

18.1

15.91

Acres

Beets
T/A

Sugar

Beets
T/A

Total
acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

6075

1031

17.0

15.54

5303

5431

3228

16.3

15.54

5076

5758

3332

17.5

15.50

5442

Gross
sugar
Lbs/A

Fine-textured soils

75 99

1238

18.0

14.85

5343

1375

18.1

15.77

5912

2613

18.1

15.33

5642

100 - 124

955

18.3

14.82

5433

1769

17.8

15.81

5632

2724

18.0

15.46

5562

125 149

221

17.6

14.99

5279

15.62

6227

15.38

5865

17.6

15.11

5312

20.0

15.76

6291

18.9

15.45

5825

175 or more

173

21.2

13.81

5815

19.4

15.73

6080

578
309
415

19.0

147

357
162
242

19.9

150 - 174

20.2

14.93

5970

0-24

73

14.9

14.39

4290

30

18.0

15.80

5700

103

15.8

14.80

4701

2 5 - 49

357

16.2

14.71

4768

247

18.5

13.60

5025

604

17.1

14.26

4873

50 - 74

583

17.0

14.63

4983

299

16.9

15.82

5347

882

17.0

15.03

5106

Coarse-textured soils

75 - 99

139

17.9

14.60

5230

409

17.6

16.01

5641

548

17.7

15.72

5537

100 - 124

315

18.6

13.76

5114

567

17.2

15.58

5360

882

17.7

14.93

5272

125 - 149

136

20.0

14.45

5785

395

16.5

15.37

5086

531

17.4

15.13

5265

150 - 174

31

18.1

13.79

5000

188

18.3

15.65

5728

219

18.3

15.39

5625

175 or m o r e

61

16.3

13.64

4453

95

19.8

15.33

6048

156

18.4

14.67

5424

Table 8.Beet yields and percent sugar as affected by nitrogen application method, 1961 and 1962.
1961

Nitrogen
application
method
Pre-plant
Side-dress
Combination

Pre-plant
Side-dress
Combination

Acres
980
3820

85
365
902
62

Beets
T/A

Sugar
%

19.1
16.8
17.0

18.4
17.4
16.0

Weighted average

1962

Gross
sugar
Lbs/A

Sugar
%

Gross
sugar
Lbs/A

14.77
14.97
15.26

Fine-textured soils
5638
1168
19-3
5041
5199
18.4
5178
8
16.1

15.7
15.9
16.1

6045
5863
5200

14.35
14.28
14.64

Coarse-textured soils
5285
589
18.5
4970
1333
17.5
4683
365
16.0

14.7
15.6
15.8

5451
5480
5045

Acres

Beets
T/A

Total
acres

Beets
T/A

Sugar
%

Gross
sugar
Lbs/A

2148
9019

19.2
17.7
16.9

15.28
15.51
15.33

5859
5515
5180

18.5
17.5
16.0

14.57
15.07
15.63

5387
5274
4992

93
954
2235

427

Table 9.Beet yields and percent sugar as affected by time of side-dressing of nitrogen. 1961 and 1962.

1961
Time of
side-dressing

Acres

Beets
T/A

1962

Weighted average

Sugar
%

Gross
sugar
Lbs/A

Beets
T/A

Sugar
%

Gross
sugar
Lbs/A

Fine-textured soils
5690
304
17.7
5462
2101
18.9
5005
2745
18.1
4773
106
16.4
4666

15.58
16.03
15.88
15.86

5515
6072
5758
5190

Acres

Prior to June 1st


June 1 - 14
June 1 5 - 3 0
July 1 - 14
After July 14

11
735
2315
802
18

19.1
18.4
16.8
15.6
14.8

14.90
14.81
14.91
15.34
15.75

Prior to June 1st


June 1 - 14
June 1 5 - 3 0
July 1 - 14

43
272
588
51

19.9

13.36

Coarse-textured soils
5306
152
19.9

15.37

6107

19.1
16.5
14.7

13.99
14.51
14.37

5337
4802
4239

15.87
15.55
16,38

5897
5027
5340

395
745
10

18.6
16.2
16.3

Total
acres

Beets
T/A

Sugar
%

Gross
sugar
Lbs/A

315
2836
5060
908
18

17.8
18.8
17.5
15.7
14.8

15.56
15.71
15.44
15.40
15.75

5556
5914
5413
4822
4666

195
667

19.9

14.93

5930

18.8
16.3
15.0

15.10
15.09
14.70

5669
4928
4419

1333

61

Table 10.Beet yields and percent sugar as affected by total pounds of nitrogen per acre, 1961 and 1962.
1961

Total N
Lbs/A

Acres

Beets
T/A

(Fine-textured soils)
Weighted average

1962

Sugar
%

Gross
sugar
Lbs/A

Beets
T/A

Sugar

Acres

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar
%

Gross
sugar
Lbs/A

20 or less

346

14.0

15.31

4276

307

14.9

16.00

4770

653

14.4

15.63

4508

20 - 39

820

15.5

15.01

4665

1026

15.9

16.01

5086

1846

15.7

15.74

4899

40 - 49

687

15.9

15.21

4841

554

17.4

16.14

5603

1241

16.6

15.63

5181

50 - 59

1230

17.2

15.06

5177

865

17.7

16.07

5676

2095

17.4

15.48

5383

60 69

668

17.1

14.91

5107

853

18.7

16.03

6006

1521

18.0

15.54

5611

70 - 79

814

17.6

15.08

5299

1286

188

15.85

5975

2100

18.3

15.55

5713

83 89

768

18.5

14.41

5317

809

18.2

15.67

5695

1577

18.4

15.06

5511

90 - 99

357

17.4

14.67

5111

. 670

20.6

15.69

6449

1027

19.1

15.34

5984

365

18.0

14.72

5292

812

19.1

15.77

6014

1177

18.7

15.44

5790

313

19.4

14.44

5602

680

18.3

15.70

5756

993

18.6

15.30

5707

100

119

120 or more

Table 11.Beet yields and percent sugar as affected by the total pound of fertilizer used on tile-drained land, 1961 and 1962.
1961
Total
fertilizer used
Lbs.

Acres

Beets
T/A

Sugar

Acres

Beets
T/A

Weighted average

1962
Gross
sugar
Lbs/A

Sugar

Gross
sugar
Lbs/A

Total
acres

Beets
T/A

Sugar

Gross
sugar
Lbs/A

6,

Fine-textured soils
635

14.4

15.11

4337

531

16.9

16.20

5482

1166

15.5

15.60

4858

599

2227

16.7

14.98

4992

2251

16.9

16.00

5410

4478

16.8

15.49

5202

600 - 799

1963

17.7

14.87

5264

3343

18.6

15.78

5864

5306

18.3

15.44

5642

800 - 999

685

18.9

14.83

5616

1275

20.1

15.78

6343

1960

19.7

15.45

6089

1000 - 1199

188

19.3

14.67

5670

134

18.2

16.34

5940

322

18.8

15.36

5782

4790

200 - 399
400

Coarse-textured soils
200 - 399

129

16.5

14.90

4916

47

13.9

16.05

4446

176

15.8

15.21

400 - 599

341

16.3

14.52

4739

578

16.9

1587

5358

919

16.7

15.37

5128

600- 799

481

18.3

14.54

5326

977

17.8

15.56

5538

1458

18.0

15.22

5468

800 - 999

405

19.4

13.67

5295

505

18.1

14.62

5306

910

18.7

14.20

5301

39

17.6

13.80

4856

105

18.0

15.27

5489

144

17.9

14.87

5316

1000 - 1199

JUL

522

JOURNAL OF THE A. S. S. B. T.

beets and gross sugar was affected by the time of application


of the nitrogen. W h e n nitrogen was applied later than J u n e 15,
there was a marked reduction in the yield of beets and gross
sugar. In the case of the coarse-textured soils, a lower percent
sugar in beet roots was produced than on the fine-textured
soils but there was no apparent effect on the percent sugar
due to time of application of nitrogen. Here, as with the finetextured soils, there was a sharp decline in yield of beets and
gross sugar when nitrogen was applied later than mid-June.
An interesting comparison was obtained with the total pounds
of nitrogen per acre (Table 10). According to these data there
was a definite trend of increased tonnage of beets and gross
sugar up to the level where 70-80 pounds of nitrogen were
applied per acre. Beyond this point levelling off of yields
occurred. T h e data indicate that the m a x i m u m point of beet
and sugar yield occurs somewhere in the range of a nitrogen
application between 70-100 pounds per acre. T h e r e was a slight
reduction in the percent sugar up to the level where 70-80
pounds of nitrogen were applied per acre. Beyond this level
of nitrogen the reduction in percent sugar was definitely lower.
T a b l e 11 illustrates the effect of total pounds of fertilizer
on tiled, fine-textured soil. T h e effect on the percent sugar due
to the pounds of fertilizer applied appears insignificant.
T h e yield of beets and gross sugar increased as the pounds
of fertilizer used increased up to the 800-1000 pounds per acre
level. T h e rate of tertilizer application (Table 11) influenced
the percent sugar on tile-drained, coarse-textured soils. A reduction in percent sucrose occurred beyond the 600-800 pound
per acre level. T h e yield of beets and gross sugar follows a
similar trend as for the fine-textured soils with maximums apparently at the 600-800 pound per acre level.
Summary
Several sugar beet production practices for 1961 and 1962
have been correlated with the yield of roots and gross sugar
per acre and percent sucrose. T h e yield data on the coarsetextured soils are very similar to the yields obtained on the
fine-textured soils. In general, the production practices on both
soils had marked effects on the yield of roots and gross sugar
with a somewhat lesser effect on the percent sucrose.
In interpreting results from a survey such as this, there are
many confounding factors to be considered. However, these
data provide some useful information and trends. Caution should
be used in assessing the various factors where the acreage is
small.

Host-Parasite Relations of Nacobbus


Batatiformis and the Sugar Beet and O t h e r Hosts 1
M . L . S C H U S T E R , R O B E R T SANDSTEDT AND L A R R Y W .

ESTES2

Received for publication September 16, 1964

Introduction
D e t a i l e d investigations of t h e parasitic b e h a v i o r a n d host
reaction have b e e n m a d e of Nacobbus batatiformis, T h o r n e a n d
Schuster in roots of sugar beets a n d o t h e r plants to e l u c i d a t e t h e
biology of this n e m a t o d e . T h i s p a t h o g e n causes an economically
i m p o r t a n t disease in western N e b r a s k a a n d elsewhere. Some
studies h a v e b e e n m a d e a n d r e p o r t e d in 1956 (7,9s)3 b u t a d d i tional i n f o r m a t i o n has b e e n o b t a i n e d o n histopatholosrv a n d
cytopathology t o m o r e fully u n d e r s t a n d t h e n e m a t o d e ' s activities.
T h i s p a p e r e m b o d i e s these studies.
Because t h e g e n e r a Aleloidogyne a n d Heterodera h a v e received m o r e a t t e n t i o n t h a n o t h e r g e n e r a a n d because certain
similarities exist, it seems desirable to c o m p a r e a n d contrast these
genera w i t h N. batatiformis. Studies on t h e cytopathology a n d
histopathology of Aleloidogyne a n d Heterodera in p l a n t roots
are well reviewed in t h e l i t e r a t u r e (1,4) a n d a d e t a i l e d review
will n o t b e p r e s e n t e d h e r e .
Materials and Methods
M. batatiformis used in these e x p e r i m e n t s was taken from a
p o p u l a t i o n collected in Scotts Bluff C o u n t v , N e b r a s k a . T h e s e
cultures h a d b e e n m a i n t a i n e d on Beta vulgaris L. g r o w n in
benches, flats, pots, or p e t r i dishes.
O t h e r n e m a t o d e species c o m p a r e d with N. batatiformis included
Meloidogyne
hapla C h i t w o o d , a n d Meloidogyne incognita
C h i t w o o d o b t a i n e d from infected sugar beets g r o w n in Scotts
Bluff C o u n t y , N e b r a s k a a n d m a i n t a i n e d for several years in t h e
g r e e n h o u s e on this host. Heterodera schachtii S c h m i d t was obtained from t h e same area a n d m a i n t a i n e d on g r e e n h o u s e g r o w n
sugar beets.
T h e effects of t h e n e m a t o d e s on p l a n t growth w e r e d e t e r m i n e d in g r e e n h o u s e a n d tissue cultures. In g r e e n h o u s e tests,
sugar b e e t a n d o t h e r seeds t r e a t e d with 1 0 % P u r e x (a commercial bleach c o n t a i n i n g 5 . 2 5 % N a O C l ) for 2 0 m i n u t e s w e r e
1
Published with the approval of the Director as paper No. 1603, Journal Series,
Nebraska
Agricultural Experiment Station.
2
Department of Plant Pathology, University of Nebraska, Lincoln.
3
Numbers in parentheses refer to literature cited.
Research funds for this study were contributed in part by T h e Public Health Service
Grant No. E-2033, National Institute of Allergy and Infectious Diseases and T h e Great
western Sugar Company, Denver, Colorado. We thank Monna J. Greenstreet for her technical assistance.

524

JOURNAL OF THE A. S. S. B. T.

sown in wooden flats of soil that had been steam sterilized at


250 F for 2 hours. Infections were obtained by adding to the
sterilized soil ample amounts of inoculum consisting of chopped
roots from the greenhouse subcultures. Seeds of sugar beets and
other crops were sown at the time of inoculation.
Tissue cultures for the nematode study were obtained from
root tips of sugar beets and other crops. T h e apices of excised
roots or intact seedlings were placed aseptically in petri dishes
containing White's m e d i u m as modified by Skoog and T s u i (8V
Each dish contained one excised root or intact seedling and
about 35 ml of the 1% agar medium. T h e seeds were treated
15 minutes with 1 0 % Purex, rinsed in sterilized distilled water,
and transferred to moist filter paper to germinate. W h e n the
seedling root was about 3 cm long, 1 cm of the apex was excised
and transferred to the artificial m e d i u m . Egg sacs of the nematodes were sterilized by agitating 4 minutes in a test tube containing 5 ml of 1 0 % Purex. T h i s procedure broke up the egg
sacs and surface sterilized the eggs (6). W h e n cysts were used,
the cyst was first broken and the released eggs were sterilized
by the method just described. T h e suspension of eggs was transferred into a separatory funnel containing 100 ml of sterilized
distilled water. T h e nematode eggs that settled in about 25
minutes were drawn off (with m i n i m u m liquid) through the
base of the funnel. These eggs were then transferred to the
tissue culture agar m e d i u m u p o n which were placed excised
root tips or intact seedlings of sugar beets or other plant species.
T e m p e r a t u r e s ranged from 70-75 F.
Standard histochemical methods were followed in processing
the infected roots (3). T h e samples for histological studies were
collected at periodic intervals. T h e n the roots were fixed in FAA
(90 ml 70%, ethanol, 8 ml glacial acetic acid and 4 ml formaldehyde) or F P A (90 ml 5 0 % ethanol, 8 ml propionic acid, and
4 ml formaldehyde). Dehydration a n d paraffin embedding were
accomplished in an Autotechnicon with tert-butanol as solvent.
Sections were cut 10-15 a n d stained with appropriate reagents.
T h e stains used with paraffin sections were Harris' haemotoxylin,
safranin and fast green, Johansen's q u a d r u p l e stain or Feulgen's
reagent. Johansen's q u a d r u p l e stain proved very useful in illustrating changes in the cell walls, nuclei, and cytoplasmic
contents.
For whole mounts, roots were boiled in lactophenol-acid
fuchsin, rinsed in r u n n i n g tap water to remove excess stain, then
cleared in lactophenol. Root portions containing the parasite
a n d considered suitable for histopathological study were re-

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1965

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m o v e d for f u r t h e r processing a n d paraffin e m b e d d i n g . I n o t h e r


instances, u n s t a i n e d roots w i t h easily recognizable s y m p t o m s
(galling, necrosis) w e r e e m b e d d e d in paraffin p r e p a r a t o r y to
sectioning a n d s t a i n i n g . O b s e r v a t i o n s a n d p h o t o g r a p h s o f p r e p a r e d m a t e r i a l s w e r e m a d e w i t h light, phase, o r u l t r a v i o l e t
microscopes. T i s s u e c u l t u r e s w e r e observed t h r o u g h a p h a s e
microscope e q u i p p e d w i t h s u i t a b l e objectives. O b s e r v a t i o n s of
e g g h a t c h i n g a n d larval d e v e l o p m e n t were m a d e i n BPI watch
glasses u s i n g t h e low or h i g h p o w e r objectives of t h e phase
microscope.
Experimental Results
In s t u d y i n g t h e life cycle of N. batatiformis, e x a m i n a t i o n of
h u n d r e d s of eggs d i d n o t reveal a n y m o l t i n g of t h e larvae p r i o r
t o h a t c h i n g . T h e s e eggs h a t c h soon after e m b r y o n a t i o n : t h e
larvae w e r e seen p u n c t u r i n g t h e egg c u t i c l e a n d eclosion resulted t h r o u g h a slit in t h e egg cuticle. T h e larvae m a k e a
circle or figure 8 d u r i n g m o v e m e n t in t h e egg before eclosion.
T h e larvae h a t c h i n g from t h e eggs m e a s u r e a b o u t 380 . in
length a n d a r e t h e infective stage. T h i s stage is c o n s i d e r e d t h e
first larval stage. In B P I dishes, this stage was seen to m o l t
a n d assume a l e n g t h of a b o u t 500 . H o w e v e r , t h e larval stage
does n o t take o n t h e b r o w n i s h c o l o r a t i o n f o u n d i n t h e r o o t
tissue after p e n e t r a t i o n a n d feeding. T h i s stage in t h e r o o t is
t h e " C " stage w h i c h p e r h a p s i s t h e second larval stage. T h i s
stage t h e n assumes a spiral stage a b o u t 800 in l e n g t h a n d
is c o n s i d e r e d t h e t h i r d larval stage; it has 1-2 1 / 2 coils. P r e s u m a b l v
the f o u r t h a n d a d u l t stages a r e sedentary, w h i l e t h e t h r e e early
stages a r e m o t i l e .
Sexual d i m o r p h i s m is p r o n o u n c e d in N. batatiformis. T h e
male r e t a i n s its eel-shape. In tissue c u l t u r e , an a d v a n c e d larval
stage was observed moltinsr w h i l e a t t a c h e d to t h e surface of an
excised sugar b e e t r o o t . U p o n m o l t i n g , t h e larva p r o v e d t o b e
male. M o l t i n g was similar to Pratylenchus r a t h e r t h a n to
Meloidogyne. In t h e latter, t h e m a l e is coiled in a sausage s h a p e
cuticle. T h e m a l e a p p a r e n t l y uses its stylet a n d p r e s s u r e o f its
body to release itself from t h e f o u r t h larval skin. Nacobbus is
closely r e l a t e d to Pratylenchus r a t h e r t h a n
Meloidogyne
so t h e
mode of m o l t i n g of t h e m a l e was n o t u n e x p e c t e d .
I n tissue c u l t u r e s a n d i n f i e l d a n d g r e e n h o u s e g r o w n sugar
beet roots, ( F i g u r e 1) N. batatiformis is f o u n d most c o m m o n l y
in t h e c o r t e x as c o n t r a s t e d to t h e stele for Meloidoryne a n d
Heterodera.
H y p e r t r o p h y of t h e e p i d e r m a l a n d cortical cells
occurred w i t h i n a few days after surface feeding or p e n e t r a t i o n .

526

JOURNAL OF THE A. S. S. B. T.

In tissue culture, the nematode larvae caused such pronounced


hypertrophy and associated loosening of cells of the surface and
cortical cells as to induce callus-like effects. In soil-grown roots
such effect is not evident because the tissue remains more compact. T h e epidermal cells tend to become spherical in infected
tissue culture roots; this causes a loosening of adjacent cells.
T h e nematode larvae frequently feed on root hairs and cause
hypertrophy of the tip, base or center part of the root hairs
(Figure 2). Root hairs in non-galled areas show these effects but
are usually inhibited in the galled areas.
T h e nematode was not restricted to the root tip as larvae
were found up and down the root a few days after inoculation.
Frequently the root tip would become swollen and further
apical growth inhibited due to entry and penetration through
the root tip. Apparently meristematic activity of the root tip
was inhibited, but some lateral swelling of the root tip resulted.
T h e resulting gall, pendulum-like, at the end of the root occurs
more frequently in the tissue culture than in soil-grown roots.
Often larvae were located near the base of the lateral root where
entry occurred probably through the separation of cortex and
epidermis resulting from emergence of the branch root.
T h e larvae tend to penetrate the root intracellularly. This
is noticeable from prepared sections and from destruction of
cells as evidenced by necrosis along the path of the larvae. This
latter effect is quite evident in whole mounts of roots boiled in
lactophenol acid fuchsin. Microscopically the injured areas
appear red to reddish brown. In this stain, the epidermal cell
walls in the necrotic spots appear brown as do the nuclei. T h e
cell walls in the gall stain a dark brown to purplish brown; the
walls fluoresce white u n d e r ultraviolet light. T h e larvae, while
migrating through the root, had apparently broken down end
walls and passed through rows of cells (Figure 3). T h e destruction of cells is more severe than that of Meloidogyne larvae
which migrate largely intercellularly. Intracellular migration
can be adequately demonstrated, for at times the larvae are
coiled within a single cell which reacts to its invader by thickened necrotic walls. These walls fluoresce with ultraviolet light.
T h e larvae can be seen passing through the cells with the nematode embedded in the cell causing a r u p t u r e due to the nematode's movement or increase in size.
Initially the larva orients itself in the cortex parallel to the
long root axis; its anterior may be directed toward or away from
the root tip. After the first larval stage, the coiled or "C" shaped
stages do not orient themselves to the root axis. Associated wita

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Figures 1-61.) Sugar beet root galls induced by N. batattfarmts.


1 X. 2 ) Effect of 2V. batatiformis feeding on root hairs of sugar beet root.
155 X. 3.) Intracellular penetration of root by larvae; note nucleus near
nematode's anterior. 1820 X. 4.) Cavities in small gall showing hypertrophy and increased granularity of cells. 366 X. 5.) Young root galls showing necrotic areas induced by the nematods. 105 X. 6A.) Advanced coiled
larva (300 X) in necrotic spots illustrated in Figure 5; note the heavy
concentration of fat globules in this and Figure 6B which shows anterior
portion of the coiled larva. 4000 X.

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JOURNAL OF THE A. S. S. B. T.

this " C " or coiled stages are the tunnels of broken cells in the
cortex of the young roots (Figure 4). These cavities were evident
by the presence of necrotic spots in small galls (Figure 5). T h e
necrotic spots readily took up acid fuchsin stain. Teasing of
the necrotic spots in young galls yielded usually more than one
" C " or coiled dark brown larva (Figure 6A & B). Upon sectioning of these small galls, larvae could be found in the cavities.
Breakdown of several cells results in formation of the cavity
which appears to be devoid of cellular material. T h e r e seems
to be little evidence of chemical dissolution of cell walls so
pronounced in older infections. T h e cavity is formed by mechanical tearing, pressure, and feeding on the cortical cells (Figure
7). T h e faces of the cell walls are indented as though pressure
were exerted centrifugally. T h e cavity containing the nematode
seems to enlarge as the nematode grows. Granular residues, precipitates from the cytoplasm, or excreta from the nematode are
deposited in necrotic cells or on the outer walls of the cavity.
T h e " C " and coiled larvae are gorged with globules (fat as
determined by Sudan III test) so that an oblique angle between
the anterior position and the intestine, so characteristic of the
hatched larval stage, is masked due to presence of globules and
the brownish cast of the advanced larvae.
T h e immediate reaction of root tissues to infection is necrosis
and hypertrophy. T h e first macroscopic change is the necrosis
and hypertrophy of the cortical and epidermal cells. In small
galls, 7-10 days after infection, this effect is noticeable. Sectioning of these galls shows that galling is due primarily to hypertrophy of the cortical cells. T h e n u m b e r of rows of cortical cells
was similar in galled and non-galled roots. Hypertrophy is not
confined to cells adjacent to larvae, but extends also to those
some distance away. Cells of the endodermis may show necrosis
and some hypertrophy but this effect is usually not in the central
cylinder.
Usually the female tends to lie parallel to the long axis of
the root and does not affect the vascular tissue as viewed through
bright light. W h e n the female lies perpendicular or obliquely
to the longitudinal axis, it affects the vascular tissue; with bright
light this appears more physical than chemical. Under such
circumstances, the endodermal cell walls fluoresce white as though
affected. T h e phloem and xylem elements, although separated
from the nematode by only a layer or two of cells, seem singularlv
free from necrosis, hypertrophy, and r u p t u r e . However, fluores
cent studies show that the alteration of cellulose that glows white

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529

a n d has b e e n seen to be characteristically associated w i t h n e m a tode-induced necrosis a n d cell wall changes, is f o u n d s u r r o u n d ing t h e n e m a t o d e ; fluorescence is a p p a r e n t in t h e p h l o e m a n d
in t h e x y l e m too, b u t difficult to assess in t h e x y l e m which
fluoresces strongly n a t u r a l l y . X h e cells l i n i n g the cavity in t h e
small galls increase in t h e i r v o l u m e of cytoplasm a n d g r a n u l a r
c o n t e n t . L a r g e n u m b e r s of larvae e n t e r a r o o t in t h e same
area, b u t a p p a r e n t l y they a r e dispersed because clusters of a d u l t
females a r e usually n o t observed. In contrast, several Meloidogyne
larvae sometimes occupy a single gall a n d cause a p r o p o r t i o n a l
increase in swelling up to a p o i n t . It has b e e n assumed that
the a d v a n c e d larval stages of N. batatiformis leave t h e necrotic
roots a n d infect o t h e r roots (9). X h e w o u n d s a n d necrosis interfere w i t h t h e f u n c t i o n of t h e smaller b r a n c h roots cr t h e t a p
roots. In t h e field, c o m p l e t e loss of stand d u e to N. batatiformis
occurs. P l a n t s t h a t r e m a i n n e v e r a t t a i n good g r o w t h , b u t a r e
stunted.
E s t a b l i s h m e n t of t h e n e m a t o d e in a p e r m a n e n t site w i t h its
entire body w i t h i n t h e cortex initiates a series of p h e n o m e n a .
O n e effect is t h e p r o d u c t i o n of lateral roots in t h e galled areas.
Xhis is of interest because t h e lateral roots arise from the pericycle
a n d t h e n e m a t o d e is localized in t h e cortex. X h e presence of
the n e m a t o d e seems to s t i m u l a t e m i t o t i c activity in t h e pericycle
which is s o m e distance away. S t i m u l a t i o n of lateral roots on
the galls is a characteristic m o r p h o l o g i c a l s y m p t o m of N. batatiformis. X h e n u m b e r s p e r gall r a n g e from several to over 50.
Xhese lateral roots arise from all p a r t s of the gall. In contrast,
lateral roots arise in two rows from t h e n o r m a l , diarch sugar
beet t a p r o o t ( F i g u r e 8). X h e presence of a n e m a t o d e in, a
nearby cell c a n cause t h e lateral r o o t to alter its original p a t h
or even to c o n t i n u e d o w n t h e cortex parallel to t h e vascular area.
In tissue c u l t u r e , N. batatiformis affects t h e r o o t in a way
somewhat a n a l a g o u s to t h e p r o l i f e r a t i o n of lateral roots. X h i s
effect is t h e f r e q u e n t i n d u c t i o n of stem b u d s (rootlings). X h e
buds arise from t h e pericycle in a m a n n e r similar to lateral
roots. X h e vascular tissue l e a d i n g to the b u d base is similar
in m o r p h o l o g y to t h a t in t h e lateral root. X h e leaves on these
buds h a v e typical leaf a p p e n d a g e s . X h e i n d u c t i o n of stem b u d s
may b e d u e t o a n increase i n t h e a d e n i n e / i n d o l e a c e t i c acid
ratio w h i c h is k n o w n to differentiate callus i n t o leaves a n d
stems (8). A n a t t e m p t i s b e i n g m a d e t o grow these b u d s t o
mature plants.

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JOURNAL OF THE A. S. S. B. T.

Another characteristic histological symptom induced by N.


batatiformis is the syncytium which is comparable to the giant
cell complex of Meloidogyne. T h e syncytium induced by the
feeding of N. batatiformis becomes a highly granular, deeply
staining, multinucleate, protoplasmic mass of what earlier had
been h u n d r e d of cells. It is formed by the merging of protoplasts
due to the gradual dissolution of cell walls. Despite the progressive incorporation of neighboring cells into the syncytium,
many cells still tend to retain their individuality. T h e changes
begin in a localized area and extended into surrounding tissues.
T h e syncytium is poorly delimited, merging gradually with the
normal tissue. T h e syncytium typically is located entirely within
the cortex and is bounded by the endodermis, or on occasion
by the xylem elements. In cases when the nematode is located
very near the stele, the syncytium is crescent-shaped in transection with the concave side toward the stele (Figure 9). T h e
general shape of the syncytium is oval or spindle with its long
axis parallel to the main axis of the root. T h e anterior portion
of the nematode is embedded in one end of the spindle about
one-fourth the total length of the syncytium. T h e greatest diameter of the syncytium is not necessarily closest to the nematode's stylet. Cell wall dissolution occurs most rapidly and in
greatest intensity adjacent to the nematode's head. T h e damage
to the cortex may extend a few mm from the feeding point.
T h e nematode, when in its permanent position with its head
in the syncytium, does not need to move for feeding. Anteriorly
from its stylet there appears to be a channel into the syncytium.
It may draw its nutrition via this route (Figure 19).
T h e syncytia, which result from hyperplasia and hypertrophy,
vary in size and content depending upon age and stage of development. T h e y have been observed to reach up to 3 mm in
length and about 2 mm in diameter six to eight weeks after
inoculation. T h e walls of the syncytial cells thicken. Upon dissolution certain portions dissolve differentially giving the appearance of scalariform type xylem tissue (Figure 10). Dissolution
of cell walls first causes perforations which enlarge until the
entire walls are dissolved.
In the earliest stages of syncytium development, the larva
causes characteristic symptoms which are retained later. The
initial cells become hypertrophied. In these cells, the nucleus
becomes hypertrophied and the nucleolus becomes enlarged and
measures 5 times larger than in normal adjacent cells. The
nucleolus stains a deep red in Johansen's quadruple stain and

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Figures 7-12.7.) Cavity showing cross sections of nematodes and


evidence of mechanical pressure on faces of cells lining the cavity. Polarized starch grains are shown in the three lower cells. 700 X. 8.) Transverse
section of a normal diarch sugar beet root. 400 X. 9.) Transverse section
of Nacobbus-infected sugar beet root showing crescent-shaped syncytium
with the concave side toward the stele. 420 X. 10.) Differential dissolution
of syncytial cell walls giving an appearance of scalariform xylem cells.
780 X. 11.) Multinucleate syncytial cells in sugar beet root; note the enlarged nuclei and nucleoli. 1720 X. 12..) Multinucleate syncytial cells in
spinach. 1720 X.

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JOURNAL OF THE A. S. S. B. T.

the nucleoplasm stains a dark green. T h e cytoplasm becomes


granular. Similar enlargement of nuclei and nucleoli, granularity in cytoplasm, and dissolution of cell walls extends gradually
outward from the point of origin of the syncytium. T h e syncytium
enlarges more rapidly longitudinally than radially. T h i s change
does not occur laterally or posteriorly to the nematode's body.
Understandably, the cell walls of the developing syncytium are
irregular and incomplete. T h e hypertrophied nuclei are confined
by the cell wall fragments; this individuality of many syncytial
cells is retained past the egg-laying stage of the female. Most
frequently the syncytial cells are uninucleate, but multinucleate
cells are also found (Figure 11). T h i s condition is also found
in other hosts, such as spinach (Figure 12).
T h e multinucleate condition in syncytial cells might arise
by mitotic division or by pooling of nuclei from adjacent cells.
In Nacobbus-induced syncytia, the individuality of many syncytial
cells is visible and a multinucleate condition does not arise by
pooling of nuclei. In sugar beet root galls, nucleoli are found
budding with concomitant invagination of the nuclei (Figure
13A, B). T h i s appears to be amitotic rather than mitotic division.
Although hundreds of sections have been examined, mitotic
figures have not been observed in syncytial areas. In Kochia
scoparia and Opuntia tortispina root galls, b u d d i n g nuclei are
found which are abnormally shaped and somewhat cylindrical
(Figure 14). T h i s may be interpreted as disintegration of nuclei,
except that this does not explain the multinucleate cells. Although it is the consensus of current investigators that the multinucleate condition in Meloidogyne and Heterodera infected
tissues results from the pooling of nuclei from coalescing cells,
it appears that in N. batatiformis-induced syncytia, the increase
in n u m b e r of nuclei is due to amitotic division.
T h e syncytium acts as a unit; cytoplasmic contents of the
syncytial cells differ from adjacent normal cells. In Opuntia
tortispina, druses (composed of calcium oxalate) are absent
in the syncytia, b u t present in the adjacent normal cells. Starch
induction in the syncytium is apparent in root galls of sugar
beets and other hosts (5). Starch is a very early histological
symptom occurring with the initial hypertrophy of cells and
nuclei (Figure 15). T h e starch grains at this early stage of infection stain faintly with crystal violet and are smaller than in
later stages of infection. T h e starch grains appear first in the
periphery of the nuclei (Figures 7, 13A, 16). As the syncytium
increases in size, starch is conspicuous near the feeding area of

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Figures 13-18.13A.) Nucleolus b u d d i n g with concomitant invagination of the nucleus in syncytium of sugar beet gall. 1100 X. 13B.) Similar
condition in syncytium of spinach a n d the resulting m u l t i n u c l e a t e condition d u e to amitotic divisions. 1100 X. 14.) A b n o r m a l cylindrical nuclei
in syncytium in Kochia showing densely stained areas. T h e long axes of
the nuclei are usually p a r a l l e l to t h a t of the root. 3500 X. 15.) Syncytium
in 5-day old gall showing presence of small starch granules occurring with
the initial h y p e r t r o p h y of cells, nuclei, a n d nucleoli; arrows p o i n t to starch
grains a n d transverse section of young larva. 970 X. 16.) Starch grains
are initiated in vicinity of the nuclei of affected cells; similar condition
is illustrated in Figures 7 a n d 13A. 800 X. 17.) Starch located n e a r
feeding a r e a of a d u l t female; starch granules are darkly stained bodies.
200 X. 18.) Starch g r a n u l e s a r e n u m e r o u s d u r i n g dissolution of syncytial
cell walls. 750 X.

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JOURNAL OF THE A. S. S. B. T.

the nematode (Figure 17) b u t absent in the areas posterior or


lateral to the parasite. T h i s is substantial evidence that the
nematode is directly responsible for starch induction due to
its feeding processes. Starch grains are large and n u m e r o u s
d u r i n g dissolution of synctial cell walls (Figures 18, 21). In
advanced stages of infection when the female is in the process
of egg-laying, there is either a cessation of starch synthesis or
starch carbohydrate may be utilized in reproduction (Figure
19). In tissue culture, males have been found to induce starch
b u t because of their mobility, it is difficult to determine if starch
is utilized.
Starch grains are spherical and range in size from 1-30 in
diameter in sugar beets (Figure 20A, B) and other hosts, such
as Portulaca oleracea (Figure 21). T h e starch granules do not
differ in optical properties from those that occur naturally in
other plants. T h e y are larger t h a n those found in the sugar
beet leaves in which the granules are about 0.1 . T h e symmetrical granules in the root galls have conspicuous concentric
layering a r o u n d a h i l u m when observed through oil immersion
u n d e r bright light (Figure 22A) or polarized light (Figure
22B). Birefringent crosses typical of starch grains are noted
when viewed through crossed polaroids.
General Considerations
It has been postulated that the giant cell complex caused
by Meloidogyne spp. acts as a nectary in providing nutriments
for the parasite. Initially the parasite modifies the host tissues
by physical a n d chemical means. Similarly, N. batatiformis alters
the metabolism of the cells of the syncytium to induce starch
formation in relatively large quantities. T h e galls or the syncytia
are nectaries or metabolic sinks upsetting the normal gradients
in a curious manner. T h e gravid N. batatiformis female, by inducing hypertrophy a n d hyperplasia of cortical parenchyma,
probably forces the plant to translocate a larger portion of plant
nutrients to this area and thus insures the permanently located
female a lasting n u t r i e n t supply. Not only does it appear that
nutrients are differentially directed to the gall, b u t wound reaction has been instigated and w o u n d phelloderm formation
is in progress. T h e presence of starch in the syncytium is unequivocal evidence that the nematode incites a chemical change
in the gall. T h i s fact, the progressive dissolution of cell walls,
hypertrophy of cells, nuclei, nucleoli, increase in granularity of
the cytoplasm, and necrosis suggests a strong enzymatic activity
of a type foreign to a normal root and therefore arising from
the nematode.

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Figures 19-24.19.) Anterior portion of gravid female in syncytium in


which starch is absent; note the channel in syncytium through which the
nematode may obtain its nutrients. 750 X. 20A.) Induced starch grains
are spherical and range in size from 1-30 u in sugar
beet galls; photographed with bright light. 720 X. 20B.) Same as 2 0 A photographed, with
polarized light. 720 X. 21.) Starch grains in syncytium of Portulaca
oleracea 1200 X. 22A.) Starch grains in sugar beet gall showing concentric
layering around a hilum photographed with bright light. 1250 X. 22B.)
Same as 22A photographed with polarized light;; note birefrigent crosses
typical of starch grains and concentric layering. 1250 X. 23.) Longitudinal
section of iodine-stained sugar beet gall showing presence of starch as
depicted by dark staining area in the syncytium as shown by arrows. 25 X.
24.) Giant cells caused by Meloidogyne incogntta show no starch grains
photographed with polarized light. 450 X.

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JOURNAL OF THE A. S. S. B. T.

T h e use of iodine staining might be a useful diagnostic


method for the detection of N. batatiformis in sugar beet roots
(Figure 23). Since none 01 the other gall-forming nematodes
(Figure 24) cause starch formation, the iodine stain of a dissected
gall may provide a quick method of identification. Care must
be exercised in selecting fairly young galls since starch is often
depleted in older galls. Usually all stages of gall and nematode
development can be found during the growing season.
Fluorescence studies indicate that the changes in cells affected
by N. batatiformis cause the cell walls to fluoresce similarly to
epidermal walls. Epidermal walls and wound phellogen develop
a layer of suberin. T h i s coloration may indicate that the plant
tissues react to the nematode as they would to mechanical wounding, by producing suberin deposits on the walls of the cavity
cells. T h i s would also explain the characteristic staining with
safranin of these walls, for epidermal cells exhibit this type
of reaction which is also true for necrotic cell walls and walls
of cells surrounding the nematode. Comparable studies with
M. incognita infected tissues show that these same staining reactions (bright light or fluorescent) do not occur. Since suberin
is impervious to water, its deposition would interfere with the
nematode's ability to obtain nutrients. Nacobbus ruptures cell
walls, but it is not known whether this occurs before or after
suberin deposition.
T h e coiled larval stages fluoresce a glowing white, mostly
obscuring their interior structures. Young larvae exhibit a yellowish fluorescence that does not entirely obscure the internal
structures. Stained with acid fuchsin, the nematode fluoresces
a cherry red. T h e stain is concentrated in the hypodermis and
cuticle. T h e esophageal b u l b is very dark and dense.
T h e female fluoresces yellow in unstained paraffin-embedded
material. A white fluorescing substance in a layer or two of plant
tissue cells surrounds the female and the canal that extends
posterior from her body to the outside of the gall. T h i s canal
may be the path of entry or perhaps the nematode exudes some
substance that dissolves a channel preparatory to egg-laying
T h i s channel is quite straight and narrow. T h e degree of fluorescence in the areas around the female and in the channel are
about the same, indicating that the channel has been there for
some time. T h e unstained areas that fluoresce cannot be distinguished from other adjacent cells when viewed through bright
light. T h u s , ultraviolet miscroscopy can be a tool to detect differences not observable through light microscopy.

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L i t e r a t u r e Cited
(1) CHRISTIE, J. R. 1936. T h e development of root-knot nematode galls.
Phytopathology 26: 1-22.
(2) ESAU, KATHERINE. 1960. Anatomy of seed plants. J o h n Wiley a n d Sons,
Inc., N . Y. 376 p p .
(3) JOHANSEN, D. A. 1940. Plant microtechnique. McGraw-Hill, New York.
523 p p .
(4) MANKAU, R. and M. B. LINFORD. 1960. Host-parasite relationships of
the clover cyst n e m a t o d e Heterodera trifolii Goffart. Illinois Agr.
Expt. Sta. Bull. 667.
(5)

SCHUSTER, M. L., R O B E R T SANDSTEDT, a n d LARRY W . ESTES.

formation induced by a p l a n t parasitic nematode.


1342-1343.

1964.

Science

Starch

143:

(6) SCHUSTER, M. L. and X. SULLIVAN. 1960. Species differentiation of nematodes through host reaction in tissue culture. I. Comparison of
Meloidogyne hapla, Meloidogyne incognita
incognita, a n d Nacobbus
batatiformis. Phytopathology 50: 874-876.
(7)

SCHUSTER, M. L. a n d GERALD T H O R N E .

1956.

Distribution, relation to

weeds, a n d histology of sugar beet root galls caused by Nacobbus


batatiformis X h o r n e and Schuster. J. Am. Soc. Sugar Beet Xechnol.
IX (3) : 193-197.
(8) SKOOG, F. a n d C. Xsui. 1948. Chemical control of growth a n d b u d
formation in tobacco stem segments and callus cultured in vitro. Am.
J. Bot. 35: 782-787.
(9)

X H O R N E , GERALD a n d M. L. SCHUSTER.

1956.

N a c o b b u s batatiformis n .

sp. (Nematoda:Xylenchidae), producing galls on the roots of sugar


beets a n d o t h e r plants. Proc. H e l m i n t h o l . Soc. Wash. 23 (2) : 128134.

Combining Ability in Autotriploid Sugar Beets,


Beta vulgaris L.
R. H. H E L M E R I C K , R. E. FINKNER AND C. W. DOXTATOR 1
Received for publication October 12, 1964

Introduction
Twenty-eight years ago plant breeders were presented with
a new and revolutionary breeding method (4,8) 2 . T h e discovery
that polyploid strains of plants could be produced in large
numbers through the action of the drug colchicine appeared
to be a valuable plant breeding tool.
Shortly following this discovery European and North American sugar beet geneticists and plant breeders began to produce
autotetraploid strains of sugar beets (1,12,13,14,15). T h e y found
that although these tetraploid sugar beets could be maintained
at this high polyploid level, the yield of tetraploid beets was
generally lower than the original diploid varieties (2,3). Several
North American plant breeders (2,12,15) produced and tested
limited quantities of triploid sugar beets. T h e y concluded that
there was little difference between the yield of the triploid
hybrids and diploid varieties. Peto and Boyes (12) reported
that the percentage of sugar did not decrease with increased size
of root as rapidly in triploids as in diploid beets. Perhaps because of the difficulty in producing tetraploids or the failure of
substantial success in polyploid beet breeding, further work on
polyploid beets was terminated in North America.
Such was not the case in Europe where today many of the
diploid varieties are being replaced by polyploid hybrids (7).
T h e European sugar beet breeders believe that better combination of high sucrose with high tonnage can be obtained at the
polyploid level than at the diploid level. T h e y attribute the
increase in yields to the triploid hybrids. Because male-sterile
diploids were unavailable, most of the commercial polyploid
hybrids now used in Europe are produced by planting tetraploid
and diploid seed together in the same field. T h e percentage of
triploids in the resulting commercial seed is controlled by unbalancing the parental populations. T h e commercial plantings
are actually a mixture of tetraploid, triploid and diploid plants.
T h e experiments reported in this paper were designed to
test the combining ability of pure triploid sugar beets produced
with male-sterile diploid parents. T h e hybrid populations are
1
Plant Breeder, Manager Research Station and Plant Breeder, respectively, American
Crystal
Sugar Company, Rocky Ford, Colorado.
2
Numbers in parentheses refer to literature cited.

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1965

a p p r o x i m a t e l y 100 p e r c e n t t r i p l o i d except for an occasional


d i p l o i d p l a n t w h i c h o c c u r r e d because of foreign p o l l e n or
r o g u i n g failures.
Materials and Methods
In 1960, the A m e r i c a n Crystal Sugar C o m p a n y received
twenty m u l t i g e r m strains of t e t r a p l o i d sugar beets t h r o u g h a
cooperative a g r e e m e n t w i t h K l e i n w a n z l e b e n e r Saatzucht of Einbeck, G e r m a n y . X h e s e t e t r a p l o i d p o p u l a t i o n s are coded in this
p a p e r as 61-4T1 t h r o u g h 61-4T20 w i t h 61-4T28 b e i n g t h e A m e r ican t e t r a p l o i d US 401(4n). Seven d i p l o i d m a l e sterile m o n o germ p o p u l a t i o n s were o b t a i n e d from U n i t e d States D e p a r t m e n t
of A g r i c u l t u r e sources. T a b l e I presents the code n u m b e r a n d
pedigree of these m a l e steriles. Male steriles 2, an A m e r i c a n
Crystal increase a n d 8, a U S D A increase, are believed to differ
only by the seed source.
Table 1.Code numbers and pedigrees of the seven diploid male-sterile lines used
as the female parents to produce the triploids in this paper.
Code N o .
2
3
4
5
6
7
8

Pedigree
SLC #129 MS (ACS 60-424)
C 9561-3HO
7-515 MS x 9561
F59-569HO
F59-507H1 (8-515 X 507 rr)
F59-507H2 (8-569HO x 8-507 rr)
SLC #129 MS (SLC 0166 rr ram MS)

T h e r m a l l y i n d u c e d stecklings of these t e t r a p l o i d a n d d i p l o i d
p o p u l a t i o n s w e r e o b t a i n e d from P h o e n i x , Arizona, i n t h e s p r i n e
of 1961. T h e t e t r a p l o i d a n d male-sterile p o p u l a t i o n s w e r e p l a n t e d
i n t w e n t y isolated g r o u p s a t C a n o n City, Colorado. T h e g r o u p s
were p l a n t e d t o p e r m i t each male-sterile p a r e n t t o b e b o r d e r e d
by the tetraploid pollinator, thus ensuring adequate available
pollen. T h e male-sterile p o p u l a t i o n s were r o g u e d t o male-sterile
plants i n o r d e r t o h e l p e n s u r e t h e p r o d u c t i o n o f 1 0 0 % t r i p l o i d
seed.
Each male-sterile p o p u l a t i o n was harvested separately w i t h
the r e s u l t t h a t 140 t r i p l o i d h y b r i d s were o b t a i n e d . H o w e v e r ,
sufficient t r i p l o i d seed to c o m p l e t e a diallel series was o b t a i n e d
on 16 of t h e t r i p l o i d isolations. T h e r e s u l t i n g 112 t r i p l o i d s
were tested at C l a r k s b u r g , California; R o c k y F o r d , C o l o r a d o ;
and East G r a n d Forks, M i n n e s o t a . T h e e x p e r i m e n t a l design a t
all locations was an 11 X 11 t r i p l e lattice r e p e a t e d t h r e e times.
T h e d a t a i n c l u d e d i n this p a p e r w e r e extracted from t h e larger
design a n d g r o u p e d as a r a n d o m i z e d c o m p l e t e block e x p e r i ment. N i n e r e p l i c a t i o n s w e r e used i n t h e C o l o r a d o a n d M i n nesota d a t a w h i l e only six r e p l i c a t i o n s w e r e used for t h e California d a t a .

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JOURNAL OF THE A.

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B.

T.

T h e data were analyzed using the component of variance


method in T a b l e 2. T h e variances associated with males and
females were considered a measure of general combining ability,
while the interaction of males X females was considered a measure of specific combining ability.
T a b l e 2.Analysis of variance, m e a n square e x p e c t a t i o n s a n d variance c o m p o n e n t s
used to analyze the d a t a for the three locations.
Source of
variation

Mean
square

d.f.
r-1

Replications

Test crosses

(c-1)

Females
Males
Females X males
Error
Total

Variance
components

Parameters
estimated

(f-1)
(m-l)
(f-1) (m-1)
(r-1) fm-1)
(rfm-1)

Experimental Results
T h e combination analysis of variance for the three locations
showed highly significant mean square values for all sources of
variation involving locations, T a b l e 3. T h i s reaction was expected due to the wide environmental differences between growing areas. T h e mean square values for the interaction of males
with locations wer^ greater than the interaction of females with
locations for both factors studied. Because of the magnitude
of these interactions it was necessary to analyze and discuss each
location separately.
T a b l e 3 . C o m b i n e d analysis of variance for the 112 triploid hybrids at three locations.
Source of
variation

d.f.

Tons/Acre
MS

222
30
12
180

19.06**
47.64**
23.88**
13.97**

Sucrose
MS
2.52**
7.36**
3.54**
1.64**

California Combining Ability Test


Highly significant differences existed between male and female entries indicating the presence of general combining ability
for tons per acre and sucrose percent in the California data,
Tables 4 and 5. T h e specific combining ability as measured by
the female X male interaction was highly significant for tons
per acre while specific combining ability for sucrose percent
was non-existant. T h e variance components of both yield factors
were considerably higher for males than females.

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Table 4.Analysis of variance and variance components for tons per acre at the three locations.
Source of
California
Colorado
Minnesota
variation

d.f.

M.S. 1

V.C. 2

M.S. 1

V.C. 2

Females
Males
Females x males

6
15
90

31.99**
112.43* *
23.26* *

.091
2.122
2.110

455.92**
985.52**
124.61*

2.301
13.665
2.977

1
2

M.S.*
47.99*
170.61**
22.49

V.C*
.1771
2.351
.0233

Mean square
Variance components
Table 5.Analysis of variance and variance components for sucrose percent at the three locations.
California

Source of
variation

d.f.

M.S. 1

Females
Males
Females x males

6
15
90

7.85**
14.52*0.52

V.C. 2
.0764
.3335
:0406

Colorado
M.S. 1
349**
9.97**
3.22**

Minnesota
V.C2

M.S. 1

V.C2

.0019
.1071
.2451

2.793**
2.082* *
0.632

.0150
.0865
.0090

Mean square
Variance components

T a b l e s 6 a n d 7 p r e s e n t t h e California yield data. T h e r e is


little statistical evidence t h a t w o u l d indicate reliable differences
between females crossed to the same male, however, t h e experim e n t m a y n o t h a v e b e e n precise e n o u g h to detect these differences. Statistical differences b e t w e e n males crossed to t h e same
female w e r e n u m e r o u s for b o t h yield factors. T h i s w o u l d indicate a g r e a t e r v a r i a t i o n b e t w e e n the tetraploid types crossed
with the same female t h a n b e t w e e n male-sterile types crossed
to t h e same m a l e . Statistical differences are e v i d e n t b e t w e e n
certain m a l e a n d female p a r e n t s w h e n averaged over all t h e i r
a p p r o p r i a t e testers.
Colorado C o m b i n i n g Ability T e s t
T h e analysis of v a r i a n c e a n d variance c o m p o n e n t s for t o n s
per acre a n d sucrose p e r c e n t of t h e C o l o r a d o data are p r e s e n t e d
in T a b l e s 4 a n d 5. H i g h l y significant differences b e t w e e n m a l e s
and b e t w e e n females a r e evident. T h e variance c o m p o n e n t s for
males a r e considerably h i g h e r t h a n those for females for b o t h
factors. T h i s r e a c t i o n was also a p p a r e n t in t h e California d a t a .
T h e i n t e r a c t i o n of females X males was significant at t h e 5 percent level for tons p e r acre a n d highly significant for sucrose
percent.
T h e actual yields o f t h e triploids a t Rocky F o r d a r e included i n T a b l e s 8 a n d 9 . T h e general t r e n d f o u n d i n t h e
California d a t a c o n t i n u e d . G r e a t e r variability is f o u n d b e t w e e n
tetraploids crossed to t h e same female t h a n b e t w e e n females
crossed to the same male.

542

JOURNAL OF THE A.

S.

S.

B.

T.

Table 6.Tons per acre for the 112 triploid hybrids produced from 16 tetraploid pollinator strains
and 7 cytoplasmis male-sterile types (California data).
Tetraploid
pollen
parents
6I-4T1

L.S.D.

Male-sterile diploid parents


3

2
3
4
8
9
10
11
13
14
15
16
17
19
20
28

23.22
18.40
18.32
21.00
19.74
24.82
18.10
17.89
22.28
20.45
19.30
22.69
19.69
18.58
22.08
19.43

20.34
22.12
18.70
22.47
19.03
22.59
25.40
20.73
25.02
22.48
24.37
19.71
22.98
21.30
22.61
20.49

19.99
20.06
19.52
21.04
18.54
25.31
21.47
20.86
22.96
17.75
22.86
22.08
21.14
21.89
21.36
21.84

20.47
18.79
17.30
22.45
20.04
22.87
22.47
18.81
24.48
20.77
21.30
18.89
20.10
21.78
22.41
20.65

19.48
16.89
19.94
22.74
21.55
24.02
20.16
17.71
22.35
21.01
23.41
22.46
19.13
22.82
21.18
20.73

23.74
17.43
18.39
22.18
18.56
25.85
22.76
19.14
23.50
24.03
25.64
20.20
20.42
23.50
22.45
18.77

19.41
16.73
21.91
22.60
18.45
24.71
23.28
18.10
23.28
18.03
23.79
19.44
17.44
21.31
19.20
19.43

20.95
18.63
19.15
22.01
19.41
24.31
21.94
19.03
23.41
20.64
22.95
20.78
20.12
21.59
21.64
20.19

Mean
LSD (0.05)
LSD (0.01)

20.38
2.43
3.23

21.90
3.97

21.17

20.85

20.98

NS
NS

NS
NS

21.66
4.25
5.64

20.42
4.14
5.49

21.04

NS
NS

NS

Mean

.05

.01
NS

3.24
NS
NS
NS
NS
3.85
NS
NS
NS
NS
NS
NS
NS
NS
NS

Yield of commercial check = 18.38 tons per acre


LSD

Between male-sterile means


Between tetraploid means
Between triploid means

.05 = .92
.05 = 1.39
.05 = 3.69

.01 = 1.22
01 = 1.84
01 = 4.86

Table 7.Sucrose percentages for the 112 triploid hybrids produced from 16 tetraploid pollinator
strains, and 7 cytoplasmic male-sterile types (California data).
Tetraploid
pollen
parents
61-4T1

L.S.D.

Male-sterile diploid parents

2
3
4
8
9
10
11
13
14
15
16
17
19
20
28

Mean
LSD (0.05)
LSD (0.01)

Mean

12.97
13.01
12.67
13.89
13.60
13.49
12.14
12.66
13.36
12.54
13.08
12-97
12.56
13.44
13.29
11.16

12.65
12.16
12.49
13.19
13.62
13.25
11.57
12.52
11.75
12.58
12.09
12.44
11.78
13.02
11.91
11.08

12.64
12.28
12.28
13.40
13.19
13.12
12.34
11.82
11.92
12.54
12.28
12.33
12.62
13.03
12.28
10.75

12.68
12.38
12.31
13.11
13.38
1 3.27
12.04
12.17
12.78
12.29
12.33
12.48
12.09
12.70
12.18
11.58

12.72
12.93
13.08
13.90
14.19
13.47
12.63
12.45
12.84
12.58
12.32
12.44
13.50
13.54
12.44
11.36

13.30
13.28
12.68
14.19
14.21
13.34
12.18
12-79
12.94
12.98
12.42
13.23
13.28
13.62
12.94
11.71

13.46
12.16
12-93
13.83
14.38
13.05
12.54
13.03
13.05
12.75
12.52
12.85
12.84
12.99
13 16
11.16

12.91
12.60
12.63
13.64
13.79
13.28
12.20
12.49
12.66
12.61
12.43
12.68
12.67
13.19
12.60
11.26

1293
1.09
1.45

12.38
1.03
1.37

12.43

12.49

12.92
1.05
1.40

12.73

.96
NS

12.90.
1.05
1.39

13.07

.96

1.28

.99

1.32

Yield of commercial check = 12.67 percent sucrose


LSD

Between male-sterile means


Between tetraploid means
Between triploid means

.05 = 0.25
.05 = 0.37
.05 = 0.99

.01 = 0.32
.01 = 0.49
.01 = 1.30

.05

.01
NS
NS
NS
NS
NS
NS
NS
NS

.78

105
NS
NS
NS

1.03

NS
NS
NS
NS

VOL.

13,

No.

6,

JULY

543

1965

Table 8.Tons per acre for the 112 triploid hybrids produced from 16 tetraploid pollinator strains
and 7 cytoplasmic male-sterile types (Colorado data).
Tetraploid
pollen
parents

Male-sterile diploid parents

51-4T1
2
3
4
8
9
10
11
13
14
15
16
17
19
20
28

Mean
LSD (0.05)
LSD (0.01)

L.S.O.

Mean

25.49
23.44
23.49
25.21
23.62
27.85
24.38
26.76
25.72
24.08
24.22
23.18
25.35
26.59:
24.36
29.17

25.10
25.12
24.38
24.34
22.35
27.16
27.09
26.91
26.64
26.16
25.19
25.51
27.44
28.06
25.86
27.76

24.39
22.92
24.74
24.97
23.72
26.06
27.63
25.96
26.53
22.95
26.97
25.05
26.45
22.09
25.25
28.85

25.32
24.26
24.43
24.08
23.32
25.72
24.99
26.71
25.11
26.19
28.04
21.81
24.07
25.47
25.57
26.46

27.82
23.29
25.05
25.12
22.46
27.05
26.88
26.80
27.45
25.57
25.18
24.98
28.64
26.33
25.14
26.56

28.20
23.96
26.71
25.47
22.79
28.73
28.42
28.48
27.03
23.72
27.02
24.25
26.58
26.49
27.08
27.19

25.86
22.44
22.99
24.63
22.45
25.15
28.13
23.68
24.51
19.81
27.37
24.14
24.65
26.25
23.71
29.15

26.02
23.63
24.54
24.83
22.96
26.81
26.78
26.47
26.09
24.07
26.28
24.13
26.11
25.90
25.28
27.87

25.17
2.78
3.68

25.93
3.11

25.27
3.12
4.11

25.08
3.04

25.88

26.37
3.33

24.62
3.10
4.09

25.49

NS

NS

NS
NS

NS

.05

.o1
NS
NS
NS
NS
NS
NS
NS
NS
NS

2.83

3.77
NS
NS

2.98

NS
NS
NS
NS

Yield of commercial check = 24.04 tons per acre


LSD

Between male-sterile means


Between tetraploid means
Between triploid means

.05 =. .77
.05 =
1.77
.05 = 3.10

.01 = 1.02
.01 = 1.54
.01 = 4.08

Table 9.Sucrose percentage for the 112 triploid hybrids produced from 16 tetraploid pollinator
strains and 7 cytoplasmic male-sterile types (Colorado data).
Tetraploid
pollen
parents

Male-sterile diploid parents

61-4T1
2
3
4
8
9
10
U,
13
14
15
16
17
19
20
28

Mean
LSD (0.05)
LSD (0.01)

Mean

.12-86
12.51
14.44
14.42
14.23
14.34
14.58
14.74
14.05
14.14
14.03
13.72
11.50
13.73
14.20
14.05

13.58
14.26
15.22
13.54
14.91
13.65
14.18
14.32
14.03
14.33
13.89
14.03
12.90
13.51
13.78
13.90

13.83
13.50
14.36
14.18
13.82
14.32
12.98
15.12
14.09
14.12
12.67
13.53
12.83
13.73
14.25
13-94

13.72
14.41
14.58
13.88
13.88
13.76
14.21
14.62
14.50
14.05
13.63
13.79
13.28
13.61
14.15
13.85

13.96
14.24
13.19
14.10
14.60
13.90
14.56
14.48
13.39
14.24
13.70
13.90
12.98
13.50
14.18
14.04

14.08
14.56
14.31
14.33
14.82
14.30
13.88
14.73
14.16
14.62
13.78
14.94
13.26
13.85
13.82
14.39

13.95
14.10
14.30
14.69
14.82
14.24
14.44
14.71
13.99
13.72
13.35
13.19
13.88
14.12
15.35
14.00

13.71
13.94
14.34
14.16
14.44
14.07
14.12
14.67
14.03
14.18
13.58
13.87
12.94
13.72
14.25
14.02

13.84
1.09
1.44

13.99
1.15

13.82
1.11
1.46

13.99

13.93

14.23

NS
NS

NS
NS

14.17
1.02
1.35

14.00

NS
NS

NS

Yield of commercial check = 14.63 percent sucrose


LSD Between male-sterile means
Between tetraploid means
Between triploid means

.05 = 0.23
.05 = 0.35
.05 = 0.93

.01 = 0.31
.01 = 0.46
.01 = 1.22

.05

L.S.D.
.

JO I

NS

1.13
1.08

1.51
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS

1.16

1.54
NS
NS

.93

NS

544

JOURNAL OF THE A. S. S. B. T.

Minnesota Combining Ability Test


T h e analysis of variance, mean squares and variance components for tons per acre and sucrose percent are included in
Tables 4 and 5. Statistical differences are evident between males
and between females for both yield factors. T h e general trend
that greater variation was present between males than between
females was noted in this data. Specific combining ability measured by the female X male interaction is not important in this
area for either tons per acre or sucrose percent.
T h e actual yields of the triploids are included in Tables 10
and 11. T h e same general combining ability trends were evident
at this location as was found at the other two locations.
Discussion and Conclusions
T h e variance components were higher at all locations for
the males than for females. T h i s would indicate that the malesterile types used in this study were genetically more alike than
the tetraploids. T h i s reaction should have been expected since
all the male-sterile parents except SLC # 1 2 9 originated from
Dr. McFarlane's project. All the tetraploid pollinators originated
from the Kleinwanzlebener Saatzucht Company b u t they had
been previously selected for certain diverse agronomic characteristics.
Oldemeyer and Rush (10) found the greater variation between females when studying combining ability in diploids.
Actually the magnitude of the general combining ability components of variance depend on the genetic diversity of the parental
material.
T h e predominance of non-significant differences for females
crossed to the same male and the occasional non-significant differences for males crossed to the same female would indicate the
need of more than one tester. Perhaps a series of three or four
genetically diverse (tonnage type, sugar type, etc.) male-sterile
lines with varying combining ability would better evaluate tetraploid pollinators than the male-sterile series reported in this
paper. Oldemeyer (9) suggested the use of several testers for
efficient evaluation of inbreds for combining ability. However,
statistical differences were evident between males and females
in this study when the diallel averages were compared.
T h e variance components for general combining ability are
greater than those for specific combining ability at all three
locations. T h u s a screening procedure for evaluating the tetraploid and male-sterile material using a common broad base
tester should be effective. Oldemeyer (9) reported that the correlation coefficients between a variety tester and the red beet

VOL.

13,

No.

6,

JULY

545

1965

Table 10.Ton per acre for the 112 triploid hybrids produced from 16 tetraploid pollinator strains
and 7 cytoplasmic male-sterile types (Minnesota data).
Tetraploid
pollen
parents
61-4T1
2
3
4
8
9
10
11
13
14
15
16
17
19
20
28

Mean
LSD (0.05)
LSD (0.01)

Male-sterile diploid parents

L.S.D.

___
8

Mean

16.16
15.49
16.72
16.28
15.22
17.51
15.64
16.11
16.78
15.54
16.89
15.79
16.34
16.44
16.80
15.28

16.14
14.82
15.06
14.68
15.16
16.97
16.16
16.24
16.18
15.52
17.03
15.68
15.46
16.54
16.31
15.89

14.95
15.39
14.58
16.48
13.90
15.40
15.54
15.63
15.24
16.16
17.40
15.80
15.47
15.86
15.97
15.98

14.64
14.74
14.84
16.19
14.95
16.21
15.53
15.52
16.00
15.42
17.39
16.67
15.23
16.29
15.91
16.46

15.23
14.15
15.48
16.38
14.90
15.83
16.74
14.78
15.52
15.04
16.68
15.50
15.87
16.30
16.49
15.90

16.10
15.62
15.39
16.32
14.12
16.37
15.60
15.31
15.96
15.93
17.61
14.97
16.72
16.08
15.42
15.49

15.64
14.95
16.48
16.91
15.60
16.55
15.66
16.00
16.41
14.39
17.56
16.45
16.03
16.03
15.84
15.27

15.55
15.02
15.51
16.18
14.84
16.40
15.84
15.65
16.01
15.42
17.22
15.84
15.87
16.22
16.11
15.75

16.18

15.86
1.47

15.60
1.58

15.74

15.66
1.55

NS

15.80
1.47
1.94

15.98
1.39
1.84

15.84

NS

NS
NS

NS
NS

NS

.05

.01
NS
NS

1.44

NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS

Yield of commercial check = 15.83 tons per acre


LSD

Between male-sterile means


Between tetraploid means
Between triploid means

.05 = 0.37
.05 = 0.56
.05 = 1.49

.01 = NS
.01 = 0.74
.01 = 1.96

Table 11.Sucrose percentage for the 112 triploid hybrids produced from 16 tetraploid pollinator
strains and 7 cytoplasmic male-sterile type (Minnesota data).
Tetraploid
pollen
parents
61-4T1
2
3
4
8
9
10
11
13
14
15
16
17
19
20
28

Mean
LSD (0.05)
L
SD (0.01)

Male-sterile diploid parents

L.S.D.

2
16.21
16.19
16.25
16.26
16.39
15.65
15.75
16.09
16.20
16.80
16.00
16.08
16.32
16.31
16.03
15.72

3
16.22
15.50
15.84
16.41
16.02
16.36
15.21
15.98
16.02
16.11
15.61
15.84
15.54
15.72
15.88
14.73

4
15.80
15.71
16.09
16.42
16.10
15.80
15.54
15.85
16.58
16.10
15.95
16.41
15.89
16.32
15.78
14.62

5
16.10
15.99
16.13
15.94
16.25
16.00
15.98
16.20
16.23
15.86
16.09
15.92
15.89
16.11
15.93
14.58

6
16.24
16.20
15.78
16.24
16.54
15.42
15.73
15.80
16.08
16.09
15.26
15.91
16.04
16.03
16.50
15.02

7
15.84
15.85
15.90
15.88
16.82
15.54
15.72
15.80
16.15
16.23
16.01
15.88
15.59
15.62
15-76
15.32

8
16.45
16.22
16.06
16.69
16.98
15.75
15.45
15.62
16.33
16.60
15.88
16.29
16.05
16.58
16.42
15.70

Mean
16.12
15.95
16.01
16.26
16.44
15.79
15.62
15.90
16.22
16.25
15.83
16.04
15.90
16.10
16.04
15.10

16.13

15.80

15.92

15.94

15-92

15.86

16.18

15.98

.74
.98

.74
.98

.71
.94

.67
.88

.66
NS

.63
.83

NS
NS

Yield of commercial check = 15.30 percent sucrose


LSD

Between male-sterile means


Between tetraploid means
Between triploid means

.05 = 0.17
.05 = 0.18
.05 = 0.69

.01 = 0.22
.01 = 0.34
.01 = 0 9 0

.05

.0L
NS
NS
NS
NS

.63

NS
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS

74

.98

546

JOURNAL OF THE A. S. S. B. T.

tester were sufficiently high to consider the red beet a reliable


tester parent. Others have considered the red beet cross an
adequate early test for combining ability (5, 6, 11).
A red beet tester would eliminate the need of an isolation
for each tetraploid population. Hybrids would be distinguished
at thinning by the red leaf and hypocotyl color. Preliminary
screening with the red beet and a second screening of the
selected group with a male-sterile series should thoroughly evaluate a prospective tetraploid parent. A similar program for malesterile evaluation; that is, male steriles crossed to the red beet
followed by selected male steriles crossed to a tetraploid series,
should also be effective. W i t h this combining ability information
available, specific hybrid types could be combined that should
produce a superior hybrid. However, the success of such a program would depend on the true yield of these hybrids. One
difficulty may be that the use of a common tester for both
parents may result in the selection of genetically similar types
which in turn may decrease the yield in the final cross.
T h e yields of the better triploid hybrids ranged from 5 to
25 percent better than the open-pollinated commercial varieties
depending on the location. T h e diversity of germ plasm; males
from Europe, females from the United States sources, may have
contributed to the increases in yield. However, the primary
question that must be answered before the true value of triploids
are known is: " W h a t effect does the polyploid level have in this
expression of heterosis?"
T h e types and amount of disease resistance varied in both
the tetraploid and male-sterile diploid parents. T h i s factor could
have contributed greatly to the high significance of the genotype
X location interaction. T h e variances for specific combining
ability were highly significant in the California and Colorado
data where disease complexes were different. These same interactions were insignificant in Minnesota where disease problems
are negligible. T h u s , it would appear that one of the main
factors influencing the yields of the triploids was disease resistance.
Summary
T r i p l o i d sugar beet seed was produced in isolated crossing
plots using 16 tetraploid pollinator varieties and 7 cytoplasmic
male-sterile diploid strains. Yield trials were conducted in California, Colorado and Minnesota, using the 112 possible triploid
hybrids. Estimates of general and specific combining ability
were calculated by the components of variance method for
tons per acre and sucrose percent. T h e combined analysis indicated that all the interactions with locations were highly sig-

VOL.

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6, J U L Y

1965

547

nificant for b o t h factors s t u d i e d . T h u s , local a d a p t i o n was o f


m a j o r i m p o r t a n c e i n t h e y i e l d i n g ability o f t h e h y b r i d s . I n d i v i d u a l l o c a t i o n tests i n d i c a t e t h a t specific c o m b i n i n g a b i l i t y as
m e a s u r e d by t h e m a l e X female i n t e r a c t i o n was i m p o r t a n t for
tons p e r a c r e i n C a l i f o r n i a a n d C o l o r a d o . T h e m a l e X female
i n t e r a c t i o n , for sucrose p e r c e n t was significant o n l y in C o l o r a d o .
Specific c o m b i n i n g a b i l i t y was of l i t t l e i m p o r t a n c e in M i n n e s o t a .
Differences b e t w e e n m a l e s a n d b e t w e e n females was statistically
significant at all l o c a t i o n s .
L i t e r a t u r e Cited
(1) ABEGG, F. A. 1940. T h e induction of polyploidy in Beta vulgaris L.
by colchicine treatment. Proc. Am. Soc. Sugar Beet T e c h n o l . p 118.
(2) ABEGG, F. A. 1942. Evaluation of polyploid strains derived from curly
top resistant a n d leaf spot resistant sugar beet varieties. Proc. Am.
Soc. Sugar Beet T e c h n o l . pp 309-320.
(3)

ABEGG, F . A., D E W E Y STEWART a n d G. H . COONS.

1946.

F u r t h e r studies

on sugar-beet autotetraploids. Proc. Am. Soc. Sugar Beet T e c h n o l .


pp 223-229.
(4) BLAKESLEE, A. F. a n d A. AVERY. 1937. Methods of inducing doubling
of chromosomes in plants. J. Hered. 28: 393-411.
(5) D E M I N G , G. W. 1942. Use of red garden beet in sugar beet top crosses.
Proc. Am. Soc. Sugar Beet Technol. pp 337-341.
(6)

DICKENSON, D . D . a n d D . F. PETERSON.

1956.

Results of use of t h e

red marker beet as a top cross parent. J. Am. Soc. Sugar Beet
T e c h n o l . IX (3) : 217-220.
(7) K N A P P , E. 1957. T h e significance of polyploidy in sugar beet breeding. I n t e r n a t l . Genet. Symp. Proc. 1956. 300-304.
(8) N E B E L , B. R. 1937. 1937 Mechanism of polyploidy through colchicine. N a t u r e 140: 1101.
(9) OLDEMEYER, R. K. 1954. General combining ability of sugar beet
inbreds as determined with two different top-cross testers. Proc.
Am. Soc. Sugar Beet T e c h n o l . VIII (2) : 59-63.
(10)

(11)
(12)
(13)
(14)
05)

OLDEMEYER, DONALD L .

a n d GEORGE E.

RUSH.

1960.

Evaluation

of

combining ability in self-fertile lines of sugar beets using malesterile testers. J. Am. Soc. Sugar Beet Technol. XI (2) : 175-185.
PETERSON, D. F. a n d D. D. DICKENSON. 1958. Results of divergent
selection for general combining ability. J. Am. Soc. Sugar Beet
T e c h n o l . X (1) : 60-65.
P E T O , F. H. and J. W. BOYES. 1940. Comparison of diploid a n d triploid sugar beets. C a n a d i a n J. Res. Sec. C. Bot. Sci. 18: 273-282.
RASMUSSON, J. a n d A. LEVAN. 1939. Tetraploid sugar beets from
colchicine treatments. Hereditas 25: 97-102.
SCHWANITY, F. 1938. Die herstellung polyploider rassen bei beta
rviben u n d gemusearten durch b e h a n d l u n g mit colchicin. Der
Ziichter 10: 278-279.
STEWART, D. a n d J. O. GASKILL. 1952. Results of field tests with triploid sugar beets in 1951. Am. Soc. Sugar Beet T e c h n o l . Proc.
7: 452-453.

Paired-Plant Crosses in Sugar Beets


R. H. H E L M E R I C K , R. E. FINKNER AND C. W. DOXTATOR 1
Received for publication October 12, 1964

T h e maintenance of selected genotypes is one of the main


concerns of most sugar beet breeders who are using self-sterile
material. Self-fertilized seed may be obtained from some pseudoself sterile plants providing the proper environment is maintained (5) 2 . Attempts to maintain genotypes by self fertilization
and cuttings at this station have been disappointing because of
the effort involved to preserve a limited n u m b e r of plants. T h e
incorporation of the self-fertile gene into the populations has
been considered b u t not attempted because of the difficulties
involved in producing hybrids.
A scheme for plant population improvement involving a
limited amount of controlled pollination and record keeping
was initiated in 1961. One version of the scheme was suggested
by Harland (2) although no data were reported. Lonnquist
(3, 4) reported on the results of one selection cycle. T h e method
consisted of crossing plants in pairs within a varietal population.
T h e crosses are grown in performance trials and r e m n a n t seed
of the superior crosses are composited to form the next cycle
population. T h e premises on which this scheme is based are:
1. A variety of sugar beets is composed of a wide array of
yield genotypes whose mean yield is that of the variety
itself.
2. T h e yield genotypes represented will probably show a
normal distribution with half being above and half below
the population mean.
3. Crossing plants by pairs should result in high X high,
high X low, and low X low yield genotypes in a population of about 1:2:1.
4. If such crosses are grown in replicated trials the upper
yielding 25 percent of the crosses should include a good
n u m b e r of the high X high genotype combinations. Their
selection would result in an increased frequency of favorable genes in the new population with a resulting improvem e n t in performance.
Paired-Plant Breeding Schemes
Several modifications of the breeding scheme previously discussed are possible. Figure 1 shows a scheme using mass selection.
1
Plant Breeder, Manager Research Station and Plant Breeder, respectively, American
Crystal
Sugar Company, Rocky Ford, Colorado.
2
Numbers in parentheses refer to literature cited.

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Figure 1.Modified paired-plant scheme using mass selection within


the selected p l a n t s to form the next cycle.

T h e sib p o l l i n a t e d seed is p l a n t e d in short rows the same year


that t h e yield trial results a r e o b t a i n e d . M o t h e r beets can be
harvested from all t h e sib rows p r i o r to harvesting t h e yield t r i a l .
Results of t h e yield trial will indicate which rows to f u r t h e r
analyze a n d m a k e t h e selections for the n e x t p o p u l a t i o n . S h o u l d
e n v i r o n m e n t a l c o n d i t i o n s p e r m i t , t h e results of t h e yield trial
could be o b t a i n e d , first, a n d only t h e selected n u r s e r y rows d u g
for m o t h e r beet analysis. T h i s cycling m e t h o d should utilize
the a d d i t i v e genetic variance p r e s e n t in sugar beet varieties.
F i g u r e 2 shows t h e p a i r e d - p l a n t scheme designed to utilize
specific c o m b i n i n g ability a n d the non-additive g e n e effects.
T h i s modification is designed to follow the first scheme after
the a d d i t i v e g e n e t i c effects a r e utilized to their fullest. P a i r e d
plants o b t a i n e d at r a n d o m a r e crossed with male-sterile tester
plants. T h e yield trials i n d i c a t e t h e b e t t e r sib pairs w h i c h can
be f o r m e d i n t o synthetic p o p u l a t i o n s for future cross to t h e
male sterile, utilized as c o m m e r c i a l varieties or crossed by family
pairs t o t h e m a l e sterile. T h e latter scheme w o u l d p r o b a b l y b e
the m o r e p r o d u c t i v e because it w o u l d e n a b l e t h e b r e e d e r to
select w i t h i n t h e p r o g e n y of sib pairs a n d d e v e l o p a specific
hybrid.

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JOURNAL OF THE A. S. S. B. T.

Figure 2.Modified paired-plant scheme using male sterile top cross


to evaluate pairs.

Materials a n d Methods
T h e open-pollinated variety selected to be the male parent
was 61-307 an American #2 type. T w o male-sterile lines, F59507HI and the F 1 of 515 X C 9561, were chosen as the female
parents. T h e parental populations were sent to Phoenix, Arizona
in 1960 and stecklings were returned for planting in the spring
of 1961. Fifty pairs of each male sterile were paired with one
h u n d r e d pairs of 61-307. Because of faulty bolting caused by
severe hail only thirty-five pairs crossed to F59-507H1 and
twenty-one pairs crossed to 515 X C 9561 set sufficient quantities
of seed for yield trials. T h e seed from the female plants was
included in yield trials and the seed of the sib pairs was grown
in nursery rows d u r i n g the summer of 1962. Stands in the yield
trials were erratic, thus the data for tons per acre were adjusted
by regression to a uniform stand.
T h e pairing and crossing was accomplished by using plastic
covered cages. These cages are approximately twenty-four inches
square and five and one-half feet high. T h e tops are open to
prevent a heat build-up within the cage. Some crossing between
cages is possible b u t it was felt to be negligible.

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Experimental Results
T h e results r e p o r t e d i n this p a p e r are n o t conclusive a s t h e
d a t a for a c o m p l e t e cycle are n o t available. H o w e v e r , it was
thought desirable to report on the methods and the data available
to d a t e .
T a b l e 1 shows the m e a n s of the yield factors for t h e t w o m a l e
steriles, G r o u p I (F59-507H1), G r o u p II (515 X C 9561) a n d
t h e p a r e n t a l p a r e n t (61-307). N o statistical differences w e r e detectable b e t w e e n these p o p u l a t i o n means. T h u s , n o expression
of heterosis w o u l d be evident, if t h e o p e n p o l l i n a t e d p o p u l a t i o n ,
61-307, w e r e crossed w i t h these m a l e steriles. H o w e v e r , e x a m i n a t i o n of t h e i n d i v i d u a l pairs w i t h i n each g r o u p indicates considerable heterosis for c e r t a i n pairs. T h e highest p a i r was 4 . 6 %
b e t t e r t h a n t h e p a t e r n a l p a r e n t for sucrose p e r c e n t w h i l e t h e
t o n n a g e p a i r was 2 2 . 9 % b e t t e r t h a n the p a t e r n a l p a r e n t .
Table 1.Means of the yield factors for the two male-steriles (Group I and Group II)
and the paternal parent (61-307).
Yield factors
Lbs. sugar per acre
Tons per acre
Percent sucrose
n

Group I

Group II

6199
20.62
15.03
35

5853
19.72
14.81
21

Parent
6420
20.60
15.60

T a b l e 2 shows t h e n u m b e r of sib pairs that were in t h e high


X h i g h g r o u p based on t h e 1:2:1 hypothesis. L a r g e r p o p u l a t i o n s
of t h e sib pairs w o u l d h a v e given a greater l a t i t u d e for selection.
T h e use of a p a t e r n a l p a r e n t t h a t gave an i n d i c a t i o n of heterosis
when crossed to a specific m a l e sterile w o u l d also have increased
the possibility of yield increases d u e to certain specific pairs
giving h i g h e r yields.
Table 2.Number of sib pairs in the high x high class for yield factors.
Yield factors
Lbs. sugar per acre
Tons per acre
Percent sucrose
Tonnage + Sugar

Group I

Group II

7
4
7
0

5
5
3
2

C h i S q u a r e values w e r e calculated to test t h e 1:2:1 r a t i o of


the low X low, low X h i g h , a n d h i g h X high hypothesis ( T a b l e
3). T h e e x p e c t e d values for t h e calculation of t h e C h i S q u a r e
values w e r e o b t a i n e d by d i v i d i n g t h e r a n g e of yields for a particular factor i n t o f o u r t h s a n d c o m b i n i n g t h e t w o m i d d l e
quartiles. T h e observed w e r e o b t a i n e d b y c o u n t i n g t h e n u m b e r
of pairs t h a t fell w i t h i n each q u a r t i l e . As shown in T a b l e 3, all
Chi S q u a r e values e x c e p t t h e c o m b i n e d g r o u p for tons p e r a c r e

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JOURNAL OF THE A. S. S. B. T.

fit a 1:2:1 ratio. T h e hypothesis of normal distribution appears


to be functioning in the paired plant method.
Table 3.Chi square values calculated for goodness of fit to a 1:2:1 hypothesis ratio
proposed for random pairs crossed to male-sterile testers for several yield factors.
Yield factors
Lbs. sugar per acre
Tons per acre
Percent sucrose
1

Group I

Group II

3.44
3.50
1.67

0.05
0.01
0.96

Combined
groups I & II
4.33
9.671
3.76

Failed to fit a 1:2:1 ratio

5% Rejection level 5.99

Discussion
T h e population cycling data if available would be the conclusive evidence that the paired system was either working or
not working in sugar beets. T h e cycles were attempted by planting the sib seed of the pairs and taking selected mother beets
from the higher yielding pairs. For two years curly top in the
mother beets prevented the production of seed in the second
cycle, thus most of the material was subsequently lost. Since
then, the selected pairs have been propagated as stecklings, a
method that appears to be working.
T h e need of maintaining the genotype in any hybrid program
is of prime importance. T h e effort involved to obtain a small
quantity of seed on a few plants by self pollination stimulated
the search for a more adaptable method. Sib pollination is an
alternative method of genotype maintenance and has been used
to maintain sugar beet inbred lines (1). Self pollination being
quite severe fixes the genotype quite rapidly. Sib pollination
fixes the genotype at a slower rate enabling the plant breeder
to observe more combinations of genotypes while selecting the
better ones. T w o other advantages of sib pollination are reduced plant deterioration connected with self pollination and
the production of sufficient quantities of seed for yield trials
and disease nurseries.
T h e necessity of having homozygous material to produce
heterotic responses is debatable. However, a certain amount of
genotype fixing is necessary in order to approximately duplicate
each plant in the hybrid population. Hybrids selected by the
proposed cycling methods should be good yielders because of
the selection and progeny testing in each cycle and the utilization
of additive and nonadditive gene action. T h e reduced genotype
fixing involved in sib matings should give a greater yield flex
ibility in a genotype X environmental situation.

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1965

Sib p a i r i n g for preservation of type " O " beets as suggested


by O w e n (6) has b e e n used in t h e male-sterile p r o g r a m of
A m e r i c a n Crystal. T h e g e n o t y p e m a i n t e n a n c e o f t h e p r i m a r y
" O " type screening has b e e n d o n e mainly b y steckling reversion
a n d self p o l l i n a t i o n . T h e secondary screenings a n d purification
of t h e semi " O " types have b e e n d o n e by sib m a t i n g w i t h substantial success. H y b r i d i z a t i o n of " O " types a n d selection w i t h i n
t h e i r segregating p r o g e n y u s i n g t h e p a i r system described in
F i g u r e 2 is in t h e c o m b i n i n g ability phase of this p r o g r a m at
the present.
T h e only d a t a t h a t a r e available on a c o m p l e t e cycle of a
sib m a t i n g system comes from t h e Nebraska C o r n Project. L o n n quist (4) r e p o r t s t h e results of o n e cycle of paired-plant crosses
from t h e synthetic variety K r u g . T h i s synthetic h a d previously
been subjected to t h r e e cycles of r e c u r r e n t selection a n d t h r e e
increases (Syn-3) from t h e KIII selection. Plants of t h e p a r e n t
KIII Syn-3 w e r e crossed by pairs, yields o b t a i n e d a n d t h e 13 h i g h
yielding a n d 13 low y i e l d i n g pairs selected. At t h e same t i m e
the f o u r t h cycle of d i v e r g e n t r e c u r r e n t selection was b e i n g cond u c t e d . T h e results o f this e x p e r i m e n t are shown i n T a b l e 4 .
T h e 13 selected h i g h pairs exceeded t h e yield of t h e p a r e n t by
7.6 bushels a n d t h e f o u r t h cycle synthetic by 4.5 bushels. T h e
low p a i r e d selection was also m o r e effective in l o w e r i n g t h e
yields t h a n t h e r e c u r r e n t selection cycle. F r o m these d a t a it
w o u l d a p p e a r that p a i r - p l a n t selection was effective in corn.
Xable 4.Performance of selected high and low yielding parental paired-plant crosses
together with the syn-2 generation synthetic resulting from their intercrossesLincoln, 1959
(Lonnquist).
Population
Parental KIII Syn-3
Selected High 13 Crosses
Kiv (c) High Yiell Syn-2
Selected Low 13 Crosses
Kiv ( C ) LOW Yield Syn-2

Acre
yield
88.6
96.2
91.7
76.4
83.2

T h e i m p r o v e m e n t of a cross p o l l i n a t e d species is d e p e n d e n t
on t h e a b i l i t y of t h e p l a n t b r e e d e r to select on t h e basis of a d d i tive g e n e effects. T h e success of h y b r i d i z a t i o n is believed to be
largely d u e to one's ability to capitalize on n o n a d d i t i v e g e n e
effects over a n d a b o v e t h e c h a n c e a d d i t i v e p o r t i o n of t h e selected
lines. Cycling m e t h o d s of a c c u m u l a t i n g these a d d i t i v e effects
such as r e c u r r e n t selection a n d p a i r e d crossing w o u l d give t h e
hydridizer s u p e r i o r source p o p u l a t i o n s from which to d e v e l o p
lines.

JOURNAL OF THE A. S. S. B. T.

554

Summary
Methods of using paired-plant crosses were discussed. O n e
cycling method is designated to accumulate the additive genetic
effects. T h e second method which follows the first is designed
to capitalize on heterosis and the non-additive genetic effects.
Preliminary data pertaining to the expected frequency of
low X low, low X high, and high X high combinations from
pair crosses are presented.
Literature Cited
(1) DOXTATOR, C. W. and A. W. SKUDERNA. 1946. Crossing experiments in
sugar beet lines. Proc. Am. Soc. Sugar Beet Technol. pp 230-236.
(2) HARLAND, S. C. 1946. A new method of maize improvement. Trip.
Agr. 23: 114.
(3) LONNQUIST, JOHN H. 1960. El mejoramiente de las poblaciones de maiz.
Improvement of corn populations. 6a Reunion Centroamericana.
Departmento de Divulgacion Tecnica. Inst. Nal. de Inv. Agric,
S. A. G. pp 14-22.
(4) LONNQJUIST, JOHN H. 1961. Progress from recurrent selection procedures for the improvement of corn populations. Neb. Agr. Expt.
Sta. Bull. 197 July.
(5) OLDEMEYER, R. K. and H. L. BUSH. 1952. Influence of geographical
locations and type of bag on selfed-seed setting for sugar beets.
Proc. Am. Soc. Sugar Beet Technol. pp 360-363.
(6) OWEN, F. V. 194?. Utilization of male sterility in breeding superioryielding sugar beets. Proc. Am. Soc. Sugar Beet Technol. pp 158161.

Sugar Beet Breeding Lines Combining Resistance


To Bolting and Disease
J.

S.

M C F A R L A N E AND

I.

O.

SKOYEN1

Received for publication December y, 1964

D u r i n g t h e past ten years t h e California sugar b e e t i n d u s t r y


has witnessed an almost c o m p l e t e change from o p e n - p o l l i n a t e d
to h y b r i d sugar b e e t varieties. H y b r i d s are p r o d u c e d by crossing
a cytoplasmic male-sterile seed-bearing p a r e n t with a pollenfertile p a r e n t . T h e seed-bearing p a r e n t may b e t h e male-sterile
e q u i v a l e n t of an i n b r e d ; an F 1 h y b r i d between t h e male-sterile
line a n d a second i n b r e d ; or the male-sterile e q u i v a l e n t of an
o p e n - p o l l i n a t e d line. T o i n s u r e c o m p l e t e m a l e sterility, t h e seedb e a r i n g p a r e n t m u s t be d e v e l o p e d from b r e e d i n g lines w i t h t h e
T y p e O characteristic (3) 2 . T h e p o l l e n p a r e n t can be an i n b r e d ,
an o p e n - p o l l i n a t e d line, or an F 1 h y b r i d between two i n b r e d s .
T h e F 1 p o l l i n a t o r is p r o d u c e d t h r o u g h the use of M e n d e l i a n
male sterility (4).
A m a j o r objective of t h e sugar beet b r e e d i n g p r o g r a m at
the U. S. A g r i c u l t u r a l Research Station, Salinas, California, has
b e e n to d e v e l o p b r e e d i n g lines that possess characteristics n e e d e d
in h i g h - p e r f o r m i n g h y b r i d varieties. P a r t i c u l a r a t t e n t i o n has
been given to T y p e O i n b r e d s a n d m a l e steriles c o m b i n i n g resistance t o b o l t i n g a n d curly top. Selections have b e e n m a d e
for d o w n y - m i l d e w a n d virus-yellows resistance. T h e m o n o g e r m
character has b e e n i n c o r p o r a t e d i n t o h i g h - p e r f o r m i n g seed-bearing p a r e n t s . T h e s e b r e e d i n g lines have been m a d e available to
sugar beet breeders i n t h e U n i t e d States a n d C a n a d a t h r o u g h
the Beet Sugar D e v e l o p m e n t F o u n d a t i o n .
D e s c r i p t i o n of B r e e d i n g L i n e s
B r e e d i n g lines m a d e available to sugar beet b r e e d e r s from
the U. S. A g r i c u l t u r a l Research Station i n c l u d e b o t h open-pollinated selections a n d i n b r e d lines. T h e o p e n - p o l l i n a t e d selections
are all m u l t i g e r m . T h e i n b r e d s i n c l u d e b o t h r n u l t i g e r m a n d
m o n o g e r m lines. T h e b r e e d i n g lines are designated by a comb i n a t i o n o f letters a n d n u m b e r s . T h e letter " C " refers t o California, " N B " t o n o n b o l t i n g , " H " t o cytoplasmic m a l e sterility,
and " M " t o M e n d e l i a n m a l e sterility. I n t h e " C " series t h e f i r s t
n u m e r a l refers t o t h e year o f seed p r o d u c t i o n . T h e l e t t e r " C "
and t h e f i r s t n u m e r a l h a v e b e e n o m i t t e d from t h e d e s c r i p t i o n s
of t h e Fi m o n o g e r m h y b r i d s .
1
Research Geneticist and Research Agronomist, respectively, Crops Research Division,
Agricultural
Research Service, U. S. Department of Agriculture.
2
Numbers in parentheses refer to literature cited.

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JOURNAL OF THE A. S. S. B. T.

Open-Pollinated
Selections
C361.A T y p e O, bolting resistant selection from US 22/3.
T h e line was developed by making two successive T y p e O selections, followed by selections for bolting resistance (1). C361
is similar to US 75 in curly-top and bolting resistance. It has
been used to produce the male-sterile equivalent of C361 and
as a Type O, self-sterile breeding line.
C361HO.A male sterile produced by crossing C361 to a malesterile plant from US 56 and backcrossing four times to C361.
Increases of this male sterile have been used as the seed-bearing
parent to produce commercial hybrid varieties.
C366.The second successive bolting-resistant selection from
US 35/2. T h i s selection is superior to US 75 in sucrose percentage b u t inferior in root yield. It is similar to US 75 in
curly-top and bolting resistance. C366 has been used as a topcross parent in producing commercial hybrid varieties.
C663.A pollen parent derived from a cross between US 15
and US 22/3. Selections for curly-top resistance, bolting resistance, and high sucrose content were made in the F 2 and F 3
generations. Curly-top resistance is similar to that of US 75
and bolting resistance to that of US 56. Increases of C663 have
been used extensively as the top-cross parent in commercial hybrid
varieties.
C585.A bolting-resistant selection from C361. T h i s selection
combines very good bolting resistance with good curly-top resistance and has been used to produce the male-sterile equivalent
of C585.
C585HO.A bolting-resistant selection from 361 HO combining
very good bolting resistance with good curly-top resistance. Increases of C585HO have been used as the seed-bearing parent
to produce commercial hybrid varieties.
C586.A bolting-resistant selection from C366 combining very
good bolting resistance with good curly-top resistance (2). C586
has been used to replace C366 as a top-cross parent.
C671.A composite of T y p e O selections made at Salinas, California, from US 75, US 22/4, and US 56/2. T h e line has moderate
resistance to bolting and curly top. C671 has been widely used
as a T y p e O breeding line.
C681M.A Mendelian male sterile of C366 which segregates
approximately 50 percent aa. C681M has been used to produce
four-way hybrids such as (MS of NB1 X NB3) X (C681M
X NB4).
C787.A bolting-resistant selection from US 75. T h e selection
is similar in performance and disease resistance to US 75. C787

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557

has b e e n used as a p o l l e n p a r e n t to p r o d u c e t h e c o m m e r c i a l
h y b r i d (MS of N B 5 X N B 6 ) X C787.
C951.A T y p e O selection from C366, used only as a b r e e d i n g
line.
C952.A T y p e O selection from US 15, used only as a b r e e d i n g
line.
C953.A T y p e O selection from t h e E u r o p e a n Klein E variety,
used only as a b r e e d i n g line.
C264.A bolting-resistant selection from C663. Tests have shown
C264 to be s u p e r i o r to C663 in b o l t i n g resistance b u t similar
in o t h e r characteristics. C264 is b e i n g used as a r e p l a c e m e n t
for C663.
C 3 3 0 . T h e fifth successive selection from US 75 for resistance
to virus yellows. D a m a g e from virus yellows is only a b o u t onehalf as severe in C330 as in US 75. T h i s selection is similar to
US 75 in o t h e r characteristics. C330 is b e i n g used as t h e p o l l e n
p a r e n t in p r o d u c i n g e x p e r i m e n t a l q u a n t i t i e s of h y b r i d seed.
C321.A yellows-resistant selection from t h e T y p e O, selfsterile line C 6 7 1 . T e s t s at Davis, California, showed C321 to be
s u p e r i o r i n yellows resistance t o t h e p a r e n t variety a n d t o t h e
US c o m m e r c i a l h y b r i d varieties. T h i s selection has good resistance to b o l t i n g a n d curly t o p . C321 is b e i n g used as a yellowsresistant b r e e d i n g line.
Multigerm
Inbred
Lines
N B l . A n increase of S 5 (CI 179 X CI 1-707) c o m b i n i n g resistance to b o l t i n g a n d curly t o p (1). CI 179 was a T y p e O clone
with high sucrose p e r c e n t a g e from US 1. CI 1-707 was a selffertile i n b r e d h i g h i n curly-top resistance a n d d a r k g r e e n i n
color. N B l is an excellent T y p e O i n b r e d w i t h very good comb i n i n g ability a n d has b e e n utilized in t h e p r o d u c t i o n of t h e
MS of N B l a n d as a b r e e d i n g line.
MS of N B l . A m a l e sterile p r o d u c e d by crossing N B l to a
cytoplasmic male-sterile p l a n t f o u n d in US 56 a n d t h e n backcrossing four times to N B l (1). MS of N B l has b e e n used as
the seed-bearing p a r e n t to p r o d u c e t h e F 1 h y b r i d s MS of N B l
X N B 2 , M S of N B l X N B 3 , M S of N B l X N B 4 , a n d M S of
N B l X N B 5 . T h e s e F 1 hybrids are m a l e sterile a n d have b e e n
used extensively as seed-bearing p a r e n t s to p r o d u c e c o m m e r c i a l
hybrid varieties.
N B 2 . An i n b r e d d e v e l o p e d from a cross m a d e at Salt L a k e
City, U t a h , in 1943 b e t w e e n a bolting-resistant clone a n d a
self-fertile line (2). F o l l o w i n g intensive selection, a high-performing S 5 l i n e was designated N B 2 . T h e i n b r e d possesses good
vigor, m o d e r a t e l y g o o d b o l t i n g resistance, a n d fair c u r l y - t o p
resistance. It has good c o m b i n i n g ability especially from t h e

558

JOURNAL OF THE A. S. S. B. T.

standpoint of sucrose percentage. NB2 has been used to produce the seed-bearing parent MS of NB1 X NB2.
NB3.A moderately bolting-resistant segregate from the CT 9
inbred developed by Dr. F. V. Owen at Salt Lake City, Utah
(2). T h i s inbred has very good curly-top resistance but poor
downy-mildew resistance. NB3 has been used to produce the
seed-bearing parent MS of NBI X NB3.
NB4.An inbred developed from the same cross as NB2. This
inbred combines bolting and downy-mildew resistance with good
combining ability but lacks vigor and is susceptible to curly
top. NB4 has been used to produce the seed-bearing parent
MS of NBI X NB4.
NB5.A T y p e O, bolting-resistant inbred from US 56/2 X N B I .
T h i s inbred has good curly-top resistance, excellent vigor, and
good combining ability. NB5 has been used to produce MS
of N B I X NB5.
MS of NB5.A male sterile produced by crossing NB5 to MS
of NBI and backcrossing twice to NB5 (3). Increases of MS
of NB5 have been used as the seed-bearing parent to produce
the F 1 hybrid MS of NB5 X NB6.
NB6.An increase of S 4 (US 22/3 X N B I ) . This inbred combines excellent bolting resistance with good curly-top resistance.
NB6 has been used to produce MS of NB5 X NB6, the seedbearing parent in the bolting-resistant hybrid (MS of NB5 X
NB6) X C787.
NB7,An increase of S 4 (Type O US 56/2 X N B I ) . T h i s inbred
combines very good curly-top resistance with good bolting resistance and has good combining and seed-setting ability. NB7
is being used as the pollen parent to produce commercial monogerm hybrid varieties such as (515HO X 569) X NB7 and
(562HO x 569) X NB7.
C7508.A downy-mildew resistant inbred selected from S 2 (US
22/3 X N B I ) . T h i s inbred has very good bolting resistance and
moderate curly-top resistance. C7508 has been used as a Type
O, mildew-resistant breeding line.
C7508HO.A male sterile produced by backcrossing four times
to C7508. C7508HO has been used only as a breeding line.
C8503.An increase of S 5 (C453 X N B I ) . T h i s inbred has very
good downy-mildew resistance, good bolting resistance, and fair
curly-top resistance. C8503 has been used as a source of mildew
resistance.
C884.The second successive bolting-resistant selection from
US 20IB. C884 has been used as a breeding line combining
bolting and leaf-spot resistance.

Vop.

13,

No.

Monogerm

6,

JULY

Inbred

1965

559

Lines

C7507.An increase of S 4 ( U S 2 2 / 3 X SLC 101mm). T h i s inbred has very good b o l t i n g resistance, m e d i u m vigor, a n d good
seed-setting ability, b u t lacks curly-top resistance. A l t h o u g h t h e
i n b r e d i s n o t completely T y p e O , p o l l e n p r o d u c e r s d o n o t
occur in h y b r i d s w i t h good cytoplasmic m a l e steriles. C7507
has b e e n used to p r o d u c e t h e seed-bearing p a r e n t s 5 1 5 H O X
507 a n d 5 6 9 H O X 507.
C7515.An increase of S 5 ( N B 1 X S L C 101mm). T h i s i n b r e d
has g o o d b o l t i n g resistance, good vigor, a n d fair seed-setting
ability, b u t lacks curly-top resistance. It is a very g o o d T y p e O
line w i t h good c o m b i n i n g ability. C7515 has b e e n used to produce t h e male-sterile C 7 5 1 5 H O .
C 7 5 1 5 H O . A m a l e sterile p r o d u c e d by crossing C7515 to MS
of N B 1 a n d t h e n backcrossing four times to C7515. Increases
of C 7 5 1 5 H O have b e e n used as t h e seed-bearing p a r e n t to p r o d u c e
the Fi h y b r i d s 5 1 5 H O X 507, 5 1 5 H O X 569, 5 1 5 H O X 5 6 1 ,
and 5 1 5 H O X 562. T h e s e F 1 h y b r i d s have been used as seedb e a r i n g p a r e n t s to p r o d u c e c o m m e r c i a l m o n o g e r m h y b r i d varieties.
C7569.An increase of S 3 ( N B 1 X S L C 101mm). T h i s m o n o germ i n b r e d has good b o l t i n g , m e d i u m vigor, good seed-setting
ability, a n d m o d e r a t e curly-top resistance. A l t h o u g h t h e i n b r e d
i s n o t completely T y p e O , pollen p r o d u c e r s d o n o t occur i n
hybrids w i t h good cytoplasmic m a l e steriles. C7569 has b e e n
used to p r o d u c e t h e m o n o g e r m seed-bearing p a r e n t s 5 1 5 H O X
569, 5 6 1 H O X 569, a n d 5 6 2 H O X 569.
C8569HO.C7569 crossed to a cytoplasmic male sterile from
MS of N B 1 a n d backcrossed to C7569. Increases of C 8 5 6 9 H O
have b e e n used as t h e seed-bearing p a r e n t to p r o d u c e c o m m e r c i a l
m o n o g e r m h y b r i d varieties.
C9561.A bolting-resistant m o n o g e r m i n b r e d from S 4 ( N B 1 X
C7507). T h i s i n b r e d has good b o l t i n g resistance, fair curly-top
resistance, good seed-setting ability, a n d is a good T y p e O.
C9561 has b e e n used t o p r o d u c e t h e seed-bearing p a r e n t s 5 1 5 H O
X 561 a n d 5 6 9 H O x 5 6 1 .
C9561HO.A m a l e sterile p r o d u c e d b y backcrossing t o C 9 5 6 1 .
Increases of 9561 HO h a v e b e e n used as the seed-bearing p a r e n t
to p r o d u c e 561 HO X 569.
C0562.A T y p e O sister line of C 9 5 6 1 . T h i s m o n o g e r m i n b r e d
combines good b o l t i n g a n d curly-top resistance with good combining ability. C0562 has b e e n used to p r o d u c e t h e m a l e sterile
C0562HO.

560

JOURNAL OF THE A. S. S. B. T.

C0562HO.A male sterile produced by backcrossing twice to


C0562. Increases of C0562HO have been used as the seed-bearing
parent to produce 562HO X 569, 562HO X 546, and 562HO
X 549.
C1546.An increase of S 2 (C7507 X NB6). T h i s inbred has good
resistance to curly top and bolting. Although not completely
T y p e O, pollen producers do not occur in crosses with good
cytoplasmic male steriles. CI546 has been used to produce the
seed-bearing parent 562HO X 546.
C2563.A curly-top resistant selection from C0562. Curly-top
and bolting resistance are superior to that of C0562 and similar
to that of NB1. C2563 has been used to produce the male sterile
C2563HO.
C2563HO.A male sterile produced by backcrossing twice to
C2563. T h i s male sterile has been used as the seed-bearing parent
to produce F 1 hybrids high in curly-top and bolting resistance
such as 563HO X 546.
C2549.A bolting-resistant sister line of CI546. T h i s inbred is
similar to CI546 in curly-top resistance. C2549 is not completely
Type O but yields male-sterile progeny when crossed with good
male steriles such as 562HO.
C3550.A curly-top resistant selection from S 4 (C7507 X NB6).
T h i s is a sister line of CI546 and C2549. It is superior to both
CI546 and C2549 in curly-top resistance and to CI546 in bolting
resistance. C3550 has been used as the pollen parent to produce
F 1 hybrids such as 563HO X 550.
C3550HO.A male sterile produced by backcrossing to a segregate of C7507 X NB6. C3550HO is being used to produce the
male-sterile equivalent of C3550.
C3505.The second backcross of a monogerm inbred to the
high-performing NB1 multigerm inbred. T h i s T y p e O inbred
combines good resistance to bolting and curly top. C3505 is
being used as a breeding line.
C2648-3.An increase of a leaf-spot-resistant selection made at
Fort Collins, Colorado, from S 3 (673-2 X C7507). 673-2 is a Type
O plant found at Salinas, California, in polycross selections from
US 401. C2648-3 has moderate leaf-spot resistance but lacks
curly-top and bolting resistance. It is being used as a leaf-spotresistant inbred parent and as a breeding line.
C2648-11.Bolting-resistant sister line of C2648-3. C2648-11 is
being tested as an inbred parent in hybrids requiring bolting
and leaf-spot resistance.
C3534.A curly-top resistant, T y p e O, monogerm inbred selected from a cross between NB1 a n d C2563. Greenhouse tests
have shown C3534 to be equal or superior to C2563 in curly-

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t o p resistance. B o l t i n g resistance is very good. C3534 is b e i n g


tested as a curly-top-resistant m o n o g e r m i n b r e d p a r e n t .
C 3 5 3 4 H 4 . A m a l e sterile from a cross b e t w e e n C 2 5 6 3 H O a n d
C3534. C 3 5 3 4 H 4 is b e i n g used as the seed-bearing p a r e n t to
p r o d u c e t h e male-sterile e q u i v a l e n t of C3534.
Utilization of Seed Releases
B r e e d i n g lines d e v e l o p e d by the U. S. D e p a r t m e n t of Agric u l t u r e at Salinas, California, have b e e n used as p a r e n t s in t h e
p r o d u c t i o n of b o t h US a n d sugar c o m p a n y h y b r i d varieties. Seed
p r o d u c t i o n records f u r n i s h e d by M r . S. C. C a m p b e l l , M a n a g e r ,
West Coast Beet Seed C o m p a n y , show that b e t w e e n 1958 a n d
1963 a p p r o x i m a t e l y 15,283,000 p o u n d s of sugar b e e t seed w e r e
Xable 1.Utilization of sugar beet breeding lines developed by the U. S. Department
of Agriculture, Salinas, California, in the production of commercial hybrid seed from 1958
to 1963.
Seed-bearing parent

Seed
production

Pollen parent

pounds
Multigerm see<
MS of NB1 X
MS of NB1 X
MS of NB1 X
MS of NB1 X
MS of NB1 X
MS of NBI X
MS of NBI X
MS of NBI X
MS of NBI X
MS of NBI X
MS of NB5 X
C361HO
C585HO
MS of NBI X
MS of NBI X
MS of NBI X
MS of NBI X

NB2
NB2
NB2
NB3
NB3
NB3
NB4
NB4
NB4
NB5
NB6

NB2
NB3
NB4
NB5

C366
C586
C681M x NB4
C586
C663
C681M X NB4
C366
C586
C663
C663
C787
Company pollinators
Company pollinators
Company pollinators
Company pollinators
Company pollinators
Company pollinators

47,500
599,300
800

84,900
854,800
1.000
14,400
247,200
210,100
418,400
18,900
1,887,800
957,500
84,300
2,629,300
372,300
363,800
Xotal

Monogerm
515HO X
515HO x
515HO x
569HO x
562HO x
515HO x
562HO x
569HO
569HO
562HO
561 HO x
5R2HO x

seed
507
561
569
507
569
569
569

562
569

C663
C663
C663
C663
C663

1,400
2.300
49,000
2,000
37,700
39,800
34,900

NB7
NB7
NB7

Company
Company
Company
Company

8,792,300

900

pollinators
polli nators
poll nators
polli nators

1,153,600
9,800
198,200
386,900
Total

1,916,500

562

JOURNAL OF THE A. S. S. B. T.

produced for use in California and Nevada. Included were


2,665,000 pounds of US hybrid seed and 8,044,000 pounds of
sugar company hybrid seed involving male-sterile parents developed by the U. S. Department of Agriculture. Utilization of
the seed releases in commercial hybrids is summarized in T a b l e 1.
Acknowledgments
Source material for many of the breeding lines was furnished
in 1947 by Dr. F. V. Owen and associates of the U. S. Sugar Beet
Investigations Laboratory, Salt Lake City, Utah. Dr. V. F.
Savitsky of the Salt Lake City Laboratory furnished the original
SLC 101 monogerm line. Mr. Dewey Stewart and associates of
the U. S. Sugar Beet Investigations office in Beltsville, Maryland,
furnished the US 401 lines.
Curly-top resistance evaluations and selections were made by
Mr. A. M. Murphy at the U. S. Sugar Beet Field Laboratory,
T w i n Falls, Idaho, and the Crops Research Laboratory, Logan,
Utah. Dr. C. W. Bennett also made curly-top resistance evaluations in the greenhouse at Salinas. Mr. J. O. Gaskill of the
U. S. Sugar Beet Field Laboratory, Fort Collins, Colorado, made
leaf-spot resistance selections and evaluated the resistance of
C2648-3 and C2648-11.
Variety performance tests were made by Mr. Charles Price
and associates, and by Mr. K. D. Beatty, at the U. S. Southwestern
Irrigation Field Station, Brawley, California. Varieties were also
evaluated in each of the major sugar beet growing areas of California by the American Crystal Sugar Company, Holly Sugar
Corporation, Spreckels Sugar Company, and Union Sugar Division, Consolidated Foods Corporation. Seed production tests
were made by the West Coast Beet Seed Company, Salem, Oregon.
T h e Agronomy and Plant Pathology departments of the University of California, Davis, cooperated in tests to determine the
yellows resistance of C330 and C321.
Literature Cited
(1) MCFARLANE, J. S. 1954. New non-bolting and mildew-resistant seed
releases. Proc. Am. Soc. Sugar Beet Technol. 8 (2) : 88-89.
(2)

MCFARLANE, J.

S.,

F. V.

OWEN

and ALBERT M.

MURPHY.

1961.

New

hybrid sugar beet varieties for California. J. Am. Soc. Sugar Beet
Technol. 11: 500-506.
(3) OWEN, F. V. 1948. Utilization of male sterility in breeding superior
yielding sugar beets. Proc. Am. Soc. Sugar Beet Technol. 5: 156-161
(4) OWEN, F. V. 1954. Hybrid sugar beets made by utilizing both cytoplasmic and Mendelian male sterility. Proc. Am. Soc. Sugar Beet
Technol. 8(2) 64.

JOURNAL
of the
American Society of Sugar
Beet Xechnologists
Volume 13
Number 7
October 1965

Published

quarterly

by

American Society of Sugar Beet Xechnologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado 80521

Subscription
$4.50
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TABLE OF CONTENTS

Title

Author

Control of Cercospora leaf spot of sugar


beet with ultra-low-volume oil-based fungicidal mists
Liming of sugar beet cossettes

T h e host range of the sugar beet nematode,


Heterodera
schachtii Schmidt
Heterodera schachtii in relation
from root rot of sugar beets

to

Page

C. L. Schneider

563

Alan E. Goodban
Holland M. McCready

566

Arnold E. Steele

573

Charles Price
C. L. Schneider

604

S. T. Dexter
M. G. Frakes

607

Jay L. Haddock
Darrel M. Stuart

613

V. F. Savitsky
Helen Savitsky

621

Walter E. Peay

645

damage

Evaluation of sugar beet storage practices


by using the percentage purity of the
"thin juice"

Nutrient balance and concentration in sugar


beet production

Weight of fruits in self-fertile, male-sterile,


and self-sterile diploid and tetraploid
monogerm Beta vulgaris L.

Laboratory screening tests of the insecticides


for control of the beet webworm

Insect control on sugar beets by seed or


soil treatments
Howard E. Dorst

649

Control of Cercospora Leaf Spot of Sugar Beet


W i t h Ultra-Low-Volume Oil-Based Fungicidal Mists
C.

L.

SCHNEIDER1

Received for publication December 18, 1964

Introduction
T h e following a r e a m o n g t h e advantages of oil sprays over
o t h e r c o n v e n t i o n a l spray materials for the control of p l a n t p a t h o genes: they are effective spreaders a n d stickers; relatively c h e a p ;
well a d a p t e d t o aerial application; a n d n o n p h y t o t o x i c ( l ) 2 .
Fisher (2) r e p o r t s satisfactory c o n t r o l of greasy spot disease of
citrus (Cercospora citrigrisea sp. nov.) with oil e m u l s i o n spray.
Calpouzos (1) cites several instances of c o n t r o l of Sigatoka leaf
spot disease of b a n a n a (Mycosphaerella musicola Leach = Cercospora musae Zimm.) w i t h ultra-low-volume oil-mist sprays, applied e i t h e r as fungicide carriers or applied alone. E v i d e n c e
a c c u m u l a t e d by Calpouzos indicates t h a t oil acts as a fungistatic
t h e r a p e u t i c agent in c o n t r o l l i n g this disease. A c c o r d i n g to W i l s o n
et al. (3), a d d i t i o n of emulsifiable oil to fixed c o p p e r sprays for
control of sugar beet leafspot (Cercospora beticola Sacc.) is
beneficial because it a p p e a r s to r e d u c e t h e e r o d i n g a c t i o n of
w i n d a n d r a i n o n t h e c o p p e r deposit.
T h e successful c o n t r o l of t h e a f o r e m e n t i o n e d cercosporoses
of citrus a n d b a n a n a w i t h oil sprays p r o m p t e d o u r o w n tests of
the efficiency of ultra-low-volume mist applications of p e t r o l e u m
oilalone a n d as a fungicide c a r r i e r i n the c o n t r o l of sugar
beet leaf spot i n d u c e d by C. beticola.
Materials and Methods
We c o m p a r e d t h e following t r e a t m e n t s in a field test u n d e r
severe a n d sustained e x p o s u r e to C. beticola:8
1. P e t r o l e u m spray oil (Esso) 4 ; a n a p h t h e n i c spray oil of 92
U S R ( u n s u l f o n a t a b l e residue) a n d 70 second viscosity
(Saybolt Universal Seconds).
2. C o p p e r o x y c h l o r i d e (0.5 l b / g a l of e m u l s i o n c o m p r i s i n g
o n e p a r t p e t r o l e u m oil + 3 p a r t s water).
3 . M a n e b (1.25 l b / g a l p e t r o l e u m oil).
4. T r i - b a s i c c o p p e r sulfate (1 l b / g a l p e t r o l e u m oil).
5. C o n t r o l .
1
Plant Pathologist, Crops Research Division, Agricultural Research Service, U. S.
Department
of Agriculture, Logan, Utah.
2
Numbers
in parentheses refer to literature cited.
3
All materials were prepared and furnished by Esso Research and Engineering Company,4 Linden, New Jersey.
Mention of material and company name is for identification only and does not
emply endorsement by U. S. Department'of Agriculture.

JOURNAL OF THE A. S. S. B. T.

564

T h e plots, arranged in a 5 X 5 latin square, were situated


in a field of approximately 3 acres of sugar beets at the Plant
Industry Station, Beltsville, Maryland, in 1961. Each plot comprised four 20-ft. rows of commercial sugar beet variety, SL 122
MS X SP 5481-0, spaced 2 ft apart.
On June 23, inoculum consisting of dried and ground sugar
beet leaves infected with C. beticola was applied to all sugar beet
plants in the test plots and in the surrounding field.5 By July
10, leaf spot symptoms began to appear. A severe leaf spot
epiphytotic prevailed throughout the field during August and
September.
T h e treatments were applied with a "Solo Port" knapsack
type mist blower on July 10, 20, 31, August 14, 25, 28 (following
heavy rains on August 26, 27), September 7 and 22. All treatments, except copper oxychloride + oil were applied at the rate
of approximately one gal/acre. Copper oxychloride and oil was
applied at 4 gal/acre on July 10 but subsequent applications were
at one gal/acre because foliage injury was associated with the
higher rate. No injury was associated with any of the other
treatments.
Leaf spot severity readings were made on July 30, August 14
and September 25. On October 4 the two center rows of each
plot were harvested and weighed. We obtained sucrose percentage and refractometer readings from a sample comprising all
plants in the two center rows of each plot.
Results and Conclusions
Each of the 3 fungicide-oil treatments reduced leaf spot
severity and caused a proportionate increase in gross sugar, root
Table 1.Results of ultra-low-volume fungicide-oil spray test for control of Cercospora
beticola on sugar beet variety SL 122 MS x SP 5481-0 at Beltsville Maryland, 1961.
Leaf spot a / b
severity
rating

Acre yield a

Apparent"
purity
Sucrose" coefficient
Percentage
2.5
Copper oxychloride + oil
14.38 ab
80.52 a
3.2
Maneb + oil
14.51 a
79.52 a
3-6
Tri-basic copper sulfate 4- oil
13.72 bc
79.98 a
5.0
Oil alone
13.07 c
78.63 a
6.2
Control
12.14 c
77.44 a
a
Results given as 5-plot averages. Superscripts indiicate levels of significance for ranked
means. Treatments not including the same letter a r e significantly different at the 5%
level (Duncan's multiple range test).
b
Leaf spot readings based on a scale from 0 (no disease)i to 10 (all :leaves dead ). Ratings
shown are maxima for readings on 3 different dates.
Treatment

Gross
sugar
Pounds
3986 a
3832 ab
3373 bc
2887 c
2255 d

Roots
Tons
13.86 a
13.21 a
12.30 ab
11.06 b
9.29 c

5
Inoculum was prepared and applied by Dr. G. E. Coe, Crops Research Division
Agricultural Research Service, U. S. Department of Agriculture, Beltsville, Maryland.

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weight a n d sucrose p e r c e n t a g e i n comparison w i t h t h e u n t r e a t e d


c o n t r o l ( T a b l e 1). A p p l i c a t i o n of oil alone r e s u l t e d in a significant r e d u c t i o n in leaf spot d a m a g e b u t n o t so effectively
as c o p p e r oxychloride + oil or m a n e b + oil.
T h e results of this study indicate the efficacy of oil-based
ultra-low-volume fungicide mists in c o n t r o l l i n g cercosporosis of
sugar beet. T h e results also indicate the advisability of f u r t h e r
studies w i t h different oils a n d dosages to d e t e r m i n e t h e e x t e n t
to which t h e antifungal p r o p e r t y of oil can be utilized in cont r o l l i n g this disease.
Literature Cited
(1) CALPOUZOS, L. 1962. Some contributions from the tropics to p l a n t
disease control methods. Phytopath. 52: 942-945.
(2) FISHER, FRAN E. 1961. Greasy spot and tar spot of citrus in Florida.
Phytopath. 5 1 : 297-303.
(3)

W I L S O N , J . D., FRANK IRONS a n d J A M E S H E N R Y .

1963.

E x p e r i m e n t s in

the control of Cercospora leaf spot of sugar beets1962.


and Plant Pathology Series 42, Ohio Agr. Expt. Sta. 23 p p .

Botany

Liming of Sugar Beet Cossettes


ALAN E. GOODBAN AND ROLLAND M. MCCREADY 1
Received for publication December 14, 1964

T h e prospect of improving sugar beet processing by pretreating beets with lime has intrigued beet technologists for
many years. Proposed systems have included pressing limed
brei (2,3,4,12) 2 , pressing limed and neutralized brei (13,14),
diffusion of limed and neutralized beet chips (14), diffusion with
dilute lime solution (7), and diffusion of cossettes after treatment with a solution of lime and sugar (10). To our knowledge,
none of these processes has found commercial acceptance. This
paper reports our investigation of the effects of lime on beet
tissue and some results of processing limed cossettes.
T h e effects of lime on beet tissue are due primarily to the
reactions of pectin under alkaline conditions since other pulp
components are relatively inert. About 3 0 % of the water-insoluble portion of the beet is pectin. Pectin is a linear polymer of
galacturonic acid units which are partially methylated and
acetylated. In sugar beet pectin, about 5 0 % of the carboxyl
groups are methylated, and about 3 0 % of the hydroxyl groups,
which are in positions 2 and 3, are acetylated.
Previous knowledge of the effects of alkali on pectin (9,11)
may be summarized as follows: T w o reactions may occur when
pectin is treated with alkali. One reaction is ester hydrolysis,
which produces methanol and acetate as byproducts. T h e other
reaction is polymer degradation by glycosidic bond cleavage.
Figure 1 shows a portion of the pectin molecule and the reactions with hot or cold alkali. At low temperature the predominant reaction is ester hydrolysis. T h e resulting pectic acid
is highly resistant to alkaline degradation. T h e calcium salt of
pectic acid is insoluble, and, if lime is the alkali used to deesterify the pectin, the divalent calcium cross links acid groups
to form a rigid calcium pectate structure.
At high temperature, not only are the esters hydrolyzed, but
glycosidic bonds adjacent to esterified carboxylic groups are
broken (11). Since about half of the carboxyl groups are originally esterified, the pectin chain is almost completely destroyed.
T h i s pectin degradation makes the associated hemicellulose
soluble in water, and thus greatly reduces the mechanical strength
of the beet tissue. Although the degradation product shown in
1
Western Regional Research Laboratory, Western Utilization Research and Develop
ment Division, Agricultural Research Service, U. S. Department of Agriculture, Albany
California.
2
Numbers in parentheses refer to literature cited.

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567

F i g u r e 1 has a g a l a c t u r o n i c acid e n d g r o u p , it m a y actually be


u n s a t u r a t e d , as is the case w i t h enzymatic d e g r a d a t i o n (1).. In
e i t h e r case t h e glycosidic b o n d is b r o k e n adjacent to t h e esterified
carboxyl g r o u p . T h e d e g r a d a t i o n reaction occurs only a t esterified
loci, a n d t h u s previous cold de-esterification protects t h e p e c t i n
from s u b s e q u e n t h o t alkaline degradation.
Below 25 C t h e p r i n c i p a l alkaline reaction is de-esterification, b u t d e g r a d a t i o n increases rapidly a t h i g h e r t e m p e r a t u r e s .
At 75 C t h e r e is so m u c h d e g r a d a t i o n that sugar beet p u l p loses
its s t r u c t u r e a n d becomes mushy. T h e t e m p e r a t u r e d e p e n d e n c e
could be e x p l a i n e d by lower activation energy for de-esterification
t h a n for d e g r a d a t i o n , as is t h e case u n d e r acid c o n d i t i o n s (9).
In o r d e r to take a d v a n t a g e of these properties of beet p e c t i n ,
fresh cossettes s h o u l d be treated w i t h lime at low t e m p e r a t u r e .
T h i s t r e a t m e n t w o u l d stabilize the pectin by de-esterification,
a n d at t h e same t i m e form a calcium pectate p r e c i p i t a t e in situ.
T h i s i n s o l u b l e m a t r i x w o u l d h i n d e r the e x t r a c t i o n o f colloids
d u r i n g diffusion, b u t allow free diffusion of sugar.

Figure 1.Alkaline reaction of pectin. De-esterification (top) a n d


degradation (bottom). M indicates a methylated carboxyl, A indicates an
acetylated hydroxyl g r o u p . Dashed line indicates a broken glycosidic b o n d .

Lime Penetration
Esterified p e c t i n can be detected in p l a n t tissue in situ by
reaction w i t h a l k a l i n e h y d r o x y l a m i n e a n d ferric c h l o r i d e (6).
T h i s r e a c t i o n was used to follow t h e de-esterification of sugar
beet p e c t i n w i t h lime. Five c e n t i m e t e r r e c t a n g u l a r plugs w e r e
cut from a sugar beet w i t h a m i c r o t o m e , a n d soaked in 1% l i m e
slurry. Sections e x a m i n e d after ten m i n u t e s showed de-esterification to a d e p t h of 0.1 mm at 25 C a n d 0.2 mm at 45 C. T h i s
corresponds to 3 a n d 7 cells, respectively. No g r a d a t i o n of
partially de-esterified pectin was a p p a r e n t . T h e s h a r p d e m a r c a tion b e t w e e n de-esterified a n d u n r e a c t e d pectin indicates t h a t
the r a t e c o n t r o l l i n g step is diffusion of lime.

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JOURNAL OF THE A. S. S. B. T.

Similar tests with cossettes showed that after ten minutes


liming at 25 C only a narrow band of esterified pectin remained
in the center of the cossette. T h e greater penetration into cossettes can be explained by considering the cossette structure
shown in Figure 2. T h e numerous transverse cracks allow the
lime to penetrate rapidly which results in more extensive deesterification than would be predicted from the results on
microtome-cut pieces.

Figure 2.Sugar beet cossettes showing transverse cracks.

Saponification Rates
De-esterification and deacetylation rates were obtained by
measuring the residual ester and acetyl content of p u l p after reaction with lime. Cossettes were mixed with dry Ca(OH) 2 and
held at 25 C. Samples were withdrawn at timed intervals and
the acetyl and methoxyl content of the marc measured (5). The
results were calculated as percent saponification, and are shown
in Figure 3. T h e rate of demethylation is greater than the rate
of deacetylation at this temperature. Demethylation stabilizes
pectin, but deacetylation does not, and furthermore, it produces

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Time, minutes
Figure 3.Saponification of beet pectin. R a t e of de-esterification
and deacetylation. Cossettes treated at 25 C with dry C a ( O H ) 2 , 1.3%
on beets.
acetic acid as a b y p r o d u c t . Since d e m e t h y l a t i o n is m o r e r a p i d
than deacetylation, it may be possible to restrict deacetylation
a n d still s t a b i l i z e p e c t i n b y sufficient d e m e t h y l a t i o n .
Processing

Xests

Several different processing procedures were evaluated by


e x a m i n i n g diffusion juice a n d t h i n juice from treated beets.
Diffusion was c a r r i e d o u t in a Briiniche-Olsen l a b o r a t o r y diffuser
with a feed r a t e of 9 kg of beets p e r h o u r . First c a r b o n a t i o n was
c o n t i n u o u s , i n a D o r r - t y p e l a b o r a t o r y c a r b o n a t o r (8). X h i n j u i c e
was m a d e b y b a t c h s e c o n d c a r b o n a t i o n o f f i l t e r e d f i r s t - c a r b o n a tion juice.
Cossettes w e r e l i m e d in the following four ways:
1) dry C a ( O H ) 2 preliming, 2) lime slurry preliming, 3) direct
l i m i n g i n t h e diffuser, a n d 4 ) c o u n t e r c u r r e n t w a s h i n g w i t h l i m e d
diffusion j u i c e . Cossette l i m i n g was e i t h e r a t r o o m t e m p e r a t u r e
Hess t h a n 2 5 C ) o r a t 4 0 c , u s i n g 0 . 4 t o 1 . 0 % C a O o n b e e t s .
Diffusion t e m p e r a t u r e was 70 or 25 C w i t h a l k a l i n e beets, a n d
was 70 . C w i t h l i m e d b e e t s w h i c h h a d b e e n n e u t r a l i z e d w i t h

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JOURNAL OF THE A. S. S. B. T.

CO,. Liming at 40 C was unsatisfactory because of pulp deterioration. Lime slurry treatments were difficult to manage
because of the rapid extraction of sugar, even at low temperature. For continuous operation, a system of adding treatment
liquor to the diffusion juice at a constant rate would be needed.
Dry liming was easier, and still gave satisfactory treatment to
the beets. T r e a t m e n t levels above 0.4% CaO on beets were
unnecessary. Diffusion juice from alkaline beets was clear and
light colored. Juice from neutralized beets was darker, with
more colloids, but still much lighter and clearer than normal
diffusion juice. All of the diffusion juices were subjected to
first carbonation, with additional lime. Sedimentation and filtration rates were determined on the first carbonation juices, and
the color, lime salts, and purity of the second carbonation juices
were measured. T a b l e 1 is a summary of the results from the
best three treatments. T h e major differences are in the total
lime consumption and in the lime salts and total anions. T h e
final m u d volumes are a reflection of the amount of lime used.
Table 1.Processing comparisons for prelimed beets
Cossettes limed at roo:m temperature Lth
dry Ca (HO) 2 , (0.4'% CaO on beets)
Control
Alkaline
Neutral
Preliming time, min.
Diffusion temp., C.
First carbonation:
Lime added (% CaO on beets)
Sedimentation resistance
Mud volume (%)
Second carbonation:
Lime salts (CaO/100 Bx
Anions (meq./lOO Bx)
Purity (%)
Color (OD/100 Bx)

none
70

30
25

30
70

10
70

2.0
25
9.0

0.5
32
5.0

0.5
31
5.0

1.0
34
7.0

.05

.18

.39

.52

31

50

55

51

91.6

89.5

90.2

90.0

2.0

2.0

2.8

2.8

T h e total lime consumption for the alkaline-diffused beets was


0.9%, for the neutralized beets 1.4%, and for the control beets
2.0% CaO on beets. Lime salts and anions are high in the second
carbonation juice from the limed beets because of acetate liberated from the pectin. T h e increase in total acids was matched
by the increase in volatile acids, which we assume to be acetic
acid. These excess anions prevent the removal of lime by carbonation without the addition of corresponding cations. The
sodium carbonate needed to reduce the lime salts to the level
of the control is calculated to be 0.05 to 0.15% on beets. The
increase in anions, calculated as acetate, plus the increase in
lime salts, calculated as calcium ion, is 1.3 to 1.7 grams per 100

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grams of solids in second c a r b o n a t i o n juice. W i t h i n experim e n t a l error, this is t h e same as t h e observed d r o p in p u r i t y for
juices from l i m e d beets. P r e l i m e d cossettes do r e t a i n m o r e colloids a n d insoluble calcium salts t h a n do u n t r e a t e d cossettes,
b u t these are materials w h i c h w o u l d b e r e m o v e d d u r i n g carb o n a t i o n . T h e r e is no n e t change in the a m o u n t of s o l u b l e
nonsugars in second c a r b o n a t i o n juice, except for acetate prod u c e d by deacetylation of p u l p pectin.
P u l p from p r e l i m e d beets i s m u c h t o u g h e r t h a n n o r m a l
p u l p . L a b o r a t o r y tests showed that u p t o 4 0 % solids c o u l d b e
o b t a i n e d u s i n g a static press b u t that the p u l p was t o o t o u g h
to be pressed in a high-speed screw press. We w e r e u n a b l e to
test p u l p from p r e l i m e d beets in a factory press w i t h o u t int r o d u c i n g t h e system to an e n t i r e factory, b u t we h a v e m a d e
some factory tests on p u l p from a c o n t i n u o u s slope diffuser by
a d d i n g l i m e after diffusion. P u l p treated with 0 . 1 5 % C a O becomes h a r d a n d t o u g h in the same way as do t h e cossettes. D r y
h y d r a t e d l i m e or a slurry of l i m e was a d d e d to t h e p u l p in t h e
conveyor system feeding t h e presses. T h e t e m p e r a t u r e was 45
to 50 C. A l t h o u g h t h e trials were n o t extensive, it does seem
that l i m e - h a r d e n e d p u l p can b e pressed satisfactorily. I n o n e
trial, a b o u t 100 tons of p u l p were treated at the r a t e of 0 . 1 5 %
C a O on p u l p over a p e r i o d of two h o u r s , a n d t h e p u l p was
pressed o n t w o o r t h r e e Stearns-Rogers presses t o a b o u t 8 3 %
moisture, w h i c h was the same as for u n t r e a t e d p u l p . On t w o
occasions, lower moistures were o b t a i n e d for brief periods, b u t
the presses plugged, e i t h e r because t h e b o t t o m plates w e r e t o o
tight or because n o t e n o u g h pressure could be d e v e l o p e d to
force t h e p u l p t h r o u g h t h e discharge slot. T h e p o w e r c o n s u m p tion on t h e presses was n o t excessive, even w h e n they w e r e
plugged.
Summary
Cossette p r e l i m i n g at low t e m p e r a t u r e stabilizes t h e b e e t
s t r u c t u r e to p e r m i t alkaline diffusion w i t h o u t d i s i n t e g r a t i o n of
the p u l p . C o n s i d e r a b l e r e t e n t i o n of otherwise e x t r a c t a b l e colloids is observed, l e a d i n g to a r e d u c t i o n in the l i m e n e e d e d for
clarification. T h e p r o d u c t i o n of acetate by alkaline d e a c e t y l a t i o n
reduces t h e p u r i t y of clarified j u i c e a n d increases t h e a m o u n t of
lime salts.
T h e p r e l i m i n g o f cossettes m u s t b e carried o u t b e l o w 40
C a n d sufficient t i m e m u s t be allowed for t h e p e c t i n to be
stabilized before it is subjected to heat. A l k a l i n e diffusion m a y
presumably be c o n d u c t e d over a w i d e range of t e m p e r a t u r e . T h e
high p H will p r e c l u d e f e r m e n t a t i o n a t low t e m p e r a t u r e s , a n d
the stabilized p e c t i n will p e r m i t u n u s u a l l y h i g h t e m p e r a t u r e s

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JOURNAL OF THE A. S. S. B. T.

d u r i n g diffusion. A n o t h e r advantage of preliming cossettes is


the production of partially clarified juice, which might be suitable for direct ion exchange treatment.
Acknowledgment
T h e authors wish to thank the Holly Sugar Corporation for
the beets used in the laboratory trials; the N o r t h American Dehydrating Corporation and the American Crystal Sugar Company
for their cooperation in the factory trials; E. J. Barta, D. O.
Griffin, R. L. Patterson, and J. C. Goodwin for their assistance
in the processing laboratory; and R. M. Reeve for the photomicrograph of cossettes.
Literature Cited
(1) ALBERSHEIM, J., H. NEUKOM, and H. DEUEL.

1960.

Uber die bildung

von ungesattigten abbauprodukten durch ein pektinabbauendes


Enzym. (The formation of unsaturated degradation products by a
pectin-degrading enzyme.) Helv. Chim. Acta, 43: 1422-1426.
(2) BONELLI, A. 1955. Production of sugar with first defecation carried
out directly on the beets. Ital. Patent 534,077.
(3) BONELLI, A. 1959. Estrazione dello zucchero per defecazione operato
direttamente sulle barbabietole. (Sugar extraction with direct defecation of beets.) Ind. Sacc. Ital. 52: 399-411.
(4) BORGHI, M. 1946. 11 nuovo processo dei dott. Bonelli per la estrazione
dello zuchero dalle bietole. (The Bonelli process for extracting
sugar from beets.) Chimica e Industria (Milan), 28: 177-184.
(5) G E E , M., E. A. MCCOMB, and R. M. MCCREADY.

1958. A method for

the characterization of pectic substances in some fruit and sugarbeet marcs. Food Research 23: 72-75.
(6) G E E , M., R. M. REEVE, and R. M. MCCREADY.

1959.

Reaction of hy-

droxylamine with pectinic acids. Chemical studies and histochemical estimation of the degree of esterification of pectic substances in fruit. J. Agr. Food Chem. 7: 34-38.
(7) LOOF, H. G., and W. POHL. 1956. Process for extracting sugar beets,
using weakly alkaline water. French Patent 1,129,771.
(8)

MORGAN, A. I., E. J. BARTA, and G. O. KOHLER.

1959.

Development

of a sugar beet processing laboratory. J. Am. Soc. Sugar Beet Technologists 10: 563-570.
(9) SPEISER, R., C. R. EDDY, and C. H . HILLS.

(10)
(11)
(12)
(13)
(14)

Kinetics of deesterification

of pectin. J. Phys. Chem. 49: 563-579.


Susie, S. K. 1959. Study of the problem of alkaline difiusion in sugar
manufacture. Technika:l4 (Prehran. Ind. 13) (8), 113-118. Through
S.I.A. 23: abs. 257, 1961.
VOLLMERT, B. 1950. Uber den Alkalischen Pektinabbau. (The alkaline
degradation of pectin.) Makromol. Chemie 5: 110-127.
WEINRICH, MORIZ. 1905. Process of treating sugar beets. U. S. Patent
803,945.
WEINRICH, MORIZ. 1908. Process of treating sugar beets. U. S. Patent
881,641.
WEINRICH, MORIZ. 1910. Process of treating sugar beets. U. S. Patent
950,035.

T h e Host Range of the Sugar Beet N e m a t o d e ,


Heterodera schachtii Schmidt 1
ARNOLD

E.

STEELE2

T h e life cycle of t h e sugar beet n e m a t o d e is essentially typical


of all k n o w n species of t h e genus Heterodera. In t h e presence
of g r o w i n g host plants, eggs hatch a n d larvae escape from protective cysts a n d i n v a d e host-plant roots w h e r e they d e v e l o p to
m a t u r i t y . Soon after fertilization the a d u l t females can be observed as w h i t e lemon-shaped bodies attached to e x t e r n a l surfaces
of host roots (Figures 1 a n d 2). After a short p e r i o d of g r o w t h
the female dies a n d its c u t i c u l a r r e m a i n s forms a tough, sac-like
cyst ( F i g u r e 3), w h i c h m a y enclose m o r e t h a n 500 eggs.
O n e of t h e most effective a n d economical m e a n s of cont r o l l i n g t h e sugar beet n e m a t o d e is c r o p r o t a t i o n . H o s t crops a r e
n o t usually g r o w n m o r e often t h a n once in 3 to 5 years, d e p e n d ing on t h e severity of t h e infestation a n d local c o n d i t i o n s t h a t
d e t e r m i n e the persistence of this pest. However, d i s c r i m i n a n t
use of r o t a t i o n systems is n o t possible w i t h o u t c o m p r e h e n s i v e
i n f o r m a t i o n on t h e host r a n g e of the n e m a t o d e .
A l t h o u g h m u c h i n f o r m a t i o n has b e e n p u b l i s h e d , most reports are n o t readily available to agriculturalists. C o n s e q u e n t l y ,
this study was u n d e r t a k e n to p r o v i d e a d d i t i o n a l i n f o r m a t i o n on
the host r a n g e of t h e sugar beet n e m a t o d e .
Materials and Methods
C r o p , o r n a m e n t a l , a n d weed plants were tested for susceptibility to t h e sugar beet n e m a t o d e , Heterodera schachtii S c h m i d t .
Sources of seed a n d p l a n t s tested are listed in T a b l e 1. W e e d
plants collected in various localities in California a n d A r i z o n a
were sent to D r . G a b r i e l E d w i n , U. S. N a t i o n a l A r b o r e t u m ,
W a s h i n g t o n , D . C . , o r t o Dr. J u n e McCaskill, B o t a n y D e p a r t m e n t of t h e University of California at Davis, for species
identification.
Nematode-infested soil was o b t a i n e d from several fields in
M o n t e r e y a n d San B e n i t o C o u n t i e s of California. O n l y those
fields in w h i c h heavily infected sugar beets h a d recently b e e n
grown were selected. Soils from these fields were screened a n d
stored u p t o o n e year i n large m e t a l d r u m s .
1
Co-operative investigations of the Crops Research Division, Agricultural Research
Service,
U. S. Department of Agriculture and the Beet Sugar Development Foundation.
2
Nematologist, Crops Research Division, Agricultural Research Service, U. S. Department of Agriculture, Salinas, California.

JOURNAL OF THE A. S. S. B. T.

Figure 1.Beta trigyna infected with sugar beet nematode.

Figure 2.Adult sugar beet nematode on root of Beta trigyna.

V O L . 13, N o . 7, OCTOBER 1965

575

Figure 3.Mature cysts of the sugar beet nematode.

Plants were tested in g r o u p s of 8-12 species after t h e m e t h o d


of G o l d e n a n d Shafer (4). Each test g r o u p i n c l u d e d a sugar
beet {Beta vulgaris L., C u l t i v a r U. S. 75) check. Seeds of each
species were g e r m i n a t e d in sterilized sand a n d allowed to grow
for several days. Twenty-five y o u n g plants of each species w e r e
t r a n s p l a n t e d i n t o a l u m i n u m - f o i l cylinders filled with soil heavily
infested w i t h the sugar beet n e m a t o d e . F i g u r e 4 illustrates t h e
typical a r r a n g e m e n t of tested plants.

Figure 4.Typical greenhouse test for host susceptibility to the sugar


beet n e m a t o d e .

After t h e p l a n t s h a d g r o w n 50-60 days in a g r e e n h o u s e , t h e


plants were r e m o v e d from cylinders a n d the roots washed a n d
examined for n e m a t o d e s . W h i t e female n e m a t o d e s w e r e removed from roots of susceptible plants a n d e x a m i n e d microscopically t o d e t e r m i n e w h e t h e r o r n o t the n e m a t o d e s w e r e

JOURNAL OF THE A. S. S. B. T.

576

gravid. T h e results of this study, and published information


on the host range of the sugarbeet nematode, are listed in
T a b l e 2.
In some instances, two or more authors reported different
common names for a given plant species, or the same common
name for different species. These are listed unchanged.
Results and Discussion
Twenty-three out of 49 families listed in T a b l e 2 contained
host species. Of 283 genera and 535 species reported, 218 species
within 95 genera were hosts.
Families with host species differed greatly in the n u m b e r
of infected species. Approximately 80 percent of species within
Chenopodiaceae and Cruciferae were hosts; host species within
Boraginaceae and Onagraceae amounted to 17 and 14 percent,
respectively.
Host species within seven separate families were collected
in various fields in Monterey County of California. Roots of a
n u m b e r of weed species developed heavy infections of sugar
beet nematode, indicating that adequate weed control will
greatly increase the effectiveness of rotational programs.
Crop rotation is today the most satisfactory means available
for controlling the sugarbeet nematode. T h i s report provides a
basis for crop recommendations in beet growing areas.
Literature Cited
(1) DEN OUDEN, H. 1956. The influence of hosts and nonsusceptible
hatching plants on populations of Heterodera schachtii. Nematologica 1 (2) : 138-144.
(2) FRANKLIN, M. T. 1945. On Heterodera cruciferae n. sp. of Brassicas,
and on Heterodera strain infecting clover and dock. J. Helminth.
21 (2/3) : 71-84.
(3) GOLDEN, A. M. 1959. Susceptibility of several Beta species to the sugarbeet nematode {Heterodera schachtii) and root-knot nematodes
(Meloidogyne spp.) . J. Am. Soc. Sugar Beet Tech. 10(5): 444-447.
(4)

GOLDEN, A. M. and THELMA SHAFER.

1958.

Differential response of

Heterodera schachtii, the sugar-beet nematode, to selections of


Chenopodium album. Plant Dis. Reptr. 4 2 ( 2 ) : 184-187.
(5) GOLDEN, A. M. and THELMA SHAFER.

1959.

Host-parasite relationships

of various plants and the sugar-beet nematode


schachtii). Plant Dis. Reptr. 43(12): 1258-1262.
(6)

GOLDEN, A. M. and T H E L M A SHAFER.

1959.

(Heterodera

Susceptibility of tomato

(Lycopersicon esculentum) to the sugar-beet nematode (Heterodera


schachtii). Plant Dis. Reptr. 43(11): 1196-1197.
(7) H I J N E R , J. A. 1952. De gevoeligheid van wilde bieten voor het
bietencystenaaltje (Heterodera schachtii). Meded. Inst. Suikerproductie, Bergen op Zoom, 21 (1) : 1-13.

V O L . 13, N o . 7, OCTOBER 1965

577

(8) JONES, F. G. W. 1945. Soil populations of beet eelworm (Heterodera


schachtii Schm.) in relation to cropping. Annals Appl. Biol. 32 (4) :
351-380.
(9) JONES, F. G. W. 1950. Observations on the beet eelworm a n d other
cyst-forming species of Heterodera. Annals Appl. Biol. 37 (3) : 407440.
(10) MULVEY, R. H. 1957. Susceptibilities of cultivated a n d weed plants
to the sugar-beet nematode, Heterodera schachtii Schmidt, 1871, in
southwestern Ontario. J. H e l m i n t h . 31 (4) : 225-228.
(11) OOSTENBRINK, M. 1955. On the host plants of the beet eelworm
Heterodera schachtii Schmidt. Versl. PIZeikt. Dienst Wageningen,
127: 186-193.
(12) RASKI, D. J. 1952. On the host range of the sugar-beet nematode in
California. Plant Dis. Reptr. 36 (1) : 5-7.
(13) SHEPHERD, AUDREY M. 1959. Testing populations of beet eelworm,
Heterodera schachtii Schmidt, for resistance-breaking biotypes, using
the wild beet (Beta patellaris Moq.) as indicator. N a t u r e (London)
183 (4668) : 1141-1142.
(14) STEINER, G. 1931e. X h e findings of Heterodera schachtii, the sugar
beet nema, on Polygonum in Virginia. Plant Dis. R e p t r . 15 (13) :
145.
(15) T H O R N E , GERALD. 1935. T h e sugar beet nematode and other indigenous nemic parasites of shadscale. J. Agr. Res. 51 (6) : 509-514.
(16) VIGLIERCHIO, DAVID R. 1960. Resistance in Beta species to the sugarbeet nematode, Heterodera schachtii. Expt. Parasitol. 10: 389-395.
(17)

W H E A T L E Y , GEORGE W . a n d J . S.

MCFARLANE.

1964.

List of p l a n t s

tested in the greenhouse to determine the host range of the sugarbeet nematode (Heterodera schachtii). Spreckels Sugar Beet Bull.
28 (5) : 37.
(18) W I N S L O W , R. D. 1954. Provisional lists of host plants of some root
eelworms (Heterodera spp.). Annals Appl. Biol. 41 (4) : 591-605.

JOURNAL OF THE A. S. S. B. T.

578

Table 1.Source of seeds or plants tested for susceptibility to the sugar beet nematode.1
Reference No.
cited in
Table 2
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)
(15)
1

No.
Specimens
tested
Ferry-Morse Seed Company, Mountain View, California
Germain's, Inc., Los Angeles, California
Weed specimens collected in Monterey County, California
Yoder Bros, of California, Inc., Salinas, California
Rancho Santa Ana Botanic Gardens of the native plants
of California, Santa Ana, California
United States Department of Agriculture
Weed specimens collected in Western Arizona
Weed specimens collected in San Bernardino County, California
The Butchart Gardens, Victoria, B. C.
Kitezawa Seed Company, San Jose, California
Clyde Robin Seed Co., Carmel Valley, California
Floralana Company, Los Angeles, California
Specimens collected in Marshall County, Iowa
Los Angeles State and County Arboretum, Arcadia, California
Seed Laboratory, Fort Collins, Colorado

67
64
59
1
15
32
5
2
2
1
10
1
4
4
2

Listing of trade names is for identification of seed only and does not imply indorsement
by the Department of Agriculture over similar ones not mentioned.

Tabic 2.Susceptibility of cultivated and weed plants to the sugar beet nematode (Heterodera schachtii Schmidt)
Studies at Salinas, California

Commercial variety
or common name

Source
of seed
or plant

Number
of
plants

Index
range

(l)1

25-

Other host studies

Avg.
index

Host
status

References

AIZOACEAE
Mesembryanthemum
crystallinum L.
Tetragonia expansa Murr.
AMARANTHACEAE
Amaranthus blitoides S. Wats.
A. carneus Greene
A. caudatus L.
A. deflexus L.
A. graecizans L.
A. hybridus L.
A. hypochondriacus L.
A. palmeri S. Wats.
A. powellii S. Wats.
A. quitensis H.B.&K.
A, retroflexus L.
A. spinosus L.
A. tricolor L.
Celosia argentea
C. argentea L. v. cristata Kuntze

Pigweed
Spiny amaranth
Josephscoat
Patnpus plume
Cockscomb

AMARYLLIDACEAE
Allium cepa L.
A. porrum L.
A. schoenoprasum L.
Allium sp.
Allium sp.
Allium spp.

Onion
Leek
Hardy perennial chives
Leek
Onion
Garlic

Iceplant
New Zealand spinach

2.04

Tumbling pigweed
Love-lies-bleeding

+5
+
+
+
+

Tumbleweed
Green amaranth

+0
+0

(0

13

(2)

25
24

2.1
1.0

(1)

25

(1)
(2)

16
24

0
0

0
0

17
9,17

17
5
10,11,18

5
12
5
9
5
5
9

+
+0
+

5
5

10

0
0

0
0
0

5,12,17

5,9,17

12
10,12

17

Numbers refer to the sources of seed listed in Table 1.


2 Number of plants examined to determine the presence or absence of nematode.
3 Infection index: 0 = no adult females observed; 1 = few nematodes; 2 = moderately infected; 3 = heavily infected; 4 = very heavily infected.
4 Listing of average indices as fractional values is for comparative purposes only and does not indicate significance of data.
5 + = host; 0 = non-host.
6 Where conflicting reports on a given plant occurred, only reports listing the plant as a host were cited.

Table 2 (continued)
Studies at Salinas, California

Other host studies

Source
of seed
or plant

Number
of
plants

Index
range

Avg.
index

Milkweed
Milkweed

(3)

21

Fiddleneck
Borage
Chinese forget-me-not
Houndstongue

(3)
(2)
(1)

20
5
22

0
1
0

0
0.2
0

(1)

25

Commercial variety
or common name

Host
status

References

ASCLEPIADACEAE
Asclepias sp.
A. syriaca L.

10

0
0
0
0

18
9
18
9

0
0
+

18
18
10,11,18

+
+
+
+

18
11
9
10

BORACINACEAE
Amsinckia intermedia F.&M.
Borago officinalis
Cynoglossum amabile
C. officinale L.
Myosotis alpestris
M. arvensis L. Hill
M. oblongata
Myosotis sp.

Field forget-me-not
Bluebird forget-me-not

CAMPANULACEAE
Lobelia spp.

Lobelia

24

(1)

CANNABINACEAE
Humulus lupulus

L.

Wild hop

CAPPARIDACEAE
Cleome spinosa Jacq. v. rosea

Giant pink queen

(1)

25

4.0

Large flowered sweet-william (2)


Colorado white sim
carnation
(4)
Maiden pink

25

1-4

1.6

CARYOPHYLLACEAE
Agrostemma githago L.
Cerastium
perfoliatum
Dianthus barbatus L.
Dianthus caryophyllus
D. deltoides L.
D. plumarius L.
Dianthus sp.
Dianthus spp.

Corncockle

Carnation
Pinks

(2)

25

22

1-4

1-4

1.6

3.0

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Gypsophila acutifolia Fisch.
G. elegans Bieb.
Lychnis coronaria L.
L. flos-cuculi L.
Lychnis sp.
Melandrium dioicum (L.)C.&G.
M. noctuiflorum (L.) Fr.
M. rubrum (Weig.) Garcke
Saponaria cerastoides
S. ocymoides L.
5. officinalis L.
5. vaccaria L.
Scleranthus annuus L.
S*7en antirrhina
S. armeria L.
S. cucubalus Wibel.
S. gallica L.
S. maritima With.
5. noctiflora L.
S. nutans L.
5. pendula L.
S. quadrifida L.
S. saxifraga L.
S*7<?ne sp.
Spergula arvensis L.
Stellaria media (L. )Vill.
Stellaria spp.
Tunica saxifraga Scop.
Vaccaria pyramidata Med.
Viscaria vulgaris Bernh.

Covent garden market

Source
of seed
or plant

Number
of
plants

(1)

22

Index
range

14

Other host studies

Avg.
index

1.3

Ragged-robin
Campion
Red Campion
Night-flowering campion
Red campion

Soapwort
Annual Knawel
Sweet-william catchfly
Bladder campion
Windmill pink
Sea campion
Night-flowering catchfly
Nottingham catchfly

Catchfly
Corn spurry
Common chickweed
Chickweed

Viscid campion

(3)

(3)

25

25

Host
status
+
+
0
0
0
0
0
0
0
+
+
+
0
0
+
0

References
18
18
18
18
10
9
9
18
18
18
9,18
9
18
18
18
9

+
0
+
0
+
+
0
0
+
+
0
+
0

18
10
18
9,18
18
18
10
9,18
1,8,9,11,18
17
9
18
9

Table 2 (continued)
Other host studies

Studies at Salinas, California


Source
of seed
or plant

Number
of
plants

Sheep-fat spiny saltbush


Shadscale

(5)

24

Garden orache
Saltbush
Garden orache
Quail brush

(5)

25

2-4

3.1

(5)
(5)

25
25

4
4

4.0
4.0

Commercial variety
or common name

Index
range

Avg.
index

Host
status

References

CHENOPODIACEAE
A triplex confertifolia
A. confertifolia (T&F.) {5. Wats.
A. hastatum
A. hortensis L.
A. hortensis L.
A. hortensis L. v. rubra Moq.
A. lentiformis
A. leucophylla
A. littorale
A. patula L.
A. polycarpa
A. rosea L.
A triplex sp.
Beta atriplicifolia
B. atriplicifolia Rouy x B. t/u/gflrfa L. (Fa)
B. corolliflora Zoss.
B. intermedia Bunge
B. lomatogona Fisch. & Meyers
B. macrocarpa Guss.
B. macrorrhiza
B. maritima L.
B. patellar is Moq.
B. patula Ait.
B. patula Soland
B. procumbens Chrys. Sir1.
B. procumbens Moq.
B. trigyna Wald. et Kitt
B. vulgaris L.
B. vulgaris L.
H. vulgaris L.

Common orache
Cattle spinach

(5)

25

0
15
1
9
17
10

+
+

8,9

+
0
+
+

17
10
7
3

+
+
+

17
7,17
3,7,17

+
+
+
+
0
0
+
+
+

3,7,9,16,17
13
3
17
3,17
7,16
3,7,17
9,10,17
9,10,17
9

3.0

Coppery orache

Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Wild beet
Mangel
Red table beet
Spinach beet

+
+
+
+
+

(6)
(6)
(6)

25
25
25

4
4
4

4.0
4.0

(6)

25

4.0

(6)

25

0-1

0.1

(fi)
(6)

25
25

0
4

0
4.0

(1)

22

4.0

4.0

Table 2 (continued)

Studies at Salinas, California


Commercial variety
or common name
Beta vulgaris "L.
Beta vulgaris L.
Beta vulgaris L. v. cicla L,
Beta vulgaris L. v. US 75
B. webbiana Moq.
Chenopodium album L.
Chenopodium album L.
Chenopodium album L.
C. amaranticolor
C. ambrosioides v. chilensis (Schrad.) Spegaz.
C. bonus henricus L.
C. botrys L.
C. capitatum (L.) Aschers
C. ficifolium Sm.
C. glaucum L.
C. hybridum L.
C. murale L.
C. murale L.
C. polyspermum L.
C. quinoa Willd.
C. rubrum L.
C. schraderianum Roem. & Schult.
Chenopodium sp.
C. urbicum L.
C. vulvaria L.
Echinopsilon
hirsutus
Hablitzia
tamnoides
Kochia scoparia
Obione portulacoides (L.) Moq.
Salicornia herbacea
Salsola kali
Spinacia glabra Mill.
5. oleracea L.

Sugar beet
Swiss chard
Leaf beet
Sugar beet
Wild beet
Fat hen
Lambsquarters
White pigweed or goosefoot

Good-King-Henry

Source
of seed
or plant

(6)
(6)

Number
of
plants

25
25

Index
range

4
0

Other host studies

AYg.
index

4.0
0

Host
status

2,3,5,9,10,11,17
5,17
10

3,7,16,17
8,9
4
1
17
5
9,11
18
5,9,11,17
18
10
9
9
5,11,12,17
8,9,18
1,9,18
1,8,9,11
5
9,10
17
9,11
1
18
1
9
1
1
9
5,10,11,17

+
+
+
+
+
+
0

Strawberry-blite
Fig-leaved goosefoot
Oak-leaved goosefoot
Sowbane
Nettle-leaved goosefoot
Sowbane
Many-seeded goosefoot

+
+
+
+
+
+
+
0

Red goosefoot

+
+
0

Pigweed
Stinking goosefoot

References

+
+
+

+
+
0
+
0

Sea purslane

+
0
0

Summer spinach
Spinach

+
+

Table 2 (continued)
Other host studies

Studies at Salinas, California

Commercial variety
or common name
Spinacia oleracea L.
Suaeda maritima
CISTACEAE
Helianthemum

scoparium

Source
of seed
or plant

Number
of
plants

Index
range

Avg.
index

Winter spinach

Nutt.

Common rush-rose

(7)

25

Coldenhead
Yarrow
Perennial ragweed
Ragweed
Dogfennel
Lavender-Munstead strain
Burdock
Mugwort
Mugwort

(8)

12

(3)

25
22
25
14

0
0
0
0

0
0
0
0

16
24
25
23
25
23
25
25
25
25

0
0
0
0
0
0
0
0
0
0

0
0
0
0
0
0
0
0
0
0

(1)

25

(2)

25

(3)

25

9
17
10

17

9,10

0
0

10
9

73
z

A.

(3)
(5)
(1)
(1)
(6)
(2)
(2)
(3)
(3)
(2)

0
0
0

OF THE

Gold fields
Dble mixed English daisy
Gt. Crego mixed aster
Safflower
Dusty-miller
Bachelors-button
Napa thistle
Yellow star thistle
Green curled endive
Asparagus chicory or
catalogna
Chicory
Lg.-root coffee chicory
Canada thistle
Common thistle
Bullthistle

(3)
(9)
(10)

References
9
1

IAL

C. intybus L.
C. intybiw L.
Cirsium arvense (L.) Scop.
Cirsium arvense (L.) Scop.
C. vulgare (Savi) Tenore

+
0

Joui

COMPOSITAE
Acamptopappus
sphaerocephaltts
(Harv. & Gray) A. Gray
Achillea millefolium L.
Ambrosia psilostachya DC.
Ambrosia sp.
Anthemis cotula L.
Anthemis sp.
Arctium lappa L.
Artemisia sp.
A. vulgaris L.
Aster sp.
Baeria chrysostoma v. gracilis
Bellis perennis
Callistephus sinensis
Carthamus tinctorius
Centaurea cineraria L.
C. cyanus
C. melitensis L.
C. solstitialis L.
Cichorium endivia
C. intybus L.

Host
status

C/i
C/i
W

0
_ _

Table

(continued)
Studies at Salinas, California

Commercial variety
or common name
Coreopsis tinctoria Nutt.
Cosmos bipinnatus Cav.
Cynara cardunculus
C. scolymus L.
Dicoria canescens T. & G.
Dimorphotheca
aurantiaca
Erigeron glaucus Ker.
Erigeron sp.
Eupatorium coelestinum L.
Gaillardia picta
Grindelia camporum v. interioris
(Jepson) Steyermark
Helianthus annuus L.
Helianthus spp.
Helichrysum bracleatum Andr.
Heterotheca grandiflora Nutt.
Hypochoeris radicata L.
Hypochoeris radicata L.
Lactuca saligna
L. sativa L.
L. serriola L.
Layia platyglossa v. elegans
Leontodon leysseri (Wallr.) G. Beck
Madia gracilis (Smith) Keck
Palafoxia linearis (Cav.) Lag.
Parthenium argentatum Gray
Peucephyllum schottii Gray
Picris echioides L.
Senecio douglasii D. C.
S. vulgaris L.
Silybum marianum (L.) Gaertn.
Sonchus arvensis L.
S. oleraceus L.

Dble mixed calliopsis


Sensation dazzler cosmos
Lg. smooth cardoon
Greenglobe artichoke
Desert dicoria
Mixed African-daisy
Seaside daisy
Blue mink ageratum
Ann. double mixed
Gum plant
Common sunflower
Sunflower
Mixed strawflower
Telegraph-plant
Coast dandelion
Hairy cats-ear
Willow lettuce
Lettuce
Prickly lettuce
Tidy tips
Hairy hawkbit
Slender tarweed
Spanish needle
Guayule v. 593
Pigmy cedar
Bristly oxtongue
Shrubby butterweed
Common groundsel
Milkthistle
Sowthistle
Sowthistle

Other host studies

Source
of seed
or plant

Number
of
plants

Index
range

Avg.
index

(1)
(1)
(1)
(1)
(7)
(1)
(3)
(3)
(1)
(0

25
25
25
25
17
22
23
24
24
25

0
0
0
0
0
0
0
0
0
0

0
0
0
0
0
0
0
0
0
0

(3)
(3)
(2)
(1)
(3)
(3)
(3)
(6)
(6)
(3)
(5)
(3)
(3)
(7)
(6)

25
14
25
22
25
23
25
25
25
25
25
25
13
20
22

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

(3)
(7)
(3)
(3)

5
25
25
25

0
0
0
0

0
0
0
0

Host
status

References

10

10

0
0

5,9,10,12,17
17

17

10
17

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Sonchus sp.
Stephanomeria sp.
S. tenuifolia (Torr.) H. M. Hall
Tagetes spp.
Tagetes spp.
Taraxacum officinale Wiggers
Taraxacum officinale Web.
Taraxacum sp.
Tithonia
rotundifolia
Tragopogon porrifolius L.
Xanthium italicum Mor.
Zinnia elegans
CONVOLVULACEAE
Calonyction aculeatum House
Calystegia septum L. R. Br.
Convolvulus arvensis L.
Convolvulus arvensis L.
Ipomoea botata
1. cardinalis L.
/. quamoclit L.
Ipomoea sp.
CRUCIFERAE
Aethionema
grandiflora
A. pulchellum
A. stylosum
Alliaria officinalis Bieb.
^iy55m alpestre
A. argenteum
A. borzaeanum Nyar.
4. mariHmum L.
A. marilimum L.

Source
of seed
or plant

Sowthistle
Stephanomeria
Narrow-leaved stephanomeria
African crackerjack marigold
Dwarf marigold
Common dandelion
Dandelion
Dandelion
Mexican-sunflower
Salsify
Cockle-bur
Calif, giant-white
purity zinnia
Moonflower
Larger bindweed
Lesser bindweed
Morning glory
Sweet potato
Cardinal climber
Mixed cypress vine
Candy pink morning-glory

Garlic mustard

Sweet alyssum
Sweet alyssum v. violet queen

(3)
(3)
(2)
(1)
(3)

Other host studies

Number

of
plants

20
16
25
25
25

Index
range

0
0
0
0
0

Avg.
index

25
25

0
0

0
0

(2)

25

(1)

12

22
25
25

0
0
0

References

0
0

17
10

0
0

12
17

0
0
0
0

9
9
17
17

0
0
0
+
0
+
+
+
+

18
18
18
18
18
18
18

0
0
0
0
0

(0
(3)

(1)
(1)
(1)

Host
status

0
0
0

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Alyssum

maritimum

Lam.

A. montanum
A. saxatile L.
Alyssum sp.
A. spinosum
Arabis arenosa Scop.
A. bellidifolia
A. caucasia Willd.
A. muralis Bertol
A. turrita L.
A. verna R. Br.
Armoracia lapathifolia Gilib.
Aubrieta columnea Guss.
Aubrieta sp.
Barbarea longirostris
B. praecox R. Br.
B. vulgaris R. Br.
Berteroa incana (L.) DC.
Biscutella auriculata L.
B. depressa
B. laevigata L.
Brassica campestris L.
Brassica campestris L.
B. caulorapa Pasq.
B. cernua Thbg.
B. ;'unca
B. /tmcca (L.) Czern. & Coss.
B. napobrassica Mill.
B. na/7W$ L.
B. napMi L.
B, napus L.

Sweet alyssum v. carpet


of snow
Yellow saxatile
compactum alyssum
Sweet alyssum

Source
of seed
or plant

Number
of
plants

(3)

25

(2)

10

Other host studies

Avg.
index

Host
status

1-4

2.2

+
+

5
18

0-1

0.1
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
T
0
+
+
+
+
+

10,18
18
11
18
18
18
11
18
9,10
11
9
18
11
11,18
18
18
18
18
5,17
18
10
11

+
+
+
+
+

18
10
9,10,17,18
2,9,18
5

Index
range

Garden arabis

Horseradish

Yellow-rocket

Common mustard
Wild turnip
Kohlrabi
So. gt. curled mustard
Trowse mustard
Rutabaga
Coleseed or rape
Swede
Turnip

(1)

25

References

4.0

Table 2 (continued)

Number
of
plants

Black mustard
Broccoli
Brussel sprouts
Cabbage
Red cabbage
Cauliflower
Coliards
Kale
Kohlrabi

(11)

25

Kale or coliards
Dwarf curled kale
Cauliflower
Cabbage
Brussd sprouts
Early white Vienna kohlrabi
Chinese cabbage
Turnip

(11)
(2)

25
25

Commercial variety
or common name

Gold-of-pleasure
Sheoherds-purse
Cuckoo flower
Hoarycress
Siberian wallflower

Wallflower
Horseradish
Scurvy grass

Host
status

References

4.0

4
4

4.0
4.0

+
+
+
+
+
+
+
+
+
+
+

8,9,11,18
5,9,10,17,18
5,9,17,18
2,5,9,17,18
2
5,9,17,18
5
9,17,18
9
11
10

+
+
+

10
10
10

+
+
+
0
+
+
+
+

5
8,9,10,17,18
9
1
18
1,8,9,10,17,18
11
1,18

+
+
+
+
+
+
+

9
11
18
9,10,11
8
18
9,18

(2)

25

4.0

(1)

25

4.0

(3)

25

4.0

(3)

20
11

3-4
4

3.9
4.0

(1)

C4

OUR]

B. nigra (L.) Koch.


Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L.
Brassica oleracea L. v. acephala DC.
B. oleracea L. v. acephala DC.
B. oleracea L. v. botrytis L.
B. oleracea L. v. capitata L.
B. oleracea L. v. gemmifera Zenk.
B. oleracea L. v. gongylodes
B. pekinensis (Lour.) Rupr.
B. rapfl L.
Brassicella erucastrum (L.) 0. & E. Schultz
Cakile maritima
Camelina sativa (L.) Crantz
Capsella bursa-pastoris (L.) Medic.
Cardamine impatiens L.
C. pratensis L.
Cardaria pubescens (C. A. Mey) Roll.
Cheiranthus allionii Hort.
C. alpinus L.
C. annuus
C. chetrit L.
Cochlearia armoracia
C. glastifolia L.
C. o/jit-ijia/ii- L.

Avg.
index

Index
range

00

Other host studies

Studies at Salinas, California


Source
of seed
or plant

>
r
0

>
C/3
C/3

Table 2 (continued)Studies at Salinas, California

Commercial variety
or common name
Conringia orientalis (L.) Dum.
Coronopus squamatus (Forsk.) Asc.
C. ruellii
Descurainia sophia (L.) Prantl.
Diplotaxis erucoides (L.) DC.
D. muralis (L.) DC.
D. tenuifolia (L.) DC.
Erysimum
allionii
E. capitatum
E. cheiranthoides L.
E. cheiranthoides L.
E. heiraciifolium L.
E. insulate v. grandifolum
Hesperis matronalis L.
Hesperis matronalis L.
Hesperis sp.
/fecrw coronaria Hort.
/ t e r i j sp.
Jfcem spp. (amara & umbellata)
I. umbellata L.
Isatis tinctoria L.
Lepidium graminifolium L.
L. latifolium L.
I. nitidum Nutt.
L. sativum L.
Lobularia maritima (L.) Desv.
Lunaria annua L.
JL biennis
L. rediviva L.
Malcolmia littorea (L.) DC.
Af. maritima (L.) R. Br.
Matthiola annua

Hares-ear-cabbage
Swinecress
Swinecress
Flixweed
White-wall rocket
Wall rocket
Perennial wall rocket
Siberian wallflower
Wallflower
Treacle-mustard
Wormseed-mustard
Wallflower
Dames-violet
Sweet rocket
Rocket
Candytuft
Mixed candytuft
Candytuft
Woad
Broad-leaved pepperwort
Common peppergrass
Garden cress
Sweet alison
Hon; sty
Money plant

Source
of seed
or plant

Other host studies

Number

of
plants

Index
range

Avg.
index

Host
status

18
1,18
8
9,18
1,18
18
18
18

'+
+
+

8,9,18
10
18

1,5,18

0
+
+

10
18
9,10

+
-f0
0

11,18
9,11,18
18
18

+
0
+
+
+
0
0
(5)

21

1-2

1.0

(5)

25

3-4

3.6

(1)

24

0-1

0.1

(2)

21

1-4

2.3

(3)
(2)

15
20

0
4

0
4.0

(1)

24

2-4

3.5

+
0
+
+
0

Virginian stock

References

(2)

24

3-4

3.6

+
0

8,9,10,18
18
9,10,18
18
18
1,2,11,18
1

Tabic 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
M. bicornis (Sib. & Sm.) DC.
M. incana (L.) R. Br.
Af. sinuata (L.) R. Br. v. glabra
Matthiola sp.
Moricandia arvensis (L.) DC.
M. sonchifolia
Myagrum perfoliatum L.
Nasturtium microphyllum (Boenn.)
N. officinale R. Br.
Peltaria alliacea Jacq.
Raphanus maritimus Sm.
R. raphanistrum L.
R. sativus L.
R. sativus L.
Rapistrum perenne All.
R. rugosum (L.) All.
Rorippa amphibia (L.) Besser
jR. islandica (Oeder) Borbas
R. nasturtium
R. sylvestris (L.) Besser
Saxatile citrinum
Sinapis alba L.
S. arvensis L.
S. kaber (DC.)
Sisymbrium austriacum Jacq.
S. irio L.
S. officinale (L.) Scop.
S. orientale L.
S. sophia L.
Teesdalia nudicaulis
Thlaspi arvense L.

Source
of seed
or plant

Number
of
plants

Index
range

Other host studies

Avg.
index

Night-scented stock
Stock
Sea stock
Garden stock

Rchb.

One-rowed watercress
Watercress
Sea radish
Wild radish
Radish
Wild radish

Great yellowcress
Marsh yellowcress
True watercress
Creeping yellowcress
Alyssum
White mustard
Charlock
Wheeler charlock
London rocket
Hedgemustard
Eastern rocket

Field pennycress

(1)
(3)

25
25

14
4

2.6
4.0

(11)

20

4.0

(9)

10

Host
status

References

0
0
0
0
0
+
+
+
+
+
+
+
+

18
5
18
9,10
18
18
18
18
18
18
18
9,17
2,5,9,10,17

+
+
+
+

11
18
18
18

18

+
+
+
+
+
+

2,9,18
1,9,18
10
18
18

+
+
+
+

1,8,9,11
18
11
1
1,9,11

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Thlaspi arvense L.
Tropaeolum spp.

Mithridate mustard
Calif, giants mixed
nasturtium

Avg.
index

1-3

1.3

(2)

25

(2)
(2)
(2)
(2)

25
25
25
25

0
0
0
0

0
0
0
0

(12)

19

(3)

(11)

21

(1)

25

Sun spurge
Caper spurge
Castor bean
Castor oil bean

(11)

(2)

25

Redstem filaree

(3)

23

CYPERACEAE
Cyperus eragrostis Lam.

Tall umbrella-plant

DIPSACACEAE
Dipsacus fullonum
D. lacineatus L.
Scabiosa caucasia

Fullers teasel
Teasel
Pincushion flower

GERANIACEAE
Erodium cicutarium L'Her.
. mancescavi Coss.
Geranium rotundifolium L.
Pelargonium hortorum Bailey

Index
range

Other host studies

25

Kleckley's sweet watermelon


Muckmelon
Hale's best 936 cantaloupe
Cucumber
Banana-squash
Halloween pumpkin
Marrow
Squash
Loofa sponge

L.

Number
of
plants

(2)

CUCURBITACEAE
Citrullus vulgaris
Cucumis melo L.
C. melo reticulatus
C, sativus L.
Cucurbita maxima
C. pepo L.
C. pepo L.
Cucurbita sp.
Luffa cylinrica

EUPHORBIACEAE
Euphorbia helioscopia
E. lathyrus L.
Ricinus communis L.
Ricinus communis L.

Source
of seed
or plant

Round-leaved cranes-bill
Geranium

Host
status

References

10

10,12,17

0
0

9
10

0
0
0

9
9
12.17

0
0
0
0

17
9
9
17

Table 2 (continued)
Other host studies

Studies at Salinas, California

Commercial variety
or common name
GRAMINEAE
Agropyron sp.
Alopecurus sp.
Arrhenatherum
elatius (L.)
Arrhenatherum
elatius (L.) J. & C. Presl.
Avena byszantina L.
A. fatua L.
A. saliva L.
A. sterilts
Briza maxima L.
Bromus inermis Leys
B. mollis L.
B. secalinus L.
Cynodon dactylon L.
Cynosurus cristatus L.
Dactylis glomerata L.
Dactylis glomerata L.
Echinochloa crusgalli |[L.) Beauv.
Festuca elatior L.
Festuca elatior L.
F. pratensis Huds.
Hordeum
jubatum
H. sativum Pers.
H. vulgare L.
H. vulgare L.
Lolium multiflorum Lam.
L. perenne L.
Phalaris canariensis L.
Phleum pratense L.
Poa annua L.
P. pratensts agg.
Polypogon mott.speh"enstj L.

Couchgrass
Foxtail
Tall oatgrass
False-oatgrass
Red cultivated oa s
Wild oats
Oats
Oats
Quaking grass
Smooth bromegrass
Soft bromegrass
Bermuda grass
Crested dogs-tail
Cocks-foot
Orchard Grass
Watergrass
Meadow fescue
Tall fescue
Meadow fescue
Wild barley
Barley
Atlas barley
Barley
Italian ryegrass
Ryegrass
Canarygrass
Timothygrass
Annual meadowgrass
Smooth stalked meadowgrass
Rabbitfootgrass

Source
of seed
or plant

'(13)
(3)

Number

of
plants

25
25

Index
range

0
0

Avg.
index

25

(2)
(3)

24
25

0
0

0
0

25

References

0
0
0
0
0
0
0

9,10
10
17
9
17
17
9,10,12,17

0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0

17
9
9
17
9
9
17
17
17
17
9
17
9
12
10,17
9
9

0
0
0

9,10
9
9

0
0

(3)

(3)

Host
status

Table 2 (continued)Studies at Salinas, California

Commercial variety
or common name
Secale cereale L, Bieb.
Setaria glauca L.
Sorghum halapense L. Pers.
S. vulgare Pers.
S. vulgare Pers.
Triticum aestivium L.
T. vulgare Vill.
lea mays L.
lea mays L. v. saccharata
loysia

japonica

HYDROPHYLLACEAE
Nemophila
insignis
Phacelia campanularia
P. minor
P. tennuifolia Harv.
LABIATAE
Ajuga chamaepitys (L.)
Galeopsis dubia Leers
G. pyrenaica Bartl,
G. speciosa Mill.
G. tetrahit agg.
Hyssopus officinalis L.
Lamium album L.
/.. mo//
L. purpureum L.
Lavandula sp.
Leonurus sibiricus
Majorana hortensis
Marrubium vulgare L.
Melissa officinalis L.

Rye
Yellow foxtail
Johnson grass
Sweet sorghum v. honeydrip
Milo
Wheat
Wheat
Corn
Stowell's evergreen
sweet corn
Japanese lawngrass

Baby-blue-eyes
Desert bluebell
Calif, bluebell

Source
of seed
or plant

(2)

Number
of
plants

25

Index
range

Other host studies

Avg.
index

(1)

22
25

0
0

0
0

(1)
(5)
(5)

7
17
19

0
0-1
0

0
0.7
0

Large-flowered hemp-nettle
Common hemp-nettle
Hyssop
White deadnettle
Red deadnettle
Lavender
(1)
(3)
(1)

25
25
16

0
1
0

References

0
0
0

9,17
17
17

0
0
0
0

17
10,12
9,17
9,10,17

0
0
+
+
+
0
0
0
0
0
0

9
18
9,18
2,8,9,18
18
9
9,18
18
9,18
10
18

10

Groundpine
Downy hemp-nettle

Sweet marjoram
Horehound
Lemon balm

Host
status

0
1.0
0

Table 2 (continued)
Other host studies

Studies at Salinas, California

Commercial variety
or common name
Mentha arvensis L.
M. piperita L.
Molucella laevis
Ocimum basilicum L.
Ocimum basilicum L.
Salvia lanigera
S. officinalis L.
S. sclarea L.
Salvia sp.
Scutellaria orientalis L.
Stachys annua (L.) L.
S. officinalis (L.) Trev.
S. sylvatica L.
Thymus sp.
T. vulgaris
LEGUMINOSAE
Anthyllis vulneraria L.
Arachis hypogaea L.
Arachis hypogaea L.
Arachis hypogaea L.
Crotalaria mucronata
C. pumila
C. spectabilis
Cyamopsis tetragonoloba
Glycine max (L.) Merr.
Glycine max (L.) Merr.
Lathyrus odoratus L.
Lathyrus odoratus L.
Lens sp.
Lespedeza stipulacea Maxim.

Corn mint
Peppermint
Bells of Ireland
Ornamental basil
Sweet basil

Source
of seed
or plant

Number

(2)
(1)
(1)
(14)

25
24
25
19

of
plants

Index
range

Avg.
index

0
0
0
0-1
0
0

0
0.1
0
0

Garden sage
Sage
Skullcap
Woundwort
Betony
Hedge woundwort
Thyme
Thyme

Kidneyvetch
Argintine-peanut
(Spanish-type)
Southeastern runner
Virginia bunch peanut

Rattlebox
Texel guar
Soybean
Soybean v. Earlyana
Winter-flowering
spencer sweetpea
Sweetpea
Lentil
Korean lespedeza v. auburn

(2)

25

Host
status

References
9
10

10

0
0
0
0
0
0
0
0

9
9
10
9
9,18
18
18
10

9,18

(6)
(b)
(6)
(14)
(14)

20
25
25
9
25

0
0
0
0
0

0
0
0
0
0

(6)

25

(6)

25

1-3

2.0

(2)

25

(6)

23

0-1

0.3

12

5,10,17

+
0

12
18

Table 2 (continued)
Studies at Salinas, California
Number
of
plants

Koren lespedeza v. climax


Korean lespedeza v. rowan
Common lespedeza
Giant striata
Commercial Kobe type
Birdsfoot trefoil

(6)
(6)
(6)
(6)
(6)

15
21
12
22
19

Lupine
Annual lupine

(5)

Burclover
Hairy medick
Alfalfa
Caliverde alfalfa
Chilean alfalfa
German alfalfa
Lahontan alfalfa
Lucerne
Ranger alfalfa
White swectclover
Common melilot
Yellow sweet clover
Sweetclover
Sanfoin

(3)

21

0-2

0.6

(2)
(2)
(2)
(2)

25
25
25
25

0
0
0
0

0
0
0
0

(2)
(3)

25
25

0
0

0
0

(2)

25

Scarlet runner bean


Fordhook pole lima bean
Runner bean
Scarlet runner bean
Black bean

(1)
(2)

22
25

0
0

0
0

22

Commercial variety
or common name

Lespedeza stipulacea Maxim.


Lespedeza stipulacea Maxim.
L. striata (Thunb.) H. & A.
L. striata ( T h u n b . ) H. & A.
L. striata ( T h u n b . ) H. & A.
Lotus corniculatus L.
L. tetragonolobus L.
Lupinus
albo-coccineus
L. nanus
Lupinus sp.
Medicago ciliaris Willd.
M. hispida Gaertn.
Af. hispida Gaertn.
Af. sativa L.
Af. sativa L.
Af. sativa L.
Af. sativa L.
Af. sativa L.
Af. sativa L.
Af. sativa L.
Melilotus alba Desr.
Af. officinalis (L.) Lam.
Af. officinalis (L.) Lam.
Melilotus Mill. sp.
Onobrychis viciifolia Scop.
Ornithopus compressus L.
O. safii/tw Brot.
Phaseolus coccineus
P. lunatus L.
Phaseolus multiflorus Willd.
Phaseolus multiflorus Willd.
Phaeseolus sp.

Other host studies

Source
of seed
or plant

(1)

12

Index
range

0-2
0-1
1-2
1-2
1-3

Avg.
index

Host
status

0.7
0.6
0.9
1.5
2.3
0
0
0

17,18
18
18

+
0
0
0
0

12
18
17
18
5,10,12,17

9,18

0
0
0
a
0
0
0

10
18
17
12
9,18
18
18

0
0

17
9,18

10,12

Table 2 (continued)Other host studies

Studies at Salinas, California

Commercial variety
or common name
P.
P.
P.
P.
P.
P.

vulgaris
vulgaris
vulgaris
vulgaris
vulgaris
vulgaris

L.
L.
L.
L.
L.
L.

P. vulgaris L.
P. vulgaris L.
P. vulgaris L.
Pisum sativum L.
P u m sativum L.
Piswm sativum L. v. arvense
Sesbania exaltata (Ref.) Rydb.
S. macrocarpa
Trifolium hybridum L.
T. pratense L.
7. repens L.
T. repens L.
T. rejbens L.
Trifolium sp.
Trigonella caerulea (L) Ser.
Fjda americana Muhl.
V. atropurpurea
V. faba L.
V. faba L.
V. sativa L.
Vicia sp.
Vicia sp.
V. villosa Roth.
Vigna sinensis Endl.
Vigva sinensis L.
Figna sp.

Black wax bean


Blue Lake #21 bean
Dwarf bean
French bean
Golden wax bush bean
Kentucky wonder #191
white bean
Navy bean
Pencil pod wax bean
White-seeded pole bean
Dwarf telephone pea
Garden pea
Papago pea
Sesbania
Alsike clover
Redclover
Whiteclover
White Dutch clover
Wild whiteclover
Berseem clover
American vetch
Purple vetch
Broad bean
Small-seeded horse bean
Winter vetch
Lana wooly pod vetch
Purple vetch
Hairy vetch
Blackeye cowpea
Cowpea
Iron cowpea

Source
of seed
or plant

(2)

Number
of
plants

25

Index
range

Avg.
index

24

(2)
(2)
(2)
(2)
(6)

25
25
25
25
25

0
0
0
0
0

0
0
0
0
0

24

1-3

1.6

(2)

15

(2)

25

25

(2)

References

17

0
0
+

18
9
12

0
0

17
5,17

(2)

(6)

Host
status

(3)
(2)
(2)

25
25
12

0
0
0

0
0
0

(2)

25

(1)

25

12

+
+
0
0
0

11
12
10
9,10,12,17.18
17,18

18

0
0
0
0

17
9
17
9

+
0
+
0
+

12
10
11
17
12

Table 2 (continued)

$
Studies at Salinas, California
Source
of seed
or plant

Number
of
plants

Index
range

Avg.
index

Asparagus

(6)

25

Tritoma

(1)

20

Perennial flax
Scarlet flax
Blue flax

(1)
(2)

23
25

0
0

0
0

Kudzu-vine

(1)

25

(13)

25

(1)
(11)

16
14

0
0

0
0

Commercial variety
or common name
LILIACEAE
Asparagus officinalis L.
Bulbine annua Willd.
Kniphofia uvaria
LINACEAE
Linum anglicum Mill.
L. grandiflorum rubrum
L. perenne
Linum sp.
Pueraria thunbergiana Benth.
MALVACEAE
Abutilon theophrasti
Althaea rosea

Medic.

Callirhoe involucrata ( T . & G.) Gray


Gossypium spp.
Malva parviflora L.
M. rotundifolia L.
M. sylvestris L.

Velvetleaf
Double newport pink
hollyhock
Poppy-mallow
Cotton
Cheeseweed
Cheese-mallow
Common mallow

NYCTAGINACEAE
Mirabilis jalapa L.

Mixed four-o'clock

ONAGRACEAE
Clarkia biloba v. Brandegeae
Clarkia sp.
Fuchsia sp.
Godetia amoena
Lopezia coronata Andr.
Oenothera biennis L.
O. hookeri
0. hookeri Montereyensis
0. lamarckiana Ser.
0. ttiicrantha (Nels.) Munz

Other host studies

Clarkia
Fuchsia
Double-mixed godetia
Evening
Evening
Evening
Evening
Evening

primrose
primrose
primrose
primrose
primrose

(2)

21

(5)

15

(1)

24

(11)
(11)
(11)
(3)

14
7
14
25

0
0
0
0

0
0
0
0

Host
status

References

0
0

5,9
9

0
0
0
0

12,17
5
17
9

0
0

10
10

+
0

9
9

T a b k 2 (continued)Other host studies

Studies at Salinas, California

Commercial variety
or common name
PAPAVERACEAE
Argemone mexicana L.
Eschscholtzia californica Cham.
Eschscholtzia californica Cham.
Eschscholtzia sp.
Papaver rhoeas L.

Mexican-poppy
Extra golden Calif.-poppy
True deep orange
Calif-poppy

Papaver rhoeas L.
Papaver sp.

Double-mixed Shirleypoppy
Field poppy
Poppy

PHYTOLACCACEAE
Phytolacca acinosa Roxb.
P. americana L.

Pokeweed

Source
of seed
or plant

Number
of
plants

Index
range

Avg.
index

(1)

18

(2)

23

(1)

25

0-1

0.6

PLANTAGINACEAE
Plantago coronopus L.
P. lagopus L.
P. lanceolata L.
P. lanceolata L.
P. major L.
P. major L.

(3)

25

Buckhorn plantain
Narrow-leaf plantain
Common plantain
Greater plantain

(3)

25

PLUMBAGINACEAE
Limonium sinuatum Mill.

Statice-mixed straw-flower

(1)

25

(5)
(11)

25
1

0
4

0
4.0

Cut-leaved plantain

POLEMONIACEAE
Gilia abrotanifolia Nutt.
G. achilleaefolia
G. capitata
POLYGONACEAE
Emex spinosa Campd.
Fagopvrum esculentum Moench
Fagopyrum sagittatum Gilib.

Buckwheat
Buckwheat

(3)

23

Host
status

References

0
0

17,18
9

0
0

18
10

+
+

18
17

17

0
0
0

9,18
1,10.18
9

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
F. tartaricum Gaertn.
Polygonum amphibium L.
P. aviculare agg.
P. convolvulus L.
P. lapathifolium L.
P. orientate L.
P. pensylvanicum L.
P. persicaria L.
P. punctatum
Polygonum sp.
Rheum rhaponticum L.
Rumex acetosa
R.
J?,
.R.
R.
J?,
i?.
R.
jR.
ii.
JR.
i?.

acetosella L.
alpinus L.
confertus
crispus L.
hydrolapathum Huds.
maritimus
obtusifolius L.
palustris Sm.
patientia L.
pulcher L.
sanguineus L.

Source
of seed
or plant

Number
of
plants

Index
range

Other host studies

Avg.
index

Amphibious polygonum
Knotgrass
Willow smartweed
Princesfeather

(3)

25

0-1

(1)

21

4.0

(0

25

Monks-rhubarb
Curled dock
Great water dock

References

0
0
0
+

9
9
9,18
11

0
+
+
+
+
+

10
14
8,9,18
14
10,17
5,9.10,11

+
+
+
+
+
+
+
+
+

11
18
18
2,8,9,10
11,18
1
9
18
11

18

10,11,17

0.6

Persicaria
Smartweed
Rhubarb
Large-leaved
French sorrel

Host
status

(3)

25

1-4

1.7

Fiddle dock
Red-veined dock

(3)

25

4.0

PORTULACACEAE
Montia
gypsophiloides
Portulaca grandiflora Hook.
P. oleracea L.

Indian lettuce
Moss rose portulaca
Purslane

(6)
(1)
(3)

22
22
25

PRIMULACEAE
Anagallis arvensis L.
Anagallis arvensis L.

Common pimpernel
Scarlet pimpernel

(3)

Broad-leaved dock
Marsh dock

0
0-1
0-1

0
0.7
0.4

T a b k 2 (continued)
Studies at Salinas, California

Commercial variety
or common name

Other host studies

Source
of seed
or plant

Number
of
plants

(14)
(1)
(2)

25
25
25

0
0
0

0
0
0

(1)

22

Index
range

Avg.
index

Host
status

References

RANUNCULACEAE
Aquilegia vulgaris
Aquilegia vulgaris
Delphinium ajacis
Delphinium sp.
Nigella damascene
Nigel la sp.

Columbine
Imperial mixed columbine
White supreme larkspur
Dwarf delphenium
Love-in-a-mist
Love-in-a-mist

0
0
0
0

18
9
18
17

Agrimony
Strawberry
Salad burnet

0
0
0

9
17
9

Goosegrass

0
0

9
9

RESEDACEAE
Reseda alba L.
Reseda alba L.
R. lutea L.
R. odorata
R. odorata

Upright mignonette
White mignonette
Wild mignonette
Mignonette
Red goliath mignonette

(1)

14

4.0

ROSACEAE
Agrimonia eupatoria L.
Fragaria chiloensis Duchesne
Poterium sanguisorba L.
RUBIACEAE
Crucianella angustifolia L.
Galium aparine L.
RUTACEAE
Dictamnus albus L,
Larrea divaricata Cav.

Creosote bush

(8)

15

Snapdragon
Snapdragon
Small toadflax

(6)

25

SCROPHULARIACEAE
Antirrhinum majus L.
Antirrhinum sp.
Chaenorrhinum minus (L.) Lange

18

18

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Collinsia heterophylla
Linaria sp.
L. vulgaris Hill
Nemesia compacta
Scrophularia nodosa L.
Veronica agrestis L.
V. persica Poir.
V. spicata L.

Toadflax
Butter-&-eggs
Triumph mixed nemcsia
Figwort
Procumbent speedwell
Buxbaum's speedwell
Spiked speedwell

Other host studies

Source
of seed
or plant

Number
of
plants

(5)

21

1-2

0.5

(1)

23

Index
range

Avg.
index

Host
status

References

9
5

0
0
0
0

9
9
18
18

SOLANACEAE
Capsicum annum
Capsicum annum
C. frutescens L.
Datura stramonium L.
Datura stramonium L.
Hyoscyamus niger L.
Lycopersicon esculentum Mill.
Lycopersicon esculentum Mill.
Lycopersicon esculentum Mill.
L. pimpinellifolium Mill.
Nicotiana affinis
N. glauca Graham
N. tabacum L.
N. tabacum v. Samsun
Petunia parviflora Juss.
Petunia spp.
Physalis longifolia
P. pubescens
Physalis sp.
Physalis sp.
Salpiglossis sinuata

Bell pepper
Pimiento pepper
Pepper
Jimson weed
Thornapple
Black henbane
Pearson A-l tomato
Pearson XL tomato
Tomato
Wild tomato
Affinis hybrid nicotiana
Tree tobacco
Tobacco
Turkish tobacco
Petunia seaside
Colorama mixed petunia
Perennial groundcherry
Yellow husk tomato
Golden queen tomato
Yellow pear tomato
Painted tongue

(2)
(2)

25
25

0
0

0
0

(3)

25

(3)
(2)

25
25

0
2

0
2.0

(6)
(2)
(3)

25
25
25

2
0
0

2.0
0
0

(6)
(3)
(2)
(15)
(13

25
25
25
25
10

0
0
0
0
0

0
0
0
0
0

(13)

25
19

2
1-3

2.0
2.3

(I)

17

0
0

9
9

40

6
17

0
0

17
17

12

Table 2 (continued)
Studies at Salinas, California

Commercial variety
or common name
Solarium douglasii
S. dulcamara
S, eleagnifolium Cav.
S. melongena
S. nigrum L.
S. rostratum Dunal
S. sarrachoides Sendt.
Solarium sp.
S. tuberosum L.

Nightshade
Bittersweet nightshade
White horsenettle
N.Y. improved egg plant
Black nightshade
Kansas-thistle buftalo-bur

Source
of seed
or plant

Number
of
plants

(6)

25
25
22
20
25
11

(7)
(2)
(15)

Index
range
1-3
0
0
0-1
0-1
0

Other host studies

Avg.
index
1.6
0
0
0.1
0.4
0

Nightshade
Potato

Host
status

References

12

0
+
0

9
12
9,10,17

18

0
0

9
9

0
0
0

9
5,9,12,17
10

0
0

17
9

5,9,12

10,17

TROPAEOLACEAE
Tropaeolum

peregrinum

UMBELLIFERAE
Aethusa cynapium L.
Anethum graveolens L.
Anthriscus
cerefolium
Apium graveolens L.
Apium graveolens L.
Apium graveolens L. v. dulce Pers.
A. graveolens L. v. rapaceum
Carum carvi L.
Cicuta douglasii (PC) C & R.
Conium maculatum L.
Conium maculatum L.
Coriandrum sativum
Daucus carota L.
Daucus carota L.
Daucus carota L. v. sativa DC.
Foeniculum dulce
F. vulgare

Fools-parsley
Dill
Chervil
Celeriac
Celery
Celery
Smooth Prague celeriac
Caraway
Water hemlock
Hemlock
Poison-hemlock
Coriander
Carrot
Imperator carrot
Carrot
Anise
Sweet-florence fennel

(2)
(1)

(2)
(1)

25
15

25
25

0-3
1-2

0
0

1.0
1.0

0
0

(3)
(1)

25
25

0-3
0

0.6
0

(1)

22

(1)
(2)

25
25

0
0

0
0

Table 2 (continued)Studies at Salinas, California

Commercial variety
or common name

Pastinaca sativa L.
Pastinaca sativa L.
Pastinaca sp.
Petraselinum crispum L. (Mill.)
P. hortense crispum
Scandix pecten-veneris L.
Trachymene caerulea

Nym.

Hollow crown parsnip


Parsnip
Wild parsnip
Parsley
Plain-leaved parsley
Shepherds-needle
Blue lace flower

Source
of seed
or plant

Number
of
plants

(1)

25

Index
range

0-1

Other host studies

Avg.
index

Host
status

References

0.1

(2)

25

(1)

25

0
0
0

9,10
10
9,12,17

0
+

9
17

0
0

9
9

URTICACEAE
Vrtica dioica L.
U. gracilis Ait.
U. holosericea Nutt.
U. pilulifera L.
U. urens L.

Common nettle
Stinging nettle
Hoary nettle
Roman nettle
Small nettle

(3)

Red valerian
Large-leaved corn-salad

(3)
(1)

25
25

0
0

0
0

Swiss giants pansy


Heartsease

(2)

25

Redstem filaree

(3)

25

VALERIANACEAE
Centranthus ruber (L.) DC.
Valerianella olitoria (L.)

VIOLACEAE
Viola tricolor
Viola tricolor agg.

ZYGOPHYLLACEAE
Erodium

cicutarium

L'Her.

23

Heterodera Schachtii in Relation to Damage from


Root Rot of Sugar Beets
CHARLES PRICE AND C. L. SCHNEIDER 1
Received for publication July 13, 1964

Introduction
Damping-off is one of the principal causes of poor stands of
sugar beets in some areas. T h e suddenness of damping-off attack
is often impressive, inasmuch as one day the seedlings may look
healthy and the next day they may be dying in large patches in
the field. Damping-off fungi are almost universally present in
the soil and early planted sugar beets are subject to attack,
especially in wet soil. Preemergence damping-off is perhaps
the most serious aspect of the disease because seedlings are
attacked before they reach the surface of the soil and nothing
can be done to remedy the situation except to replant.
In commercial sugar beet production postemergence rotting
of sugar beets also causes serious losses because rotting sometimes
continues throughout the entire period of growth resulting in
low quality or final death of the plant. It has been observed
by the senior author that the incidence of root rot is greater
and the disease more severe in fields of sugar beets in which
the sugar beet cyst nematode is also present in the soil than
when the pathogen is absent. It is concluded, therefore, that
there is a relationship between injury by Heterodera schachtii
and infection by soil-borne fungi that causes damping-off and
root rotting of sugar beets. T h e increased damage may in part
be due to openings of infection courts to the fungi by the feeding
punctures of the nematode.
Powell (1) cites proven examples of association between
plant parasitic nematodes and root-rotting fungi.
In connection with breeding sugar beets for resistance to
Heterodera schachtii it has been observed in greenhouse tests
at Salinas, California, that damage to sugar beets from rootrotting fungi is greatly increased if grown in soil infested with
the nematode. It was, therefore, important to determine the
amount of reduction in root weight caused by the nematode
alone and in combination with root-rotting fungi. This paper
reports results of experiments with sugar beets grown under
controlled conditions. A vigorous program is in progress at the
1
Research Agronomist and Plant Pathologist, respectively, Crops Research Division.
Agricultural Research Service, U. S. Department of Agriculture, Salinas, California and
Logan, Utah.

VOL.

13, No. 7, OCTOBER

605

1965

U. S. Agricultural Research Station at Salinas, California, in


which sugar beets are being bred for tolerance to combination
of root-rotting fungi and Heterodera schachtii.
Plan and Procedure
T h e variety US 41 was used in these tests. Damage caused
by the nematode and root-rotting fungi was measured on the
basis of root weights of the sugar beets. Seeds of sugar beets
were planted in sterile sand and the seedlings were transplanted
to the soil in three-gallon crocks with one seedling to each
crock. T h e r e were 3 replications of each of the four treatments
and 10 crocks in each replication. Each individual sugar beet
root was weighed and the average weight was taken as basis
for assessment of the relative damage of the treatments. T h e
treatments were as follows: T r e a t m e n t 1) sugar beets were
grown in soil in which nematode cysts were added; T r e a t m e n t
2) pathogenic fungi added; T r e a t m e n t 3) both nematodes a n d
pathogenic fungi added; and T r e a t m e n t 4) untreated (control).
T h e light-texture soil used in this test was steam sterilized.
Heterodera schachtii cysts which were produced on sugar
beets growing in sterilized soil were added to the sterilized soil
in crocks of treatments 1 and 3. Small amounts of soil k n o w n
to contain root-rotting fungi were added to the sterilized soil
in crocks of treatments 2 a n d 3. T h e fungus most commonly
associated with root rot of plants grown in soil infested with
Heterodera schachtii a n d root rotting fungi ( T r e a t m e n t 3) was
Rhizoctonia solani, although other fungi were also associated to
a lesser degree. T h e fungi isolated from pieces of diseased roots
that were surface disenfected and planted out on p r u n e agar
included:

Fungus genus
Rhizoctonia
Fusarium
Pythium
Unidentified
Total number of plants bioassayed

Number of plants yielding


each fungus genus
111
11
5
24
130a

Some plants yielded more than one fungus genus, therefore sum of plants yielding
fungi exceeds total number of plants bioassayed.

Results
Results given in the following table show comparisons between
sugar beets grown in nema tode-free soil a n d soil infested with

606

JOURNAL OF THE A. S. S. B. T.

Heterodera schachtii and root-rotting fungi. T h e reduction in root


weight was 12.74% for those grown in soil with root-rotting
fungi alone, 26.9% in soil with H. schachtii alone, and 45.8%
in soil in which both nematodes and root-rotting fungi were
added.
Tabic I.Relation between Heterodera schachtii and root rot of sugar beets.
Treatment
Sugar beets grown
I n soil w i t h r o o t - r o t t i n g fungi a l o n e
In soil infested w i t h H. schachtii alone
In soil w i t h b o t h H. schachtii a n d rootr o t t i n g fungi
N o disease ( c o n t r o l )
LSD 5 %
LSD \%

Avg. wt.
of beets

Difference between
control and disease

Loss due
to disease

Grains

Grams

Percent

586.0
491.0

85.6
180.6

12.726.9

364.0
671.6
6.7
8.8

307.6

45.8

Summary
In this test it is evident that reduction in weight of the
sugar beet roots was lowest in the sugar beets grown in soil
with root-rotting fungi alone, next lowest with the nematode
alone and highest with both nematodes and root-rotting fungi
present. Sugar beets exposed to both nematode and root-rotting
fungi suffered more damage than the sum of losses due to
nematodes alone and root rot alone.
Literature Cited
(1) POWELL, N. P. 1963. T h e role of plant parasitic nematodes in fungus
diseases. Phytopathology 53 (1) : 28-35.

Evaluation of Sugar Beer Storage Practices by Using


the Percentage Purity of the "Thin Juice" 1
S.

T.

D E X T E R 2 AND

M.

G.

FRAKES 2

Received for publication July 16, 1964

In 1954 Brown a n d Serro (2) 3 described a rapid laboratory


method for simulating the purity determination of the " t h i n
juice" of the sugar factory. T h e results of this m e t h o d were, for
every practical purpose identical with those of the factory after
liming and carbonation. Carruthers (3) corroborated their results a n d added a modification of his own. He states "if the
ratio of sugar to total solids is known in the clarified (thin)
juice, then the proportion of sugars which the factory can expect
to obtain in crystal form is predictable." T h e Great Western Sugar
Company has provided a formula for this prediction, together
with a full explanation, on page 136 of "Sugarbeet Research"
compiled by C R D , A R S and USDA. T h e sugar factories in
Michigan seem in agreement with the formula, with slight modifications.
T h e authors of this paper make no claim to expert knowledge
of factory practice or the complications of the application of this
formula. However, since the thin juice methods and the formula
are presented by the e m i n e n t authorities cited above, they may
carry considerable authority to agronomists.
T h e particular purpose of this paper is to call attention to
the tool that has been furnished the agronomist. By this m e t h o d
and formula, agronomists in a reasonably well e q u i p p e d a n d
mechanized laboratory can determine how much sugar will be
bagged p e r ton of beets that have been grown u n d e r various
practices. Most past a n d c u r r e n t papers report tons of beets
per acre, a n d perhaps percentage sucrose, from which gross sugar
per acre may be computed. However, with beets of various
sucrose percentages a n d various thin juice purities, the percentage
recovery as bagged sugar may readily range from less than 70
to more t h a n 9 0 % in beets of any one variety, harvested in any
one season (See T a b l e 1). Results based on total gross sugar are
likely to lead to recommendation of uneconomic practices.
For example, a new beet variety was reported to yield 34
tons per acre in contrast to 21 tons for a commercial one. Gross
sugar p e r acre was 2 0 % higher for the new variety. A l t h o u g h
1

Contribution from the Michigan State University Experiment Station, Journal paper
No.
3414
2
Professor of Crop Science, MSU, and Research Director, Michigan Sugar Company.
3
Numbers in parentheses refer to literature cited.

608

JOURNAL OF THE A. S. S. B. T.

Table 1.Percentage recovery as bagged sugar from beets of various qualities, according
to The Great Wesetern formula.
Clear juice
purity %

12

13

14

15

16

17

18

85
86
87
88
89
90
91
92
93
94
95
96
97

68.7
71.1
73.2
75.3
77.4
79.4
81.4
83.4
85.3
87.1
89.0
90.8
92.5

68.9
71.2
73.4
75.5
77.6
79.6
81.6
83.5
85.4
87.3
89.1
90.9
92.6

67.0
71.3
73.5
75.6
77.7
79.7
81.7
83.7
85.6
87.4
89.3
91.1
92.9

69.1
71.4
73.6
75.7
77.8
79.8
81.8
83.8
85.7
87.6
89.4
91.2
93.0

69.2
71.5
73.7
75.8
77.9
79.9
81.9
83.9
85.8
87.7
89.5
91.3
93.1

69.2
71.6
73.8
75.9
78.0
80.0
82.0
84.0
85.9
87.8
89.6
91.4
93.2

69.3
71.7
73.9
76.0
78.1
80.1
82.1
84.1
86.0
87.9
89.7
91.5
93.3

no
to
be
34
of

figures were given for bagged sugar per acre, it was possible
estimate from purities given that about 5,100 pounds would
bagged per acre in each case, although it required processing
vs. 21 tons for the two lots. Similar examples could be given
the effects of other agronomic variables.
T h e difficulties that would be involved in attempting to determine bagged sugar per ton by actual factory processing of small
samples seem evident. By the use of this new technique, we have
been able to evaluate a wide range of agronomic practices in
terms of bagged sugar per ton and per acre.
In this paper, sucrose loss, in terms of bagged sugar per ton
of original beets, will be reported as affected by storage conditions and original quality of beets. In the remainder of this
paper, the juice clarified in the laboratory by Carruthers' modification will be designated "clear juice", to distinguish it from
the factory "thin juice" with which we occasionally work.
Literature Review
T h e literature on storage of beets is so voluminous that only
a few papers particularly pertinent to this study will be mentioned. In general, respiration of beets increases rapidly with
the temperature of storage, Barr et al. (1). Although there is
considerable variation in the results of the experiments in which
carbon dioxide production per pound of beets was determined,
there is remarkably good agreement in some cases between the
sugar loss by computation and that lost by actual analysis. In
some cases loss of sugar from undetermined causes is very considerable, but in many cases this loss is related to growth or to
the quality of the beets. Larmer (5) studied the keeping quality

VOL.

13, No. 7, OCTOBER

1965

609

as influenced by n u t r i t i o n a l factors and found that adequate


phosphate was particularly helpful. In Michigan, it has commonly been thought that beets high in sugar a n d / o r purity store
better than do beets inferior in these regards. Desiccation d u r i n g
storage has been shown by Pack (6) to lead to accelerated loss
of sugar. Freezing and thawing has been shown to be detrimental,
but storage in the frozen condition has repeatedly been found
to essentially stop sugar loss. Storage of beets submerged in
water at a b o u t 35F has been shown by Dexter (4), to lead to
small losses in total sugar up to about 4 weeks of storage, with
appreciable losses thereafter. Pack (6) found sugar losses of
about 5 0 % when beets were submerged over 100 days in cold
water.
Method
In the first four experiments, beets from one variety were
used. Except as indicated, they were machine topped from a
field with an excellent stand of beets. T h e y were washed thoroughly a n d surface dried. From a pile of such beets, samples
were selected, in turn, as follows:
1. Six half-bushel mesh bags of the largest beets and six of
the smallest beets were taken and weighed. On two lots
of each, analyses were made at once. T w o of the six bags
were b u r i e d in a company pile for storage, and two were
stored in the frozen condition for ten weeks. After reweighing, analyses were made to compare the storage characteristics of large and small beets.
2. F r o m the r e m a i n i n g middle-sized beets, b e e t s h i g h in
specific gravity (over 17 Brix) and those of m e d i u m specific
gravity (15-17 Brix) were selected. By harvesting an o u t side row, a few beets of low specific gravity (below 14
Brix) were obtained. These were weighed, stored, reweighed and analyzed as in the first experiment. It has
been suggested that beets high in sugar store with very
little difficulty or loss in sugar. Since small beets or those
high in specific gravity are likely to be high in sugar,
a n d since it should be a simple matter to separate such
beets, on a large scale, their storage characteristics would
be of interest.
3. Similarly, from the same variety of beets, samples were
prepared w i t h o u t removal of the crown, and some even
had traces of leaf petioles left; others were topped near
the first leaf scar; while other samples consisted of the
removed crown tissue. T h e s e were similarly weighed, stored
in the pile or frozen condition, reweighed and analyzed.

JOURNAL OF THE A. S. S. B. T.

610

4. From the remaining machine-topped beets, samples were


prepared for storage at about 32F in air, in ice water,
in 2% salt brine, in the company pile, and in the frozen
condition. In all cases, weights before and after storage
were taken to correct bagged sugar recovery in terms of
the original weight of the samples.
Results and Discussion
Table 1 gives an idea of the percentage of recovery of gross
sugar in sugar beets of varying percentages of sucrose and clear
juice purity. It may be noted that the percentage recovery as
bagged sugar is relatively uniform for any thin juice purity, increasing less than 1% in the sucrose range from 12 to 18% in
the beets.
Table 2 compares the bagged sugar per ton of original beets
and the clear juice purity of the samples after storage for 10
weeks under two conditions. In general, the beets of the highest
quality, over 17 Brix and small beets, appeared to store as well
in the company pile as in the frozen condition.
Table 2.The bagged sugar per ton and clear juice purity are compared for beets
stored 10 weeks in the company pile or in the frozen condition.
Bagged sugar per ton of original beets
Sample
L a r g e beets
Small beets
Over 17 B r i x
15-17 B r i x
Below 14 B r i x
W h o l e beets
T o p p e d beets
Crowns

Clear juice purity

In Pile
lbs.

Frozen
lbs.

In Pile
%

Frozen
%

258
301
293
267
202
240
265
122

276
293
284
281
237
285
307
158

91.1
92.9
92.1
91.1
86.1
88.2
89.7
80.2

90.2
92.4
91.1
91.3
88.6
90.6
92.6
81.7

With beets of originally poorer quality, storage in the frozen


condition appeared to preserve the sugar considerably better
than did storage in the company piles. In beets below 14 Brix,
in untopped beets, and in crowns only, recoverable sugar was
notably higher in samples that were stored in the frozen condition. It would appear that in the pile these beets lost about
0.5 pound of sugar per ton per day more than when frozen.
T a b l e 3 compares loss in percent sugar in the company pile
versus storage in the frozen condition. In the "Ratio calculated
from shrinkage", the weights after shrinkage were compared
for storage in the pile or frozen. T h e beets stored in the pile

VOL.

13, No. 7, OCTOBER 1965

611

Table 3.Effect of 10 weeks storage in the pile or in the frozen condition on the loss
in percentage sugar in beets, corrected to the original weight of each sample. The ratio
% sugar in beets stored in pile
of
. is given in the table.
% sugar in beets stored in frozen condition
Ratio calculated from Actual ratio from
Change in
Sample
weight shrinkage
sugar analysis
ratio of sugar %
Large beets
Small b e e t s
O v e r 17 B r i x
15-17 B r i x
Below 14 B r i x
W h o l e beets
T o p p e d beets
Crowns

0.981
0.951
0.956
0.990
0.961
0.986
0.998
1.002

0.081
0.025
0.020
0.050
0.077
0.127
0.084
0.183

0.900
0.976
0.976
0.940
0.884
0.859
0.914
0.819

shrunk slightly less (except for crowns) than did the frozen
samples. T h u s , from consideration of shrinkage alone, one would
expect the beets stored in the pile to contain a slightly lower
concentration of sugar than those stored frozen. Since in most
cases the shrinkages were slightly different in the pile a n d in
the frozen condition, the ratios in the first column a p p r o x i m a t e
1.00. T h e ratio of sugar percentages, determined by actual
analysis, however, (corrected to the original weight of each
sample) shows that losses in percent sugar were appreciably
greater in the samples stored in the pile than in those stored
frozen, except in the case of the two samples, over 17 Brix a n d
small beets. T h u s , 0.081 (for example) more of the gross sugar
in the large beets disappeared in the pile than in the frozen
condition.
If there were no loss of sugar, the values in the two columns
would be identical. If there were no loss of sugar in the frozen
samples, one must conclude that samples slightly higher in sugar
were accidentally used in the case of "over Brix 17" a n d "small
Table 4.Bagged sugar per ton of original beets and clear juice purities of beets stored
at near freezing temperatures under water, under 2% brine, or in air, frozen, and in the
company pile.
Storage Method
Storage
period

Under water
32F.

Under 2% brine
32F.

In air
32F.

Frozen

Company pile

Bagged sugar per ton, corrected to original weight


4 weeks
7 weeks

260
231

244
215

277
276

269
272

258
265

Clear juice purities


4 weeks
7 weeks

90.5
87.0

89.1
. 86.7

91.8
90.1

91.6
90.3

89.6
89.8

612

JOURNAL OF THE A. S. S. B. T.

beets," stored in the pile. In general, however, the conclusions


agree with those from the previous tables, namely, that storage
in the frozen condition was particularly helpful in the case of
the beets cf poorer quality. Since, in general, both percentage
sugar and clear juice purity were maintained better in beets
stored in the frozen condition, bagged sugar per ton of beets
was inevitably improved.
Table 4 compares storage under water with other storage.
For the first 4 weeks, storage losses in beets under cold water
were almost identical to those in the company pile. After this
period, lcsses were appreciable in submerged storage. Storage
under 2% brine was not helpful in preserving sugar, but did
almost eliminate uptake of water by the sample.
Summary
Beets of various qualities were obtained by selecting samples
of large beets; small beets; beets high, medium or low in specific
gravity, and beets with and without crowns. They were stored,
for 10 weeks, until January 10, in the company pile or in the
frozen condition. Loss of recoverable sucrose in the company
pile was much less in the case of high quality beets than in
lower quality beets, as judged by the difference between sugar
losses in the pile and in the frozen condition.
Beets stored under near-freezing water or 2% brine for 4
weeks lost slightly more bagged sugar, in terms of the weight
of the original sample, than did beets stored in air at 32 or
frozen. After 7 weeks of storage, the loss was appreciable in beets
stored under water.
Literature Cited
(1) BARR, C. GUINN, E. M. MERVINE and R. A. RICE.

(2)
(3)
(4)
(5)
(6)

1940. A preliminary

report on the effect of temperature and beet conditions on respiration and loss of sugar from beets in storage. Proc. Am. Soc. Sugar
Beet Technol. 2 ( 1 ) : 52-65.
BROWN, R. T. and R. F. SERRO. 1954. A method for determination of
thin juice purity from individual mother beets. Proc. Am. Soc.
Sugar Beet Technol. 8 (2) : 274.
CARRUTHERS, A. and J. F. T. OLDFIELD. 1960. Methods of assessment
of beet quality. Proc Xlth Session of the commission internationale
technique de sucrerie. Frankfort, 1960. Elsevier Pub. Co. 1962.
DEXTER, S. T. 1963. Storing sugarbeet roots under cold water. Proc.
Eastern Regional Meeting Am. Soc. Sugar Beet Technol.
LARMER, F. C. 1937. Keep ; ng qualities of sugar beets as influenced by
growth and nutritional factors. J. Agr. Res. 54: 185-198.
PACK, DEAN A. 1926. The effect of moisture on the loss of sugar from
beets in storage. J. Agr. Res. 32: 1143-1152.

Nutrient Balance and Concentration in


Sugar Beet Production1
JAY L. HADDOCK AND B A R R E L M. STUART 2
Received for publication July 24, 1964

Eighty percent of all commercial fertilizer used on the sugar


beet producing land in Western United States is applied without
the guidance of soil or plant tissue analysis. Many students
(1,4,6,7,9,10,11,12,15) 3 of plant production requirements indicate
that mineral n u t r i e n t content and balance in plant tissue strongly
affect yield and quality.
Ulrich et al. (13) have proposed and successfully used the
theory of "critical concentrations" of nitrogen, phosphorus, a n d
potassium in sugar beet petioles as a guide to crop fertilization.
Ulrich (14) has defined the critical nutrient level as that range
of concentrations within which the growth of the plant is restricted in comparison to plants maintained at higher n u t r i e n t
levels. He proposes 1,000 p p m of nitrate-nitrogen, 750 p p m
phosphorus, and 10,000 p p m potassium as constituting "critical
levels" in sugar beet petioles. T h e s e criteria however, give little
consideration to total n u t r i e n t concentration or balance. F u r t h e r more, the authors (2) have presented data which indicate the
superiority of quantity-quality factor over critical levels as a
basis for identifying nutritional disturbances in sugar beet plants.
Sugrar beet plants are exposed to a wide range of n u t r i e n t ratios
and concentrations in commercial suq;ar beet fields. Little information is now available on the influence of these factors on
sugar beet production. It appeared desirable to extend the
former study and comparison to include a wide range of n u t r i e n t
ratios a n d total n u t r i e n t concentrations in the n u t r i e n t m e d i u m
and in plant tissue.
A survey of the literature indicates that serious consideration
has not b e e n given to the statement of Shear and C r a n e (8)
that each essential element must occur in the leaf in proportion
to every other essential element, nor to that of Lucas et al. (5)
that a balanced n u t r i t i o n does not imply an adequate n u t r i t i o n
but an a d e q u a t e n u t r i t i o n does imply a balanced n u t r i t i o n .
Yield, quality, and chemical composition data derived from
analyzing sugar beets grown in seven n u t r i e n t culture treatments
1
Contribution from the Southwest Branch Soil & Water Conservation Research Division.
Agricultural Research Service USDA in cooperation with the Utah Agricultural Experiment
Station.
2
Research Soil Scientist and Soil Scientist respectively. USDA, Logan, Utah.
3
Numbers in parentheses refer to literature cited.

614

JOURNAL OF THE A. S. S. B. T.

are presented below. T h e experiment had been designed to test


the hypotheses that:
1. "Critical levels" do not constitute a sensitive measure of
nutrient conditions in the sugar beet.
2. T h e relative concentrations of the components of a nutrient
medium may by important factors in yield and quality of sugar
beets.
3. T h e balance of nutrients attained in the plant tissues exerts
an important influence on yield and quality of sugar beets.
Experimental Methods and Procedure
Ten-gallon cans (each with a 14-inch diameter and a 15-inch
depth) painted inside with asphaltic interior water tank coating
and with five holes punched in the bottom for free drainage were
filled with No. 2 vermiculite. They were buried in moist soil
to within 1 inch of the top rim in order to maintain plant roots
at normal soil temperatures. T h e cans were spaced on 40-inch
centers with two sugar beet plants growing in each can.
T h e compositions of the nutrient solutions used are shown
in Table 1. Nutrient concentrations were unchanged throughout
the experiment. Each treatment was replicated 12 times. One
gallon of each nutrient solution was applied to its respective
can daily except during hot weather in mid-July and August
when a total of one and one-half gallons were used in two
applications.
Table 1.Nutrient concentration in various nutrient solutions, 1962.

No.

Nutrient
Solutions

NOsN

Ca

Mg

Na

pH

Salinity
E . C . X 103
@ 25 C.

1
2
3
4
5
6
7

Check
2 X Check
Field**
2 X Field
Vt- Field
Vi Field
Field + K

100
200
60
120
30
15
60

16
32
16
32
8
4
16

100
200
20
20
20
20
40

150
250
80
160
70
60
100

40
60
50
100
25
15
50

12
24
40
80
20
15
50

7.4
7.4
7.6
7.5
7.5
7.5
7.6

1.60
2.60
1.00
1.53
0.70
0.55
1.10

Various nutrients * in solution p p m

*Minor
M i n o r eelements
l e m e n t s aadded
d d e d to all nnutrient
u t r i e n t s osolutions:
l u t i o: n s
B == 0.25,
= 0.25,
0.25 , Mn
Mn =
0.25 Zn = .028,
Cu = .01, Mo = .004, and Fe = 4.5 ppm.
** Field solution modified from check so as to produce beets chemically typical of those
found in commercial fields.

Leaf petiole samples were taken from the most recently


matured leaves on each test plant. A sample was taken from
each plant J u n e 25, July 16, August 6, September 20, and October
15. These tissues were rinsed in deionized water, dried rapidly
at 70 C, ground to pass a 40-mesh screen, and examined by

VOL,.

13, No. 7, OCTOBER 1965

615

standard chemical procedures to determine acetic-acid-soluble


nutrients.
Hoagland's (3) n u t r i e n t solution No. 1 was used at one-half
strength as a check solution. Modifications of the check solution
as shown in T a b l e 1 constituted the other six treatments referred
to in this study. T h e solution designated Field (F) 4 was devised
to produce sugar beet plants typical of those found in commercial
fields.
T h e concentrations of the n u t r i e n t components of each solution per se were not the primary concern in these experiments.
Rather, the n u t r i e n t concentrations in the tissues of sugar beet
plants growing in these solutions were the major interest. T h e
objective of these studies was to relate plant tissue composition
to productivity.
Experimental Results
Gross sugar yields are shown in Figure 1. T h e electrical
conductivity of each n u t r i e n t solution indicates both the total
concentration of soluble salts and the intensity of n u t r i t i o n .
T h e differential responses of roots and tops to a range of
nitrogen a n d phosphorus concentrations are shown in Figure 2.
Critical levels are not shown here for phosphorus and potassium. In no instance, however, did the phosphorus concentration
fall below 1500 p p m nor the potassium below 30,000 p p m in
the petiole tissues examined. T h e seasonal levels for nitratenitrogen are shown in Figure 3. N o n e of the plants contained
nitrogen below the critical level.
T h e senior a u t h o r has previously used quality factors as a
means of characterizing nutritional status of sugar beets (2).
T h i s t e c h n i q u e is used in presenting the data in Figures 4 a n d
5. Data in Figure 3, used here to characterize the nitrogen n u t r i tional status of sugar beet plants by the "critical level" technique,
can be compared with the data on quality of nitrogen presented
in Figure 4.
Discussion
Yields of sugar, from treatments involving solutions 3, 4, 5,
and 6 (left half of Figure 1), indicate that the concentrations of
nitrogen a n d phosphorus in the root m e d i u m significantly affect
yield. W h e n the ratio of nitrogen to phosphorus was held constant, concentrations higher or lower than those in solution F
depressed yields significantly. T h i s situation prevailed only rela* Field survey in 1961 of 48 high-producing commercial fields indicated plant tissue
high in N, high in Na, and low in K relative to composition of ideal nutrient-cultured
beets.

616

JOURNAL OF THE A. S. S. B. T.

tive to the four nutrient solutions 14-F to 2-F. Sugar yield was
not necessarily depressed as a result of increasing the total concentration of nutrients in the root medium. T h e Ck solution
contained higher concentrations of nitrogen and potassium than
did solution F. T h e Ck solution also had a high electrical conductivity, yet it did not depress yields. However, when the Ck
solution ratio among nitrogen, phosphorus, and potassium was
maintained but the total concentration of salts was increased
(as in 2-Ck), sugar yield was depressed significantly relative to
Ck treatment.

Figure 1.Yield of gross sugar as affected by nutritional environment, 1962.

Figure 2.Yield of sugar beet roots and tops as related to nitrogen,


phosphorus and potassium concentration in nutrient solutions, 1962.

These observations suggest that the concentration of nutrients


in the root medium plays a significant role in sugar beet production. T h e balance or ratio among the three primary nutrients
appears to be important in the growth of sugar beets in the

VOL.

13,

No. 7, OCTOBER

1965

617

range of concentrations studied. Undoubtedly, n u t r i e n t elements


other than nitrogen, phosphorus, and potassium exert their individual and combined influence on growth. It is impossible
to avoid completely a confounding of some cause and effect
relationships when one is working with such complex mediums.
It has often been said that one cannot grow a crop of beets
without a good yield of tops, b u t top growth is not a good indicator of root yields. W h i l e growth response of both roots a n d
tops is similar over the lower portion of the n u t r i e n t concentration range, data in Figure 2 indicate that root growth is less
sensitive to continued increases in nutrient concentration than
is top growth. T h i s is because top growth continues to respond
favorably to higher intensities of nutrition and higher concentrations of nitrogen which have no additive effect on root
growth (Figure 2).
T h e ranges in yields of roots, tops, and sugar shown in Figures
1 and 2 would seem to indicate the existence of significant differences in chemical composition among plants grown in these
nutrient cultures. N o n e of the plants studied, however, were
deficient in nitrogen, phosphorus, or potassium on the basis of
critical levels. "The nearest approach to a critical level was associated with the July 16 sampling for the 1/4-F treatment (Figure
3). O n e must assume from these observations either that nitrogen,
phosphorus, a n d potassium are not related to the yield variations
shown in Figures 1 and 2, or that proposed "critical levels" are
inadequate as a measurement of nutritional status.

Figure 3.Seasonal nitrate-nitrogen content of sugar beet petioles as


influenced by nutritional environment, 1962.

Data on the chemical composition of the sugar beet petiole


samples' were calculated to obtain quality factors for nitrogen,
md are represented graphically (Figure 4). T h e most productive

JOURNAL OF THE A. S. S. B. T.

618

plant is assumed to be the best nourished. Seasonal ranges in


the nitrogen quality factor for an adequately nourished plant
are selected as tolerance limits, within which such a plant can
be identified. These arbitrary limits are shown by horizontal
solid lines in Figure 4. T h e extent of departure from these
tolerance limits is assumed to indicate the nature and extent
of nutritional disturbance resulting in unsatisfactory growth
performance. T h e 2-F and 2-Ck treatments produced plant tissue
high in nitrogen quality (Figure 4). Four of the treatments resulted in plant tissue too low in nitrogen quality for optimum
growth.

6/25

7/16

8/6

9/20

IO/I5

Figure 4.Seasonal quality factor for nitrogen in sugar beet petioles


as influenced by nutritional environment, 1962.

Figure 5.Seasonal quality factor for potassium in sugar beet petioles


as influenced by nutritional environment, 1962.

VOL.

13, No.

7, OCTOBER

1965

619

Since the quality technique involves a calculation of the


ratio N : P : K, it follows that when nitrogen quality is low,
potassium quality tends to be high. T h i s is illustrated by the
data in Figure 5. W h e n this analysis was extended to phosphorus,
it was evident that n o n e of the plants was deficient in phosphorus.
T h e influence of the three primary, as well as other plant
nutrients, has been poorly defined with respect to their potential
excess a n d deficiency relative to sugar beet production. T h e s e
interrelations need clarification and more precise definition.
Summary and Conclusions
T h e yield of sugar from sugar beets was influenced significantly by various concentrations of nitrogen and phosphorus in
n u t r i e n t cultures that did not approach critical levels.
U n d e r the conditions of n u t r i e n t culture used in this study,
as total n u t r i e n t concentration increased, n u t r i e n t balance became a more i m p o r t a n t factor influencing the yield of sugar.
T h e m a x i m u m peak in the growth curve for sugar beet tops,
appears to occur at a higher concentration of nitrogen in the
growth m e d i u m than does the comparable peak in root growth.
Both curves are markedly and differentially affected by n u t r i e n t
balance in the growing m e d i u m .
N u t r i e n t culture studies using the buried pot, out-of-doors
technique have shown that intensity of sugar beet n u t r i t i o n a n d
the balance a m o n g nutrients are important factors in o b t a i n i n g
high yields of sugar beets.
W h i l e critical levels have been widely used to identify n u t r i e n t
deficiencies in commercial sugar beet fields, they appear to be
an inadequate measure of good nutritional status of the sugar
beet plant. Some modification of definition or levels should be
made to increase the value of this technique.
T h e use of quality factors for nitrogen, phosphorus, a n d
potassium in sugar beet petioles appears to provide a good means
of appraising the quality of n u t r i t i o n . T h i s and techniques other
than "critical levels" need further study and more precise definition for commercial field application.
Literature Cited
(1) GREGORY, F. G. 1937. Mineral nutrition of plants. Ann. Review Biochem. 6: 557-578.
(2) HADDOCK, JAV L. and DARREL M. STUART. 1964. Critical levels versus
quantity-quality factors for assessing nutritional status of sugar beet
plants. J. Am. Soc. Sugar Beet Technol. 13 (1) : 42-58.

620

JOURNAL OF THE A. S. S. B. T.

(3) HOAGLAND, D. R. and D. I. ARNON. 1938. The water-culture method


for growing plants without soil. California Agric. Expt. Sta. Circ.
347.
(4) LUNDEGARDH, H. 1951. Leaf analysis, p. 46. Hilger and Watts Ltd.
London, England.
(5) LUCAS, R. E., G. D. SCARSETH and D. H. SIBLING.

(6)
(7)

(8)
(9)
(10)
(11)
(12)

(13)

1942.

Soil fertility

level as it influences plant nutrient composition and consumption.


Purdue Univ. Agr. Exp. Sta. Bui. 468.
MACY, PAUL. 1936. The quantitative mineral nutrient requirements
of plants. Plant Phys. 11: 741-764.
SCARSETH, G. D. and R. E. LUCAS. 1947. Potassium calcium and magnesium balance and reciprocal relationship in plants. Agron. J.
39: 887-896.
SHEAR, C. B. and H. L. CRANE. 1946. Nutrient-element balance. A
fundamental concept in plant nutrition. Proc. Am. Soc. Hort. Sci.
51: 319-326.
SHEAR, C. B. and H. L. CRANE. 1943-1947. Nutrient-element balance.
Yearbook of Agriculture pp. 592-601.
SHIVE, J. W. and W. R. ROBBINS. 1939. Mineral nutrition of plants.
Ann. Rev. Biochem. 8: 503-520.
STEWARD, F. C. 1935. Mineral nutrition of plants. Ann. Review Biochem. 4: 510-544.
TYNER, E. H. and J. R. WEBB. 1946. The relation of corn yields to
nutrition balance as revealed by leaf analysis. J. Am. Soc. Agron.
38: 173-185.
ULRICH,

ALBERT,

DAVID

RIRIE,

F.

J.

HILLS,

ALAN

G.

GEORGE

and

MORTON D. MORSE. 1959. Plant analysis, a guide for sugar beet


fertilization. Calif. Agr. Exp. Sta. Bui. 766.
(14) ULRICH, A. 1948. Plant analysis methods and interpretation of results.
Diagnostic Techniques for Soils and Crops. Chap. IV American
Potash Institute, Washington, D. C.
(15) ZHURBITSKII, Z. I. and WEN-HANG HUANS.

1962.

Influence of nutrient

solution concentration on absorption of mineral elements by plants.


(Toane) Am. Institute of Biolog. Sci. 8: 468-474.

W e i g h t of Fruits in Self-Fertile, Male-Sterile,


and Self-Sterile Diploid and Tetraploid
Monogerm Beta Vulgaris L.
V.

F.

SAVITSKY AND H E L E N SAVITSKY 1

Received for publication October 13, 1964

Introduction
T h e replacement of multigerm sugar beet varieties by m o n o germ varieties is approaching completion in the USA (23) 2 . T h e
first phase of developing monogerm varieties, usually based on
application of the backcross method for incorporation of the
gene m (monogermity) into multigerm varieties is almost accomplished. At the present time, a second phase of breeding of m o n o germ varieties is starting. T h i s phase is the routine improvement
of monogerm sugar beets in different agronomic characters, including quality of the monogerm fruits themselves.
T h e weight of fruits and germs of monogerm beets has a
practical importance in agriculture as one of the factors d e t e r m i n ing the quality of seed (18). D u r i n g the past few years, utilization
of differences in the size of fruits were used in breeding a n d
seed p r o d u c t i o n in the U n i t e d States and in E u r o p e for different purposes. In the USA male-sterile monogerm beets
have been planted together with the multigerm pollinator to
obtain commercial monogerm hybrids. T h e harvested m o n o g e r m
seed (hybrid seed) has then been separated from the m u l t i g e r m
seed by screening (25,26). In Europe, seedballs of the m u l t i g e r m
sugar beets have been screened for size to increase the percent
of triploids in the commercial polyploid (anisoploid) varieties
(1,3,4,24).
Because the seedballs of tetraploids are larger, screening o u t
the small seedballs does eliminate a portion of the diploids. In
the variety Polybeta, for instance, seedballs 2 to 3 mm in diameter
contained 3 9 . 4 % diploids, whereas larger seedballs (5 mm in
diameter) contained only 16.3% diploids (22). Schneider (21)
reported that small seed contained 6 8 % diploids and the large
seed, 10.9%. According to Belgian data, small seed of Polybeta
contained 4 9 . 0 6 % diploids a n d the large seed (5 mm in diameter),
15.93% diploids (6,7). Therefore, retention of larger seedballs
decreased the percent of diploid beets in the anisoploid varieties.
1
Geneticists, Crops Research Division, Agricultural Research Service, U. S. Department
of Agriculture,
Salinas, California.
2
Numbers in parentheses refer to literature cited.

622

JOURNAL OF THE A. S. S. B. T.

F 2 and F 3 generations of hybrids between monogerm beets


and multigerm varieties give monogerm plants differing in the
weight of fruits and germs. Some monogerm segregates develop
small fruits (15,16). This is a natural consequence of inability
to control the genes responsible for the weight of fruits in multigerm populations.
In the first phase of backcross selections for monogerm beets,
the variability of stored genes in multigerm varieties responsible
for the weight of fruits was used. T h e value of such stored genes
in determining weight of monogerm fruits is based on the homeostatic mechanism which controls the interaction of these genes
with the gene m in monogerm sugar beets (8).
Selection for desirable characters in monogerm fruits is necessary for further seed improvement. Selection for weight of fruits
and germs has not been attempted in multigerm populations.
Monogermity has made possible selection for improvement of
the many characters of fruits and germs (2,15,16,18). No multigerm sugar beet population of proven high quality of fruits
(size of fruits and germs, shape of fruits, etc.) was available which
could be used as a recurrent parent for improvement of monogerm fruits. Further improvement of monogerm fruits is possible only by hybridization and selection of monogerm beets,
and not by finding stabilized valuable characters of fruits and
germs in the multigerm populations, which could be used for
incorporation in the monogerm beets.
Information concerning the variability of weight of the monogerm fruits and germs under different environment and genetic
factors, types of matings, selection, and ploidy levels is presented
in this report.
Materials and Methods
T h e following monogerm and multigerm breeding materials
developed by the authors at the Salt Lake City station were used
for a study of the weight of fruits: 10 diploid monogerm selfsterile backcross populations; 12 diploid monogerm self-fertile
inbreds; 18 tetraploid monogerm self-sterile brother-sister lines;
18 tetraploid monogerm self-fertile inbreds; 1 diploid and 1
tetraploid male-sterile equivalent of the monogerm inbred SLC
9 1 ; 1 tetrapoid multigerm open-pollinated population of US 104;
1 tetraploid monogerm population US 35/2; and some other
populations.
All diploid self-fertile inbred lines were derived from hybridization of the original monogerm SLC 101 with different American multigerm varieties (15,19). T h e diploid monogerm selfsterile stocks were obtained after three backcrosses of monogerm

VOL-

13, No. 7, OCTOBER 1965

623

SLC 3 to different self-sterile multigerm open-pollinated varieties


(11,12,15). In every backcross the multigerm pollinator was r e p resented by 30 to 100 plants. T h e male-sterile monogerm equivalents of SLC 91 carried male-sterile cytoplasm of the variety
Janasz (19).
T h e tetraploid monogerm beets were obtained in two ways.
Some of them, as SLC 15 mm self-sterile, SLC 91 mm self-fertile
and its male-sterile equivalent SLC MS 91 mm, were obtained
by colchicine treatment of the corresponding diploid stocks (10).
All other m o n o g e r m self-fertile and self-sterile stocks were obtained by selection of monogerm tetraploid plants from hybrids
between m o n o g e r m tetraploids SLC 15 and SLC 91, with the
tetraploid multigerm population derived from US 35/2 (19,20).
All experiments were conducted near Salt Lake City, Utah,
during 1958, 1959, a n d 1960. Seed beets were grown in 1-row
plots 30 to 50 ft long in overwinter plantings, or by planting of
stecklings in spring. T h e seed plants were not damaged by curly
top, leaf spot, or virus yellows. Stecklings destined for each
experiment were harvested the same day from plots grown in
one field.
O n e sample for d e t e r m i n a t i o n of the weight of 1000 fruits 3
was taken from each plot, except in two experiments where the
weight of 1000 fruits was studied in tetraploid self-sterile a n d
self-fertile m o n o g e r m lines. In each of these plots, seed of 10
plants was harvested individually (Tables 9,10).
T h e n u m b e r of replications in the experiments is indicated
in tables giving the analysis of variances. To study the weight
of germs 3 , fruits were treated by 4-5% hydrochloric acid (HC1)
and dried to a constant weight in the oven at 50 C (13).
Experimental Results
Weight of fruits of multigerm beets
T h e fruit (seedball) of a multigerm sugar beet is a phylogenetically c o m p o u n d structure which accomplished several
functions in the evolution of B. vulgaris L. In wild Beta species
the m u l t i g e r m seedball provided certain advantages of survival
value. T h e presence of several germs in one fruit increased t h e
number of seedlings. T h e presence of several flowers in a cluster
extended t h e time of flowering of a cluster and increased the
amount of pollen produced, which is of importance for t h e
cross pollination of species.
Variability of the weight of germs within a seedball is characteristic of m u l t i g e r m fruits. Up to a certain o p t i m u m , an in3

Weight of fruits and/or germs always designate weight of 1000 fruits or/and 1000
germs

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JOURNAL OF THE A. S. S. B. T.

crease in the number of flowers in a cluster not only increases


the variability of the weight of germs, but as shown in our experiments, one particularly large germ appears.
Natural and artificial selection controls the weight of seedballs in multigerm beets by affecting the number and the size
of individual fruits in the seedball. T h i s in turn influences the
weight of germs in the seedballs.
T h e size and number of individual fruits within a seedball
essentially are controlled by different genetic factors. T h i s is
indicated by the fact that the weight of seedballs and the number
of individual fruits in them are only partially correlated genetically and environmentally. T h e environment coefficient of correlation for these characters equals +0.42, while the genetic
coefficient of correlation is lower and equals +0.24 (14). The
weight of seedballs and number of individual fruits within a
seedball correlate differently with other characters of seed beets
(14). Also, genetic and environment variability is different for
these 2 characters (14). A joint action of these 2 different groups
of genes may increase the weight of the seedballs by increasing
the number or the weight of individual fruits and sometimes
by simultaneous augmentation of both.
Doubling of chromosomes increases the weight of seedballs
(Table 1) but influences differently the two elements conditionTable 1.Weight of fruits (1000) in diploid and tetraploid open-pollinated sugar beet
populations.
Variety
US 35/2 multigerm
US 401 multigerm
SLC 15 monogerm

Diploid

Tetraploid

grams
15.70
26.60
11.17

grams
34.00
36.20
19.70

ing them. Increase of the weight of seedballs in multigerm


tetraploids is caused by the increased weight of individual fruits
within a seedball rather than by the greater n u m b e r of flowers
per cluster. T h e tetraploids usually develop fewer flowers per
cluster than the original diploids (Table 2).
Table 2.Percent of seedballs with different number of individual fruits in diploid
and tetraploid sugar beet populations of US 401.
Level of ploidy
Diploid US 401
Tetraploid US 401

Number of fruits in the seedball


1
%

4 and more

Percent of seedballs
...
48
11
67

47
22

VOL.

13, No. 7, OCTOBER

1965

625

T h u s , the effect of these two characters on the weight of


seedballs cannot be attributed solely to the simple pleiotropic
action of the same set of genes on the n u m b e r and size of individual fruits in the seedball of the multigerm beets.
Variability

of

weight of germs within a multigerm seedball


and in monogerm fruits
W e i g h t of germs in the individual fruits within a seedball
is usually very different. In a Russian sugar beet variety
"Uladovka" the large multigerm seedballs sifted on 3 mm sieves
contained large, m e d i u m , and small germs. T h e weight of 1000
large germs equaled 3.7 gms, of 1000 m e d i u m germs 2.6 gms,
and a 1000 small germs weighed 2.2 gms (27,28).
We studied the variability of the weight of germs within
the seedballs taken at r a n d o m in an open pollinated population
of the variety US 3 5 / 2 . T h e ratio of the weight of the largest
and the smallest germs within three germ seedballs equaled 7:1
(Table 3). Such a big variation in the weight of germs was
caused by a high reduction of the weight of the small germ, as
well as by the poor development of some small germs. A considerable n u m b e r of the small germs is inviable, although other
small germs develop normally within a seedball and are able to
germinate.
Table 3.Weight of large, medium and small germs (1000 germs) in the seedballs
with 2 and 3 fruits in the diploid open-pollinated sugar beet population US 35/2.
Size of germs

Number of flowers
per cluster

Large

Medium

Small

3
2

grams
4.75
4.26

grams
2.50

grams
0.65
3.45

In the p o p u l a t i o n of US 35/2 the range of variation of the


weight of germs was greater for seedballs with 3 fruits t h a n for
seedballs with 2 fruits ( T a b l e 3). T h i s type of variability in
the weight of germs within a seedball, which is caused by different n u m b e r of fruits in seedballs, is more or less similar for
different sugar beet varieties and for seed grown in different
environments (years, soil, location). Therefore, seedballs containing 3 or 4 fruits usually have 1 germ of greater weight a n d
1 germ lower in weight than the weight of the largest a n d
smallest germ in the double-fruited seedballs of the same seed
sample.
Since there can be no variability of the weight of germs
within a fruit of m o n o g e r m beets, the genes responsible for

626

JOURNAL OF THE A. S. S. B. T.

monogerm and multigerm characters exhibit different pleiotropic


effect on the variability of the weight of germs. Screening of
seedballs of the multigerm beets according to size, or segmentation
of seedballs, does not alter the variability in weight of germs
within a seedball. Segmentation of seedballs leads to separation
or to destruction of germs and results only in production of segments which contain one germ of any weight.
In monogerm beets the variability of weight of germs is determined by the variability of weight of fruits only. T h e coefficient of correlation between weight of fruit and germ equals
0.949 (16). Therefore, in monogerm beets the weight of germs
may be controlled by separation of seeds of different size (18,19).
Because of the absence of variability within a fruit, the range
of variability of weight of germs in monogerm beets is less than
in the multigerm beets (Table 4). Weight per 1000 germs in
small and in large monogerm fruits varied from 2.9 to 4.70 gms.
In the multigerm parental variety US 35/2 the range of variation in the weight of small and large germs was 0.65 to 4.75 gms
(Table 3). T h e effect of transition from the multigerm to the
monogerm condition is manifested in a sharp decline of variability in the weight of germs in the new monogerm population
as compared to the previous multigerm populations.
Table 4.Weight of fruits (1000) and germs (1000) in the diploid open-pollinated
self-sterile monogerm sugar beet population SLC 15 derived from backcrosses of monogerm
beets to multigerm variety US 35/2.
Size of monogerm
fruits

Fruits

Germs

Large
Small

grams
20.10
12.59

grams
4.70
2.90

T h e monogerm character acts as a stabilizing agent in the


genetic composition of populations in M-m allele and in the
corresponding linkage group (17,19). T h e different influence
of individual genes members of the multiple allele M-m on
weight of germs peculiar to multigerm beets disappears in the
monogerm beets. In connection with this bias of all multigerm
alleles in estimation of weight of fruits and germs also is eliminated. Accumulation of genes favorable for the weight of fruits
and germs becomes simpler. A new stable interaction between
gene m and all other genes causing variability of the weight of
fruits is established. T h e former gene pool for the weight of
fruits in the multigerm populations acquires a new level of
expression in the monogerm populations.

VOL.

13, No. 7, OCTOBER 1965

627

Weight of fruits in diploid backcross monogerm open-pollinated


populations and in monogerm self-fertile inbred lines
Backcrossing is the usual method for breeding of the monogerm beets (15,19). For the evaluation of the effect of backcross
method on the variability of the weight of monogerm fruits, the
monogerm self-sterile populations obtained after repeated hybridization of self-sterile monogerm beets with different multigerm varieties (US 35/2, US 216, US 2 2 / 3 , Klein E, Klein Z etc.)
were used. In the following generations, which were segregating
for the m o n o g e r m character, the multigerm segregates were
discarded a n d the monogerm plants were crossed again with t h e
corresponding recurrent parents in open-pollinated populations.
W h e n the backcross pollination was discontinued, the m o n o g e r m
plants were intercrossed. In 10 such open-pollinated backcross
populations the weight of 1000 monogerm fruits varied from
16.3 to 19.0 gms, i.e., the difference was 2.7 gms. T h e difference
in weight of monogerm fruits in the backcrossed populations.
however, was not statistically significant, because the standard
error obtained on the basis of analysis of variances in 10 populations in 3 replications almost equaled this value (2.7882 gms).
T h e calculated F-value was 1.09, whereas the tabulated F-value
should be a m i n i m u m of 2.46 (Table 5).
Table 5.Weight of fruits (in grams) in 10 sugar beet diploid monogern self-sterile
populations derived from backcrosses of monogern beets to different multigerm varieties.
Analysis of variaance.
F

Factors
Total
Between populations
Between reps.
Experimental error

I>egree of
freedom
29
9
2
18

Calculated

5%
Level

1%
Level

1.0951
0.4768

2.46
3.55

3.60
6.01

Variance

3.0533
1.3295
2.7882

Absence of significant differences in the weight of m o n o g e r m


fruits in backcrossed self-sterile populations is caused by the
absence of differences in weight of fruits in different parental
sugar beet m u l t i g e r m populations. Multigerm populations have
not been exposed to selection for this character. Therefore, t h e
multigerm sugar beet varieties do not differ in the average
weight of the fruits (14). T h i s is one of the reasons why t h e
weight, or the size of fruits, were never used as distinctive characteristics of the m u l t i g e r m sugar beet varieties. T h e r e are no
indications in the literature concerning hereditary varietial differences in the weight of fruits in multigerm sugar beets. At

628

JOURNAL OF THE A. S. S. B. T.

the same time the individual seed samples, even the seed samples
of the same variety, may differ in the weight of fruits, but these
differences are usually caused by the locations, or by the conditions of seed growing.
Other studies of variability were made of monogerm fruits
using self-fertile inbred lines obtained from selfing of backcross
hybrids between multigerm US 35/2 and the self-fertile monogerm inbred SLC 101. It was found that inbred monogerm lines
isolated from the backcross progeny of repeated backcrosses to
the same multigerm population differed in weight of fruits.
Presence of monogerm inbreds with different fruit weights
indicates that the original multigerm populations were heterogeneous in the genes determining fruit weight. Multigerm openpollinated varieties carry an intravarietial storage of genetic variability in genes determining weight of fruits. These genes are
maintained in a certain equilibrium in open-pollinated multigerm populations (14). By backcrossing of monogerm beets to
the multigerm populations this system of stored variability is
recovered in the same equilibrium in the backcross monogerm
populations.
Inbreeding applied to such backcross populations destroys
their genetic equilibrium and leads to isolation of genetic combinations and to the production of inbred lines differing in the
weight of fruits. T h e grade of differences in the weight of monogerm fruits, fixed by inbreeding, will be proportional to the
heritability of multigerm pollinators.
Comparison of the weight of fruits in 12 monogerm diploid
inbreds shows that the value of variance among inbreds eouals
19.46, whereas the variance between the blocks is only 2.37
(Table 6). T h e calculated value of F among inbreds is 9.25,
which is statistically significant. A high value of variance among
inbreds indicates the presence of significant stored heritability
in the diploid multigerm populations and the genetic differences
between monogerm inbreds isolated from these populations.
Table 6.Weight of fruits (in gram') in 12 sugar beet diploid monogerm self-fertile
inbreds. Analysis of variance.
F

Factors
Total
Between populations
Between reps.
Experimental error
* significant

Degree of
freedom

Variance

59
11
4
44

19.4601
2.3656
2.1026

Calculated

5%
Level

Level.

9.2553*
1.1251

2.01
2.58

2.68
3.78

VOL.

13, No. 7, OCTOBER

1965

629

W e i g h t of fruits of 12 diploid inbreds studied varied from


12 to 18 gms. Of these 12 lines, only 3 significantly exceeded
(in the weight of fruits) the standard monogerm i n b r e d SLC 9 1 ,
by D u n c a n ' s m u l t i p l e range test. T h e s e 3 lines were o b t a i n e d
as a result of selecting for the weight of fruits, whereas the rem a i n i n g lines were selected for other characters. Some of these
lines developed small fruits. T h u s , breeding between m o n o g e r m
diploid stocks for control of fruit weight was effective.
Environmental

variability of the weight of monogerm diploid


and
tetraploid fruits
Effectiveness of b r e e d i n g for the weight of m o n o g e r m fruits
and germs d e p e n d s u p o n the degree of variability of this character; it d e p e n d s u p o n the effect of action of genes a n d t h e
effect of ploidy levels, as well as on influence of the environmental factors that affect variability.
Influence of the e n v i r o n m e n t on the variability of weight
of m o n o g e r m fruits was studied in diploid and tetraploid malesterile equivalents of the m o n o g e r m inbred SLC 91 d u r i n g 3
years (1958, 1959, 1960) on 10 different isolated plots ( T a b l e
7,8).
Table 7.Variability of the weight of fruits (in grams) in diploid and tetraploid sugar
beet monogerm inbred SLC 91 during 3 years in 10 different locations.
F

Factors
Total
Reps.
Populations
(Ploidy level)
Isolations
Years
Ploidy - Isolations
Ploidy - Years
Error

Degree of
freedom

Variance

Level

1%
Level

5%

Calculated

179
2

33.28

8.88*

3.06

4.75

1
9
2
9
2
154

3281.60
22.48
171.12
6.60
6.70
3.75

875.70*
6.00*
45.66*
1.76
1.78

3.91
1.94
3.06
1.94
3.06

6.81
2.53
4.75
2.53
4.75

* significant

Variability in different years. Analysis of variances showed t h a t


the value of calculated F exceeded by 1% the value of the tabulated F. In the 3 years the average weight of 1000 m o n o g e r m
fruits r a n g e d from 13.88 to 17.23 gms; in other words, difference
between weight of fruits in separate years reached 24.14% a n d
was significant ( T a b l e 8).
Variability in different locations. Fluctuations in the weight of
fruits in different locations d u r i n g the 3 years were also significant. T h e value of the experimental F equaled 6.00 a n d of
tabulated F at 0.01 point, 2.53. T h e different isolations were

Table 8.Range of variation in weight of fruits (1000) in the sugar beet monogerm male-sterile equivalent of SIX 91 in different years, locations,
and at different ploidy levels.
During 3 years
Materials

grams
Diploid MS 91
Tetraploid MS 91
For all strains

9.9067
17.8467
13.88

In 10 locations
Minimum

Maximum

Minimum

grams
100.00
100.00
100.00

12.5267
21.9300
17.2300

%
126.45
122.88
124.14

grams
9.5111
18.7111
14.09

Average for years


and locations

Maximum

grams

grams

100.00
100.00
100.00

12.4889
22.8889
17.6900

131.31
122.33
125.55

11.1689
19.7044

100.00
176.42

VOL.

13,

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1965

631

grown on soils of different types a n d the weight of fruits ranged


from 14.09 to 17.69 gms (Tables 7,8).
F o u r of the 10 isolations were always overwinter plantings.
T h e r e m a i n i n g 6 isolations were grown from stecklings p l a n t e d
in spring. T h e weight of fruits was 15.4 gms for both types of
plantings.
T h e F-values for the interaction ploidy level a n d years, a n d
for the interaction ploidy level a n d isolations was n o t significant.
Effect of colchicine-induced tetraploidy on the weight of
monogerm
fruits produced
in
different
environments
Anaysis of variance showed that variability in weight of fruits
was caused by t h e differences between the two p o p u l a t i o n s
studiedmale-sterile diploid SLC 91 and male-sterile tetraploid
SLC 9 1 r a t h e r than by the environmental factors associated
with years of p r o d u c t i o n or location of plots ( T a b l e 7).
T h e value of variance between diploid a n d tetraploid populations was 3281, whereas the value of variance between years was
171 a n d between locations only 22.
T h e differences in weight were as follows: the weight of
fruits in the diploid m o n o g e r m male-sterile equivalent of SLC
91 d u r i n g 3 years in 10 different locations was 11.1689 gms a n d
the weight of fruits in the tetraploid m o n o g e r m male-sterile
equivalent of SLC 91 u n d e r the same conditions was 19.7044
gms, or an increase of 76.42% for the tetraploid ( T a b l e 8). T h u s ,
colchicine-induced tetraploidy greatly increased the weight of
monogerm fruits in the i n b r e d line SLC 9 1 .
To verify the universality of this appearance, a comparative
study of t h e weight of fruits in different diploid a n d tetraploid
self-fertile a n d self-sterile lines was made.
Weight

of

fruits

in

tetraploid

monogerm

self-sterile

lines

Eighteen tetraploid self-sterile lines were planted in overwinter p l a n t i n g in 2 replications together with the diploid check
and tetraploid male-sterile equivalent of m o n o g e r m i n b r e d SLC
91. T h e 18 self-sterile lines were derived from individual tetraploid plants isolated in an open-pollinated m o n o g e r m p o p u l a t i o n
and propagated by brother-sister m a t i n g for 2 generations.
W e i g h t of fruits was studied on 10 plants of each line in
each replication. Analysis of variance of the weight of fruits
shows that t h e m a i n factor of variability is the difference in
weight between d i p l o i d a n d tetraploid populations (Figure 1).
T h e calculated value of F highly exceeds its tabulated value at
the 1% p o i n t ( T a b l e 9).

632

JOURNAL OF THE A. S. S. B. T.

Figure la.Monogerm fruits of a diploid self-sterile population. X 1.3

Figure lb.Monogerm fruits of a tetraploid self-sterile population. X 1.3

VOL. -13,

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633

Table 9.Weight in fruits (in grams) in 18 sugar beet tetraploid monogerm self-sterile
lines and in diploid and tetraploid SLC 91. Analysis of variance.
F
Factors
Total
Between populations
Between blocks
Experimental error
(Interaction:
blocks-population)
Monogerm: 2n versus 4n
Among tetraploids

Degree of
freedom
399
19
1
19

Variance
Calculated
530.91
21.07

18.96*
Not

5%
Level

1%
Level

2.18
3.06
s i g n i f i c a n t

27.9950

Grouping of population comparison


1
6488.33
231.77*
18
199.95
7.14*

4.38
2.21

8.18
3.12

* significant

G r o u p i n g of p o p u l a t i o n comparison shows that the presence


of a diploid p o p u l a t i o n a m o n g the tetraploids is the 'main factor
causing variability. T h e value of F is 231.77. Differences in the
weight of fruits a m o n g separate tetraploid lines are also significant; the value of the calculated F is 7.14 whereas the value
of the t a b u l a t e d F is only 3.12.
For evaluation of differences in the weight of fruits a m o n g
many populations, the principle of Duncan's new m u l t i p l e r a n g e
test was used. In 18 tetraploid self-sterile m o n o g e r m lines the
weight of fruits fluctuated from 20 to 31 gms. T h e weight of
fruits in the diploid male-sterile equivalent of SLC 91 was 8
gms, a n d in the tetraploid male-sterile equivalent of SLC 91
it was 18 gms.
In this e x p e r i m e n t , the weight of fruits of all tetraploid lines
significantly exceeded the weight of fruits in the diploid malesterile e q u i v a l e n t of SLC 9 1 . A m o n g the tetraploids, only 2 lines
with the largest fruits significantly exceeded in weight the tetraploid male-sterile e q u i v a l e n t of SLC 9 1 .
Weight of fruits in the tetraploid monogerm self-fertile inbreds
Eighteen tetraploid i n b r e d lines derived from the hybridization of tetraploid i n b r e d SLC 91 m o n o g e r m with tetraploid
multigerm p o p u l a t i o n US 3 5 / 2 , were studied for variability of
the weight of fruits.
Analysis of variances for 18 monogerm tetraploid self-fertile
inbreds a n d for the d i p l o i d a n d tetraploid male-sterile e q u i v a l e n t
of SLC 91 ( T a b l e 10) indicated that in the inbreds. as in selfsterile lines, the m a i n factor of variability is the difference in
weight of fruits a m o n g different populations. T h e calculated
value of F is 19.48 a n d the tabulated value of F is 2.18 or 3.06.
G r o u p i n g of p o p u l a t i o n comparison also shows that the m a i n

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JOURNAL OF THE A. S. S. B. T.

T a b l e 10.Weight of fruits (in grams) in 18 sugar beet tetraploid m o n o g e r m self-fertile


inbred lines and in diploid a n d tetraploid SLC 91. Analysis of variance.

Factors
Total
Between p o p u l a t i o n s
Between blocks
Experimental error
(Interaction:
blocks-population )
M o n o g e r m : 2n versus 4n
Among tetraploids

Degree of
freedom

Variance
Calculated

399
19
1

237.4123
1.4641

19

12.1856

5%
Level

1%
Level

19.4830*
2.18
3.06
N o t s i g n i f i c a n t

G r o u p i n g of p o p u l a t i o n c o m p a r i s o n
1787.1280
146.6590*
1
18
151.3170
12.4177*

4.38
2.21

8.18
3.12

* significant

factor of variability is the difference in weight of fruits between


a diploid SLC 91 and tetraploid inbreds. T h e value of the mean
squares between a diploid and tetraploids is 1787, whereas the
value cf the mean squares among tetraploid lines is 151. In
spite of this, the calculated value of F among tetraploids is 12.42,
while the tabulated value of F is 2.21 or 3.12.
T h e monogerm tetraploid inbred lines differed in the weight
of fruits and these differences were statistically significant. In
the individual lines, the weight of fruits fluctuated from 7 to
21 gms.
All tetraploid inbreds had larger fruits than the diploid
inbred SLC 91. But, according to Duncan's new multiple range
test, only 13 tetraploid inbreds significantly exceeded the diploid
inbred SLC 91 in fruit weight. One tetraploid inbred line had
significantly larger fruits than 7 other lines.
Thus, genetic variability in the weight of fruits was recorded
for tetraploid inbreds. Tetraploidy not only increased the weight
of fruits, but a new genetic variability of this character arose at
this new level.
Variability of the weight of fruits, estimated for the diploid
strains only, will be highly increased if diploid and tetraploid
strains are studied together in one experiment, even if these
strains are closely related.
Coefficient of variation for weight of fruits estimated in this
experiment in diploid self-sterile monogerm populations was
only 10%, while coefficient of variation for tetraploid self-sterile
monogerm populations and 1 diploid line rose to 90.2%. The
same was recorded for the inbreds. W h e n only diploid inbreds
were studied, the coefficient of variation was 28.74%. For tetraploid inbreds studied together with 1 diploid line, the coefficient
of variation increased to 96.55%.

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Weight of germs in diploid and tetraploid monogerm sugar beets


W e i g h t of fruits a n d weight of germs are highly correlated in
diploid m o n o g e r m beets (16). T h e present investigation was
conducted to study the relationship of these correlations in related diploids a n d tetraploids. Increased weight of fruits in
m o n o g e r m tetraploids may be i m p o r t a n t only if it is correlated
with the increased weight of germs. To study the correlation
between weight of fruits a n d germs, samples were taken from
29 m o n o g e r m diploid a n d 29 m o n o g e r m tetraploid strains ( T a b l e
Table 11.Correlation between weight of fruits and germs in monogerm diploid and
tetraploid sugar beet strains.

* The tetraploids are in blackface italic type.

T h e average weight of fruits of the diploid m o n o g e r m strains


was 13.12 0.78 gms, a n d in tetraploid m o n o g e r m strains,
21.43 0.64 gms, or an increase of 6 3 . 3 % . Difference in weight
of fruits b e t w e e n diploid a n d tetraploid strains was significant.
Average weight of 1000 germs in the diploid m o n o g e r m strains
was 2.169 0.073 gms a n d in the tetraploid m o n o g e r m strains,
3.631 0.078 gms, or an increase of 6 7 . 4 % . T h i s difference is
significant. T h u s , tetraploid m o n o g e r m beets developed larger
fruits a n d larger germs t h a n diploids (Fig. 2).
W e i g h t of 1000 germs in percent of weight of fruits was
16.53% in diploids a n d 16.94% in tetraploids; therefore, in
monogerm tetraploid strains with different weight of fruits t h e
increase in g e r m weight is proportional to the increase in fruit
weight.

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JOURNAL OF THE A. S. S. B. T.

Figure 2.Germs excised from: a) a monogerm diploid strain (upper


row), b) a monogerm tetraploid strain (lower row) X 1.5.

Tetraploids exhibit the same correlation between weight of


fruits and weight of germs as the diploids. For 29 diploid strains
studied, this correlation equaled 0.9500; and for 29 tetraploid
strains, 0.9244. T h e t-value for the diploids was 31.03, and for
tetraploids, 23.79. Value of t for diploids and for tetraploids is
significant at a probability of above the 1% level; therefore, both
correlations are significant.
Hence, selection for the larger fruits in monogerm diploids
and in tetraploids automatically increases the weight of germs.
We have not observed monogerm diploid or tetraploid strains
with large fruits containing small germs nor strains with small
fruits and large germs.
Fruits and germs in monogerm beets containing

twin

germs

Some monogerm fruits produce double, triple, or multiple


seedlings. Presence of two or more germs in a locule complicates
the relation between weight of fruits and germs in mcnogerm
beets, because the increase in the n u m b e r of germs generally
is not followed by a corresponding enlargement of a fruit. The
total weight of 2 or 3 germs developed in 1 fruit almost corresponds to the weight of a single germ in a fruit of a given
size. T h e weight of the individual germs is naturally much
reduced.
T h e appearance of twins is caused by an abnormal flower
development. H. Savitsky observed different deviations from the
normal embryological development of flowers and ovules in
multigerm beets which lead to formation of twins. These abnormalities consisted of 1) development of 2 ovaries in the same
flower with 1 ovule in each (Figure 3); 2) development of 2
ovules in the same cavity of the ovary (Figure 4)sometimes in
the flower cluster 2 ovaries contained double ovules; and 3) development in 1 ovule of 2 nucelli surrounded by the internal
integuments, each nucellus containing a normally d e v e l o p e d

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N O . 7, OCTOBER 1965

637

Figure 3.Two ovaries in 1 flower, each containing 1 ovule, X 58.

Figure 4 T w o ovules in 1 cavity of the ovary, X 58.

embryo sac (Figure 5). Each of these types of deviations produces

non-identical twins. T h e identical twins are formed as a result of a splitting of a


porembryo at an early stage of development or after a cleavage
of a zygote a n d formation of a g r o u p of embryogenic cells from
which several embryos develop. O t h e r manifestations of polyembryonydevelopment of adventive embryos from o t h e r n u c l e i
of the same e m b r y o sac (mostly from cynerg.ds), or from t h e
cells outside t h e e m b r y o sac (from nucellus or integument) can
not result in f o r m a t i o n of identical twins because such embryos

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JOURNAL OF THE A. S. S. B. T.

Figure 5.Ovule with 2 nucelli, each containing an embryo sac, X 125.

will be genetically unlike the embryo developed from the fertilized egg cell. Only if a twin pair will arise from somatic cells
outside the embryo sac (nucellus, integument) in a result of
partogenesis, such twins will be identical. Polyembryony resulting from splitting of an embryo, or from a cleavage of a fertilized
egg cell is a rare appearance in angiosperms. A development of
a twin pair from somatic cells is also extremely rare. An intensive embriological study of sugar beets indicates that occurrence of identical twins in sugar beets should be extremely rare.
In his intensive study of twins in beets, Fisher (3) , using the
morphological analysis, found that twin seedlings grew from
2 ovules of the same flower as well as from a single ovule. Triple
seedlings were also observed. Some twin pairs grew from the
ovules which contained 2 separate perisperms, and some of the
ovules with a common perisperm. T h e latter twin pairs may
be represented by the identical twins. However, the majority
of twin pairs, even in this group, did not appear to be identical,
although the morphological resemblance of some twin plants
lead the author to the conclusion that identical twins are also
produced in Beta.
In monogerm beets the appearance of twin seedlings is caused
mainly by the development of 2 ovules in the ovary of a flower.
Some twins grew from the same ovule. In the majority of monogerm strains the percent of twins is not higher than in the multigerm strains. In self-sterile monogerm populations twin seedlings

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are rare. B u t in some m o n o g e r m i n b r e d lines, up to 3 0 % of


fruits p r o d u c e twin seedlings. Variation in percent of twins
produced in the individual i n b r e d lines indicates that formation
of twins is hereditarily controlled. In fruits c o n t a i n i n g m a n y
small germs, the germs are of a low a n d variable weight. It is
i m p o r t a n t that the increase in weight of the m o n o g e r m fruits
in tetraploid strains is not followed by an increase in the n u m b e r
of fruits that p r o d u c e twin or triple germs. In all b r e e d i n g
materials investigated, tetraploid beets developed fewer twin
germs t h a n the diploid beets. It is self-evident that the p r o b l e m
of twins is m u c h m o r e i m p o r t a n t for the m o n o g e r m than for
the m u l t i g e r m beets. T h e genetic n a t u r e of their formation a n d
the grade of influence of the e n v i r o n m e n t on their p r o d u c t i o n
are not k n o w n at present. T h e supposition can be made, however, that in a d d i t i o n to genetic effects, environmental factors
greatly influence the development of surplus ovules and twin
seedlings in m o n o g e r m beets.
Discussion a n d Conclusion
In m u l t i g e r m sugar beets, the n u m b e r of flowers in clusters
is controlled by the genes which are members of one m u l t i p l e
allele M-m (partially also by modifying genes); therefore, segregation for m u l t i g e r m a n d m o n o g e r m plants is evident in the F 2
and b 1 generations (15,17,19).
It was established that the embryological d e v e l o p m e n t of
the inflorescence is different in m u l t i g e r m a n d m o n o g e r m beets.
In the m o n o g e r m beets only one flower develops on the peduncle.
Subsequent flowers which develop in m u l t i g e r m beets on t h e
same p e d u n c l e are absent in the m o n o g e r m beets. T h e i r absence
is not caused by the arrested development, or degeneration of
the r u d i m e n t a l flowers, b u t because they are not formed in the
ontogenetic d e v e l o p m e n t of the homozygous m o n o g e r m plants
(10). T h i s explains why the m o n o g e r m character is n o t subject
to a wide r a n g e of variability u n d e r different genetic or environmental conditions.
Segregation for weight of fruits or germs is m o r e complicated
than segregation for t h e n u m b e r of flowers in a cluster (monogermity or m u l t i g e r m i t y ) . T h e limits between large a n d small
fruits are easily changeable u n d e r different e n v i r o n m e n t .
T h e availability of m o n o g e r m self-sterile a n d self-fertile
populations m a d e it possible to obtain information c o n c e r n i n g
environmental (years, soils, locations) a n d genetic variability of
the weight of fruits a n d germs a n d also p e r m i t t e d a study of
the variability of this character caused by polyploidy. An adequate study of the variability of the weight of fruits a n d germs

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JOURNAL OF THE A. S. S. B. T.

is impossible in the multigerm races of B. vulgaris L., because


of the bias action of genes determining the number of flowers
in the clusters.
Environment is an important factor which causes variability
of the weight of monogerm fruits. Variability of weight of 1000
monogerm fruits was proved by cultivation of a diploid and a
tetraploid monogerm inbred line during 3 years in 10 different
locations. At the same time, interaction between level of ploidy
and years, and between level of ploidy and locations was statistically insignificant.
Mutation which led to the development of a single flower
on a peduncle (monogerm character) was not accompanied by
increase in fruit size; therefore, improvement of seed size must
be accomplished by selection.
In spite of their different origins, the monogerm diploid
populations obtained from repeated backcrosses of monogerm
beets to the open-pollinated multigerm varieties, did not differ
from each other in the weight of monogerm fruits. But if inbreeding or selection for the weight of fruits is started within
such monogerm population, the monogerm lines with different
weight of fruits and germs may be isolated. Inbreeding or selection fixes the genetic variability in the weight of fruits derived
from heterogeneous multigerm populations. Stored heterogeneity
for weight in fruit may be transmitted to the monogerm populations from the multigerm populations used in backcrosses. Every
one of these populations carried diverse genes determining the
weight of fruits, but these genes were not expressed in the multigerm fruits and, therefore, the multigerm populations did not
differ greatly among themselves in the weight of fruits.
Open-pollinated populations of sugar beet are heterogeneous
in the multiple allele M-m (17) . A multiple allelic system of a
locus may give rise to genotypes of different viability in the
population. If these genes remain in equilibrium in the population, the population will exhibit a balanced polymorphism in
the number of flowers per cluster. A stable equilibrium is reached
when the viability of homozygotes is less than the mean viability
of a population (a kind of generalized heterosis) . Under such
conditions no heterozygote will be less viable than either of its
associated homozygotes (9) .
T h e new monogerm panmictic populations did not receive
a ready system of variability of the weight of fruits based on the
gene pool obtained as a result of previous selection for the
breeding for the weight of monogerm fruits is based on the
weight of fruits in the multigerm populations. T h e temporary

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13, N o . 7, OCTOBER

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641

breeding for the weight of monogerm fruit is based on the


genes that were present in the multigerm populations. T h e s e
genes could n o t be controlled by the natural or artificial selection
in the interaction with the gene m in the homozygote. T h u s , t h e
stored genetic variability of multigerm varieties furnishes the
genes which produce variations in weight of fruits in m o n o g e r m
beets. Variability in the weight of fruits may occur in different,
and sometimes undesirable, directions. T o obtain the optimal
weight of fruits a n d germs, selection of m o n o g e r m lines w i t h
desirable expression of this character is necessary, a n d such
selection was successful in our experiments.
H e r i t a b i l i t y in the direction of increased weight of fruits
and germs appears to be low in different m o n o g e r m openpollinated populations; b u t because of heterogeneity of populations in the genes controlling the weight of fruits a n d the absence
of bias caused by the variation of the different n u m b e r of flowers
in the clusters, selection for increased weight of m o n o g e r m fruits
is effective. Such selection results in a rapid change of t h e
frequency of genes controlling the weight of fruits a n d germs
in the m o n o g e r m populations. A former type of variability of
weight of fruits a n d germs, which was created in the process of
evolution of the m u l t i g e r m populations on the basis of 2 hereditarily different characters (size a n d n u m b e r of individual fruits
within a seedball), is completely replaced in the m o n o g e r m
populations by the variability of genes controlling the weight
of fruits. T h i s d e t e r m i n e s the selective advantage of m o n o g e r m
beets for the i m p r o v e m e n t of the weight of fruits.
Occurrence of twins is a factor which increases the variability
of the weight of germs, w i t h o u t changing the variability of the
weight of fruits. T w i n germs reduce the weight of individual
germs w i t h i n a fruit. T w i n seedlings appear in m u l t i g e r m , as
well as in m o n o g e r m sugar beets. T h e i r appearance is caused
by the a b n o r m a l development of flowers a n d ovules. In m o n o germ sugar beets twin seedlings grow mainly from 2 ovules
which developed in the same cavity of the ovary. As indicated
by embryological study appearance of identical twins should
be extremely rare in beets. T h e tetraploid strains develop fewer
twins t h a n t h e i r d i p l o i d ancestors.
T h e effectiveness of b r e e d i n g for larger fruits in the monogerm p o p u l a t i o n s may be increased if the tetraploid m o n o g e r m
strains or the hybrids between tetraploid m o n o g e r m a n d tetraploid m u l t i g e r m beets are used. D o u b l i n g of chromosomes increases the weight of fruits a n d also the weight of germs a b o u t
60 to 7 0 % .

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JOURNAL OF THE A. S. S. B. T.

Presence of tetraploid monogerm strains raises the comparative genetic variability of monogerm diploid beets in the weight
of fruits and germs. This phenomenon was observed in selfsterile open-pollinated populations and in self-fertile inbred lines.
Tetraploidy results in an increase in weight of fruits in
populations, regardless of their origin. A new genetic variability
arises within autotetraploid populations. Therefore, tetraploid
lines with different weight of fruits were selected from the same
population.
Of the 3 factorsenvironment (years, locations), genetics,
and ploidy levelwhich determine the degree of variability in
weight of fruits in monogerm beets, ploidy level appeared to
be the most significant and universal. Increase in weight of
fruits in tetraploid beets may be caused by an increase in the
volume of nuclei and cells, a phenomenon manifested in all
tetraploid crops. Universality of the increase in the weight of
fruits and germs in tetraploid sugar beets may be caused by
this reason.
However, a new genetic variability of weight of fruits and
germs in monogerm tetraploid sugar beets is caused as directly
by chromosome doubling, as also by the shift from diploid to
tetraploid heredity (new mode of gene action, changes in
equilibrium of genes in populations, in segregation of hybrids,
in linkage and inbreeding effect).
Self-fertile and self-sterile tetraploid monogerm strains permit
breeding for the weight of fruits on a higher level than is possible
for the diploid monogerm beets. Diploid monogerm populations
did not provide as wide a range of genetic variability as tetraploid
monogerm populations in breeding for the weight of fruits.
Literature Cited
(1)

(2)
(3)
(4)

(5)

BARTL, K., P. CURTH, H. E. FISHER, and H. S. SCHNEIDER.

1957.

Unter-

suchungen liber die Keimfahigkeit von Polyploidem Zuckerrubensaatgut. Zucker 7: 142-147.


DOXTATOR, C. W. and R. H. HELMERICK. 1962. Selection for seed size
in monogerm varieties. J. Am. Soc. Sugar Beet Technol. 2 (3) : 268272.
FISHER, H. E. 1956. Untersuchungen an Zwillingen von Beta vulgaris L. Ziichter 26: 136-152.
FURSTE, W. 1958. Massnahmen zur Erhohung des Anteiles triploider
Pflanzen in polyploiden Zuckerriiben. Deutsche Akademie der
Landwirtschaftswissenschaften zu Berlin. Wissenschaftliche Abhandlungen 34: 28-36.
Knapp, E. 1958. Beta-Ruben bes. Zuckerriiben. In Roemer, T. and
W. Rudolf. Handbuch der Pflanzenzuechtung. 2d ed. 3: 197-284

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13,

No. 7,

OCTOBER

1965

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(6) LAZAR, O. 1961. Metabolisme respiratoire de la betterave sucriere,


IX. Activite de la polyphenoloxydase et des decarboxylases au
cours du reveil des plantules diploides et tetraploides. La Sucrerie
Beige. 80(10) : 409-419.
(7) LAZAR, O. 1962. Germination des glomerules de la betterave sucriere
en fonction du lavage, du faconnage at des activities enzymatiques
I. La Sucrerie Beige 81 (5) : 175-196.
(8) LERNER, I. M. 1954. Genetic homeostasis. Wiley, New York. 134 p p .
(9) MANDEL, S. P. H. 1959. T h e stability of a multiple allelic system.
Heredity 13: 289-302.
(10) SAVITSKY, H. 1950. Embryology of mono- and multigerm fruits in the
genus Beta L. Proc. Am. Soc. Sugar Beet Technol. 6: 160-164.
(11) SAVITSKY, H . 1952. Selective fertilization studies and recovery of selfsterile from self-fertile races of monogerm sugar beets. Proc. Am.
Soc. Sugar Beet Technol. 7: 339-343.
(12) SAVITSKY, H. 1954. Self-sterility and self-fertility in monogerm beets.
Proc. Am. Soc. Sugar Beet Technol. 8 (2) : 29-33.
(13) SAVITSKY, H. 1954. Obtaining tetraploid monogerm self fertile, self
sterile, and male sterile beets. Proc. Am. Soc. Sugar Beet Technol.
8 (2) : 50-58.
(14) SAVITSKY, V. F. 1930. A study of the variability of characters of Beta
vulgaris L. in the second year of life. Bull. Belaya Tserkov PlantBreed. Sta. Bull. 5: 69 pp.
(15) SAVITSKY, V. F. 1952. Methods and results of breeding work with
monogerm beets. Proc. Am. Soc. Sugar Beet Technol. 7: 344-350.
(16) SAVITSKY, V. F. 1954. Relation between the weight of fruit and weight
of germ in mono- and multigerm beets. Proc. Am. Soc. Sugar Beet
Technol. 8(2) : 16-22.
(17) SAVITSKY, V. F. 1954. Inheritance of the number of flowers in flower
clusters of Beta vulgaris L. Proc. Am. Soc. Sugar Beet Technol.
8(2) : 3-15.
(18)

(19)
(20)

(21)

(22)

SAVITSKY, V. F.,

G.

K.

RYSER,

G.

E. RUSH,

and C.

P.

PARRISH.

1954.

Interrelation between weight of seed and fruit and utilitarian characters in inbred lines and hybrids of monogerm sugar beets. Proc.
Am. Soc. Sugar Beet Technol. 8 (2) : 399-403.
SAVITSKY, V. F. 1958. Genetische Studien und Ziichtungsmethoden
bei monogermen Ruben. Zeitschr. fur Pflanzenzuchtung. 40: 1-36.
SAVITSKY, V. F. 1962. Sucrose and weight of root in tetraploid monogerm and multigerm sugar beet populations under different mating
systems. J. Am. Soc. Sugar Beet Technol. 1 1 ( 8 ) : 676-711.
SCHNEIDER, H. 1960. Untersuchungen iiber die Beziehungen zwischen
Knauelgrdsse, Reifegrad, Keimfahigkeit und Ploidiestufe bei Zuckerriibensaatgut. Deutsche Akademie der Landwirtschaftswissenschaften
zu Berlin. Wissenschaftliche Abhandlungen. 48: 39-46.
SEDLEMAYR, T. 1961. Untersuchungen uber den Einfluss verschiedener
Faktoren auf die Ploidiestufenanteile einer anisoploiden Zuckerrubensorte. Zuchter. 31 (7) : 310-317.

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(23) STEWART, D. 1961. Development of monogerm varieties of sugar beets


in North America. Report of first joint meeting Am. Soc. of Sugar
Beet Technol. with Inst. Int. de Recherches Betteravieres. London,
p p . 119-122.
(24) STAUDE, H. 1959. Toleranzen bei Stichprobenpriifungen von Ploidiegradbedingungen im Handel mit anisoploidem Zuckerrubensaatgut.
Deutsche Akanemie der Landwirtschaftswissenschaften zu Berlin.
Wissenschaftliche Abhandlungen. 41: 29-50.
(25) T O L M A N , B. 1961. Genetical monogerm seed and its practical application. Report of first joint meeting Am. Soc. of Sugar Beet Technol.
with Inst. Int. de Recherches Betteravieres. London, p p . 115-119.
(26) T O L M A N , B. and R. JOHNSON. 1956. Materials and methods used in
producing commercial male-sterile hybrids. Proc. Am. Soc. Sugar
Beet Technol. 9 ( 2 ) : 153-160.
(27) ZOSSIMOVITSCH, V. 1935. T h e weight and quality of sugar beet germs
in different flowers of a seed ball. Scientific Notes for Sugar Industry. 5(6) : 128-131.
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flowers of a sugar beet ball. Selekts. i Semenovodsvo. 5: 90-92.

Laboratory Screening Tests of


Insecticides for Control of the Beet Webworm 1
W A L T E R E.

PEAY2

Received for publication October 26, 1964

W i t h the growth of the livestock industry, sugar beet tops


are b e i n g utilized m o r e for livestock feed. T h u s , pesticide residues on beet foliage have become an i m p o r t a n t p r o b l e m . Considerable work is b e i n g d o n e in an effort to find materials that
will control the beet w e b w o r m (Loxostege sticticalis (L.)) witho u t leaving harmful or illegal residues on the foliage. H a g e n ( l ) 4
found that carbaryl a n d endosulfan gave as good control as e n d r i n
u n d e r field conditions. McDonald (2) found trichlorfon to be
the most effective of 17 c o m p o u n d s tested in the laboratory a n d
field for beet w e b w o r m control. In 1962, laboratory screening
tests of m a n y new materials were started at T w i n Falls, I d a h o .
T h i s is a r e p o r t of these tests and not a r e c o m m e n d a t i o n of any
of t h e materials m e n t i o n e d .
Experimental

Procedure

Beet w e b w o r m adults were collected in black light traps a n d


the healthy ones transferred to a cylindrical cage, 16 inches high
and 8 inches in diameter, m a d e of 8-mesh galvanized screen wire.
T h e t o p contained a 1-inch hole for inserting the moths, a n d
the b o t t o m was placed in a 10-inch diameter tray c o n t a i n i n g
l 1/2 inches of moist sand. T h e s e adults were fed a 5% sugar
solution, d r i p p e d by gravity flow o n t o toweling placed on the
top of the cage. Eggs were deposited by the moths on w h i t e
paper toweling tightly wrapped a r o u n d the outside of the cage
and secured with masking tape.
As the eggs hatched, the larvae were collected by placing
leaves of sugar beets or lambsquarters (Chenopodium album
L.) a r o u n d the u p p e r edge of the cage. T h e y were t h e n transferred to ice cream cartons covered with cheesecloth a n d fed
fresh l a m b s q u a r t e r s daily. W h e n they were in t h e t h i r d a n d
fourth instar, they were used for the insecticide screening tests.
Small sugar beet plants were used in evaluating t h e insecticides. T h e beet seed was sprouted in a light sandy soil a n d single,
uniformly vigorous seedlings were transplanted to 4-inch flower
pots. W h e n the beets were in the 6- to 8-Ieaf stage, foliar sprays
1
Published with the approval of the Director of the Idaho Agricultural Experiment
Station
as Research Paper No. 590.
2
Entomology Research Division, Agricultural Research Service, USDA, Twin Falls,
Idaho.
3
Robert D. Berard, temporary Research Helper, assisted in some of these tests.
4
Numbers in parentheses refer to literature cited.

646

JOURNAL OF THE A. S. S. B. T.

were applied to individual plants as they revolved on a turntable. All materials were tested at the rates of 1/4, 1/2 1, a n d 2
p o u n d s of toxicant in 35 gallons of water. T h e plants were
sprayed with a DeVilbiss 5 spray g u n at 10 psi, with 5 cc of spray
p e r plant, which is approximately 35 gallons per acre. F o u r
plants treated with each of the four concentrations of each
material a n d four u n t r e a t e d plants constituted a test, a n d each
test was replicated twice.
O n e day and again 1 week after spraying, five t h i r d to fourthinstar beet w e b w o r m larvae were caged on each plant. Larval
mortality counts were m a d e 4, 8, 24, 48, a n d 72 h o u r s after
larvae were placed on the plants.
T h e r e were 33 materials tested, of which 17 were proprietary
products, as follows:
Bayer 25141
0,0-diethyl
o [p-(methylsulfinyl)phenyl] phosphorothioate
Bayer 37289 0-ethyl 0-2,4,5-trichlorophenyl ethyl phosphonothioate
Bayer 38156
0-ethyl
S-p-tolyl e t h y l p h o s p h o n o d i t h i o t e
Bayer 41831 0,0-dimethyl #~4-nitro-m-tolyl phosphorothioate
Bidrin
3-hydroxy-iV, i V - d i m e t h y l - c i s - c r o t o n a m i d e dimethyl phosphate
Ciodrin
alpha-methylbenzyl 3 - h y d r o x y c r o t o n a t e d i methyl phosphate
EPN
0-ethyl 0-p-nitrophenyl p h e n y l p h o s p h o n o t h i o a t e
Ethyl Guthion 0,0-diethyl S - ( 4 - o x o - l , 2 , 3 - b e n z o t r i a z i n - 3 ( 4 H ) ylmethyl) p h o s p h o r o d i t h i o a t e
Guthion
0,0-dimethyl S-(4-oxo-l,2,3-benzotriazin-3(4H)ylmethyl) p h o s p h o r o d i t h i o a t e
Perthane
a m i x t u r e of 1, l-dichloro-2,2-bis(p-ethylphenyl)
e t h a n e (95%) a n d related reaction products
(5%)
Shell SD-8280 2-chloro-l-(2,4-dichlorophenyl)vinyl d i m e t h y l
phosphate
Shell SD-8436 2-chloro-l-(2,4-dibromophenyl)vinyl d i m e t h y l
phosphate
Shell SD-8447 2-chloro-l-(2,4,5-trichlorophenyl)vinyl dimethyl
phosphate
Shell SD-8448 2-chloro-l-(2,4,5-trichlorophenyl)vinyl d i e t h y l
phosphate
5

Mention of this proprietary product does not necessarily imply endorsement by


the USDA.

VOL.

647

13, N o . 7, OCTOBER 1965

Telodrin
Zectran
Zinophos

l,3,4,5,6,7,8,8-octachloro-l,3,3a,4,7,7a-hexahydro-4,7-methanoisobenzouran
4-dimethylamino-3,5-xylyl methylcarbamate
0,0-diethyl 0-2-pyrazinyl phosphorothioate

Table 1.Materials, minimum dosages that gave the best control, and the mortality
of the beet webworm in 48 a hours when the larvae were caged on the plants 1 day anil
1 week after spraying. Twin Falls, Idaho, 1963.
Larvae caged on plants

1 day after s p r a y i n g
Material

Bayer 25141
Bayer 37289
Bayer 38156
Bayer 41831
Bidrin
Diazinon
Dimethoate
EPN

Guthion
Guthion-Ethyl Guthion
Mevinphos
Naled
Phosphamidon
Shell SD-8280
Shell SD-8436
Shell SD-8447
Shell SD-8448
Telodrin
Trichlorfon
Zinophos
Untreated check
DDT

Endrin

Toxicant
per acre
Pounds
.50
.25

Mortality

Mortality

Percent

Pounds

Percent

.50
.25
1.OOb
1.00
.25
2.00b
.25
.50
1.00
1.00
.25
.25
.50
.25
.25
.25
.25

100
100
95
100
100
100
100
100
100
100
100
100
95
100
100
100
95
100
95
100

2.00
.25

95
100

1.00

1 week after s p r a y i n g
Toxicant
per acre

.50
1.00
2.00
2-00
2.00
2.00b
1.00
.50
2.00b
2.00
2.00
2.00b
2.00
2.00
.25
2.00
.25
.50
2.00
2.00

98
100
70
85
100
95
100
100
93
95
85
20
40
100
100
100
100
100
93
98
1

2.00
1.00

95
100

Fortv-eight hours seemed to be the optimum time to use. In many cases the larvae
were moribund but not dead in 24 hours; and if they lived more than 48 hours, they did
considerable feeding.
b
T h e materials at these dosages caused plant burning.

Discussion of Results
T h e materials that gave as good control o the beet webworm as D D T and endrin, which were used as standards, are
shown in Table 1.
Several materials gave 100-percent mortality with as little as
0.25 pound per acre 1 day and also 1 week after the beets were
sprayed. T h e quickest kills were obtained with Shell SD-8436
and EPN. They gave good control in 8 hours. Bayer 37289,
Bidrin, mevinphos, naled, Shell SD-8436, and Telodrin gave
very good control in. 24 hours.

648

JOURNAL OF THE A. S. S. B. T.

Materials that gave the best control in these tests a n d that


have been registered for use on sugar beet tops to be used for
feed are naled, p h o s p h a m i d o n , a n d trichlorfon. Materials that
gave only fair control b u t are registered for use on sugar beet
tops are carbaryl, c a r b o p h e n o t h i o n , a n d p a r a t h i o n .
Malathion, menazon, phorate, schradan, a n d Bacillus thuringiensis var thuringiensis Berliner were ineffective in these tests.
Literature Cited
(1) HAGEN, ARTHUR F. 1961. Evaluation of T h i o d a n and sevin for control
of webworms in sugar beets. J. Econ. Ent. 54 (4) : 799-800.
(2) MCDONALD, S. 1963. Chemical control of the beet webworm on sugar
beets in southern Alberta. J. Econ. Ent. 5 6 ( 3 ) : 248-51.

Insect Control on Sugar Beets by Seed


or Soil Treatments 1
H O W A R D E.

DORST2

Received for publication January 27, 1965

Control of certain insects in sugar beets by seed or soil treatm e n t has b e e n reported by various workers. For control of sugar
beet root maggot [Tetanops myopaeformis ( R o d e r ) ] , Jones et
al. (1) 3 r e p o r t e d that aldrin a n d heptachlor were effective, applied
dry at 1/2 a n d 14 p o u n d respectively per 100 p o u n d s of seed.
Callenback et al. (3,4) found these insecticides were not effective
unless seed was pelleted with wettable powder of the insecticide,
to give 1 p o u n d of toxicant per 100 p o u n d s of seed. A l l e n et al.
(6) in 1957 showed that heptachlor seed treatments increased
yields, b u t a l d r i n a n d d i e l d r i n did not.
Morrison (5) reported soil treatments to be superior to seed
t r e a t m e n t s for control of the garden symphylan [Scutigerella
immaculata
(Newport)].
Hills et al. (2,8) a n d Dorst (7) reported the effectiveness of
p h o r a t e a n d Di-Syston [0,0-diethyl S-[2-(ethylthio)ethyl] phosp h o r o d i t h i o a t e ] on sugar beet seed for beet leafhopper control
on sugar beets grown for seed.
M o n o g e r m sugar beet seed, which has almost entirely replaced the larger m u l t i g e r m seed, was developed by the p l a n t
breeders to p r o d u c e single plants to permit mechanical t h i n n i n g .
Precision p l a n t i n g involved in this operation has sometimes req u i r e d that the seed be pelleted to make a more u n i f o r m size.
Insecticides a n d fungicides added to the exterior of seed as
dry material or slurries for insect or fungus control, sometimes
fail to a d h e r e to the seed when handled a n d their value is often
lost. Because of the smaller size of the m o n o g e r m seed, it is
difficult .to m a k e e n o u g h of the insecticide adhere to the seed
surface.
Pelleting sugar beet seed provides an inexpensive m e t h o d of
a d d i n g insecticides. In 1962 studies were conducted in n o r t h e r n
U t a h to d e t e r m i n e whether the sugar beet root maggot, t h e
garden symphylan, a n d the beet leafhopper [Circulifer tenelhis
(Baker)], could be controlled on sugar beets a n d w h e t h e r such
control w o u l d increase yields a n d reduce curly top disease. A
factor considered was w h e t h e r effective concentration of t h e
1
2
3

In cooperation with the Utah Agricultural Experiment Station.


Entomology Research Division, Agricultural Research Service, USDA, Logan, Utah.
Numbers in parentheses refer to literature cited.

650

JOURNAL OF THE A. S. S. B. T.

systemic insecticide could be i n t r o d u c e d i n t o the p l a n t w i t h o u t


causing excessive phytotoxicity. T h i s is a r e p o r t of the results
of these studies a n d n o t a r e c o m m e n d a t i o n of any material used.
T h e two organophosphorus systemic insecticides, phorate, a n d
Di-Syston, formulated in carbon powder, were included in the
coating material used in p r o d u c i n g the pellets. V-C 13 (0-2,
4-Dichlorophenyl 0,0-diethyl phosphorothioate), a n o t h e r organophosphorus systemic insecticide was added to a portion of the
pellets as a wettable powder; aldrin, wettable powder, was added
to a n o t h e r portion. A b o u t two-thirds of the e x p e r i m e n t a l work
had been completed w h e n it was discovered that a b a n d or insulating layer between the seed a n d the phorate, Di-Syston, or
V-C 13 insecticide provided a pellet that caused markedly less
toxicity and damage to the planted seed t h a n formulations witho u t the insulating layer. Effectiveness of insecticide granules
mixed in the t o p 2 to 3 inches of the soil in a 6-inch b a n d along
the row p r i o r to p l a n t i n g was c o m p a r e d with that of using the
pelleted seed.
T h e sugar beet seed (curly-top resistant) was furnished by
T h e Amalgamated Sugar Company 4 . T h e seed was pelleted a n d
treated with insecticide formulations by G e r m a i n ' s Incorporated 4 .
T r e a t m e n t plots were 8-rows wide a n d the length of the field
or 72 rods. T h e seed was planted with a 4-row drill. U n t h i n n e d
stands were compared by e x a m i n i n g 100 inches of row in each
plot a n d c o u n t i n g the n u m b e r of such inches c o n t a i n i n g plants.
T h i n n e d stands were c o m p a r e d by c o u n t i n g the plants in pred e t e r m i n e d sections of rows 33 a n d 34, a n d sections 33 feet long
in t h e 2nd, 4th, a n d 6th rows, respectively, of the u p p e r , center,
a n d lower thirds of each plot. In d e t e r m i n i n g stand losses, plants
were c o u n t e d in the sample areas at 3-week intervals from Tune
to August. At each c o u n t dead plants were pulled up a n d exa m i n e d to d e t e r m i n e w h e t h e r mortality was d u e to root maggot
damage.
Symphylans feed on the g e r m i n a t i n g seed a n d roots of young
sugar beets, w e a k e n i n g or killing the seedlings, and t h u s r e d u c i n e
the emerged or u n t h i n n e d stand. T h e y c o n t i n u e to m u l t i p l y on
the roots d u r i n g the season, further r e d u c i n g the yield of the
crop. In late August or September symphylans were counted
on seven beets taken at r a n d o m from the top, center, a n d lower
thirds of each plot. Soil samples were o b t a i n e d by removing
one h e a p i n g tablespoon of moist soil (approximately 50cc) from
the root zone of t h e sugar beet a p p r o x i m a t e l y 6 inches below
t h e surface. T h e soil was placed in 6-inch pans, m i x e d with

VOL.

13, N o . 7, OCTOBER

1965

651

water, a n d the containers set on a slope. T h e symphylans were


c o u n t e d as they came to the surface in an effort to escape excess
moisture.
C u r l y top counts were obtained in August by e x a m i n i n g
100 plants p e r plot. Yield records were o b t a i n e d by harvesting
the beets by h a n d in 15 feet of row in the second, fourth, a n d
sixth rows of the u p p e r , center, a n d lower thirds of each plot.
T h e sampled sugar beets from the best leafhopper control plots
were sacked in labeled r u b b e r i z e d bags, weighed, a n d a composite
of the p u l p o b t a i n e d for a sucrose reading by the C e n t r a l Laboratory of the U t a h - I d a h o Sugar Company*.
T h e results of sugar beet root maggot a n d symphylan control
are r e p o r t e d in T a b l e 1. T h e s e data show substantial r e d u c t i o n s
in the n u m b e r of plants killed by root maggots from pelleted
seed c o n t a i n i n g aldrin, Di-Syston, phorate, a n d V-C 13. B a n d
treatments of phorate, V-C 13, a n d p a r a t h i o n granules at m u c h
h i g h e r dosages were comparable to the pelleted seed t r e a t m e n t
in protecting the seed from the sugar beet root maggot. In t h e
field where 4.0-ounce rate of V-C 13 in pelleted seed was applied,
the t r e a t m e n t resulted in phytotoxicity a n d reduced t h e emerged
stand of beets by 6 0 % a n d the t h i n n e d stand by 3 5 % . Seed
pelleted with the o t h e r insecticides did not cause r e d u c t i o n of
the t h i n n e d stand a n d w h e n the seed was treated with aldrin,
the stand increased by 2 4 % .
Table 1.Sugar beet root maggot and symphylan control on sugar beets in field plots
with insecticide incorporated in the pelleted seed and with row band applications of
insecticide granules. Stevenson Field, Lewiston, Utah. 1962.
Insecticide and dosage
in ounces per acre in
pelleted seed and pounds
per acre in granules
In Pelleted Seed
Phorate
2.0
V-C 13
4.0
Aldrin
0.8
Di-Syston
0.9
Granules in Soil
Phorate
2.0
V-C 13
2.0
Parathion
2.0
Check (Unpelleted Seed
LSD at 5 percent

Number of
thinned
plants per
100 feet

Number of plants
killed by maggot
per 100 feet

Number of
symphylans
per 7 samples
each plot

69
69
75
65

12
1.7
0.7
1.0

1.5
1.0
3.7
4.7

27
31
26
25

0.2
2.2
2.5
19.7
3.3

4.0
3.2
3.0
6.5
1.8

24
24
26
22
4

66
70
68
60
14.6

Yield
tons
per
acre

At the t i m e of emergence, the y o u n g seedlings were a p p a r e n t l y


protected from symphylan damage by b o t h the pelletized seed
a n d t h e row-band treatments. Seed pelleted with p h o r a t e a n d
* Mention of a company name does not necessarily imply endorsement of this company's
product by the U. S. Department of Agriculture.

JOURNAL OF THE A. S. S. B. T.

652

V-C 13 were superior to those pelleted with a l d r i n a n d Di-Syston,


b u t the dosages were higher. T h e b a n d treatments with insecticide granules were inferior to the pelleted seed for symphylan
control.
Even on resistant sugar beets, curly top transmitted by the
beet leafhopper causes some r e d u c t i o n in yield. Results of experiments to control the leafhopper a n d reduce curly t o p with
systemic insecticides in pelleted seed a n d p h o r a t e granules are
given in T a b l e 2. P h o r a t e pelleted seed t r e a t m e n t at 2 ounces
per acre reduced the stand of t h i n n e d beets 2 1 % . T h e o t h e r
pelleted seed treatments also t e n d e d to reduce the t h i n n e d stand
as compared with that in the u n t r e a t e d check. Curly t o p incidence was reduced a n d the total sugar p e r acre increased by
all treatments. T h e percentage of sucrose was similar for all
treatments a n d ranged from 15.2 to 15.5.

Table 2.Beet leafhopper control on sugar beets in field plots with insecticide incorporated in the pelleted seed or granules applied in a 6-inch band of row. Two fields,
Gardner and Johnson. Delta, Utah. 1962.
Insecticide and dosage
in ounces per acre in
pelleted seed and pounds
per acre in granules

Number of
thinned
plants per
100 feet

Percentage
obvious
curly top

In Pelleted Seed
.25
Phorate
.5
Phorate
1.0
Phorate
2.0
Phorate
.9*
Di-Syston
2.0
V-C 13
4.0
V-C 13

76
76
77
58
75
72
67

7
7
7
5
9
6
5

69
69
69
74
67
70
74

2.80
2.78
2.88
2.92
2.72
2.93
3.02

Ganules in Soil
2.0
Phorate
Check
LSD at 5 percent

81
73
8

7
33
3

70
63
5

2.89
2.56
0.07

Pounds of
raw

sugar beets
per 45 feet

Net sugar
tons
per acre

Summary
In field e x p e r i m e n t s in U t a h in 1962 low dosages of V-C 13,
Di-Syston, a n d p h o r a t e i n c o r p o r a t e d i n t o the coating m a t e r i a l of
pelleted sugar beet seed gave p r o m i s i n g results in the control
of the sugar beet root maggot, the garden symphylan, a n d the
beet leafhopper on sugar beets. Similar treatments with aldrin
were also promising against the root maggot a n d symphylan.
Indications were t h a t phytotoxicity may be a l i m i t i n g factor in
this type of t r e a t m e n t .

VOL.

13, N o .

7, OCTOBER

653

1965

Literature Cited
(1)

JONES, E. W., J. R. DOUGLASS, C. P. PARRISH and VERNAL JENSEN.

1952.

Experiments on control of the sugar beet root maggot. Proc. Am.


Soc. Sugar Beet Technol. p p . 490-496.
(2)

H I L L S , O R I N A., F. H . HARRIES a n d A. C. VALCANCE. 1956. T r e a t m e n t

of sugar beet seed with systemic insecticides for control of the beet
leafhopper. J. Am. Soc. Sugar Beet Technol. 9 ( 2 ) : 124-128.
(3)

CALLENBACK, J . A., W . L. G O J M E R A C a n d D. B. OGDEN. 1957. T h e sugar

beet root maggot in North Dakota. J. Am. Soc. Sugar Beet Technol.
9 (4) : 300-304.
(4) CALLENBACK, J. A. and W. L. GOJMERAC. 1957. Biology and control
of sugar beet root maggot. North Dakota Agr. Exp. Sta. Unpublished
Rept.
(5) MORRISON, H. E. 1957. Controlling symphylins. Oregon Agricultural
Experiment Station Circular 574.
(6)

ALLEN, W . R., W . L. ASKEW a n d K. SCHREIBER. 1959. R e l a t i o n of sugar

beet root maggot control by insecticides on sugar beet yields. J. Am.


Soc. Sugar Beet Technol. 10 (4) : 330-334.
(7) DORST, H. E. 1960. Experimental control of the beet leafhopper on
sugar beets grown for seed. J. Am. Soc. Sugar Beet Technol. 11 (1) :
12-14.
(8)

HILLS,

ORIN

A.,

A.

C.

VALCARCE,

H.

K. JEWELL

and D.

C.

COUDRIET.

1960. Beet leafhopper control in sugar beets by seed or soil treatments. J. Am. Soc. Sugar Beet Technol. 11 (1): 15-24.

JOURNAL
of the
American Society of Sugar
Beet Technologists
Volume 13
Number 8
January 1966

Published

quarterly

by

American Society of Sugar Beet Technologists


Office of the Secretary
P. O. Box 538
Fort Collins, Colorado 80521

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Made in the United States of America

TABLE OF CONTENTS
Title

Author

Effectiveness of selection for tetraploid plants


in C 0 generation on the basis of the
n u m b e r of chloroplasts in stomata

Recent advances in molasses exhaustion

Colorimetric micro determination of calcium


in sugar-house products

Helen Savitsky

K. W. R. Schoenrock
J. R. Johnson

G. V. Heyndrickx

A method for positive selection of 4N sugar


beet plants in the vegetative C 0 generation G. H. Rank
L. E. Evans

On the nature
schachtii. I I I .
activity

655

... 662

681

687

of hatching of Heterodera
P r i n c i p l e s of h a t c h i n g

Odor in refined sugar

Probability analysis of herbicide response

A regression study on tare samples of sugar


beets in relation to factors influencing
productivity a n d quality

Notes Section

Page

D. R. Viglierchio
P. K. Yu

698

Robert R. West
Robert S. Gaddie

716

E. F. Sullivan
H. L. Bush

721

George K. Ryser

727

.. 748

Effectiveness of Selection for Tetraploid Plants in


Co Generation on the Basis of the Number
of Chloroplasts in Stomata
HELEN

SAVITSKY 1

Received jar publication September 3, 1964

Introduction
Study of some characteristics of leaves such as chromosome
n u m b e r (7,11,19) 2 , size of stomata cells (2,12,14,16), n u m b e r of
nucleoli chromocenters (15), a n d n u m b e r of chloroplasts (3,4,5,
7,10,12) is widely used at present time for d e t e r m i n a t i o n of ploidy
levels. Except for e x a m i n a t i o n of chromosome n u m b e r , the most
used m e t h o d involves d e t e r m i n a t i o n of the n u m b e r of chloroplasts in the g u a r d cells of stomata.
Mochizuki a n d Sueoka (13) first indicated that diploid, triploid a n d tetraploid sugar beets differ in the n u m b e r of chloroplasts in the stomata cells of leaf epidermis ( T a b l e 1).
Table 1.Average number of chloroplasts per stoma.
According to
Mochizuki
and Sueoka

Ploidy
levels
Diploids
Triploids
Tetraploids

14
20
25

(12-16)
(17-22)
(22-28)

Butterfass
14.23 0.10
20.34 0.07
25.36 0.17

Graf

Margara
and Touvin

Savitsky

17.09 0.25
20.74 0.19
24.42 0.36

14.74 + 0.32
18.97 0.36
25.98 0.54

14.57 0.28
20.87 0.37
25.90 C.57

F u r t h e r investigations by Butterfass (3,4,5), comprising m o r e


than 900 plants, lent m o r e definition to this method. T h e k i n d
and age of the leaves that are best for examination were indicated. A l t h o u g h the average n u m b e r of chloroplasts is different
for the diploid, triploid a n d tetraploid populations, the variation
in the n u m b e r of chloroplasts in the individual plants makes it
difficult, in some cases, to d e t e r m i n e the ploidy grade. T h e limits
of the n u m b e r of chloroplasts between diploids a n d triploids a n d
between triploids a n d tetraploids are overlapping.
Modification of the n u m b e r of chloroplasts at different ploidy
levels was the m a i n subject of the investigations of Butterfass
(5). He indicated that the limits of variation between diploids,
triploids a n d tetraploids should be established for every plot
studied, by using the frequency curve. He concluded that c o u n t s
of chloroplasts in 10 stomata cells p e r m i t t e d d e t e r m i n a t i o n of
ploidy grades for the majority of plants; however, this d e t e r m i n a 1
2

Geneticist, Crops Research Division, ARS, USDA, Salinas, California.


Numbers in parentheses refer to literature cited.

656

JOURNAL OF THE A. S. S. B. T.

tion was w r o n g for 1 to 1.5% of the plants a n d could n o t be


performed for 16 to 1 8 % of the plants. According to Graf (10)
d e t e r m i n a t i o n of ploidy grades in individual plants was w r o n g
for 1.4% of diploids, for 7.0% of triploids a n d for 5.6% of
tetraploids. Dudley (6), basing on study of chloroplasts in one
g u a r d cell of stomata in eight diploid a n d eight tetraploid sugar
beets, concluded that diploids a n d tetraploids can be accurately
separated by c o u n t i n g chloroplasts in the g u a r d cells. Because
of a certain convenience which t h e chloroplast m e t h o d offers,
it may be used (but with some limitations) a n d its applicability
varies, d e p e n d i n g on the objectives. Especially difficult is identification of triploid plants, because the limits for chloroplast
n u m b e r in triploids also comprise some diploids a n d tetraploids.
In i n d u c i n g polyploidy in sugar beets, a study of leaf characteristics (one of which is n u m b e r of chloroplasts) is a c o m m o n
m e t h o d for selection of tetraploid C 0 plants after colchicine treatm e n t . T h e results of a study of reliability of this m e t h o d for
selection of tetraploid plants in the m i x o p l o i d C 0 generation are
presented in this report.
Materials and Methods
Seed of the following sugar beet strains were exposed to
colchicine t r e a t m e n t : 2 self-sterile m o n o g e r m p o p u l a t i o n s (573
mm high in curly-top resistance a n d 202 mm N-type strain); 4
self-fertile m o n o g e r m i n b r e d lines (171 mm E-type, 127 mm
Z-type, 200 mm leaf-spot resistant, a n d 537 mm which is highly
leaf-spot resistant); a n d 1 self-fertile m u l t i g e r m i n b r e d line (509
M M ) which is curly-top resistant.
After t r e a t m e n t , seed was p l a n t e d in soil in a greenhouse.
Seedlings c o m i n g up were classified as affected (with short,
thickened hypocotyls, thick a n d dissected leaves), or unaffected.
All unaffected seedlings were discarded a n d the affected ones
were transplanted i n t o cylinders a n d k e p t in the cold frame during t h e winter for t h e r m a l i n d u c t i o n . In t h e spring they were
transplanted to the field for further growth a n d selection for
tetraploid C 0 plants.
T r e a t m e n t of seed is an effective m e t h o d of i n d u c i n g polyploidy (17). A b o u t 4 0 % of affected seedlings develop completely,
or almost completely, tetraploid m a i n inflorescence in which
h u n d r e d s of flowers carry diploid gametes.
T e t r a p l o i d plants were selected on the basis of the size of
pollen grains in flowers developed in the m a i n inflorescence. It
is well k n o w n that tetraploidization increases the size of pollen
grains in many c r o p plants as well as in sugar beets. Abegg (1),
Artschwager (2), Schlosser (18) a n d V o n Rosen (19) indicated

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larger size of pollen in tetraploid beets in comparison with t h e


diploid. E r n o u l d (8) found that mean size of pollen in diploid
beets equals 19.86m a n d in tetraploid 26-27m. Feltz (9) r e p o r t e d
that t h e average size of pollen for diploids was 21.87m, a n d for
tetraploids 26-27m. According to Savitsky (16) the size of m a t u r e ,
normally developed pollen varies in diploid beets from 19 to
22m, a n d in tetraploid beets from 28 to 33m.
In several plants studied in this experiment, the chromosome
n u m b e r was d e t e r m i n e d in meiosis. T h e b u d s on branches carrying large pollen grains had 36 chromosomes in the Pollen M o t h e r
Cells at prophase, a n d those on the branches with small pollen
grains h a d 18 chromosomes.
Plants which developed large pollen grains (28-33m) in all,
or in almost all, flowers (some flowers with smaller pollen grains
may have been missed) of the m a i n inflorescence were considered to be tetraploids. Plants with smaller pollen grains (1922m) were considered to be diploids. Actually, all plants in the
treated generation (C 0 ) are to some extent mixoploids, carrying
both diploid a n d tetraploid tissues, b u t those plants which develop
prevailingly a majority of diploid gametes in flowers of the m a i n
inflorescence may be considered tetraploids, for practical purposes, because they produce tetraploid progeny after being intercrossed (17).
T h e n u m b e r of chloroplasts was counted in b o t h g u a r d cells
of stomata in the epidermis of the lower surface of leaves. Leaves
were collected from t h e same seed stalk from which the anthers
were e x a m i n e d for the size of pollen grains. Chloroplasts were
counted in 10 stomata cells (20 guard cells) of each plant. Silver
nitrate (1%) was used for dyeing.
On the basis of average n u m b e r of chloroplasts per stoma
cell, plants were classified as falling into 1 of 3 groups as follows:
1) plants having the n u m b e r of chloroplasts corresponding: to
diploids. Such plants h a d mostly 13, 14, 15 or 16 chloroplasts
per stoma cell; 2) plants with a n u m b e r of chloroplasts corresponding to tetraploids (24 or higher); a n d 3) plants with an
i n t e r m e d i a t e n u m b e r of chloroplasts (19 to 24). T h e last srroup
might i n c l u d e diploid, as well as tetraploid plants, since the
n u m b e r of chloroplasts in this g r o u p does not give a clear indication as to t h e level of ploidy of individual plants. T h i s srroup
arose partly because of the chimeral n a t u r e of the C 0 plants. In
some leaves stomata lying side by side were observed with n u m b e r
of plastids corresponding to diploids and tetraploids, or close
to their limits.

JOURNAL OF THE A. S. S. B. T.

658

Data concerning the number of chloroplasts in stomata cells


of diploid and tetraploid plants are presented in the Tables 2
and 3. Photomicrographs of chloroplasts in stomata of diploid
and tetraploid beets are presented (Figures 1 and 2).
Table 2-Number of chloroplasts in stomata cells of diploid Co sugar beet plants.
Plants
Strain

with number of chloroplasts

Corresponding to

designation

Diploids

Tetraploids

537
201
171
127
509
573
202

17
5
8
1
3
43
23

4
5
13
15
3
45
19

Total
Percent

Intermediate-Between
Diploids and Tetraploids
2
3
3
4
7
1
21
9-33

104
46.22

100
44.44

Table 3.Number of chloroplasts in stomata cells of tetraploid Co sugar beets plants.


Plants

with number of chloroplasts

Corresponding to
designation

Diploids

Tetraploids

Intermediate-Between
Diploids and Tetraploids

537
201
171
127
509
573
202

8
2
4
0
1
16
6

42
46
57
30
9
69
22

14
6
10
3
3
7
6

37
10.2

275
76.2

Total
Percent

49
13.6

Figure 1.Chloroplasts in stomata leaf cells in diploid sugar beets X 42.

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659

Figure 2.Chloroplasts in stomata leaf cells in tetraploid sugar beets X 42.

E x p e r i m e n t a l Results
O n l y 44.4 percent of diploid plants (with small pollen grains)
h a d the n u m b e r of chloroplasts characteristic of diploids ( T a b l e
2). Forty-six percent (46.2%) of plants in this g r o u p showed
chloroplast n u m b e r s corresponding to tetraploids a n d in 9 . 3 %
of plants t h e n u m b e r of chloroplasts exceeded t h e n u m b e r of
chloroplasts peculiar to diploids. T h u s , 5 5 . 5 % of the diploid
p o p u l a t i o n h a d larger n u m b e r of chloroplasts in leaves t h a n
should be present in diploids. T h i s indicated that the epidermal
tissue was affected by colchicine to a greater degree than the
s u b e p i d e r m a l tissue.
As a consequence, d o u b l i n g of chromosomes occurred m o r e
often in the epidermis t h a n in the tissues which produce sexual
cells.
A m o n g tetraploid plants (with large pollen grains) 7 6 . 2 %
of plants h a d the n u m b e r of chloroplasts corresponding to tetraploids ( T a b l e 3). O n l y 10.2% had the n u m b e r of chloroplasts
corresponding to diploid plants. Also, 13.6% of the plants showed
lower n u m b e r s t h a n tetraploids h a d to have. T h e r e f o r e , if selection of tetraploids h a d been made on the base of chloroplasts
n u m b e r , 23.8 percent of the tetraploid plants w o u l d have been
discarded.
At t h e same time, the g r o u p of tetraploid plants, selected
from the whole p o p u l a t i o n (586 plants), w o u l d have consisted
of 275 t r u e tetraploids a n d 104 (27.4%) diploids. T h i s g r o u p
of d i p l o i d plants would very highly contaminate the tetraploid
p o p u l a t i o n . Instead of o b t a i n i n g a C 1 progeny with a high percent of tetraploid plants, the C 1 generation will contain many,
if n o t a majority, of diploid a n d triploid plants which w o u l d
necessitate extensive screening to eliminate plants with undesirable c h r o m o s o m e n u m b e r s .

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JOURNAL OF THE A. S.

S.

B.

T.

Leaf characteristics, such as size of stomata cells, n u m b e r of


chloroplasts, a n d even n u m b e r of chromosomes, often used for
selection of tetraploids, should be used in C 0 generation only for
preliminary test a n d screening, if such a screening is really needed
in some circumstances or climates. Final selection for tetraploids
in the C 0 generation should be based on direct selection of diploid
gametes which will produce a tetraploid progeny. T h e ploidy
level of gametes is d e t e r m i n e d either by t h e size of pollen grains
or by chromosome n u m b e r in the pollen m o t h e r cells. T h i s
gamete selection is highly effective a n d reduces the work of controlling chromosome n u m b e r s in the following generations.
Conclusion
Selection for tetraploid plants in the treated m i x o p l o i d C 0
generation based on the leaf characteristics, n u m b e r of chloroplasts, n u m b e r of chromosomes, size of stomata, etc. is an unreliable m e t h o d .
Selection for tetraploids in C 0 generation should be based on
selection of diploid gametes (pollen grains) which will p r o d u c e
a tetraploid progeny.
Literature Cited
(1)

(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)

ABEGG, F. A., D E W E Y STEWART a n d G. H . COONS.

1946. F u r t h e r studies

on sugarbeet autotetraploids. Proc. Am. Soc. Sugar Beet Technol.


4: 223-229.
ARTSCHWAGER, E. 1942. Colchicine-induced tetraploidy in sugar beets:
Morphological effects shown in progenies of a n u m b e r of selections.
Proc. Am. Soc. Sugar Beet Technol. 3: 296-303.
BUTTERFASS, T H . 1958. Die praktische Ermittelung des Ploidiegrads
von Zuckerruben durch Zahlen der Schliesszellen-Chloroplasten.
Zuchter. 28(7) : 309-314.
BUTTERFASS, T H . 1961. Die Chloroplasten Zahl als Merkmal. Berichte
der Deutsche Botanische Gesellschaft. 7 4 ( 6 ) : 217-218.
BUTTERFASS, T H . 1961. Das Verhalten der Chloroplastenzahlen in den
Schliesszellenpaaren von Zuckerruben verschiedener Ploidiestufen
vom Keimling bis zur bluhenden Pflanze. Zuchter. 31 (2) : 62-71.
DUDLEY, J. W. 1958. N u m b e r of chloroplasts in the guard cells of
inbred lines of tetraploid and diploid sugar beets. Agr. J. 50: 169170.
ELLERTON, S. and A. J. T H . HENDRIKSEN. 1959. Note on the probable
cause of the occurrence of tetraploid plants in commercial triploid
varieties of sugar beet. Euphytica. 8: 99-103.
ERNOULD, L. 1946. L'autopolyploidie experimentale chez la betterave.
Cellule. 50 (3) : 363-430.
FELTZ, H. 1953. Untersuchungen an diploiden u n d polyploiden Zuckerruben. Zeitschr. fur Pflanzenzucht. 32 (3) : 275-300.

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661

(10) G R A F , A. 1959. Bestimmung des ploidiegrades in Z u c k e r r i i b e n Gebrauchssaatgut. Zucker. 12: 344-349.


(11) KLOEN, D. and G. L. SPECKMANN. 1953. T h e creation of tetraploid
beets. Euphytica 2: 187-196.
(12) MARGARA J. and H. TOUVIN. 1958. La polyploidie chez la betterave
sucriere. Acad. Agricult. France. Compt. Rend. 44: 172-176.
(13) MOCHIZUKI, H. and N. SUEOKA. 1955. Genetic studies on the number
of plastid in stomata. 1. Effects of autopolyploidy in sugar beets.
Cytologia (Tokyo). 20: 358-366.
(14) PETO, F. H. and K. W. H I L L . 1942. Colchicine treatment of sugar
beets and the yielding capacity of the resulting polyploids. Proc
Am. Soc. Sugar Beet Technol. 3: 287-295.
(15) REITBERGER, A. 1956. Ruhekernuntersuchungen bei gesunden und
viruskranken Diploiden und Polyploiden von Beta vulgaris. Ziichter.
26: 106-117.
(16) SAVITSKY, H. 1952. Polyploid sugar beetsCytological study and
method of production. Proc. Am. Soc. Sugar Beet Technol. 7:
470-476.
(17) SAVITSKY, H. A method of inducing autopolyploidy in sugar beets by
seed treatment. J. Am. Soc. Sugar Beet Technol. (In press).
(18) SCHLOSSER, L. A. 1940. Untersuchungen an autoploiden Zuckerrueben
(1). Z. d. Wirtschaftsgruppe Zuckerindustrie. 90: 88-106.
(19) VON-ROSEN, G. 1949. Problems and methods in the production of
tetraploids within the genus Beta. Socker Handl. 5: 199-217.

Recent Advances in Molasses Exhaustion


K . W . R . SCHOENROCK AND J . R . JOHNSON 1
Received for publication September 24,

1964

Background and History


T h e extraction of sugar from sugar beets has been very
closely related to t h e d e v e l o p m e n t of beet varieties with a high
sugar content. T h e early pioneers of this industry realized the
i m p o r t a n c e of this relationship. F r o m the annals of history it
is evident that m e n such as A r c h a r d placed as their m a i n objective first the d e v e l o p m e n t of beet varieties which m a d e industrial sugar extraction from sugar beets economically feasible.
Results of the efforts in the direction of high sugar c o n t e n t
seem to have reached a plateau a l t h o u g h t r e m e n d o u s gains have
recently been m a d e in other directions of beet development.
M o r e recently some areas even show a slow b u t a l a r m i n g
deterioration of beet processibility a n d a declining sugar extraction. A n u m b e r of investigators have studied a n d r e p o r t e d
on the subject of beet processibility a n d sugar extraction as a
function of beet quality. T h e reader is referred to the extensive
literature coverage on this subject.
According to this literature the extraction of sugar begins
with beet b r e e d i n g a n d is followed by a p p r o p r i a t e agricultural
practices. In some E u r o p e a n areas t h e level of so-called melassigenic constituents such as potassium a n d sodium is considered
in t h e beet contract. Farmers are advised as to t h e agricultural
practices favoring low potassium a n d sodium c o n t e n t a n d are
rewarded or penalized according to the deviation from the
established standard.
However, the processor cannot alwavs change the basic raw
material to comply with the concept of ideal beet quality. Processing of the beet m u s t consequently be a d a p t e d to prevailing
beet quality to attain o p t i m u m results from the material available. In this light molasses exhaustion begins in the diffusion
process.
Schneider, Reinefeld a n d Schliephake discuss the fundamentals a n d the practical application of sugar extraction from
beet cossettes (24) 2 . T h e b e h a v i o u r of nonsugars d u r i n g extraction was investigated by Schneider a n d Schliephake (IV).
R o u n d s a n d Rawlinsrs (15) r e p o r t e d their findings on t h e change
in nonsugar e l i m i n a t i o n in diffusion d u e to t e m p e r a t u r e a n d
1
Research Chemist and Research Laboratory Manager, respectively, The Amalgamated
Sugar
Company, Twin Falls, Idaho.
2
Numbers in parentheses refer to literature cited.

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draft variations. H e n r y , Vandewijer a n d Piek (7) evaluated t h e


effect of diffusion t e m p e r a t u r e a n d pH u p o n beet juice quality.
Asselbergs, Van Der Poel, Verhaart a n d Visser (1) studied t h e
extraction of nonsugars as a function of sucrose extraction a n d
its relationship to sugar losses in molasses.
T h e p r i m a r y objective of beet juice purification via the
t r e a t m e n t with lime is to prepare a clear filtrate of sufficiently
high p u r i t y from which sucrose can readily be crystallized. A
v o l u m i n o u s l i t e r a t u r e testifies to the vigorous activity in the
field of beet juice purification. It is commonly agreed that 1
p o u n d of nonsugars n o t eliminated in purification will prevent
a b o u t 1.5 p o u n d s of sugar from being recovered as salable
sugar. A n o n s u g a r elimination of about 3 0 % is very c o m m o n
for most good defecation systems. Strenuous efforts are cont i n u i n g to improve nonsugar elimination a n d increase sugar
extraction by this r o u t e .
I o n exchange has also been revitalized in the beet sugar
industry in recent years to increase extraction of salable sugar.
Reportedly, at least one ion exchange installation in J a p a n
features nearly total nonsugar removal with complete elimination of molasses p r o d u c t i o n .
Some ion exchange processes such as the Q u e n t i n process
are designed to replace so-called highly melassigenic cations such
as potassium a n d sodium with less objectionable cations such as
magnesium.
T h e sole purpose of all processing operations in a beet sugar
factory is to increase sugar extraction a n d molasses exhaustion
at a reasonable cost level.
Purification via crystallization is the final step in a refinery
processing sugar from sugar beets. Research activities in the
field of crystallization have been especially lively in recent years.
An extensive l i t e r a t u r e coverage of work performed prior to
1957 in the field of sugar technology with particular emphasis
on crystallization has been prepared by H o n i g (8).
M o r e recently Brieghel-Mueller (2) reported on super-saturation a n d t h e velocity of crystallization. Moller a n d Schmidt (12)
developed a laboratory m e t h o d to d e t e r m i n e velocity of crystallization u n d e r conditions allegedly adapted as accurately as possible to those prevailing in the factory. Schliephake (16) investigated the structure of aqueous sucrose solutions. T o n n (27)
published his assertions of empirical n a t u r e concerning the rate
of crystallization in supersaturated sugar/water systems.
A l t h o u g h t h e findings of E m m e r i c h a n d F o r t h (5) with

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JOURNAL OF THE A. S. S. B. T.

crystallization of sucrose from weakly supersaturated solutions


are based on p u r e solutions its value m i g h t well e x t e n d i n t o the
range of i m p u r e solutions. T h e i r observations with damaged a n d
so-called healed crystal surfaces may explain in p a r t the reason
for Carolan's (3) failure to exhaust molasses w h e n starting with
a higher purity m o t h e r l i q u o r (compare c h a p t e r 7, T a b l e s 7
a n d 8 with chapter 6, T a b l e s 5 a n d 6) in spite of sufficiently
high supersaturations.
Wagnerowski et al. (30); P i d o u x (14); a n d Vavrinecz (28)
a m o n g others reviewed the solubility of p u r e sucrose in water.
A large share of a t t e n t i o n has been focused on the influence
of the various nonsugars represented in beet liquors u p o n the
solubility of sucrose (salting o u t effect). In a series of publications Schneider et al. (18, 19, 20) r e p o r t e d on sucrose solubility
a n d crystallization as a function of individual n o n s u g a r fractions.
T h e y also published their work on t h e influence of beet nonsugars on molasses formation (21, 22, 23). M a n t o v a n i investigated
the individual effect of potassium chloride (9), b e t a i n e (10) a n d
raffinose (11) on sucrose solubility a n d crystallization. He found
a significant salting out effect for potassium chloride a n d a small
decrease in sucrose solubility with either betaine or raffinose.
T h e results M a n t o v a n i o b t a i n e d with raffinose were confirmed in o u r studies. According to M a n t o v a n i sucrose crystallization was only i m p a i r e d to t h e extent of the salting o u t effect
for potassium chloride or betaine while raffinose i n h i b i t e d sucrose
crystallization to a m u c h larger extent t h a n its salting o u t effect
w o u l d justify. Carolan (3) essentially confirmed Mantovani's
findings for potassium chloride a n d betaine, respectively, alone
b u t found a decrease in sucrose crystallization for a c o m b i n a t i o n
of the two m a i n nonsugars in molasses in excess of the c o m b i n e d
salting o u t effect.
G r u t (6) described a m e t h o d for the d e t e r m i n a t i o n of molasses exhaustion. Wagnerowski a n d coworkers (29) proposed
a simple laboratory m e t h o d for the estimation of specific sucrose
solubility in p u r e a n d i m p u r e systems. In s u b s e q u e n t publications (31, 32, 33) Wagnerowski et al. proposed a system for
evaluating a n d g u i d i n g molasses exhaustion u n d e r o p t i m u m
conditions. Dobrzycki (4) r e p o r t e d also on these findings.
Development and Summary
Inconsistencies in low p u r i t y crystallization on a factory level
p r o m p t e d t h e a u t h o r s to suspect a variable deviation of sucrose
solubility from the values in established solubility tables.
T h e m e t h o d of Wagnerowski et al. a p p e a r e d to be well suited
to allow the quick a n d simple d e t e r m i n a t i o n of sucrose solubility

VOL. 13, No. 8, JANUARY 1966

665

in specific low p u r i t y liquors. T h e basic findings of Wagnerowski


et al. m a y be summarized as follows:
1. T h e saturation e q u i l i b r i u m for sucrose in low p u r i t y
liquors is approached in a b o u t two hours. Schneider
a n d coworkers have published similar results.
2. T h e coefficient of saturation 3 remains constant u n d e r
t e m p e r a t u r e change for a given n o n s u g a r / w a t e r ratio.
T h i s discovery has been credited to W i k l u n d (34).
3. For low purity liquors with a n o n s u g a r / w a t e r r a t i o >
1.5 the coefficient of saturation is a linear function of
t h e n o n s u g a r / w a t e r ratio.
T h e s e findings were basically confirmed by o u r work. Solubility diagrams such as proposed by Wagnerowski et al. were
p r e p a r e d from time to time to see whether a shift in sucrose
solubility d i d occur as campaign advanced. Sucrose solubility
in molasses from different factories as well as sucrose solubility
in molasses from different processing periods was d e t e r m i n e d a n d
c o m p a r e d against each other, as well as against the values from
established solubility tables. It was a p p a r e n t that differences in
sucrose solubility d i d occur between different beet liquors. T h e s e
differences were however, relatively small for the test p r o g r a m
thus far. A calculated least square straight line relationship between t h e coefficient of saturation a n d t h e n o n s u g a r / w a t e r ratio
representing all solubility values thus far established appears to
suffice for n o r m a l factory operation.
Observed differences between d e t e r m i n e d values a n d values
taken from established solubility tables were however very large.
An accurate knowledge of actual solubility minimizes the chance
for e i t h e r u n d e r s a t u r a t i o n or excessive supersaturation d u r i n g
low raw massecuite curing. D i l u t i o n and reheating d u r i n g the
various crystallizer stages may be scheduled to conform with the
actual solubility range. T h i s will minimize danger of redissolving
already crystallized sugar or of inadequate exhaustion of t h e
molasses.
A l i g n m e n t charts are presented which show t h e relationship
between brix, purity, t e m p e r a t u r e a n d supersaturation on t h e
one h a n d a n d t h e relationship between massecuite brix, purity,
final n o n s u g a r / w a t e r ration, p a n size a n d total water a d d i t i o n
on the o t h e r h a n d . A d d i t i o n a l alignment charts may aid operators
to further optimize the operation of the entire low raw side by
k n o w i n g t h e total available t i m e for operation u n d e r a specific
set of conditions.
8
T h e coefficient of saturation expresses the ratio of dissolved sugar between impure
and pure solutions at the same temperature and at saturation.

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JOURNAL OF THE A. S. S. B. T.

A large n u m b e r of complete crystallizer cycles


tinuously sampled. Viscosity, b r i x a n d p u r i t y were
o n t h e massecuite a n d the respective m o t h e r l i q u o r
the cycle to d e t e r m i n e the rate of crystallization. T h e
crystallization rate 4 was found to be a log function
of time. Only the slope varied with a change in the

were condetermined
throughout
cumulative
of the log
conditions.

In a n o t h e r phase of this investigation the role of viscosity


in low raw crystallization was studied. O p i n i o n s are somewhat
divided on the relationship between viscosity a n d r a t e of crystallization. For all practical purposes in factory operation, however,
viscosity is the controlling factor in low raw massecuite curing.
F o r t h e p r a c t i t i o n e r it is of no i m m e d i a t e concern w h e t h e r
viscosity effects the crystallization rate directly or indirectly. Viscosity dictates the cooling rate, r e q u i r e d d i l u t i o n of m o t h e r
liquor, a n d the conditions forand results offinal p u r g i n g
on the low raw side. A t h o r o u g h u n d e r s t a n d i n g of the role of
viscosity in crystallization is therefore vital.
Recently Schneider, Schliephake a n d Klimek (25) r e p o r t e d
their findings on the viscosity of p u r e sucrose solutions. T i k homiroff, P i d o u x a n d F i l i p p i evaluated the effect of viscosity
on crystal formation in super-saturated aqeous sucrose solutions
(26). Wagnerowski, Dabrowska a n d Dabrowski (33) investigated
the relationship between foaming, viscosity a n d molasses exhaustion. P i d o u x (13) m a d e a significant c o n t r i b u t i o n w h e n he
developed a simple formula for expressing the t e m p e r a t u r e / v i s cosity relationship in sucrose solutions. H i s discovery has been
applied to o u r work with molasses exhaustion. In so d o i n g it
has b e e n possible to predict p r o p e r viscosities for p u r g i n g by
adjusting conditions to conform with o p t i m u m results. Massecuite
as well as m o t h e r l i q u o r viscosity was found to be a linear function of t e m p e r a t u r e a l t h o u g h air e n t r a i n m e n t a n d n u c l e a t i o n
of fines may be responsible for certain deviations. In any event,
cooling a n d d i l u t i o n rates may be p r o g r a m m e d on the basis of
anticipated viscosities r a t h e r t h a n solely on the i m m e d i a t e power
r e q u i r e m e n t s of the crystallizer.
Experimental Methods
Solubility d e t e r m i n a t i o n s were carried o u t according to the
procedures suggested by Wagnerowski et al. A fully jacketed three
neck flask with o n e liter capacity was used for o u r determinations. T h e center neck received the stuffing b o x w i t h stirrer
4
Cumulative crystallization rate is here expressed as % sugar crystallized per minute
on total sugar in original mother liquor and disregarding available crystal surface area.

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assembly. The two side necks were used for a reflux condenser
and a thermometer, respectively. Constant temperature water
was circulated through the jacket of the flask. The flask was
placed in a full size electric heating mantle to minimize radiation
loss, and to support the output of the constant temperature
water bath if needed. Temperature of recirculation water was
controlled through a thermistor regulated constant temperature
water bath.
T h e flask was charged with the appropriate molasses sample
of a predetermined nonsugar/water ratio. Temperature was maintained at 0.01 C. When thermal equilibrium was reached an
adequate amount of coarse sugar, 16 + 34 mesh, was added
to provide a safe excess of sugar at saturation. The molasses/sugar
mixture was then agitated for at least two hours at the test
temperature. The necessary high nonsugar/water ratio in the
molasses was either obtained by distilling off excess water from
normal molasses or by sampling a crystallizer early in the cycle
when nonsugar/water ratios were still sufficiently high to avoid
evaporation of excess water. The latter method was finally adopted
since a host of problems was associated with the former procedure.
A Carver Press fitted with a home made jacketed cylinder
assembly was used to separate the mother liquor from excess
crystals. The Carver press assembly was equipped with controlled
electric heating platens. Constant temperature water from the
water bath was also circulated through the jacket of the cylinder
to keep the massecuite at temperature equilibrium during the
pressing operation. The heavily supported screening section
allowed the use of any filtering media. Tightly woven linen cloth
was normally employed as a primary filter. The expressed mother
liquor was collected in a jar and aliquot parts were used for
the purity and viscosity determinations. Viscosity was determined
with the Brookfield-Synchro-Lectric Model RVT viscometer and
purities were determined via the single acid true purity method.
Results and Discussion
T h e differences in solubility between the values of Grut;
Brown & Nees; and Twin Falls material for the 62-63 campaign
are illustrated in Figure 1. It is easily seen that the Twin Falls
material dissolves more sugar for a given nonsugar/water ratio
than either the Grut or Brown & Nees values would indicate.
The solubility values were applied to an actual crystallizer run
to show the difference in supersaturation. Figure 2 illustrates
the difference in supersaturation when using either Grut's solubility values and actually determined solubilities respectively.

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Figure 1.Difference in sucrose solubility between the data of Grut,


Brown and Nees and actual values for low purity liquors. Xwin Falls
area, 1962.

Figure 2.Difference in supersaturation between Grut's tables and


actual values. Twin Falls 1964, complete cycle.

Xhe interesting part of this comparison is that the high supersaturation when based on Grut's tables made it difficult to properly schedule crystallizer operation and to justify in particular
the purity rise in mother liquor which is usually experienced
during the reheating period at the end of the crystallizer cycle.
On the assumption of being sufficiently safe, operators raised
temperatures and added water to improve purging. As we see
from the actual supersaturation the mother liquor was already
undersaturated at that point and sugar was redissolved. A second
deception is the belief that a supersaturation of 1.5 or so should
be maintained in low raw massecuite to secure acceptable crystallization velocity. It was found by testing a large number of
crystallizers that a supersaturation of 1.25 was maximum for
smooth operation.
The results for sucrose solubility in beet molasses from different areas are shown in Figure 4.

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A l t h o u g h variations of sucrose solubility in n o n s u g a r / w a t e r


systems from different areas have been observed w i t h i n the territory of the Amalgamated Sugar Company, these variations were
relatively small w h e n c o m p a r e d against the differences with
established solubility tables heretofor used.
A least square line representing all values for the 62-63
campaign was identical to a least square line calculated from
the solubility values of the 63-64 campaign. Figure 3 shows these
results.
O n l y 2 solubility d e t e r m i n a t i o n s at different n o n s u g a r / w a t e r
ratios are r e q u i r e d to attain a complete solubility spectrum for
the sample in question. However, more than 2 such determinations were carried o u t to gain confidence in the m e t h o d a n d
to provide sufficient data for statistical analysis. T h e 1962-63
campaign values in Figure 3 represent 49 different solubility
d e t e r m i n a t i o n s from 12 different m o t h e r liquor samples gathered
from 4 factories at up to 4 different sampling periods. T h e same

Figure 3.Difference in sucrose solubility


between 1962-63 a n d 1963-64. T w i n Falls area.

for

low

purity

liquors

Figure 4 Difference in sucrose solubility for low purity liquors from


different areas. 1962-63 campaign.

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JOURNAL OF THE A. S. S. B. T.

4 factories are represented in the 52 solubility determinations


carried out for 2 sampling periods during the 63-64 campaign.
An average of 4 solubility determinations at different nonsugar/
water ratios were performed for each molasses sample. Linearity
between the coefficient of saturation and the nonsugar/water
ration was evident in all cases. Table 1 shows the statistical correlation coefficients and least square equations for each set of data.
Table 1.Solubility regression equations and correlation coefficients.
Equation

Factory and sample time


Rupert 1962-63
Rupert Early 1963-64
Rupert Late 1963-64

Y
Y
Y

Rupert (All Combined)

0.713

Twin Falls 1962-63


Twin Falls Early 1963-64
Twin Falls Late 1963-64

Y
Y
Y

0.704
0.697
0.809

Twin Falls (All Combined)

0.722

Nampa 1962-63
Nampa Early 1963-64
Nampa Late 1963-64

Y
Y
Y

0.777
= 0.750

0.722
0.724
0.699

0.804

Nampa (All Combined)

0.780

Nyssa 1962-63
Nyssa Early 1963-64
Nyssa Late 1963-64

Y
Y
Y

= 0.802
0.702

Nyssa (All Combined)

Y = 0.741

All Data Combined 101 =

0.717
0.745

+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+
+

r (xy;

0.230 X
0.232 X
0.240 X

0.984
0.998
0.996

0.234 X

0.991

0.228 X
0.241 X
0.167 X

0.976
0.989
0.987

0.220 X

0.955

0.212 X
0.218 X
0.201 X

0.988
0.995
0.943

0.210 X

0.984

0.186 X
0.225 X
0.218 X

0.952
0.998
0.992

0.211 X

0.989

0.217 X

0.979

Differences are not significant @ 19:1 odd in Y-intercept or slope between factories or
sample time.
Observe Twin Falls for 1963-64 which has a lower slope than normal and the same for
Nyssa 1962-63. Even with these values included we find the curve for combined data to possess
a high degree of correlation C vs A. According to this we can assume that a single curve
Will suffice for solubility work without a high degree of error.

T h e most stable relationship in crystallizer o p e r a t i o n is the


n o n s u g a r / w a t e r ratio. W h i l e sucrose in solution, b r i x a n d percent p u r i t y of t h e m o t h e r l i q u o r as well as crystallization rate
a n d viscosity are continuously changing, t h e n o n s u g a r / w a t e r
r a t i o may be controlled at will t h r o u g h water a d d i t i o n . N e e d e d
water d i l u t i o n may be easily calculated for a final n o n s u g a r / w a t e r
ratio in t h e molasses if p a n brix, p a n p u r i t y a n d p a n size are
k n o w n . An a l i g n m e n t chart has b e e n p r e p a r e d in F i g u r e 5 to
illustrate graphically the r e l a t i o n s h i p b e t w e e n these variables.
It was necessary to assign an average density to t h e massecuite
to convert v o l u m e values i n t o its respective weight. T h e changes
in w a t e r a d d i t i o n d u e to a small change in t h e massecuite

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Figure 5.Nomograph showing the relationship between p a n size, p a n


RDS, p a n SAXP, desired nonsugar/water in molasses and required total
water addition in gallons to the crystallizer.

specific gravity are small and may be neglected.


A simplified nomograph may be used according to Figure 6
by stabilizing the pan size and the nonsugar/water ratio in final
molasses. For most factories this latter alignment chart is
adequate.
In the example shown in Figure 5 a straight line is projected
between the single acid true purity of the massecuite at pan
drop (in this case 77.5%) on the far left scale through the
selected nonsugar/water ratio in molasses (in this case 2.5), to
obtain a K value of 1.09. A like K value is found on the adjacent
K1 scale and connected via a straight line over the determined
massecuite RDS (in this case 95) to find a water addition of
8.5 gallons/tons massecuite. This value of 8.5 may be connected
from the second scale graduated in gallons water/ton fillmass
with the pan volume on the scale at the far right (in this case
1200 cu ft) in the third part of the nomograph by a straight
line to show a total water addition of 480 gallons. Or the value
of 8.5 may be multiplied mentally with the total tons of fillmass
at pan drop to obtain total water addition.

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Figure 6.Nomograph illustrating the relationship between p a n RDS,


p a n single acid true purity a n d required total water addition to the
crystallizer in gallons. P a n size 1300 cu ft. Projected final n o n s u g a r / w a t e r
ratio in molasses - 2.5.

Of primary importance to successful crystallizer control is the


proper timing for needed water addition. The scheduling for
water addition must be a compromise between desirable supersaturation and tolerated viscosity. Supersaturation may be easily
calculated for any set of conditions of brix, percent purity,
temperature and coefficient of saturation for a specific mother
liquor. Figure 7 illustrates this relationship graphically for the
average of all determined solubilities as shown in Figure 3.
The example given in Figure 7 shows a K value of 0.355 for
a mother liquor purity of 59% and an RDS of 86. This K value
of 0.355 is projected in the second part of the nomograph through
a temperature of 57 C to reveal a supersaturation of 1.0. Water
addition may thusly be properly scheduled and purging temperatures may be conveniently selected without the danger of either

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673

Figure 7.Nomograph illustrating the relationship between single


acid true purity, RDS, temperature, and supersaturation for low purity
liquors. T h e Amalgamated Sugar Company.

dissolving sugar or exceeding the u p p e r safe limit for supersaturation respectively.


U n d e r n o r m a l conditions of factory operation it is impractical
to o b t a i n m o t h e r l i q u o r b r i x a n d purity d u r i n g the crystallizer
cycle. A u n i f o r m p a n boiling procedure will, however, p r o d u c e
very consistent m o t h e r l i q u o r composition at pan d r o p . We
found a stable 92.5 R D S a n d 6 3 % purity for the m o t h e r l i q u o r
at a massecuite R D S of 95 at p a n d r o p for one of o u r factories.
U n d e r these conditions a n d at a pan d r o p t e m p e r a t u r e of 8 5 C
the m o t h e r l i q u o r has a supersaturation of only 1.14. Supersaturation is the d r i v i n g force for crystallization a n d should be
held as high as clean crystallization permits.
T h e crystallization rate was determined from the change in
sucrose c o n c e n t r a t i o n for the m o t h e r l i q u o r of low raw massecuite
subjected to carefully controlled curing. Figure 8 illustrates the
declining crystallization rate towards the lower t e m p e r a t u r e scale.

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Figure 8.Relationship between rate of crystallization and time for


low-purity massecuite in commercial crystallizer, T w i n Falls factory, campaign 1960-61. Mother liquor purity 65-70% (AP), supersaturation - 1.2.

It remains to be established though, to what extent the declining


rate may be charged to temperature and to what extent to other
influences such as proposed by Emmerich and Forth (5).
The values in Figure 8 represent a total of 160 different
purity determinations from 23 complete crystallizer cycles. Linearity of the relationship as expressed in Figure 8 was observed
in all cases although a shift in the slope was evident with a
change in the massecuite and curing conditions respectively.
The influence of surface area was purposely neglected for practical reasons.
To approach ideal conditions one would have to program
the cooling rate and water addition to the crystallization rate
within a favorable supersaturation limit to maintain an acceptable viscosity. The dominating role of viscosity is perhaps best
illustrated by comparing the change in viscosity and supersaturation respectively during the cooling cycle. Below 70 C
the viscosity increases more than twofold for every ten degree
drop in temperature and triples below 40 C for every 10 degree
centigrade drop. If a solution of sugar in molasses is assumed
to be saturated according to the coefficient of saturation for
Twin Falls material at 70 C, its supersaturation will be 1.13
at 60C, assuming that sucrose crystallization was dormant during that period. The values are from 1.0 to 1.09 supersaturation
for a temperature drop from 40 to 30 C. The inference is

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675

that the rate of gain in supersaturation decreases during progressive cooling while the rate of gain in viscosity increases on
a nearly logarithmic scale over the same temperature range.
Thusly, viscosity and not supersaturation becomes the controlling factor in the lower temperature range because continued
crystallization does not allow a significant build up of supersaturation during a normal cooling cycle. Water addition need

To solve for the unknown: Connect points on the inside scales via a straight line; A second
straight line connecting points of the outside scales must intersect the first straight line on
the reference line.
Example:
Number of vessels
=
4
(Scale M)
Maximum available
=
1300
cu ft
(Scale L)
Daily workload
=
9500
cu ft
(Scale O)
time

13.1

hours

(Scale T)

Figure 9.Nomograph illustrating the relationship between available


crystallizer vessels, capacity for each vessel, daily fillmass from pan in
cu ft, and total time available for each complete cycle.

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JOURNAL OF THE A. S. S. B. T.

n o t be started t h e n u n t i l the viscosity increase d e m a n d s a dilution for reasons of h a n d l i n g a n d p u r g i n g limitations. T h e only


other factor which m u s t be considered in scheduling the beginn i n g a n d the rate of water a d d i t i o n once the total a m o u n t of
water to be added has been established according to Figure 5
or 6, is the available c u r i n g time which in t u r n dictates the
cooling rate.
T h e a l i g n m e n t chart in Figure 9 illustrates the relationship
between daily low raw massecuite workload discharged from
the pans, n u m b e r of vessels, vessel capacity a n d m a x i m u m available time for each respective cycle. To solve for the u n k n o w n ,
points on the inside scales are connected via an imaginary straight
line. A n o t h e r straight line is projected between points on the
outside scales to intersect with the first straight line on the reference line. T h e chart may be used for establishing m a x i m u m
crystallizer or p a n boiling time, or for finding the n u m b e r of
crystallizers or pans needed if a p r e d e t e r m i n e d cycle time or
daily workload or vessel capacity is to be m a i n t a i n e d . Figure 10
shows an a l i g n m e n t chart at constant pan capacity, in this case
1300 cu ft. Similar a l i g n m e n t charts may be p r e p a r e d for the
centrifuge station.
After fixing the total time available for each respective
crystallizer cycle it is feasible to project a specific cooling rate.
Figure 11 shows an a l i g n m e n t chart which predicates the cooling
rate in C / h o u r if the available cooling t i m e a n d the t e m p e r a t u r e

Figure 10.Alignment chart showing the relationship between daily


fillmass workload, number of vessels and total operating time.

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Figure 11.Alignment chart illustrating the relationship between cooling rate in C/hours, available cooling time, and temperature spread
between highest and lowest temperature.

spread between the highest and the lowest desired temperature


has been assigned.
On the basis of these findings a complete program may be
projected for low raw massecuite curing under optimum conditions. T h e fundamental requirements of any such program are
predicated upon reliable control equipment. This step by step
program may be outlined as follows:
1. Obtain daily workload in cuft low raw massecuite from
the laboratory. For a uniform operation this value is
relatively stable from day to day and fluctuates only significantly with the daily slice.
2. Obtain total pieces of equipment such as low raw pans,
crystallizers and low raw centrifuges.
3. Secure equipment load capacity for the facilities under 2.

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4. Establish characteristic composition of m o t h e r l i q u o r at


p a n d r o p . T h i s composition is relatively specific for t h a t
particular factory a n d need be re-established only if a
deviation from n o r m a l boiling practices occur.
5. Decide on the final n o n s u g a r / w a t e r r a t i o in molasses. An
o p t i m u m r a t i o of 2.5 is r e c o m m e n d e d for most operations.
6. Consult Figure 9 to establish t h e m a x i m u m time available
for p a n boiling. If the p a n size is consistent use Figure
10. To m a i n t a i n uniformity stick to the t i m e schedule
thusly found.
7. Again consult F i g u r e 9 or 10 to establish total available
crystallizer time a n d adhere to the found value.
8. Resolve the lowest t e m p e r a t u r e to be reached d u r i n g the
crystallizer cooling cycle. If the operation u n d e r 7 projects
a d e q u a t e time e.g. 15 hours or m o r e t h a n the massecuite
may be cooled as low as 40 C, provided that the reheating may be accomplished in the m i x e r r a t h e r t h a n
in t h e crystallizer. However, if the total cooling t i m e
is limited to 12 h o u r s or less, t h e n do n o t cool below 50 C.
9. (a) Subtract from t h e total t i m e as found u n d e r 7 t h e
c o m b i n e d time n e e d e d for filling a n d d r a i n i n g of the
crystallizer to arrive at a fixed cooling time.
(b) Subtract the value established u n d e r 8 (lowest temperature) from the t e m p e r a t u r e of the massecuite in C at
p a n d r o p t o f i x the t e m p e r a t u r e spread i n C.
10. Project an imaginary straight line in F i g u r e 11 between
t h e values as f o u n d u n d e r 9a a n d b respectively to establish a fixed cooling rate.
11. O b t a i n p a n R D S , p a n single acid t r u e p u r i t y a n d total
p a n size from the laboratory. T o g e t h e r w i t h the value
resolved u n d e r 5 ( n o n s u g a r / w a t e r r a t i o in molasses) det e r m i n e total water a d d i t i o n to the crystallizer by consulting F i g u r e 5. If p a n size a n d n o n s u g a r / w a t e r ratios
in molasses r e m a i n constant for every strike use the simple
n o m o g r a p h in F i g u r e 6.
12. D e t e r m i n e supersaturation at p a n d r o p by u s i n g values
derived as u n d e r 4 ( m o t h e r l i q u o r b r i x a n d % purity)
in Figure 7. If supersaturation thusly d e t e r m i n e d does
n o t exceed 1.2, do n o t a d d water u n t i l the srradual viscosity
increase d e m a n d s d i l u t i o n . F o r crystallizer with 12 or
m o r e h o u r s cooling time the water addition need n o t
begin above 70 C. T h e rate of water d i l u t i o n should be
held as low as total r e m a i n i n g crystallizer t i m e permits.
T o t a l water m u s t dissipate in t h e massecuite before t h e
fillmass has to be d u m p e d . T h i s is an area w h e r e indi-

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679

v i d u a l j u d g m e n t of t h e o p e r a t o r is still t h e d e c i d i n g factor
u n t i l a satisfactory viscosity a n d b r i x sensing device may
control this function automatically. T e m p e r a t u r e of dilut i o n water should be at least as high as t h e t e m p e r a t u r e
of t h e massecuite.
13. D e t e r m i n e p u r g i n g t e m p e r a t u r e b y u s i n g b r i x a n d perc e n t p u r i t y of molasses in F i g u r e 7. F o r a u n i f o r m operat i o n these values will r e m a i n fairly constant from day
to day. Massecuite should be r e h e a t e d in t h e r a w m i x e r
to the t e m p e r a t u r e thusly found p r i o r to p u r g i n g .
Acknowledgment
T h e a u t h o r s wish to acknowledge the p a r t i c i p a t i o n of P a u l
K u n k e l , w h o performed most solubility d e t e r m i n a t i o n s for t h e
63-64 c a m p a i g n samples, a n d J a m e s M. Peterson, w h o assisted
in t h e c o n s t r u c t i o n of t h e a l i g n m e n t charts.
Literature Cited
(1)

(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
(13)
(14)

ASSELBERGS,

C.

P., P.

W. VAN

DER POEL,

M.

L.

A.

VERHAART and

N.

H. M. DEVISSER. 1963-64. Sugar losses in molasses in connection


with diffusion. Int. Sugar J. 65, 66 (780, 781): 363-366, 9-12.
BRIEGHEL-MUELLER, A. 1962. Supersaturation and velocity of crystallization. Zucker. 15 (23): 596-600.
CAROLAN, R. 1960. Laboratory experiments on molasses exhaustion.
Report No. 43. Irish Sugar Co. Ltd.
DOBRZYCKI, J. 1961. About molasses exhaustion. Zschrt. Zuckerind.
14 (4) : 202-205.
EMMERICH, A. and H. FORTH. 1962. Crystallization of sucrose from
weakly supersaturated solutions. Zucker. 15 (24) : 626-637.
GRUT, E. 1962. Evaluating the desugaring of molasses. Zucker. 1 2 ( 9 ) :
492-494.
HENRY, J., R. VANDEWIJER and R. PIECK. 1963-64. Reports concerning the effect of temperature and pH during diffusion upon the
quality of the diffusion juice. Sucr. Beige. 83: 1-14, 57-65.
H O N I G , P. 1959. Principles of Sugar Technology, Vol II crystallization. Elsevier Publishing Co., New York.
MANTOVANI, G. 1961. Investigations about sucrose crystallization in
the presence of potassium chloride. Ind. Sacc. Ital. 54 (73) : 3-4.
MANTOVANI, G. 1963. Investigations about sucrose crystallization in
the presence of betaine. Zschrt. Zuckerind. 3 ( 1 0 ) : 559-563.
MANTOVANI, G. and F. FAGIOLO. 1964. Investigations concerning
sucrose crystallization in the presence of raffinose. Zschrt. Zuckerind.
14 (4) : 202-205.
MOLLER, C. and N. O. SCHMIDT. 1963. Rate of crystallization. Zschrt.
Zuckerind. 1 3 ( 9 ) : 501-504.
PIDOUX, G. 1961. Expression of viscosity between 0 and 100C. Zucker.
14 (20) : 523-532.
PIDOUX, G. 1962. Some remarks on the solubility of sucrose in water.
Zucker. 15 (7) : 162-163.

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(15) ROUNDS, G. H . a n d F. N . RAWLINGS. Theory a n d economics of diffusion. 1954. Proc. Am. Soc. Sugar Beet Technol. VIII (2) : 289-297.
(16) SCHLIEPHAKE, D. 1963. On the structure of aqueous sucrose solution.
Zucker. 16(19) : 523-528.
(17) SCHNEIDER, F. and D. SCHLIEPHAKE. 1963. Behaviour of nonsugars
during sugar extraction. Zucker. 16(18): 503-509.
(18)

SCHNEIDER, F., A. EMMERICH a n d K. JOERN.

1959.

A b o u t the effect of

electrolytes upon crystallization and dissolution rate


Zucker. 12 (6) : 118-122.
(19)

SCHNEIDER, F., A. EMMERICH a n d K. JOERN.

1960.

of sucrose.

Investigations con-

cerning the velocity of dissolution for sucrose in aqueous electrolyte


solution. Zucker-Beih. 4 ( H ) : 1-11.
(20)

SCHNEIDER, F., E. REINEFELD a n d F. AMDING.

1962.

C o n c e r n i n g the

influence of individual nonsugar fractions upon the solubility of


sucrose. Zucker-Beih. 4 (H.3) : 55-71.
(21)

SCHNEIDER, F., A. EMMERICH, E. REINEFELD, E. W A L T E R and W. K E L M .

(22)

SCHNEIDER, F., E. REINEFELD a n d F . AMDING.

1961. Influence of beet nonsugars especially on the formation of


molasses. I. Zucker. 1 4 ( 9 ) : 208-216.
1961.

Influence of beet

nonsugars on the formation of molasses. II. Zucker. 14 (10) : 234-238.


(23)

SCHNEIDER, F., E. REINEFELD a n d A. EMMERICH.

1961.

Influence of

beet nonsugars on the formation of molasses. III. Zucker. 1 4 ( 1 2 ) :


307-311.
(24)

SCHNEIDER, F., E. REINEFELD a n d D. SCHLIEPHAKE.

1963. F u n d a m e n t a l s

and technical execution of sugar extraction from beet cossettes.


Chem. Ing. Techn. 35 (8) : 567-576.
(25)

SCHNEIDER, F., D. SCHLIEPHAKE and A. K L I M M E K .

1963.

On the viscosity

of p u r e sucrose solutions. Zucker. 16 (17) : 465-473.


(26) T I K H O M I R O F F , N., G. PIDOUX a n d R . F I L I P P I .

1963.

Viscometric in-

vestigations on crystal formation of supersaturated aqueous sucrose


solutions. Zucker. 16(22): 617-619.
(27) T O N N , H. 1964. Assertion of empirical nature concerning rate of
crystallization in supersaturated sugar-water solutions. Z u c k e r .
15(24) : 626-637.
(28) VAVRINECZ, G. 1962. A new table on the solubility of pure sucrose
in water. Zschrt. Zuckerind. 1 2 ( 9 ) : 482-487.
(29) WAGNEROWSKI,

K.,

D.

DABROWSKA

and

Cs.

DABROWSKI.

1960.

Fast

method for the determination of the effect of crystallization for


low raw massecuite. Gaz. Cukr. 62: 68.
(30) WAGNEROWSKI, K., D. DABROWSKA a n d Cs. DABROWSKI.

1961. Solubility

of sucrose in water. Gaz. Cukr. 62: 357.


(31)

WAGNEROWSKI, K., D. DABROWSKA a n d Cs. DABROWSKI.

1961.

Quanti-

tative relationship of molasses constituents. Gaz. Cukr. 63: 97, 63:


262.
(32) WAGNEROWSKI, K., D. DABROWSKA a n d Cs. DABROWSKI.

1962.

Problems

in molasses exhaustion. Zschrt. Zuckerind. 1 2 ( 1 2 ) : 664-671.


(33)

WAGNEROWSKI, K., D. DABROWSKA, a n d Cs. DABROWSKI.

1964.

Influence

of foaming of low grade massecuite on the viscosity and exhaustionof molasses. Zschrt. Zuckering. 1 4 ( 3 ) : 133-135.
(34) WIKLUND, O. 1946. Melasproblemet. Socker. 2: 65-140.

Colorimetric Micro Determination of Calcium in


Sugar-House Products
G.

V.

HEYNDRICKX1

Received for publication January 14, 1965

Introduction
T h e quantitative estimation of calcium in sugar-house products, such as raw juice, clarified juice, syrup, molasses a n d raw
sugar is very i m p o r t a n t in sugar manufacturing. Many m e t h o d s
for this estimation exist in the literature p o i n t i n g to t h e importance of the calcium content, b u t also to the complexity
of the p r o b l e m . Since the standard analytical oxalate m e t h o d
is c u m b e r s o m e , many r o u t i n e methods have been proposed. T h e
eldest of these is the soap m e t h o d of Spengler a n d Brendel (4) 2
which gives an estimate of the total q u a n t i t y of calcium a n d
magnesium, with doubtful accuracy, however.
Several workers have developed in recent years m o r e elegant
methods based on the E D T A (ethylenediamine tetra acetate) reaction. In these methods the sugar solution is titrated with a
solution of E D T A in the presence of eriochrome black [ H o n i g
(1), Saunier a n d L e m a i t r e (3)]. T h e a m o u n t of E D T A employed
is a measure of the calcium plus magnesium content. W h e n t h e
calcium c o n t e n t is desired, a second titration is carried o u t on
the sugar solution after precipitation of the calcium, or a n o t h e r
indicator such as m u r e x i d e [Ramaiah, Vishnu a n d C h a t u r v e d i
(2)] is used, which, however, requires a photometric titration
a p p a r a t u s . T h e great disadvantage of the E D T A titrations, even
with a p h o t o m e t e r , is the troubles which occur on e x a m i n i n g
stark colored solutions like raw juice and molasses. It is significant that in the 1964 edition of the I C U M S A methods of
sugar analysis, a m e t h o d for determination of calcium is n o t
mentioned.
T h e present c o m m u n i c a t i o n reports a simple, sensitive a n d
accurate m e t h o d for estimation of calcium, which is elaborate
by T r i n d e r (5) for blood, b u t so far is not applied to sugarhouse products.
Materials and Methods
In the proposed hydroxamate method, the calcium is precipitated as its naph-thalhydroxamate. After centrifugation, t h e
unwashed precipitate is dissolved and the color intensity of the
1
2

State Agricultural University, Gent, Belgium.


Numbers in parentheses refer to literature cited.

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obtained solution after addition of ferric n i t r a t e is measured in


a simple colorimeter.
ReagentsCalcium reagent solution m a d e
by dissolving
( u n d e r heat) 20 mg naph-thalhydroxarnic acid in 100 ml of water
containing 5 ml of e t h a n o l a m i n e a n d 2g of D tartaric acid.
After cooling, 9g of sodiumchloride dissolved in water is added
a n d d i l u t e d with water to 1 liter.
Alkalin solution is m a d e by dissolving 2g E D T A (disodiumsalt) in 1 liter of 0.1 N sodium hydroxide.
Color solution is m a d e by dissolving 60g ferric n i t r a t e n i n e
hydrate in 1 liter of water c o n t a i n i n g 15 ml nitric acid (D = 1.4).
Standard Calcium solution is m a d e by dissolving 100.1 mg
analytical reagent dry calciumcarbonate in 0.1 N hydrochloric
acid a n d d i l u t i n g to 1 liter with water.
Procedure0.5 ml of sugar solution is m i x e d in a centrifuge
t u b e with 5 ml of calcium reagent solution a n d after 30 m i n u t e s
is spun in an ordinary laboratory centrifuge for 5 m i n u t e s .
T h e s u p e r n a t a n t liquid is carefully p o u r e d off and, while the
t u b e is still inverted, it is placed in a rack to d r a i n on filter
paper. After a few m i n u t e s , the m o u t h of the t u b e is wiped
dry with filter p a p e r a n d 1 ml of alkaline solution is added.
T h e t u b e is covered with a glass stop or an a l u m i n i u m cap a n d
heated in a boiling waterbath, shaking the t u b e at intervals to
ensure complete solution of the precipitate. After 10 m i n u t e s ,
the t u b e is removed a n d w h e n cool 3 ml of the color solution
is added a n d the content of the t u b e mixed. T h e optical density
of the solution is t h e n measured at 450 mm against a blank prep a r e d by carrying o u t the same procedure, b u t with omission
of t h e sugar solution. T h e calcium c o n t e n t of the sugar solution
is calculated from a graph, o b t a i n e d with standard calcium solutions c o n t a i n i n g 4 to 40 m-g calcium in the 0.5 ml, analyzed together with the test.
T h e above procedure is carried o u t on u n d i l u t e d raw a n d
clarified juices, while syrup a n d molasses samples were d i l u t e d
to a concentration of a b o u t 20 mg calcium in 0.5 m l . In order
to study the possible influence of organic m a t t e r a n d complexes,
the above p r o c e d u r e is simultaneously carried o u t on t h e samples
after digestion with a m i x t u r e of concentrated nitric acid and
perchloric acid.
T h e results obtained with the h y d r o x a m a t e m e t h o d were
c o m p a r e d with those o b t a i n e d with the standard oxalate method.
H e r e w i t h , the calcium in the sugar solution, digested or not,
is precipitated at pH 5 with saturated a m m o n i u m oxalate. After

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centrifuging, washing with ammonia water and again centrifuging, the precipitate is dissolved in warm dilute sulfuric acid
and titrated with 0.01 N potassium permanganate.
Results and Discussion
The results obtained with the hydroxamate method and with
the oxalate method on 5 samples of raw juice, 3 samples of
clarified juice, 3 samples of syrup, 3 samples of molasses and
1 sample of raw sugar, with and without digestion, are mentioned in Table 1. In order to establish if the difference between
the calcium content found with the four methods is significant,

Table 2.P levels at which the difference between the mean calcium content, obtained
with four methods, is significant.

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t h e t test is applied a n d the values o b t a i n e d are m e n t i o n e d in


T a b l e 2. F r o m these two tables, it appears that the m e a n calcium
c o n t e n t found on the raw juice with the four m e t h o d s is 22.5 13.0 - 23.6 a n d 16.9 m g / 1 0 0 Brix respectively. T h e s e four
values differ significantly in all combinations, except for the
hydroxamate m e t h o d on t h e digested a n d undigested samples.
It is striking that the results o b t a i n e d with the hydroxamate
m e t h o d on t h e digested a n d undigested samples are significantly no
different, while digestion increases significantly the calcium content found with the oxalate m e t h o d . In some cases the content
increases with digestion 2 to 3 times a n d reaches almost the
values found with the hydroxamate m e t h o d .
T h e m e a n values for the calcium content of the clarified
juices, o b t a i n e d with the four m e t h o d s are 31.0 - 32.6 - 31.7
a n d 33.1 respectively, which are n o t significantly different.
T h e same conclusion is valid for the m e a n calcium c o n t e n t
of the syrup a n d the molasses samples, which have the values
22.0 - 22.8 - 22.5 - 23.1 a n d 118.8 - 120.4 - 124.2 a n d 125.2
m g / 1 0 0 Brix respectively. Summarizing it may be concluded
that, except for the raw juice samples, the results o b t a i n e d with
the h y d r o x a m a t e m e t h o d do n o t differ significantly from those
o b t a i n e d with the oxalate m e t h o d , w h e t h e r or n o t the samples
are or are n o t digested.
In regard to the precision or the reproducibility of the
methods applied, the analysis on each sample is m a d e in triplicate
a n d the coefficient of variation as a measure of the reproducibility
is calculated by dividing 100 times the standard deviation obtained with each sample by the m e a n of the three results. F r o m
t h e results m e n t i o n e d in T a b l e 1 a n d 3 it appears that the reproducibility does n o t significantly differ between the four
methods applied on the different samples, except for t h e oxalate
m e t h o d applied on digested a n d non-digested clarified juice
samples. T h e co-efficient of variation o b t a i n e d with the hyTable 3.P levels at which the difference between the coefficient of variation,
obtained with the four methods, is significant.

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d r o x a m a t e a n d with the oxalate m e t h o d applied on t h e 15 u n digested a n d digested samples attains the value of 3.0 - 3.3 - 3.6
a n d 3.7 respectively, which are only slightly different. F r o m all
these results it can be concluded that there are no great differences between the reproducibility of the four m e t h o d s applied.
Since the hydroxamate m e t h o d gives results which are n o t
significantly different from these obtained with the oxalate
m e t h o d a n d with a practical equal precision, this m e t h o d may
be r e c o m m e n d e d . T h e hydroxamate m e t h o d is m u c h less time
c o n s u m i n g t h a n the oxalate m e t h o d and at least 25 times m o r e
sensitive. T h e hydroxamate m e t h o d can be applied on the various
sugar-house products w i t h o u t digestion or defecation, even on
raw juices, which is n o t the case for the oxalate m e t h o d . It is
interesting to r e m a r k here that the complexometric titration
with eriochrome black or with m u r e x i d e is also m u c h less sensitive t h a n the hydroxamate m e t h o d and gives troubles with d a r k
solutions like raw juice a n d molasses. W i t h these solutions it
is necessary to clarify with lead reagent, with subsequent precipitation of the excess of lead. W i t h molasses this procedure is
even n o t sufficient a n d decolorization with adsorptive carbon
is necessary.
All these cumbersome manipulations are superfluous with
the h y d r o x a m a t e method, which is so sensitive that the molasses
may be d i l u t e d to 2 Brix a n d the resultant color has no influence
on the precipitated calcium. T h e simple complexometric t i t r a t i o n
on clarified juice is somewhat less time consuming t h a n t h e
h y d r o x a m a t e method, b u t this advantage disappears w h e n defecation a n d decolorization is necessary. Finally, a high sensitive m e t h o d like the hydroxamate m e t h o d is very interesting
on e x a m i n i n g samples with low calcium content like refined
sugars. It is o u r conviction that the hydroxamate m e t h o d is a
reliable a n d convenient m e t h o d a n d may be successfully applied
in the sugar industry.

Summary
A simple colorimetric micro m e t h o d for calcium determination in sugar-house products is described. T h e m e t h o d consists
essentially in a precipitation of the calcium with n a p h t h a l hydroxamic acid, which is separated by centrifuging, dissolved
in an alkaline solution, after which the color reaction with
ferric ions is measured in a simple colorimeter. F r o m the
statistical calculations, it appears that the hydroxamate m e t h o d

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gives no significantly different results in comparison with the


standard oxalate method and with practical equal precision or
reproducibility. Since the hydroxamate method is more sensitive and less time consuming than the oxalate or complexometric
method and can be carried out directly on the samples without
digestion or defecation, even on raw juices or molasses, it is a
reliable and convenient method, which can be successfully applied in the sugar industry.

Literature Cited
(1) HONIG, P. 1952. Calcium and magnesium in cane juices and sugars.
Repts. West Indies Sugar Corp. No. 2.
(2)

RAMAIAH,

N.

A.,

VISHNU

and

R.

K.

CHATURVEDI.

1958.

simple

method for estimation of calcium in technical sugar solutions using


ethylenediamine tetraacetic acid. Sharkara, 1: 124-136.
(3) SAUNIER, R. and A. LEMAITRE. 1951. Dosage volumetrique de calcium
et du magnesium dans les jus de sucrerie. Sucr. franc. 92: 231-233.
(4) SPENGLER, O. and C. BRENDEL. 1928. Die Kalkbestimmung in Zuckerfabrikprodukten. Z. Ver. Dtsch. Zuckerind. 78: 175-189.
(5) TRINDER, P. 1960. Colorimetric micro-determination of c a l c i u m in
serum. Analyst 85: 889-894.

A Method for Positive Selection of 4N Sugar Beet


Plants in the Vegetative Co Generation 1
G. H. R A N K 2 AND L. E. EVANS 3
Received for publication January 25, 1965

T h e contingent superiority of polyploid sugar beets owkx


diploids a n d the possible use of male sterility to facilitate the
p r o d u c t i o n of h y b r i d triploids has created the need to p r o d u c e
tetraploid strains efficiently. It is obvious that a selection criterion
that w o u l d readily p e r m i t selection, within the vegetative C 0
generation, of plants that produce only 4n seed would be of
practical value.
Many of the selection criteria found effective in later generations have proven useless in the C 0 generation d u e to the prevalence of cytochimeras a n d to colchicine induced morphological
variations that are n o t associated with polyploidy. Savitsky (6)
found p l a n t morphology a n d chloroplast n u m b e r s a poor selection criteria in the C Q generation while Deneuche (1) a n d
Varga (7) found stomata size unreliable. O t h e r criteria including pollen diameter, pollen pore n u m b e r , a n d n u m b e r of nucleoli
in resting nuclei of epidermal cells have been used with limited
success (2,7).
Kloen a n d Speckman (5) using the chromosome n u m b e r of
floral heart leaves were only moderately successful in selecting
C 0 plants that p r o d u c e d only 4n seed. T h e y used a rapid, nonstaining, phase microscopy m e t h o d which D e n e u c h e (1) later
found to be unreliable.
In this study the effect of colchicine treatment on chromosome
duplication at various growth stages was determined. T h i s was
related to pollen m o t h e r cell (PMC) analyses a n d progeny classification in o r d e r to ascertain if certain chromosome n u m b e r
characteristics in the vegetative stage could be used to select C 0
plants that w o u l d p r o d u c e only 4n seeds.
Methods and Materials
T h e material used was an open pollinated m o n o g e r m strain
designated as 6210, obtained from the Sugar Beet Breeding
Station, T a b e r , Alberta. T h e seeds were g e r m i n a t e d o n d a m p
blotting p a p e r a n d when 1 0 % of the seeds showed p r o t r u d i n g
root tips the entire sample was placed in a solution of . 3 %
colchicine for 6 hours at r o o m temperature, washed twice a n d
planted in the greenhouse.
1
2
3

Contribution No. 97 of the Department of Plant Science, University of Manitoba.


Research Agronomist, Manitoba Sugar Company, Winnipeg, Manitoba.
Assistant Professor, Department of Plant Science, University of Manitoba.

JOURNAL OF THE A. S. S. B. T.

Figures 1-6.Growth stages from which samples for cytological


analyses were collected. 1. two leaf stage, 2. four leaf stage, 3. twelve
leaf stage, 4. twenty leaf stage, 5. floral leaf apex stage, 6. inflorescence
PMC stage.

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As the seedlings grew, an attempt was m a d e to sample each


of the 2nd, 4th, 12th, 13th, 19th a n d 20th heart leaves (Figures
1-4). T h e heart leaves were excised when they attained a length
of four to six millimeters a n d collected directly i n t o cold water
for a 24-hour p r e t r e a t m e n t at 0 to 2C. T h e y were t h e n fixed
in Farmer's solution (3 parts C 2 H 5 O H : 1 part C H 3 C O O H ) for
a m i n i m u m of two days. H e a r t leaves then received an 8-minute
hydrolysis in 1 n o r m a l H C 1 at 60C prior to staining in Feulgen.
Squash p r e p a r a t i o n s were m a d e using acetocarmine as the c o u n t e r
stain. F r o m each heart leaf two r a n d o m samples were taken
from opposite sides of the leaf. Chromosome counts of twentyfive cells p e r sample were m a d e to give a total of fifty counts
per h e a r t leaf.
On t h e basis of the chromosome n u m b e r s e n c o u n t e r e d in
the last heart leaf sampled, the plants were divided into 3 groups;
diploids, tetraploids a n d chimeras. These plants then received
a photo-thermal i n d u c t i o n period of c o n t i n u o u s light at 4 0 F
for three m o n t h s . After the induction period one heart leaf was
taken from the floral apex of each plant w h e n the apex was
approximately six inches high (Figure 5). Fifty cells per floral
heart leaf were counted. Also at this stage one root tip was
excised a n d the chromosome n u m b e r of 25 cells obtained.
Pollen m o t h e r cell analyses were made on all plants that
bolted (Figure 6). T w o r a n d o m samples were taken from each
inflorescence a n d fixed directly i n t o Carnoy's solution (6 parts
C 2 H 5 O H : 3 parts C H C 1 3 : 1 part C H 3 C O O H ) . W h e n a c o u n t
was m a d e all five stamens were included in the squash preparation. Twenty-five counts were made per slide for a total of fifty
counts per inflorescence.
T w e n t y pollen diameters were recorded on each p l a n t that
bolted. Also the n u m b e r of plastids contained in the two g u a r d
cells s u r r o u n d i n g each of 10 stomata was recorded for ten plants
of each g r o u p .
Results and Discussion
Pre-bolting
Growth
D u e to the deliterious effect of colchicine on early p l a n t
growth only 10 plants were analyzed at all six growth stages a n d
22 plants at 5 growth stages. T h e data from cytological analysis
of these 32 plants are regrouped i n t o three growth stages a n d
presented in T a b l e 1. T h e first stage includes data from all the
heart leaves sampled prior to the 12th heart leaf. Stages two a n d
three have the c o m b i n e d data from the 12th a n d 13th, a n d 19th
and 20th h e a r t leaves, respectively.

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Table 1.The frequency of 2n, 4n and greater than 4n cells in heart leaves at three
growth stages.

* Cells other than 4n were 2n.

As indicated in Table 1, the heart leaves have a high average


chromosome number in early growth due to the occurrence of
8n and 16n cells. The mitotic configuration of a typical 4n - 8n
chimera is shown in Figure 7. Figures 8 and 9 show mitotic
metaphases of 135 and 72 chromosomes respectively,
which presumably have arisen through successive C-mitoses4. The occurrence of such cells decreases rapidly with plant maturity and
seldom are cells greater than 4n observed beyond the 4th heart
leaf. In most cases the chromosome number has stabilized by
the 12th heart leaf. In general, the percent 4n cells decreased
and the percent 2n cells increased from stage one to stage two,
* Chromosome duplication without cell division.

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Figures 7-9.High chromosome number cells in chimeras. 7. thirty-six


and. seventy-two chromosome cells in close proximity, 8. a 16n cell, 9. a
8n cell.

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b u t occasionally the opposite also occurred. T h e r e were some


plants (Nos. 3, 12, 14, 17, 19, 23, 26 a n d 27) which had only
4n or greater t h a n 4n cells in stage 1 that h a d 2n cells at later
stages. Yet there were no instances where stage one h a d all cells
of a lower chromosome n u m b e r t h a n that subsequently found
in stages two a n d three. T h u s it does n o t seem logical to a t t r i b u t e
the observation of cells n o t previously e n c o u n t e r e d to a sectorial
chimera in the corpus of t h e apical meristem. T h e p h e n o m e n o n
is m o r e adequately explained by assuming that the p r i m o r d i a
of the heart leaves in stage one were already formed in the
e m b r y o at the time of colchicine t r e a t m e n t a n d t h u s received a
colchicine t r e a t m e n t totally u n r e l a t e d to that of the p l a n t apex.
T h u s the leaves of stage one could have a higher chromosome
n u m b e r d u e to increased susceptibility to colchicine a n d the
effect of colchicine on the chromosome n u m b e r of these leaves
could be completely different t h a n that of the corpus which
later gives rise to t h e h e a r t leaves of stages two a n d three.
T a b l e 2 - D a t a f r o m t h e a n a l y s i s o f t h e floral h e a r t l e a v e s , p o l l e n m o t h e r cells, r o o t
tips, pollen diameter a n d chloroplast n u m b e r from 26 plants f o u n d to be totally 4n in the
last h e a r t l e a f c o u n t e d .
F r e q u e n c i e s of 4 n : 2 n Cells

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Post-bolting
Growth
T h e 32 plants m e n t i o n e d plus 26 plants that were analyzed
at less t h a n 5 growth stages received the photo-thermal i n d u c t i o n
treatment. On the basis of the last heart leaf sampled 26 plants
were classified as 4n, 20 as chimeras a n d 12 of the 2n plants
were included for a control. T h e data obtained from the floral
heart leaves, pollen m o t h e r cells, pollen measurements, chloroplast counts a n d root tip analysis are presented in T a b l e s 2, 3
a n d 4. T h e last heart leaf analysis for the chimera g r o u p is also
included.
T h e floral heart leaves of 25 of the 26 4n plants were completely 4n ( T a b l e 2). Only one plant (No. 49) was a c h i m e r a
at this stage a n d it p r o d u c e d a diploid inflorescence. T h i s p l a n t
was eliminated from the 4n g r o u p before anthesis on the basis
of floral leaf cytology. T h e P M C ' s of the other 24 plants were
all 4n. Seed set on the 4n plants was generally poor a n d very
variable. However, of the 250 progeny analyzed all were at the
4 n level, 2 0 % b e i n g aneuploids ranging i n chromosome n u m b e r
from 34 to 38.
T h e floral h e a r t leaves of 16 of the 20 chimera plants ( T a b l e
3) were chimeral, three were totally 4n (Nos. 10, 26 and 52)
a n d one was diploid (No. 22). PMC's of 16 of these plants were
analyzed. O n e inflorescence was chimeral (No. 24), 12 were 2n
Table 3.Data from the analysis of the last heart leaves, floral heart leaves, pollen
mother cells, root tips, pollen diameter and chloroplast number from 20 plants whose last

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Table 4.Data from the analysis of the floral heart leaves, pollen mother cells, root
tips, pollen diameter and chloroplast number from 12 plants found to be totally 2n in the
last heart leaf counted.
Frequencies of 4n:2n Cells

and three were 4n (Nos. 10, 12 a n d 52). T h e 12 plants in the


diploid class r e m a i n e d diploid in the floral heart leaves a n d
P M C ' s ( T a b l e 4). Only o n e of the 58 plants analyzed ( T a b l e
3, N o . 24) was a chimera in the inflorescence as it h a d one 2n
a n d one 4n floret. N o n e of the individual flowers sampled was
a chimera as j u d g e d by the P M C ' s .
T h e s e data indicate t h a t t h e floral h e a r t leaves of the p l a n t
will have t h e same chromosome n u m b e r as its pre-bolting h e a r t
leaf. Also if the floral heart leaf is 2n or 4n t h e n the inflorescence
will be 2n or 4n respectively. However, if the floral h e a r t leaf
is chimera t h e n t h e inflorescence will likely be completely 2n
although occasionally it may be 4n or a chimera.
D u r i n g t h e analysis of P M C ' s a range from complete bivalent
to complete q u a d r i v a l e n t chromosome association was observed.
Generally the 4n plants, h a d 2 to 4 q u a d r i v a l e n t s at metaphase
b u t some individual plants h a d a very high n u m b e r of multivalent associations (Figures 10-12).
It is evident from the data on r o o t t i p analyses in T a b l e s
2, 3 a n d 4, t h a t there is no correlation b e t w e e n t h e effect of
colchicine on t h e r o o t apex a n d t h e floral apex. Of t h e 49 plants
whose r o o t tips were analyzed all were totally 2n except 2 which
were 4n.
T h e r e was only one 4n p l a n t (No. 42) whose pollen d i a m e t e r
was smaller t h a n t h e largest 2n diameter. In general pollen
d i a m e t e r is an a d e q u a t e selection criterion of the ploidy level
of t h e inflorescence as one could discard the few plants whose
pollen measurements were i n t e r m e d i a t e b e t w e e n the two extremes. However, plants selected d u r i n g anthesis may interpollinate before they can be isolated.

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Figures 10-13.Meiotic metaphase configurations in C 0 sugar beets.


10. a 4n cell with 9 quadrivalents, 11. 8 quadrivalents plus 2 bivalents,
12. 7 quadrivalents and 4 bivalents, 13. a 2n cell with 9 bivalents included
for comparison purposes.

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On the basis of data given by o t h e r workers (3,4,6), some


plants w i t h 4n inflorescences (Nos. 37,41,43,48) w o u l d have b e e n
classified as 2n if one used the chloroplast counts as a selection
criterion. Also four plants (Nos. 14,15,17,20) which h a d 2n inflorescences would have been classified as 4n on the basis of
chloroplast counts. T h u s it is obvious that the m e a n chloroplast
n u m b e r per two g u a r d cells is n o t a good selection criterion for
ploidy n u m b e r of the inflorescence in the C 0 generation d u e to
the presence of periclinal chimeras.
Summary
Colchicine h a d a variable effect on the type of chimera a n d
chromosome n u m b e r p r o d u c e d in different plants.
It was observed that 2n initials in the corpus usually divided
at a faster rate t h a n the 4n initials b u t the opposite was also
occasionally true. However, in m a n y instances t h e chimeral cond i t i o n was m a i n t a i n e d at the time t h e floral leaf was sampled
b u t t h e majority of the inflorescences p r o d u c e d were entirely
2n with a few 4n a n d very few chimeral. H e a r t leaves of entirely
2n or 4n always p r o d u c e d 2n a n d 4n inflorescences respectively.
T h u s to select for C 0 plants with totally 4n inflorescences only
plants with totally 4n cells at t h e 20th heart leaf stage should
be induced to bolt.
T h e chloroplast counts per two g u a r d cells in conjunction
with cytology of P M C ' s indicates that periclinal chimeras were
often present which invalidates tne use of chloroplast counts as
a selection criterion for 4n inflorescences.
T h e ploidy of the inflorescence could in most cases be det e r m i n e d by the pollen diameter. However, it could be expected
that in m a n y instances such identification could n o t be m a d e
sufficiently early to remove undesirable plants from a p o p u l a t i o n
prior to flowering.
T h e root tips of the plants were seldom converted to a higher
ploidy a n d gave no indication of the type of inflorescence produced.
Acknowledgments
Financial s u p p o r t for this study from the N a t i o n a l Research
Council of Canada is gratefully acknowledged. T h e a u t h o r s are
also grateful to Mr. D. N. Fox for t h e p h o t o g r a p h i c plates.
Literature Cited
(1) DENEUCHE, J. i960. L'Analyse numerique de la composition des populations des betteraves polyploides dans le travail de selection. Genet.
Agrar. 3 1 : 268-276.

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(2) ESSAD, S. and H. TOUVIN. 1959. Techniques de production et de


controle des betteraves polyploides. Ann. Inst. Nat. Rech. Agron.
9: 553-574.
(3) FELTZ, H. 1953. Untersuchungen an diploiden u n d polyploiden zacherruben. Z. pflanzenzuchtung. 32: 275-300.
(4) FRANSDEN, K. J. 1939. Colchicininduzierte polyploide bei Beta vulgaris
L. Zuchter. I I : 17-19.
(5) KLOEN, D. and G. J. SPECKMAN. 1954. T h e creation of tetraploid beets.
II. Selection in the first generation (the C x ) from the treated material.
Euphytica 3: 35-42.
(6) SAVITSKY, H. 1952. Polyploid Sugar Beets. Cytological Study and Methods
of Production. Proc. Am. Soc. Sugar Beet Technol. 7: 470-476.
(7) VARGA, A. 1961. A poliploid cukorrepa eloallistesa es nemesitise.
Sapronhorpacs I: 67-93. 1961. (Abstract) Plant Breed. Abst. 33: 550.

On the Nature of Hatching of Heterodera schachtii.


I I I . Principles of Hatching Activity
D. R. VlGLIERCHIO AND P. K. Y u 1 '

Received for publication February 16, 1963

Introduction
T h e step, in the life cycle of animals, in which the developed
e m b r y o emerges from an egg m e m b r a n e or egg case to c o n t i n u e
its growth a n d development is a relatively c o m m o n event in the
a n i m a l world, a n d in nematodes as in o t h e r animals this transition may be a relatively inconspicuous incident in the n o r m a l
sequence of events. In p l a n t parasitic nematodes of the Heterodera
species, however, emergence from the cyst assumes a special
significance, for the cyst stage can serve as a survival form capable
of e n d u r i n g adverse e n v i r o n m e n t a l conditions.
T h i s aspect of nematology received considerable a t t e n t i o n
especially after the observation of Baunacke ( l ) 3 that leachings
of host plant roots stimulated larval emergence. T h e emergence
of larvae from cysts of Heterodera has b e e n the subject of a recent extensive review (12). T h e r e is a wealth of data concerned
with the stimulation of hatching, the h a t c h i n g factors in root
diffusates a n d the physical effects on the h a t c h i n g assay. T h e
available information is the p r o d u c t of n u m e r o u s workers in
various laboratories using different populations of animals, different sources of h a t c h i n g stimulation, a n d different conditions
a n d methods of assay. As a result n u m e r o u s apparently contradictory reports have arisen.
Investigations i n t o the h a t c h i n g process have consisted for
the most p a r t of descriptions of the physical parameters, the
testing of a n u m b e r of diverse Substances from n a t u r a l or synthetic sources for hatching factor activity a n d attempts at the
identification of a naturally occurring hatch active substance.
It was clear from previous reports (11,17,18) that if hatch data
were to be useful as a manifestation of t h e n a t u r e of hatching
it w o u l d n e e d to be o b t a i n e d from r e p r o d u c i b l e assays conducted
in a standard fashion with animals of as similar an e n v i r o n m e n t a l
history as possible a n d with hatch factor source materials prepared in a similar fashion.
T h e relative unspecificity of hatch factor sources, that is
1
Research funds for this study were contributed in part by the Beet Sugar Development Foundation.
2
Associate Nematologist and Assistant Research Nematologist, respectively, Department
of Nematology, University of California, Davis, California.
8
Numbers in parentheses refer to literature cited.

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the large n u m b e r of host a n d non-host plants whose root exudates


or plant organ extracts are capable of stimulating the emergence
of larvae from cysts of Heterodera schachtii Schmidt (11,18),
suggests that the active substance or substances in m a n y cases
could well be relatively c o m m o n products of plant metabolism.
Since p l a n t extracts a n d exudates are k n o w n to contain constituents a n d by-products of metabolism, for example, a m i n o
acids, sugars, proteins, vitamins, alkaloids, etc, it was of importance to test the hatch stimulatory activity of a n u m b e r of
representatives of the various classes of biologically active substances with cysts of H. schachtii.
Materials and Methods
X h e sugar beet nematode, H. schachtii, was reared on sugar
beets {Beta vulgaris) grown in sand culture in the greenhouse
as previously r e p o r t e d (14). At harvest time, approximately 3
to 4 m o n t h s after the initial infestation, a crude cyst concentrate
was separated from the sand by wet screening. Subsequently
cysts in t h e concentrate were separated from the associated debris
(14). T h e cysts, essentially p u r e , were then d a m p dried a n d
placed on the screen in a controlled h u m i d i t y b o x ( R H 9 8 %
at 5 C) in which the gas phase was mechanically circulated.
After 3 days the cysts were transferred to a static h u m i d i t y b o x
( R H 9 0 % at 5 C) a n d stored until used. Since the hatch tests
were c o n d u c t e d over a period of several years, cysts from a
n u m b e r of r e a r i n g lots so processed were used.
S t a n d a r d sugar beet leachate dry solids consisted of lyophilized
leachings o b t a i n e d by misting purified sugar beet germ in the
m i s t i n g a p p a r a t u s previously described (18). T h e sugar beet
g e r m was purified by the screening and w i n n o w i n g of by-products
from the commercial seed processors. T h e leachings o b t a i n e d
d u r i n g the first a n d second day of germination were discarded,
for according to preliminary tests they contained no active
material; t h e leachings of the third and fourth days were saved
a n d lyophilized. By the fifth day decomposition began a n d t h e
seed g e r m was discarded. T h e lyophilized material from a n u m b e r
of batches was thoroughly mixed, then stored at 15 C u n t i l
used. T h e chemical c o m p o u n d s used in these assays were of the
highest p u r i t y commercially available.
T h e h a t c h i n g bioassay was conducted after the m e t h o d of
Viglierchio (16). T h e cysts were suspended at the surface of
the test solution by a stainless steel screen in plastic wells containing 0.5 ml of solution sample. T h e cysts a n d solution (8
replicates p e r test solution of a concentration series) were incubated in a h u m i d i t y c h a m b e r at 25 C. At 4-day intervals the

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emerged larvae together w i t h old test solution were w i t h d r a w n


for c o u n t i n g a n d the wells refilled with freshly p r e p a r e d test
solution. Bioassays were discontinued after three collections.
T h e concentration of test materials, g r o u p e d according to class
of compounds, ranged from 0.1 mg per ml downwards. Concentration is given in terms of w e i g h t / v o l u m e to facilitate comparison with n a t u r a l hatch factor dry solids.
In figures where m o r e t h a n o n e curve is presented for a
c o m p o u n d the closed circles indicate the cyst response at maxim u m sensitivity a n d the o p e n circles, m i n i m u m sensitivity. T h e
ratio indicates the n u m b e r of tests of high sensitivity to the
n u m b e r of low sensitivity. W h e n e v e r no substantial difference in
sensitivity was observed only a representative curve is shown.
For practical considerations the curves of any one class of comp o u n d s were compiled from a n u m b e r of experiments. It was
usually impossible to test all the m e m b e r s of a class at all the
indicated concentrations at one time.
R o b i n s o n a n d Neal (8) r e c o m m e n d e d the use of a potassium,
sodium, m a g n e s i u m a n d calcium chloride solution with purified
preparations of leach material for increased sensitivity in H.
rostochiensis hatch tests. T h e suggested salt concentration was
m a i n t a i n e d constant t h o u g h test materials were diluted.
Results
Cyst Sensitivity
Cysts of H. schachtii reared, purified a n d
stored as described in the previous section responded to standard
hatch factor materials in a continually changing fashion througho u t their useful life. At t h e time of harvest there was usually
no hatch stimulation i.e., the hatch response to water was essentially the same as that to s t a n d a r d leachate. T h i s c o n t i n u e d

4 5 6 7 8
9
CYST AGE (month*)

10

II

Figure 1.The cumulative larval h a t c h response of H. schachtii to


standard solutions of the dried hatch factor materials a n d water as a
function of cyst age.

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for 1 to 3 m o n t h s (generalized curve F i g u r e 1). T h e r e a f t e r


there was a slight m a x i m u m in the leachate response before it
decreased slowly as indicated. T h e water response, however,
decreased m o r e rapidly so as to effect a differential response.
Leachate usually provided a three- to four-fold increase in hatching over water controls; however, u n d e r certain conditions a
ten-fold increase was possible. Some 9 m o n t h s after the t i m e
of harvest the sensitivity as well as the differential response decreased to a value too low to be useful for emergence assay
purposes. It was evident, therefore, that absolute values of hatch-

j^3

tZ

JT

Concentration

IOO

20

(mg/ml. x I0" 2 )

.8

IOO

ZO

.8

Figure 2.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of sugars and related compounds. The horizontal dotted lines indicate the larval emergence in distilled water.

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ing could be misleading. A relative measure w o u l d be m o r e


useful, consequently the hatch response curves have b e e n given
as percent relative hatch w i t h respect to water.
Sugars and Related Compounds
Of the compounds of this
class tested, i-inositol, sucrose a n d possibly fructose a n d lactose
possessed stimulatory activity (Figure 2). Dextrose, maltose,
sorbitol, m a n n i t o l a n d possibly lactose w i t h salts t e n d e d to be
inhibitory. T h e concentration of salt solutions beneficial for
hatching in H. rostochiensis a p p e a r e d to be inhibitory in t h e
hatching of H. schachtii. Except at the high concentration of
i-inositol a n d sucrose the effect of salt solution w h e n manifest
was to reduce larval emergence. In this class of c o m p o u n d s ,
i-inositol a n d sucrose approached the stimulatory activity of sugar
beet leachate.
Organic Acids
F r o m the observed results (Figure 3) citric,
maleic, succinic, glutaric, malonic, fumaric a n d oxalic acids could
be as active as leachate. O n l y 2-oxoglutaric acid a n d tartaric acid
were found inactive. A d d i t i o n a l tests with succinic, glutaric,
malonic, fumaric a n d oxalic acids showed that activity was not
consistent.

Figure 3.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of organic acids. T h e horizontal dotted lines
indicate the larval emergence in distilled water.

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Vitamins and Related Compounds Biotin, d-calcium pantothenate, folic acid, ascorbic acid, and thiamin could possess
stimulatory activity comparable to leachate (Figure 4). Riboflavin
and taurine appeared to be less active whereas glutathione,
pyridoxine and nicotinic acid were completely inactive in these
experiments. Repeated tests with biotin, d-calcium pantothenate
and folic acid confirmed that whereas biotin could be inactive,
d-calcium pantothenate and folic acid could be very inhibitory.
The hatch curves indicated that encysted larvae could be sufficiently sensitive to detect concentrations of vitamins on the
order of 0.1 ppm.

Figure 4.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of vitamins and related compounds. The horizontal dotted lines indicate the larval emergence in distilled water.

Amino Acids The L-amino acids as a group appeared relatively inactive in hatch stimulation tests (Figure 5 and 6). Of
some 25 amino acids tested only L-lysine, L-tryptophan, Laspartic acid and perhaps L-glutamic acid possessed some ability
to stimulate larval emergence from cysts. Some of the more
active acids appeared to possess stimulatory power at concentrations on the order of 0.01 ppm. L-lysine was the only amino
acid consistently active.

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Figure 5-6.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of amino acids. T h e horizontal lines indicate
the larval emergence in distilled water.

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Nucleosides and derivatives The hatch curves (Figure 7)


showed that though cytosine, adenine, guanine, a d e n o s i n e ,
guanosine and inosine were capable of stimulating larval emergence as much or more than standard leachate, they could also
be completely inert. There is some indication, e.g. adenosine
and inosine, that the coupling of a sugar to the nitrogen base
increased stimulatory power; however, when the adenosine wtis
phosphorylated to give the coenzymes of higher phosphate content, activity appeared to be lost. Coenzyme A and diphdsphopyridine nucleotide were not active.
Olphosphopyridine nucleotide

IOO

20

Concentration (mg./mi. x I0" 3 )

Figure 7.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of nucleosides and derivatives. T h e horizontal
lines indicate the larval emergence in distilled water.

Fatty Acids The higher fatty acids C12 and above, whether
saturated or unsaturated, were inactive in hatch test (Figure 8).
The inhibitory activity at the higher concentration could be
attributed to the high alkalinity necessary to keep the fatty acid
in solution. The lower members of the fatty acid series, acetic,
propionic, and butyric acids could be inert and/or inhibitory
to larval emergence. Caproic, caprylic and capric acids were
able to stimulate a larval emergence comparable to leachate.

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Figure 8.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of fatty acids. T h e horizontal lines indicate the
larval emergence in distilled water.

Alkaloids In tests of some 30 alkaloids, representatives of


principal structure groups, there was a general tendency to inactivity, over a wide range of concentrations (Figure 9, 10 and
11). Morphine, quinine, digitoxigenin, strychnine, codeine,
coniine could be inhibtory of larval emergence. Digitoxigenin
sometimes demonstrated inhibitory activity at concentrations of
0.01 ppm.
Dyes The dyes selected (Figure 12) were the most active of
those reported in the literature for the stimulation of larval
emergence from cysts of Heterodera schachtii (12). Methyl red
was comparable in activity to leachate; picric acid was more
active.
Miscellaneouis Biologically Active Compounds Camphor,
benzoic acid, indole-3-acetic acid, salicylic acid, and 2-naphthoxyacetic acid were comparable to leachate in ability to stimulate
larval emergence from cysts (Figure 13). At high concentrations
-mercaptoethylamine and salicylic acid were more active than
leachate, whereas, camphor appeared to be more active at greater
dilutions. Of the plant growth regulators, naturally occurring
indole-3-acetic acid and synthetic 2-naphthoxyacetic acid were
very active but gibberellic acid, 2,4-dichlorophenoxyacetic acid
and 1-naphthaleneacetic acid were only slightly active. Thiol

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r DL-Coniine*H(
took-^^J

I F

Figure 10.

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Figure 9-11.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of alkaloids. T h e horizontal lines indicate the
larval emergence in distilled water.

Concentration (mg./ml. x IO~*)

Figure 12.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of synthetic dyes. T h e horizontal lines indicate
the larval emergence in distilled water.

compounds could be very active in high concentrations as /?mercaptoethylamine, mildly active as thiodiglycolic acid and
mercaptoacetic acid at low concentrations or inhibitory at higher
concentrations as mercaptoacetic acid. Cholic acid, inert at 100
ppm, increased in activity with dilution approaching the maximum o leachate at 0.1 ppm.

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IOO

20

Concentration (mg./ml. xlCT 5 )

Figure 13.The percent larval hatch response, with respect to water,


of H. schachtii to solutions of miscellaneous compounds. T h e horizontal
lines indicate the larval emergence in distilled water.

Discussion
The cumulative larval hatch curves indicating emergence
due to water and leachate with respect to cyst age (Figure 1) are
a function of rearing conditions, method of purification, technique of drying and conditions of storage. The ordinate intercept
can be varied greatly by the technique of drying; if the conditions are drastic and the desiccation takes place quickly, cumulative larval hatch can be reduced to an insignificant value. Gradual
drying over a period of 3 to 5 days maintains a high initial
cumulative hatch. The drying is conducted at 5 C to prevent
the high emergence normally occurring at higher temperatures.

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T h e r e usually was an initial lag period d u r i n g which it was n o t


possible to detect a difference between water a n d leachate hatch.
T h i s observation suggests that t h e differential response was an
induced effect giving rise to hatch factor physiology.
Since the leachate hatch t e n d e d to rise while the water hatch
decreased after the initial lag period the differential response
was other than merely an a c c u m u l a t i o n of waste toxic materials
or inhibitors, since these w o u l d have leached o u t in water as
well as root e x u d a t e solution. Clearly, the root e x u d a t e solution
provided s o m e t h i n g which p r o m o t e d larval emergence. T h e
gradual decrease in h a t c h i n g can be visualized in part, as a
resultant of the toxic effects of accumulated waste products a n d / o r
the depletion of metabolic reserves n o t supplied either by water
or root leachings. T h e p r i m a r y interest was n o t in the longevity
of hatching b u t in the n a t u r e of the differential response.
T h e change in differential response w i t h age of cysts suggests
that the r e q u i r e m e n t s of the larvae that p e r m i t h a t c h i n g also
change or that the larvae are n o t all e q u i v a l e n t so t h a t there
is selection in the h a t c h i n g response. For purposes of comparison
it was advantageous to employ a frame of reference such that
i n h i b i t i o n a n d stimulation were measurable on the same scale.
Water, an indispensable solvent of the biological system, served
as a suitable base for the simultaneous estimation of solvolysis
effects a n d comparison of addenda. F u r t h e r m o r e t h e use of the
water eliminated t h e c o n f o u n d i n g effect of the additional param e t e r necessary with the root exudate base.
It was n o r m a l practice to include a standard leachate test
in every bioassay as an indicator of cyst response; representative
compilations were presented in the figures of data for each class
of c o m p o u n d s . It was evident that at one concentration the
activity could range from a m a x i m u m , to intermediate, to inertness, to i n h i b i t i o n ; since t h e hatch factor source, t h e same
t h r o u g h o u t , h a d n o t been observed to deteriorate appreciably
it could reasonably be assumed that there were o t h e r larval req u i r e m e n t s for emergence n o t supplied by the leachate.
T h e data presented in this r e p o r t have shown that m a n y
biologically active c o m p o u n d s at the a p p r o p r i a t e concentration
a n d u n d e r the p r o p e r conditions were as capable of s t i m u l a t i n g
larval emergence as the c r u d e leachate preparations. Sucrose
a n d i-inositol of t h e polyalcoholic c o m p o u n d s could be good
hatch stimulants. It was of interest to observe t h a t sucrose was
a good hatch s t i m u l a n t whereas dextrose or fructose were not.
T h i s suggested that sucrose activity was n o t d u e to its n o r m a l
glycolytic role, i.e. its b r e a k d o w n to fructose a n d glucose followed
by s u b s e q u e n t phosphorylation etc. b u t perhaps to its structural

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entity as a a-D-glucopyranoside-B-D-fructofuranoside. T h e salt


solutions which were i m p o r t a n t cofactors in the hatch test of
H. rostochiensis h a d an inhibitory effect when used in association
with sugars in H. schachtii hatching tests. For practical considerations their use in association with other synthetic c o m p o u n d s
was omitted.
T h e high hatching activity of a n u m b e r of the organic acids
suggested the importance of the tricarboxylic acid cycle. Citric,
succinic a n d fumaric acids are members of the cycle and their
activity may be explained as a supplement to a deficiency of
substrates. T h e inactivity of a-keto-glutaric acid, a constituent
of the cycle, could be explained by the presence of sufficient
substrate a n d the nontoxicity of a surplus. Maleic acid was active,
p e r h a p s because it isomerized to the trans active fumaric acid.
Malonic acid activity may have been the result of competitive
i n h i b i t i o n in the succinate to fumarate step in the tricarboxyclic
acid cycle.
M a n y of the vitamins tested were able to stimulate larval
emergence. W a t e r soluble, nonstorable vitamins present at low
levels could be consumed in a relatively short time. T h e i r addition to correct a deficiency would permit enzymatic reactions,
in which they were cofactors, to proceed thereby eliminating a
block in the sequence of events leading to larval emergence.
In general the a m i n o acids did not appear to be especially
active in hatch stimulation. L-tryptophan a n d L-aspartic acid
could be mildly stimulatory; only L-lysine appeared to be consistently active. A consideration of the processes involved in
h a t c h i n g suggests no need for active protein synthesis such as
could be expected in the m o l t i n g process. Normally tissue proteins w o u l d be expected to be catabolized in preference to enzymatic proteins, a n d inasmuch as the enzymatic c o m p l e m e n t was
present initially for larval emergence it could be expected to
r e m a i n . T h e active a m i n o acids, especially L-lysine, could be
involved in reactions other t h a n protein synthesis, e.g. lipid
metabolism a n d m e m b r a n e physiology.
T h e hatch stimulation effected by p u r i n e a n d p y r i m i d i n e
bases a n d their pentose derivatives would be explicable in terms
of nucleic acid synthesis or coenzyme activity. Since h a t c h i n g
was unlikely to involve a burst of new cell formation or o t h e r
vigorous g r o w t h activity, low nucleic acid synthesis w o u l d be
expected. On the other h a n d it was difficult to reconcile t h e
lack of hatch activity of polyphosphorylated nucleosides with this
n o t i o n (Figure 7).

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In view of the habitat a n d the e n v i r o n m e n t a l conditions in


which the plant parasitic n e m a t o d e lives, it is n o t surprising
that the depot lipids are in the l i q u i d phase. It is of considerable
interest, however, that the l i q u i d short chain saturated fatty
acids C 6 , C 8 a n d C 10 n o t only were m o r e active in the stimulation
of larval emergence t h a n the l i q u i d u n s a t u r a t e d long chain fatty
acids b u t that with the short chain saturated acids caproic increases in activity with d i l u t i o n , caprylic reaches a m a x i m u m
a n d decreases a n d capric decreases with d i l u t i o n . It is striking
to find that acetic acid, p r o p i o n i c acid a n d butyric acid could
be inert or completely inhibitory to hatching.
T h e m o d e of action of alkaloids is n o t u n d e r s t o o d b u t no
survey of biologically active c o m p o u n d s w o u l d be complete witho u t them. T h e y appeared to be relatively inactive except p e r h a p s
at very high concentrations. T h e r e were anomalous reactions,
for example, digitoxigenin a n d m o r p h i n e inert at higher concentrations were inhibitory at extremely low ones. T h e r e seems
to be little e x p l a n a t i o n for this o t h e r t h a n a difference between
cyst lots in ability to detoxify a particular alkaloid.
T h e hatch s t i m u l a t i n g ability of certain dye c o m p o u n d s
would a p p e a r to be a result of competitive reactions. Picric
acid a n d methyl red, for example, are n o t naturally occurring
c o m p o u n d s b u t synthetic preparations; their structure needs
correlation with that of a naturally-occurring stimulatory material for confident speculation on m o d e of action. T h i s action
is to be differentiated, however, from that of acids, enzymes,
etc. of n o n - n e m a t o d e origin which induce emergence by chemical
r u p t u r e of the egg m e m b r a n e , a m o d e of emergence irrelevant
to this report.
Organic bases differing in structure a n d strength as illustrated
by choline chloride, betaine, creatinine a n d creatine, appear
to have little stimulatory power, for example, b e t a i n e could
stimulate slightly or i n h i b i t slightly. N a t u r a l l y o c c u r r i n g indole3-acetic acid a n d the synthetic analog 2-naphthoxyacetic acid
were as active or perhaps m o r e active t h a n leachate. O t h e r synthetic auxins, however, 2,4-dichlorophenoxyacetic acid a n d 1naphthaleneacetic acid were essentially inactive. N a t u r a l l y occurr i n g gibberellic acid was also inactive.
T h i o l c o m p o u n d s differed markedly in activity. Mercaptoacetic acid for example was i n h i b i t o r y at high concentrations
a n d mildly active at the m o r e d i l u t e concentrations. T h i o diglycolic acid was mildly active at all concentrations. On the
other h a n d B-mercaptoethylamine was very active at high concentration b u t mildly so at d i l u t e ones. Cystine a n d cysteine are

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essentially inactive. T h e r e seemed to be no particular p a t t e r n


in hatch stimulation activity of these s u l p h u r c o m p o u n d s . Gholic
acid a p p e a r e d to increase in activity with dilution. In view of
the interesting results with the fatty acids additional tests w i t h
sterol c o m p o u n d s would have been especially useful; unfortunately no others were included. Benzoic acid, c a m p h o r a n d
especially salicylic acid were very active compounds. T h e n a t u r e
of t h e hatch stimulatory powers of these c o m p o u n d s is u n k n o w n .
In view of c u r r e n t understanding, hatch stimulatory activity
appears to take three forms with respect to general modes of
action: metabolicconsisting of substances serving as substrates,
cofactors, or coenzymes of conventional physiological systems, inc l u d i n g anabolism, catabolism and endocrinology; pseudometabolicconsisting of substances reacting competitively in physiological reaction systems; a n d ametabolicconsisting of substances with non-physiological modes of action. T h e diversity
of c o m p o u n d s manifesting a p p a r e n t ametabolic hatching stimulation illustrates the complexity of the hatching p h e n o m e n o n .
It is t e m p t i n g to believe that the naturally occurring stimulation is of the metabolic type since this would be m o r e compatible w i t h evolutionary or survival value. Inorganic salts,
carbohydrates, organic acids, vitamins, a m i n o acids, as well as
o t h e r unidentified substances, are present in t h e root leachings
of plants (6,7,10,13). T h e r e is evidence (17) that: stimulatory
substances in n a t u r a l leachings can be consumed or utilized as
substrate d u r i n g the process of hatching, inhibitors can be present
in cysts (17) a n d in n a t u r a l leachings together with activators
from host a n d non-host plants (18). T h e r e is great n u m b e r a n d
diversity of: plants whose leachings can stimulate larval emergence (11,18), synthetic c o m p o u n d s capable of inducing hatching
(2,3,12,19,20,21), physical a n d environmental parameters imp o r t a n t in the h a t c h i n g process (11,15,17,18), a n d in cyst lots in
readiness a n d capability to hatch. It would appear that the sum
of these observations would be consistent with the visualization
of eclosion in H. schachtii as being effected by the successful
completion of a complex series of physiological reaction any of
which could be limiting. T h e limiting steps would d e p e n d on
the c u m u l a t i v e effect of the environmental influences in the rearing a n d storage of cysts up to the m o m e n t of hatching.
T h e stimulation of larval emergence of a particular cyst lot
effected by root e x u d a t e would be a reflection in part of its
ability to supply t h e metabolic requirementssubstrate, organic
a n d inorganic cofactors, inhibitors, etc. as suggested by D r o p k i n
(4). T h o u g h c r u d e leachate can be expected to contain i n e r t

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substances in varying proportions, t h e results of this investigation


show that in some cases activity of the leachate could essentially
be accounted for by k n o w n synthetic biologically active compounds.
It w o u l d be unlikely however that hatch stimulation of active
root exudates could always be explained in terms of the b e t t e r
k n o w n metabolic reactions; one m i g h t expect that the likelihood
w o u l d decrease with increasing specificity of parasitism (5). If
hatching occurs after the fashion suggested by Rogers (9) t h e n
the search for the m o r e specific blocks in Heterodera o u g h t
necessarily to take cognizance of the simultaneous occurrence
of c o n f o u n d i n g blocks of m o r e general metabolic reactions. It
w o u l d be desirable to be certain that the same reaction was
b e i n g measured t h r o u g h o u t a hatch test series, a n d w h e t h e r the
larvae o b t a i n e d by diverse synthetic stimulants were equivalent
in physiological potential i.e. penetration, development, reprod u c t i o n etc. It is a p p a r e n t that for a clearer insight i n t o the
n a t u r e of hatching, e x p e r i m e n t a t i o n needs to be c o n d u c t e d with
m o r e finesse t h a n in the past.
Literature Cited
(1) BAUNACKE, W. 1922. Untersuchungen zur Biologie und Bekarapfung
des Riibennematoden Heterodera schachtii Schmidt. Arb. biol. Abt.
(Anst. Reichsanst.) Berl. 11: 185-288.
(2) BAUSERMAN, H. M. and R. F. OLSON. 1957. Nematode cyst hatch rate
as influenced by fractions of beet root juice. J. Am. Soc. Sugar
Beet Technol. 9: 387-392.
(3) CLARKE, A. J. and A. M. SHEPHERD. 1964. Synthetic hatching agents
for Heterodera schachtii Schm. and their mode of action. Nematologica 10: 431-453.
(4)

DROPKIN, V. H., G. C. MARTIN a n d R. W . JOHNSON.

1958.

Effect of

osmotic concentration on hatching of some parasitic nematodes.


Nematologica 3: 115-126.
(5)

(6)
(7)
(8)
(9)

MARRIAN, D. H., P. B. RUSSELL and A.

R. T O D D .

1949.

T h e potato-

root eelworm hatching factor. 6. Attempts to prepare artificial


hatching agents. Part II. Some active arylidene-Aj3"y-butenolides
and related compounds. Biochem. J. 45: 533-537.
MILLER, E. D. 1938. Plant physiology. McGraw-Hill Book Company,
Inc. New York and London. 1201 p p .
PEARSON, R. and D. PARKINSON. 1961. T h e site of excretion of ninhydrin-positive substances by broad bean seedlings. Plant and Soil
13: 391-396.
ROBINSON, T. and A. L. N E A L . 1959. T h e influence of certain mineral
elements on emergence of golden nematode larvae. Proc. Helminthol.
Soc. Wash. 26: 60-64.
ROGERS, W. P. 1960. T h e physiology of infective processes of nematode
parasites; the stimulus from the animal host. Proc. Royal Society
of London, Series B 152: 367-386.

VOL. 13, N o . 8, JANUARY 1966

715

(10) ROVIRA, A. D. and J. R. HARRIS. 1961. Plant root excretions in relation to the rhizosphere effect. Plant and Soil 14: 199-214.
(11) SHEPHERD, A. M. 1962. T h e emergence of larvae from cysts in the
genus Heterodera. Technical Communication No. 32 of the Commonwealth Bureau of Helminthology.
(12) SHEPHERD, A. M. 1962. Dyes as artificial hatching agents for beet
eelworm, Heterodera schachtii Schm. Nature 196: 391-392.
(13)

(14)

(15)

(16)
(17)
(18)

(19)

(20)

(21)

STOTZKY, G., W . CULBRETH a n d L. B. MISH.

1962. A p p a r a t u s for grow-

ing plants with aseptic roots for collection of root exudates and
C 0 2 . Plant Physiol. 37: 332-341.
VIGLIERCHIO, D. R. 1959. Collection and selection of cysts of the
sugar beet nematode, Heterodera schachtii. J. Am. Soc. Sugar Beet
Technol. 1 0 ( 4 ) : 318-329.
VIGLIERCHIO, D. R. 1960. Heterodera schachtii, hatching properties
of field importance. J. Am. Soc. Sugar Beet Technol. 11 (4) :
294-301.
VIGLIERCHIO, D. R. 1961. A simplified technique for hatching tests
of Heterodera schachtii. Phytopathology 51: 330-332.
VIGLIERCHIO, D. R. 1963. On the nature of hatching of Heterodera
schachtii. Proc. Helminthol. Soc. Wash. 30: 195-198.
VIGLIERCHIO, D. R. and P. K. Yu. 1965. On the nature of hatching
of Heterodera schachtii. II. Natural sources of hatching stimuli.
J. Am. Soc. Sugar Beet Technol. 13(4): 354-361.
WALLACE, H. R. 1956. T h e emergence of larvae from cysts of the
beet eelworm, Heterodera schachtii Schmidt, in aqueous solutions
of organic and inorganic substances. Ann. appl. Biol. 44: 274-282.
WALLACE, H. R. 1957. T h e stimulatory properties of some organic
substances on cysts of the beet eelworm, Heterodera schachtii
Schmidt. Ann. appl. Biol. 45: 251-255.
WINSLOW, R. D. 1959. A note on anhydrotetronic acid as a hatching
agent of the beet eelworm, Heterodera schachtii Schmidt. Nematologica 4: 237-238.

Odor in Refined Sugar


R O B E R T R . W E S T AND

R O B E R T S.

GADDIE1

Received for publication March 15, 1965

O n e of the most precise methods for q u a n t i t a t i v e evaluation


of odor is the threshold procedure. T h e threshold n u m b e r represents the extent to which an odor-bearing sample m u s t be
d i l u t e d with odor-free water in order to m a k e its odor barely
perceptible. T h e threshold-odor test used for m a n y years in the
water t r e a t m e n t field was modified by the Suchar Sales Corporation for sugar work a n d published in Sugar Journal, N o v e m b e r
1961. In applying this p r o c e d u r e to the analysis of refined beet
sugars, it soon became evident that the m e t h o d lacked sufficient
sensitivity for o u r purpose. A modification of this Suchar m e t h o d ,
however, was developed which shows considerable promise. For
the past operating season this m e t h o d has been used to d e t e r m i n e
threshold o d o r n u m b e r s on daily composite sugar samples from
the six U t a h - I d a h o Sugar C o m p a n y factories.
Procedure
T h e test with o u r modifications follows:
Reagents
and
Equipment
Odor-free (carbon treated) water.
C.P. citric acid, 1 0 % solution in odor-free water.
Six 500 ml glass-stoppered E r l e n m e y e r flasks.
W a t e r b a t h controlled at 60 C.
Determination
Dissolve 200 gm of the sugar to be e x a m i n e d in 200 ml of
odor-free water at r o o m t e m p e r a t u r e . T h i s provides the test
solution (Solution A) which will be used for further dilution.
T a b l e 1 shows the aliquots of this odor-bearing Solution A to
be a d d e d to freshly rinsed flasks for the d e t e r m i n a t i o n of the
odor range.
Table 1.Milliliters of solution A to be used in first dilution to determine range.

1
Head Chemist, General Laboratory and General Chemist, respectively.
Sugar Company, Salt Lake City, Utah.

Utah-Idaho

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free water plus 5 ml of 1 0 % citric acid solution to a n o t h e r flask


as a reference. H e a t all flasks to 60 C in a hot-water b a t h .
Shake the flask containing the reference solution, remove t h e
stopper, a n d sniff the vapors. Replace the stopper. Do the same
with the flask containing the 200 ml of sample u n d e r test a n d
observe w h e t h e r by comparison it contains an odor differing
from the slight characteristic odor of the citric acid. R e p e a t this
p r o c e d u r e with the flask containing the next lower concentration, always sniffing the reference solution first. Record which
flasks present differences a n d which do not. Based on the results
of the p r e l i m i n a r y test prepare one of the following sets of
dilutions:
Range
Range
Range
Range

1:
2:
3:
4:

Difference
Difference
Difference
Difference

with
with
with
with

200 ml b u t not with 50 ml


50 ml b u t not with 12 ml
12 ml b u t not with 2.8 ml
2.8 ml.

T a b l e 2 shows the dilutions of Solution A to be used with each


of these four ranges.
Table 2.Milliliters of solution A to be used for determination of threshold odor number.

If necessary, make up a n o t h e r Solution A and proceed as


described previously d i l u t i n g each to 200 ml, a d d i n g citric acid
solution, a n d h e a t i n g to 60 C. Arrange the flasks so that their
identity is u n k n o w n , a n d compare each with the reference solution. Place the flasks in which odor difference was observed in
one g r o u p a n d those w i t h o u t odor difference in another. T h e
d i l u t i o n c o n t a i n i n g the smallest volume of sample which gives
a positive test d e t e r m i n e s the threshold odor. By reference to
T a b l e 3 the corresponding threshold n u m b e r is determined.

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Table 3.Dilution conversion table.

1 2 4 817

I 2 4 8 17 35 1 2 4 8 1735 I 2 4 81735 12 4 81735

1 2 4 8 1735

THRESHOLD ODOR NUMBER

To
o b t a1.Results
i n consistent
results approximately
in odor measurements,
is necesFigure
of testing
100 days ofit production
at six that
factories
1963.
sary
the during
analyst
always observe a t e c h n i q u e somewhat as
follows:
1. Some practice is necessary to develop consistent threshold
sensitivity. T h i s consistency can be readily developed in most
individuals. An acute sense of smell is n o t essential.
2. A reliable a n d a d e q u a t e supply of odor-free
carbon treated) water is r e q u i r e d .

(activated

3. All glassware m u s t be clean a n d free from odor. It is


necessary to rinse all glassware several times with odor-free water
p r i o r to each test a n d b e t w e e n dilutions.

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4. Tests should be r u n in a room as free from foreign odors


as possible.
5. Each d i l u t i o n should be compared with the reference solution to check j u d g m e n t a n d minimize odor memory.
6. Care should be taken never to declare a d i l u t i o n as positive
in odor unless there is sufficient odor present to justify this
declaration.
T h e results of testing approximately 100 days of p r o d u c t i o n
of sugar at each of our six operating factories d u r i n g 1963 are
shown in Figure 1.
To simplify presentation, the results have been g r o u p e d so
that the results for threshold n u m b e r 1 include values of 0 a n d
1, while those for threshold n u m b e r 2 include values greater t h a n
1 b u t not greater than 2, etc. It is quite apparent that there is a
characteristic p a t t e r n for each factory, though the differences are
m o r e obvious w h e n presented as in Figure 2. Figure 2 shows
for each factory the percentage of sugar samples tested which
would pass a theoretical threshold odor n u m b e r specification.

THRESHOLD ODOR NUMBER SPECIFICATION

Figure 2.Percentage of sugar samples tested which would pass a


theoretical threshold odor number.

At this date we have found only one positive correlation between odor a n d operating conditions at the six factories. In every
case, as the campaign progressed and as storage periods increased,
u n d e r the same average operating conditions, the o d o r in sugar
increased in direct proportion. Figure 3 illustrates this a n d while

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Probability Analysis of Herbicide Response1


E.

F.

SULLIVAN AND H.

L.

BUSH2

Received for publication April i, 1965

Introduction
In many herbicide screening experiments, visual numerical
ratings of effectiveness are used to evaluate the relative differences
a m o n g treatments. T h e accuracy of these visual estimates of weed
control is d e p e n d e n t on the skill and objectivity of the investigator, particularly regarding the assessment of the density
a n d species composition of the sample. N u m e r o u s investigators
acknowledge the fact that visual estimates of weed control contain errors as high as 20 percentage points, and that valid interpretation of subjective data is difficult unless consistently large
differences are apparent.
T h e results of experimentation conducted at the Great
Western Agricultural Experiment Station (3) 3 indicated the need
for a statistical m e t h o d of interpretation of plant-count data
obtained from variable-dosage experiments. It was proposed that
a suitable statistical method existed which would allow predictions and practical recommendations to be made from variable-dosage data, particularly when small differences were expected. T h e probit analysis method was examined as a possible
tool for the interpretation of dosage-response relationships in
weed control. Heretofore, the direct method of variance analysis
was applied to variable-dosage data with reservations regarding
the validity of the interpretations.
Materials and Methods
Field
Procedure
In this study, the method of direct graphical interpretation
of the dosage-response relationships of P E B C (n-propyl ethyl-nbutylthiolcarbamate), diallate (2-3-dichloroallyl diisopropylthiolcarbamate) a n d the mixture, PEBC + diallate, was examined.
T h e introductory studies on probit theory and practice by Finney
(2) a n d Gowing (1) were consulted to formulate method.
A 12 l b / A initial dosage of active ingredient was used for
each herbicide, although the PEBC + diallate combination was
i Contribution of the Great Western Agricultural Experiment Station, Longmont,
Colorado. From a paper given at the Proc. Thirteenth General Meeting, Am. Soc. Sugar
Beet Technol., February, 1964.
"Agronomist and Agronomist-Statistician, respectively.
8
Numbers in parentheses refer to literature cited.

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a p p l i e d at the 8 + 4 l b / A dcsage. T h i s c o m b i n a t i o n h a d shown


superior b r o a d s p r e c t r u m activity w i t h o u t kochia (Kochia
scoparia) in earlier studies (3) a n d s u b s e q u e n t l y (4).
T h e t r e a t m e n t s were a p p l i e d p r e p l a n t , soil-incorporated a t
the 1.5-inch d e p t h on J u n e 13, 1963. A silt loam soil o h i g h
fertility, located at W i n d s o r , Colorado, was sampled. Soil temp e r a t u r e s at establishment a n d at t h e i n c o r p o r a t i o n d e p t h averaged 72 F. T h e e x p e r i m e n t received 2.75 inches of p r e c i p i t a t i o n
d u r i n g the e x p e r i m e n t a l p e r i o d which e x t e n d e d u n t i l July 8
w h e n the final observations were m a d e .
T h e t r e a t m e n t s were a r r a n g e d i n r a n d o m i z e d c o m p l e t e blocks
with 3 replications. P l a n t counts of pigweed (Amaranthus retroflexus) a n d foxtail m i l l e t (Setaria italica) were m a d e in each
t r e a t m e n t a n d replicate at the initial c o n c e n t r a t i o n a n d at each
half-dosage distance of 25 feet. T h e counts were taken w i t h i n a
wire rectangle which m e a s u r e d 4 inches by 36 inches. P l o t size
m e a s u r e d 44 inches by 125 feet w i t h t h e herbicides a p p l i e d in
7-inch b a n d s to two rows spaced 22 inches apart.
Graphical
Probit
Analysis
T o t a l control percentages (pigweed + foxtail) were calculated from the observed values at each dosage distance. T h e s e
percentages were c o n v e r t e d to e m p i r i c a l p r o b i t s as described by
F i n n e y (2).
In practice, t h e expected probits (Y) were p l o t t e d against
the l o g a r i t h m of the dosage (X). A straight line or w e i g h t e d
regression line was d r a w n by eye to fit t h e control p r o b i t on
the l o g a r i t h m of t h e dosage. T h i s weighted regression line was
used to calculate t h e fitted line by regression analysis. T h e r e b y ,
t h e n o r m a l sigmoid response curve was transformed to a straight
regression line, w h e n the o r d i n a t e s were m e a s u r e d on a linear
scale of p r o b i t s instead of percentages.
T h e chi s q u a r e test was used to test goodness of fit b e t w e e n
the theoretical regression line a n d t h e actual observations of t h e
relationship b e t w e e n dosage a n d response. T h e chi s q u a r e values
were greater t h a n P = .99 w h i c h indicated a very close fit.
F u r t h e r m o r e , t h e log L D 5 0 was estimated for each h e r b i c i d e
from t h e t r u e regression line at p r o b i t of control, Y-5 (2). L D 5 0
refers to the lethal dosage which controls 50 p e r c e n t of the
weeds. T h i s statistic was d e t e r m i n e d because e q u a l i n c r e m e n t s
of dosage m a y n o t p r o d u c e e q u a l i n c r e m e n t s of response above
a n d below t h e L D 5 0 position on t h e curve; response is nonlinear w i t h respect to a p p l i e d dosage, a l t h o u g h t h e L D 5 0 position is m o r e reliable t h a n response positions at lower or h i g h e r
dosages.

V O L . 13, No. 8, JANUARY 1966

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Results and Discussion


Percentage
Response
T h e experimental data showing the relationship between
dosage and response (elimination and competitive absence of
weeds) are given in Table 1.
Table 1.Relationship between dosage of PEBC, diallate and PEBC + diallate and
response at each half-dosage distance.

a
b

r Elimination and competitive absence of weeds.


p1 = Percentage control (100 r/n) after Finney (2).

T h e s e results show that the total control obtained from the


p r e p l a n t application of PEBC + diallate at 3 l b / A active ingredient was 24 percentage points higher than the control obtained from diallate alone (Table 1). PEBC applied at 3 l b / A
alone gave 56 percentage points control when compared to the
u n t r e a t e d controls. T h e untreated controls averaged 95 weed
seedlings per square foot or 59 pigweed and 36 foxtail seedlings
at the time the observations were made.
C o m p u t a t i o n s from these data (Table 1) showed that the
average control for PEBC and diallate alone at the 3 l b / A dosage
was 50 percentage points. T h e 50 percentage point averaee was
18 percentage points less than the percentage control obtained
from P E B C + diallate applied at 2 + 1 l b / A active ingredient,
respectively. "When results were compared further, the pnolication
of 1.5 l b / A of P E B C and 1.5 l b / A of diallate srave 20 and 25
percentage points control, respectively, or a total additive control of 45 percentage points (Table IV However, the control
obtained from a 2:1 combination of PEBC + diallate at the

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1.5 l b / A dosage was 40 percentage points. T h u s , t h e additive


c o n t r o l o b t a i n e d from the m i x t u r e was 23 percentage p o i n t s
h i g h e r at t h e 3 l b / A dosage t h a n t h e expected c o n t r o l o b t a i n e d
from single chemicals a d d e d at the 1.5 + 1.5 or 3 l b / A dosage
( T a b l e 1).
Regression
Analysis
T h e results of t h e regression analysis of dosage a n d response
are given in F i g u r e 1, shown below.

50.0 0

E x a m i n a t i o n of t h e t h r e e regression lines shows t h a t P E B C


-f- diallate c o u l d be expected to be significantly m o r e effective
in weed c o n t r o l t h a n t h e single herbicides, particularly w i t h i n
t h e dosage r a n g e below 3.9 l b / A active i n g r e d i e n t (Figure 1).
At dosages in excess of 3.9 l b / A , P E B C a p p l i e d alone was m o r e
effective, a l t h o u g h c r o p selectivity w o u l d l i m i t usage at dosages
above the 5 l b / A rate. Diallate was shown to be less effective
t h a n P E B C + diallate o r P E B C above t h e 1.75 l b / A rate, b u t
diallate h a d m o r e effectiveness t h a n P E B C b e l o w t h e 1.75 l b / A
dosage of active i n g r e d i e n t (Figure 1). It is k n o w n t h a t dosages
of diallate b e t w e e n 1.5 to 2 l b / A are sufficient for t h e c o n t r o l
of wild oat (Avena fatua).
Calculation a n d i n t e r p o l a t i o n showed t h a t 8 4 percentage
points c o n t r o l o c c u r r e d at 4.9, 5.9 a n d 8.75 l b / A active i n g r e d i e n t

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for P E B C , PEBC + diallate and diallate, respectively. In Figure


1, the probit of control (Y) at 5 is equivalent to 50 percentage
points of control and (Y) at 6 is equivalent to 84 percentage
points.
Inspection of the plotted LD50 positions on individual regression lines shows that the mixture was significantly more
effective in the control of weeds than the single chemicals. For
instance, the LD50 dosage for PEBC + diallate was 1.75 l b / A
while the 50 percentage response point for PEBC a n d diallate
occurred at 2.5 and 3.1 l b / A active ingredient, respectively.
T h e s e dosage comparisons show that an additive response was
obtained by applying PEBC + diallate at dosages up to 3.9
l b / A in combination.
Apparently, these data indicate that PEBC + diallate would
be more active u n d e r broad spectrum weed conditions and u n d e r
the equilibria present in some soils than the single chemicals.
Conversely, although of significant effectiveness per unit of active
ingredient above 2.5 lb/A, PEBC had a relatively narrow activity
range u n d e r the conditions of the experiment (Figure 1).
T h e s e interpretations and inferences were derived from the
relative characteristics, slope and position, of the regression lines.
For example, the relatively flat diallate regression line suggests
that increased dosages of diallate alone would have less effect
on species control than that of PEBC. T h e line intersection of
P E B C with diallate at approximately the LD50 position of
the m i x t u r e showed that a positive interaction occurred for
species a n d chemicals. On the other hand, parallelism of two
regression lines indicates that the chemicals act independently
of each other or exhibit proportionate species activity. Likewise,
Gowing (1) inferred that parallel dosage-response regression
lines were directly comparable perhaps because of similar modes
of activity of the chemicals.
Summary and Conclusions
T h e probit method of statistical analysis after Finney (2)
was employed, to evaluate the relative effectiveness of three
herbicides applied variable-dosage.
T h e results showed: 1) T h e herbicide combination, PEBC
+ diallate, was more effective in weed control than the single
chemicals P E B C and diallate within the dosage range from 1.75
to 3.9 l b / A active ingredient; and 2) inspection of the dosageresponse regression lines and the LD50 dosages indicated that
an additive effect in weed control was obtained from the herbicide combination.

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T h e s e specific h e r b i c i d e results gave evidence t h a t t h e m e t h o d


of p r o b i t analysis was a d a p t a b l e to variable-dosage h e r b i c i d e
studies to define: 1) X h e significance of small differences; 2)
p r o b a b l e d e p e n d e n t or i n d e p e n d e n t activity of a h e r b i c i d e ; 3)
relative effectiveness of a h e r b i c i d e c o m b i n a t i o n ; 4) t h e p r o b able dosage a n d response r e l a t i o n s h i p of a h e r b i c i d e ; 5) t h e
L D 5 0 value; 6) c r o p selectivity; a n d 7) t h e e r r o r of t h e dosageresponse estimate.
L i t e r a t u r e Cited
(1) GOWING, D. P. 1959. A m e t h o d of comparing herbicides a n d assessing
herbicide mixtures at the screening level. Weeds. 7: 66-76.
(2) FINNEV, D. J. 1952. Probit analysis. Cambridge U n i v e r s i t y P r e s s .
Second Edition.
(3)

SULLIVAN, E. F., R. L. ABRAMS a n d R. R. W O O D .

1963.

W e e d control

in sugar beets by combinations of thiolcarbamate herbicides. Weeds.


11: 258-260.
(4)

SULLIVAN, E. F., R. L. ABRAMS, W. R. WAGNER, and R. R. W O O D .

Preplant and postemergence herbicide


1964. Res. R p t . N C W C C 2 1 : 73-80.

screening on

1964.

sugar beets,

A Regression Study on Tare Samples of Sugar Beets


In Relation to Factors Influencing Productivity
and Quality
GEORGE

K.

RYSER 1

Received for publication April 12, ig6<j

Introduction
Many factors affect the yield and quality of the sugar beet.
Researchers have studied various factors separately and others
collectively. T h e ultimate in sugar production cannot be attained
until all the important factors of growth and their interactions
are recognized, measured and controlled.
Multiple regression procedures can be used to establish an
equation for the prediction of maximum sugar yields. Modern
computers can easily handle complicated multiple models involving large matrices including linear and nonlinear relationships a n d interaction terms (7) 2 .
In 1958 a n d 1959, the Nyssa-Nampa Beet Growers Association
made sugar determinations on tare samples of roots from each
load delivered to the Doles Station, Nyssa, Oregon. T h i s was
an u n d e r t a k i n g of the growers in the district with the major
objective of determining some factors influencing the sugar percent of the sugar beet.
Materials and Methods
For the harvest seasons of 1958 and 1959, Dr. J. H. Roblyer 3
set up a laboratory close to the Doles station and each tare sample
of roots was analyzed for sucrose percent. A sample of 26 gm
of p u l p was digested in 177 mm of water, cleared with H o m e ' s
lead subacetate and filtered. T h e sugar percent of the clear solution was determined by use of a saccharimeter 4 . Average suo^ir
percent was calculated for each field from these root samples.
Yield of roots was obtained from factory records. Acreable yield
of roots and gross sugar were calculated for each grower.
All growers were asked to supply information on each field
of sugar beets delivered to the station. In 1959 the questioni Research Agronomist, Crops Research Division, Agricultural Research Service, U. S.
Department of Agriculture, Logan, Utah.
2
Numbers in parentheses refer to literature cited.
3
Head, Chemistry Department, College of Idaho, Caldwell, Idaho, and Chemist for
Nyssa-Nampa Growers Association.
* Loaned to the Nyssa-Nampa Growers Association under Memorandum of Understanding with the Crops Research Division, Agricultural Research Service, U. S. Department of
Agriculture.

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729

naire covered information as to acres planted, date planted, soil


type, stand, type of spring work, units nitrogen a n d phosphate
applied, previous cropping history, dates of first and last irrigation, including n u m b e r of irrigations and evidence of considerable field flooding. In 1958 the questionnaire did not include
plant stand, type of spring work or n u m b e r of irrigations.
For both years Dr. J. H. Roblyer supplied data on sugar a n d
gross yield, together with all information in the grower's questionnaire, for statistical analysis. T h i s information was coded,
when needed, a n d punched on IBM cards (12). Tables of means
were obtained from various card sortings (Tables 1 to 6). T h e s e
tables vary in the n u m b e r of observations, because complete Information on the variables was not reported by all the growers.
Table 2.Means of groups on five different cultural treatments, Doles Station, 1958.

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Table 3.Means of groups on soil type and crop rotation, Doles Station, 1958.

Six variables t h a t seemed to be the most accurate a n d apparently to have some influence on t o n n a g e or sugar p e r c e n t
were selected for the regression study. C o n s i d e r a t i o n also was
given each year to only those fields w h e r e i n c o m p l e t e information for t h e six variables was available. T h i s r e d u c e d the observations to 85 for 1958 a n d to 83 for 1959. T h e six i n d e p e n d e n t
variables w e r e : (A) acres harvested; (B) u n i t s n i t r o g e n applied;
(C) u n i t s P 2 O s a p p l i e d ; ^D) n u m b e r of irrigations, coded 1 to
4 in increasing o r d e r (1959) or d a t e of first i r r i g a t i o n coded 1
to 4 earliest d a t e first (1958); (E) date of p l a n t i n g g r o u p e d 0 to
7, 0 assigned to earliest d a t e a n d 7 to the latest; a n d (F) total
n u m b e r of g r o w i n g days. T h e g r o w i n g days were estimated as
t h e days from p l a n t i n g to the date t h a t the greatest n u m b e r of
loads of beets was delivered to t h e w e i g h i n g station.
Xo o b t a i n a regression m o d e l , the first step w o u l d be to
study two-dimensional a n d three-dimensional scatter diagrams.
A m o r e realistic m o d e l can be w r i t t e n w h e n these are observed,
a n d a t t h e same t i m e m u c h a d d i t i o n a l i n f o r m a t i o n can b e obt a i n e d a b o u t t h e i n d e p e n d e n t variables.
In o r d e r to project a regression m o d e l as precisely as possible,
a three-dimensional g r a p h i c analysis was m a d e 5 (8). T h e p u n c h e d
cards t h a t carried values for t h e six i n d e p e n d e n t variables a n d
for the d e p e n d e n t variables (Yx, gross sugar p e r acre, Y 2 , tons of
roots p e r acre a n d Y 3 , sugar p e r c e n t ) were sorted i n t o ascending
5
Machine method developed by Dr. Rex Hurst, Head, Applied Statistics Department,
Utah State University, Logan, Utah.

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Table 5.Means of groups on five different cultural tretatments, Doles Station, 1959.

o r d e r for each of t h e i n d e p e n d e n t variables a n d recoded by


thirds. N u m b e r 1 was assigned to the lowest t h i r d ; 2, to t h e
n e x t lowest; a n d 3, to t h e highest. T h e n by c o n t r o l l i n g for all
possible two-way c o m b i n a t i o n s of t h e r e c o d e d i n d e p e n d e n t variables, cell totals of the d e p e n d e n t variables were o b t a i n e d for
each two-dimensional, i n d e p e n d e n t variable c o m b i n a t i o n . T h i s
r e d u c e d t h e g r a p h i c analysis to p l o t t i n g only n i n e p o i n t s for
each two-dimensional c o m b i n a t i o n a n d 15 three-dimensional
graphs for each d e p e n d e n t v a r i a b l e (8).
After s t u d y i n g t h e graphs, several m u l t i p l e regression models
w e r e selected for t h e 1959 d a t a a n d R 2 values w e r e o b t a i n e d .
S i mple correlation values, m e a n s , s t a n d a r d deviations a n d coefficients of variation were also calculated.

VOL. 13, No. 8, JANUARY 1966

73!

Table 6.Means of groups on six additional cultural treatments, Doles Station, 1959.

E x p e r i m e n t a l Results
General observations from means for 1958
T h e scatter diagram of average tons roots per acre and average sugar percent shows the production of each field with respect
to the general m e a n of the two variables in 1958 (Figure 1).
Of the 5 0 % of the fields that produced below-average tons of
beets p e r acre (quadrants 2 and 3), 3 2 % had the highest sugar
percent (mean 16.9) b u t produced only 3.963 tons of gross sugar
per acre ( T a b l e 1). T h e other 1 8 % had a mean sucrose of 15.7%
sugar a n d 4.210 tons of gross sugar per acre. T h e fields (quadrants
2 a n d 3) with the highest and lowest average sugar percent also
p r o d u c e d the lowest and next to lowest average gross sugar per
acre.
Of the 5 0 % of all the field that produced above-average
tons of roots per acre, 2 9 % (quadrant 4) produced below-average
percent (15.68) a n d produced next to the highest gross sugar
per acre (5.235 tons). T h e other 2 1 % (quadrant 1) produced
the highest tons of roots p e r acre with above-average sugar
percent (16.72) and highest gross sugar per acre (5.384 tons).

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TONS PER ACRE

Figure 1.Scatter diagramDoles Station, 1958.

Gross sugar p e r acre was influenced largely by tons o roots p e r


acre, n o t by sugar percent. Above-average yields d i d n o t t e n d
to show a decrease in sugar p e r c e n t u n t i l applications of n i t r o g e n
exceeded 150 u n i t s of N p e r acre, as in q u a d r a n t 4 ( T a b l e 1).
Q u a d r a n t 3, w i t h n e x t to t h e highest n i t r o g e n a p p l i c a t i o n (161
u n i t s N ) , p r o d u c e d below-average tons p e r acre a n d , as expected,
below-average sugar percent.
Acres harvested 1958
Fields of 5 to 10 acres averaged 29.06 tons roots p e r acre,
a n d fields above 30 acres averaged 30.57 tons roots p e r acre.
T h e small difference, p r o b a b l y is n o t significant b u t shows t h a t
field practices may have b e e n different ( T a b l e 2).
Fertilizer applied
1958
U n i t s of n i t r o g e n a n d u n i t s of p h o s p h a t e p e r acre showed
very little differences because t h e lowest a p p l i c a t i o n seemed
to be as effective as the highest ( T a b l e 2). T h e residual fertility
in t h e various fields was n o t k n o w n a n d could have b e e n a
factor. T h e association o f high t o n n a g e w i t h h i g h P 2 0 5 was
n o t significant, because only two fields fell i n t o this g r o u p .
Date of last irrigation or number of irrigations
I n 1958 t h e r e were only slight differences d u e t o t h e n u m b e r
of irrigations or to late watering. Average sugar p e r c e n t d i d
show increases in favor of late irrigations ( T a b l e 2).

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Planting
date
T h e dates of planting in 1958 ranged from March 1 to
April 25. T h e earliest plantings, however, did show some improvement over the latest in average tonnage a n d average gross
sugar. Intermediate dates of planting, March 21 to April 3,
gave the highest yield of roots and gross sugar ( T a b l e 2).
Other
observations
Soil type was difficult to judge from simple observations.
M e d i u m to heavy soil types with 32.6 tons of roots per acre
and 15.83% sugar produced the highest gross sugar or 5.134
tons ( T a b l e 3).
Beets following grain produced 30.7 tons of roots per acre
with 16.32% sugar. Beets following potatoes ranked next with
30.4 tons of roots per acre and 16.15% sugar (Table 3).
General observations from means for 1959
T h e scatter diagram (Figure 2) shows how the respective
fields placed in relation to the mean yield of tons per acre and
sugar percent of all the fields in 1959. Q u a d r a n t 1, which is
above average for both sugar percent and tons per acre, yielded
5.160 tons of gross sugar per acre and 31.2 tons roots per acre
with 16.5% sugar (Table 4). Eighty-seven percent of the acres
in q u a d r a n t 1, which comprised 20% of the total in 1959, was
planted before March 21. T h i s gave an average growing season
of 229 days, or the longest growing season of all quadrants. A
good stand of beets was also maintained in these fields. T h e

; 26 >27 28 29 30 31 32 33 34 33 36 37 38 39 40 4) 42
TONS PER ACRE

Figure 2.Scatter diagramDoles Station, 1959.

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average stand of 88 plants p e r 100 feet of r o w is h i g h e r t h a n


t h a t of any o t h e r q u a d r a n t .
Q u a d r a n t 2, with above-average sugar percentage a n d belowaverage tonnage, yielded 3.815 tons of gross sugar a n d 23 tons
of roots p e r acre with 16.6% sugar. T h e s e fields r e p r e s e n t e d
2 4 . 6 % of the total acres p l a n t e d . T h e average stand was 80
plants p e r 100 feet of r o w for this g r o u p . T h e g r o w i n g season
was 217 days, a comparatively short g r o w i n g p e r i o d i n a s m u c h
as 4 0 % of the fields were p l a n t e d before M a r c h 2 1 .
Q u a d r a n t 3, w i t h b o t h low t o n n a g e a n d low sugar percentage
w h e n c o m p a r e d with the m e a n s of all fields, r e p r e s e n t e d 2 4 . 9 %
of t h e total acres p l a n t e d . O n l y 4 8 % h a d b e e n p l a n t e d by
M a r c h 2 1 , which could be t h e m a j o r factor in t h e low yields.
Stands were d o w n to 81 plants p e r 100 feet of row, a n d the
g r o u p p r o d u c e d 3.961 tons gross sugar a n d 23.8 tons of roots
p e r acre with 1 5 . 5 % sugar d u r i n g an average g r o w i n g season
of 210 days.
Q u a d r a n t 4, characterized by high t o n n a g e b u t low sugar
w h e n c o m p a r e d with the m e a n of all fields, p r o d u c e d 4.760 tons
of gross sugar p e r acre, 30.2 tons of roots p e r acre a n d 1 5 . 5 %
sugar. T h i s q u a d r a n t h a d 218 g r o w i n g days, with 6 1 % o f t h e
fields p l a n t e d by M a r c h 2 1 , a n d r e p r e s e n t e d 2 8 . 4 % of t h e total
acres planted. An average of 178 u n i t s of N p e r acre, t h e highest
for all the q u a d r a n t s , could be t h e c o n t r i b u t i n g factor for the
low sugar percentage, especially if t h e n i t r o g e n level in the soil
was h i g h before a p p l i c a t i c n a n d late rains or irrigations m a d e
the n i t r o g e n available late in t h e season. T h e comparatively
good stand of 82 beets per 100 feet of row h e l p e d m a i n t a i n tons
of roots p e r acre.
Acres harvested 1959
Small fields ( u n d e r 20 acres) p r o d u c e d , on an average, 2
tons of roots p e r acre less t h a n fields larger t h a n 20 acres, w i t h
a n increase i n sugar p e r c e n t ( T a b l e 5). T h i s could b e d u e t o
some i n h e r e n t field practice on small fields.
Fertilizer applied
1959
A decrease in sugar percentage was associated w i t h an increase in n i t r o g e n application ( T a b l e 5). T h i s decrease o c c u r r e d
w i t h o u t an increase in average tons of roots p e r acre; in fact,
the highest t o n n a g e was o b t a i n e d from fields with t h e lowest
application of n i t r o g e n . However, t h e initial level of n i t r o g e n
fertility in these fields was n o t k n o w n .
T h e lowest P 2 0 5 a p p l i c a t i o n averaged t h e highest gross sugar
a n d tons of roots p e r acre ( T a b l e 5). T h i s m u s t be d u e to the
level of soil fertility before the applications of P 2 0 5 a n d N
were used.

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Date of last irrigation or number of irrigations


T h e n u m b e r of irrigations was associated with gross sugar
and root yield. T h e highest n u m b e r of irrigations increased
the average yield of gross sugar and roots 0.881 tons and 5.9
tons per acre, respectively, over the lowest n u m b e r of irrigations.
N u m b e r of irrigations had little or no effect on sugar percent
( T a b l e 5).
Date of planting
Early planting, March 1 to March 20, increased production.
T h e s e early plantings averaged 6.71 more tons of roots and
1.165 m o r e tons of gross sugar per acre than the later plantings.
Other
observations
Heavy soils were associated with highest yield of roots per
acre, b u t sugar percent was somewhat higher in sandy soils (Table
6). Slight increases in yield were obtained when the preceding
crops were forage or potatoes (Table 6). An average increase
of one ton per acre was obtained when the sugar beets were
irrigated before thinning, b u t when it was necessary to irrigate
for germination, there was a reduction in yield of 2.3 tons per
acre ( T a b l e 6). Fields that received mechanical work early in
the spring had a one-ton advantage over fields that did not receive this practice (Table 6). T h e r e were indications that late
side dressings in July decreased tonnage, sugar percentage, and
gross sugar (Table 6).
Date of harvest
T h e effect of date of harvest on sugar percent d u r i n g the
delivery period at the Doles station for 1958 and 1959 was
evidenced by the increased sugar percent obtained at the later
harvest dates (Table 7). T h e scatter diagram between date of
harvest a n d daily sugar percent indicates that the relation was
n o t linear over the time included in the study (Figure 3). A

Figure 3.Percent sugar by daysDoles Station, 1959.

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Xable 7.Means of daily sugar percent.

regression model, using dates a n d dates s q u a r e d as i n d e p e n d e n t


variables a n d sugar p e r c e n t as t h e d e p e n d e n t variable, gave an
R 2 value of 0.26. T h i s R 2 v a l u e a n d b o t h t e r m s w e r e highly
significant, giving significance to a c u r v i l i n e a r r e l a t i o n s h i p (3).
Statistical Results
T y p i c a l three-dimensional g r a p h s for t h e 1959 data, s h o w i n g
t h e effect of field practices on yield of roots p e r acre, a r e given
in F i g u r e 4. F i g u r e 4a shows a p r o b a b l e c u r v i l i n e a r r e l a t i o n s h i p
b e t w e e n yield of beets a n d u n i t s of n i t r o g e n a p p l i e d p e r acre.
T h i s p r o b a b l e c u r v i l i n e a r r e l a t i o n s h i p is affected by u n i t s of
P 2 O s a d d e d p e r acre. T h i s suggests t h a t b o t h l i n e a r terms, u n i t s
of n i t r o g e n a n d u n i t s of P 2 O s , a n d t h e i n t e r a c t i o n s (units of

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Figure 4.Typical three-dimensional graphs showing acre yield of


sugar beets, 1959 data.

nitrogen times units of P 2 O 5 and units of nitrogen squared times


units P 2 O 5 ) should be included in the mathematical model. T h e
curvilinear relationship with units of N is not so clear in Figure
4b a n d 4c; however, the interactions units of N times n u m b e r
of irrigations and units of N times length of growing season
should be included. In Figures 4d and 4e, an uncertain relationship is shown between yield and units of P 2 0 5 , b u t the
interaction, units of P 2 0 5 times number of growing days, should
be included in the model.
Figures 4g and 4h show a curvilinear relationship between
yield a n d n u m b e r of irrigations and that the relationship is
affected by date of planting and number of growing days. These
i n d e p e n d e n t variables should be included in the model as linear
a n d quadratic forms, together with all their interactions. T h e
negative relationships are due to the coding of planting date
with n u m b e r one being the earliest date (Figures 4e and 4g).
T h e three-dimensional graphs with Y 1 (gross sugar per acre)
were similar to those with Y 2 (tons of roots per acre), due to
the high correlations of 0.97 and 0.08 between Y 1 and Y 2 (Table
8). T h e surfaces obtained with Y 3 (sugar percent) were quite flat,
probably d u e to the low coefficient of variation and the low
n u m b e r of points plotted.
T h e multiple-linear model gave an R 2 value of 0.270 with
Y 1 (gross sugar per acre), 0.253 with Y 2 (tons roots per acre) and
a small value of 0.184 with Y 3 (sugar percent) (Table 9).
A regression model of 18 terms was then tried (Table 10).
It was felt that the terms included in this model were the ones
most evident from the 15 possible three-dimensional graphs with

-4

Table 8.Simple correlations, 1958-1959.

w
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741

Y 2 (tons roots per acre). T h e " B " model produced an R 2 value


of 0.393, an increase in precision of 13.9% over the multiplelinear model.
Since there were 10 more interaction terms that seemed
probable, a " C " model with all 28 terms was set up a n d an
R 2 value of 0.463 was obtained, an increase in precision of
21.0% over the linear model (Table 11).
W h i l e all the R 2 values gave an increase over linear Model
A, the change to Model C did not improve the relationship
enough to justify the loss of 22 degrees of freedom. T h e F
Table 9.Regression analysis linear model, 1959 data.
Mean 1
square

* Signifirant at 5% point.
** Significant at 1 % point.
1 Single d.f. mean square is that which would not have been accounted for by regression
had the variable been omitted.

Table 10.Regression analysis model B, 1959 data.

>
* Significant at 5% point.
** Significant at 1% point.
1
Single mean (d.f. mean) square is that which would not have been accounted for by regression had the variable been omitted.

CO
C/J

* Significant at 5% point.
** Significant at 1% point.
1
Single d.f. mean square is that which would not have been accounted for by regression had the variable been omitted.

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values were all r e d u c e d for M o d e l C except w i t h Y 3 which barely


reached the 5 % level ( X a b l e 11). T h e models used w e r e n o t
expected to be the c o m p l e t e answer. To o b t a i n this, one w o u l d
have to use a stepwise p r o c e d u r e to find t h e t e r m s a n d interactions of most significance.
Discussion
M u c h has b e e n w r i t t e n o n t h e p r o b l e m s involved i n prod u c i n g a h i g h q u a l i t y sugar beet. T h e c o n t i n u a l d o w n w a r d
t r e n d of recoverable sugar is of great concern to b o t h t h e prod u c e r a n d t h e processor (1) (2) (6) (10) (13) (15). It is felt
that p a r t of t h e p r o b l e m is d u e to t h e single factor a p p r o a c h
of most investigators, w i t h n o t e n o u g h study given to t h e variations a n d interactions caused by local climate, c r o p history
a n d c u l t u r a l practices at each location. Variety differences s h o u l d
always be ionsidered, since h y b r i d s behave differently w h e n they
a r e g r o w n in different locations (9) (10) (11) (14) (15) (16)
(17).
T h e survey type o f a p p r o a c h t o t h e p r o b l e m gives a n opport u n i t y t o establish t h e i n t e r a c t i o n r e l a t i o n s h i p b e t w e e n t h e variables observed or m e a s u r e d . Very close supervision m u s t be
m a d e in o b t a i n i n g t h e data, a n d full c o o p e r a t i o n of each g r o w e r
s h o u l d b e o b t a i n e d . T h i s type o f study w o u l d n o t take t h e
place of c o n t r o l l e d e x p e r i m e n t s , b u t c o u l d give a d d i t i o n a l inf o r m a t i o n a s t o i n t e r a c t i o n s b e t w e e n variables u n d e r n o r m a l
farm c o n d i t i o n s a n d at a n c m i n a l cost, especially w h e n a large
n u m b e r of variables a p p e a r .
T h e simple m e a n s of all t h e variables in t h e Nyssa-Nampa
data d i d show trends, b u t they were n o t significant. H o w e v e r ,
simple correlations in 1958 showed significant negative association b e t w e e n u n i t s of N a p p l i e d a n d sugar p e r c e n t , a n d a positive
association b e t w e e n sugar p e r c e n t a n d d a t e o f p l a n t i n g a n d d a t e
of first i r r i g a t i o n . T h i s d i d n o t h o l d for t h e 1959 data, b u t a
significant correlation was o b t a i n e d b e t w e e n sugar p e r c e n t a n d
n u m b e r o f g r o w i n g days ( T a b l e 8). T h e a u t h o r feels t h a t t h e
correlation b e t w e e n sucrose p e r c e n t a n d beet weight or tons
beets p e r acre is very u n r e l i a b l e , since it is c o n t r o l l e d largely
by t h e season or t h e location or by b o t h . O t h e r s h a v e f o u n d
n o correlation b e t w e e n sucrose p e r c e n t a n d b e e t w e i g h t (9).
T h e most significant i n d e p e n d e n t variables of t h e six used
in t h e regression study w e r e : n u m b e r of i r r i g a t i o n s ; d a t e of
first i r r i g a t i o n ; n u m b e r of g r o w i n g days; a n d d a t e of p l a n t i n g
( T a b l e 9). H a d d o c k f o u n d similar significance (6).
T a k i n g t h e regression m o d e l in its l i n e a r form for 1959
( T a b l e 9), t h e most significant m e a s u r e m e n t s w e r e for Yi, gross

VOL.

13,

No.

8, JANUARY

1966

745

sugar p e r acre, a n d Y2, tons roots per acre, n u m b e r of irrigations


a n d date of planting. For sugar percent the most significant
measurements were planting date and n u m b e r of growing days.
Regression model " B " for 1959 data (Table 10) also gives
significance to the interactions of n u m b e r of irrigations and date
of p l a n t i n g for Y1? gross sugar per acre, or Y,, tons of roots per
acre. Model " C " (Table 11) shows the same significant trends
with Y3, sugar percent. T h e significant interactions between
P 2 O s p e r acre, planting date and growing season with sugar
percent, were surprising, b u t it is possible that the nitrogen
a n d P 2 O s balance was disturbed by the phosphate treatments.
It is clear from the regression models and three-dimensional
graphs that interactions were present, which should be considered, a n d these relationships are not strictly linear. More
work needs to be done, because the regression models used
are not the most significant ones and the F values for models
B and C increased the odds over the linear regression models
with all d e p e n d e n t variables except Y3, sugar percent. It is
evident that in addition to the six field practices evaluated in
this study the association of other factors of production (such
as weather measurements, soil type, soil preparation, cropping
history a n d genetic factors) should be appraised. Elimination
of terms by regrouping those with significant individual F values
a n d those with known correlation values in a systematic and
stepwise procedure could reduce the number of necessary terms
in the regression model (7).
In this study the effect of length of harvest season on sugar
percent was very evident. T h i s effect has also been shown by
others (5) (3) (4). T h e R 2 value obtained, although only .26,
was of very high significance (F value B 1 = 18.05 and B 2 = 15.83).
T h e low R 2 value must be due to the large variation in sugar
percentage within days.
Summary
In this study, the means clearly show that high acreable yield
of gross sugar was d u e to high tonnage of roots and not to
sugar percent. T h i s was substantiated by a highly significant
correlation between gross sugar and tons of roots.
T h e significance of cultural practices (such as units of nitrogen a n d P 2 O 5 , total n u m b e r of irrigations, length of growing
season a n d date of harvest) was established by multiple regression
a n d t h e use of three-dimensional graphs. T h e importance of
interactions of these variables and their relationship to each
o t h e r is shown.

JOURNAL

746

OF THE A.

S.

S.

B.

X.

T h e use of regression models on survey type of data, to


establish t h e variables of most i m p o r t a n c e , is suggested.
Literature Cited
(1)

DAVIS, J . F., W . B. SUNDQUIST a n d M. G. FRAKES.

1959.

T h e effect of

fertilizers on sugar beets including economic optima study of the


response. J. Am. Soc. Sugar Beet Technol. 10 (5) : 424-434.
(2)

DUDLEY,

JOHN

W.

and

LEROY

POWERS.

1960.

Population

studies on sodium and potassium in sugar beets


J. Am. Soc. Sugar Beet Technol. 11 (2) : 97-127.
(3)

FINKER, R.

E., J. F.

SWINK,

C. W.

DOXTATOR,

R.

F.

genetic

(Beta vulgaris).

OLSON

and

P.

C.

HANZAS. 1959. Changes in raffinose content and other characteristics of sugar beet varieties during six different harvest dates. J.
Am. Soc. Sugar Beet Technol. 1 0 ( 5 ) : 459-465.
(4)

FRIEHAUF, R. E., H . L. BUSH a n d E. E. R E M M E N G A .

1963.

Correlations

of preharvest samples and cultural practices with final yield and


quality of sugar beets. J. Am. Soc. Sugar Beet Technol. 1 2 ( 4 ) :
273-283.
(5)

HADDOCK, J. L., P. B. S M I T H , A.

R.

DOWNIE,

J.

T.

ALEXANDER,

B.

E.

EASTON and VERNAL JENSEN. 1959. T h e influence of cultural practices on the quality of sugar beets. J. Am. Soc. Sugar Beet Tech.
10(4) : 290-301.
(6) HADDOCK, J. L. 1963. T h e interrelationships between nitrogen a n d
potassium on the quality of sugar beets. 14th A n n u . Fert. Conf.
Proc. of the Pacific Northwest, Idaho Falls, Idaho (Mtg.) : 77-88.
(7) HURST, R. L. 1963. Model building in multiple regression. 1620
general program library 06.0084, December 1963.
(8) HURST, R. L. and M. W. PEDERSEN, 1964. Alfalfa seed production as a
function of genetic a n d environmental characteristics. Advancing
frontiers of p l a n t sciences. VIII: 41-54.
(9)

MCALLISTER, D E V E R E , R E X L. HURST, DONALD C. W O O L L E Y , H A R O L D


M. NIELSEN, L. E L M E R OLSON, DELBERT A. GREENWOOD, H O M E R M.
L E B A R O N a n d W I L L I A M H . BENNETT. 1961. T h e variability of sugar

constituents as influenced by year, location, variety, a n d nitrogen


fertilization. J. Am. Soc. Sugar Beet Technol. 11 (7) : 547-564.
(10)

OWEN,

F.

V.,

MYRON

STOUT,

ALBERT

M.

MURPHY,

C.

H.

SMITH

and

G. K. RYSER. 1960. Interaction of components of impurity and


location in hybrids from inbred lines of sugar beets. J. Am. Soc.
Sugar Beet Technol. 11 ( 1 ) : 37-43.
(11)

RYSER,

GEORGE

K.,

MYRON

STOUT, ALBERT U L R I C H

and

F.

V.

OWEN.

1958. Some chemical and physiological characteristics of inbred


lines of sugar beets. J. Am. Soc. Sugar Beet T e c h n o l . 10 (6) : 525-543.
(12)

RYSER, GEORGE K. a n d BLISS H . CRANDALL.

1952.

A d a p t a t i o n of t h e

IBM card system to analysis of sugar beet varietal data.


Soc. of Sugar Beet Technol. 7: 421-425.
(13)

STOCKINGER,

K. R., A. J .

M A C K E N Z I E a n d E. E.

CARY.

Proc. Am.
1963.

and quality of sugar beets as affected by cropping systems.


Soc. Sugar Beet Technol. 12 (6 : 492-502.

Yield

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(14) STOUT, MYRON. 1954. Determining respiration rate and sampling for
chemical analysis of sugar beets. J. Agr. and Food Chem. II (26) :
1324-1328.
(15) T O L M A N , BION. 1964. Let's grow more pure sugar per acre of beets
planted. Idaho Farmer, July 2, 1964.
(16) ULRICH, ALBERT. 1961. Variety climate interactions of? sugar beet
varieties in simulated climates. J. Am. Soc. Sugar Beet Technol.
11 (5) : 376-387.
(17) ULRICH, ALBERT. Variability of sugar beet plants grown in pots without competition for light, water and nutrients. J. Am. Soc. Sugar
Beet Technol. 11 (7) : 595-604.
(18) WOOLEY, D. G. and W. H. BENNETT. 1962. Effect of soil moisture,
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12(3) : 233-237.

NOTES SECTION

Pollen Viability Determination W i t h


Tetrazolium Bromide 1
T h e p u r p o s e of this research n o t e is to r e p o r t an i m p r o v e m e n t i n the t e t r a z o l i u m b r o m i d e t e c h n i q u e for d e t e r m i n a t i o n
of sugar beet p o l l e n viability. H e c k e r (1) originally r e p o r t e d
t h a t a 0 . 5 % s o l u t i o n of 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl
t e t r a z o l i u m b r o m i d e at 20 C p r o v i d e d a specific a n d r a p i d m e a n s
of d e t e r m i n i n g the viability of m a t u r e sugar beet pollen. H o w ever, considerable cell r u p t u r e occurred, r e s u l t i n g in difficulty in
discerning r u p t u r e d p o l l e n cells from non-stained pollen cells.
T h i s p r o b l e m of p o l l e n cell r u p t u r e is avoided by u s i n g t h e
tetrazolium b r o m i d e in a sucrose s o l u t i o n of a p p r o x i m a t e l y t h e
same osmotic c o n c e n t r a t i o n as the cytoplasm. A 0 . 5 % s o l u t i o n
of tetrazolium b r o m i d e in 4 0 % sucrose solution successfully
e l i m i n a t e d t h e p r o b l e m . T h i s allows greater l a t i t u d e i n t h e p e r i o d
of e x a m i n a t i o n , w h i c h can be m a d e a n y t i m e after 5 m i n u t e s .
T h e solution m a i n t a i n s its s t a i n i n g ability for at least 90 days.
Some p o l l e n sources stain b e t t e r at lower t e t r a z o l i u m b r o m i d e
c o n c e n t r a t i o n s (as low as 0.05%) b u t take p r o p o r t i o n a t e l y l o n g e r
to react. Refrigeration of the solution will largely e l i m i n a t e t h e
p r o b l e m of m i c r o o r g a n i s m c o n t a m i n a t i o n a n d d e c o m p o s i t i o n .
It is c o n v e n i e n t to m a i n t a i n a stock s o l u t i o n of t e t r a z o l i u m brom i d e which, p r i o r t o use, can b e m i x e d w i t h distilled w a t e r
a n d sucrose.
In use it was f o u n d most c o n v e n i e n t to d r o p t h e t e t r a z o l i u m
bromide-sucrose s o l u t i o n on t h e p o l l e n grains on a glass microscope slide, m i x slightly, cover w i t h a glass cover slip, a n d set
aside a t 2 0 C i n daylight u n t i l e x a m i n e d . M a t u r e p o l l e n assumed
to be viable was stained p u r p l e to d e e p - p u r p l e , whereas n o n v i a b l e a n d a b o r t i v e p o l l e n was n o t stained. T h e m e t h o d provides
a specific a n d r a p i d m e a n s of d e t e r m i n i n g t h e viability of m a t u r e
sugar beet pollen.
L i t e r a t u r e Cited
(1) H E C K E R , R. J. 1963. Use of tetrazolium salts in determining viability
of sugarbeet pollen. J. Am. Soc. Sugar Beet T e c h n o l . 12(6) : 521-528.
Richard J. Hecker, Research Geneticist
Crops Research Division, ARS, USDA
Salinas, California
1
Joint contribution of the Crops Research Division, Agricultural Research Service,
U.S. Department of Agriculture, the Colorado Agricultural Experiment Station, and the
Beet Sugar Development Foundation. Approved by the Colorado Agricultural Experiment
Station for publication as Scientific Series Article No. 1006.

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