INTRODUCTION

Status of Sericulture in India

India is the second largest silk producing country in the world next to China, and

accounts for 16 % of the global production of silk. More than six million people are engaged

directly or indirectly in this highly remunerative agro-based cottage industry (Dandin et al.,

2003). Though India ranks second in the world with regard to silk production, it is the highest

producer of silk among the tropical countries.

The agro climatic conditions in many parts of India are favourable for the luxuriant

growth of mulberry facilitating silkworm rearing throughout the year. The major traditional

mulberry growing states in India are Karnataka, Andhra Pradesh, West Bengal, Tamil Nadu

and Jammu and Kashmir apart from some non-traditional states like Kerala, Uttar Pradesh,

Maharashtra, etc. However, the production of mulberry raw silk is mainly confined to the

states of Karnataka, Andhra Pradesh, Tamil Nadu, West Bengal and Jammu & Kashmir,

which together account for about 98 % of the country’s total mulberry silk production (Dandin

et al., 2003).

Silkworm (Bombyx mori L.)

Silkworms are monophagous insects, which complete their life cycle only by feeding

the mulberry leaves (Plate 1). It was estimated that about 60 % of the cost of cocoon

production goes towards mulberry leaf production (Yokoyama, 1962; Anonymous, 1997). The

silkworm synthesizes the silk proteins, fibroin and sericin, in the silk glands. These are directly

derived from the proteins and amino acids available in the mulberry leaf, on which the worm
feeds during the larval period lasting 25 - 30 days (Ullal and Narasimhanna, 1987). Similarly,

65 - 70 % of the silk protein produced by the silkworm is directly derived from the protein of

the mulberry alone (Fukuda et al., 1959; Rangaswamy et al., 1976).

It is estimated that the silkworms emerging from 100 diseased free layings (50,000

larvae), consume more than 1.5 tonnes of mulberry leaf to produce 65 - 70 kg cocoons

(Kawakami, 2001).

Mulberry leaves of selected maturity and quality are repeatedly harvested for feeding

the silkworms in accordance with the requirements at different instars of the silkworm larvae.

Mulberry leaf is a major economic component in sericulture since the quality and

quantity of leaf produced per unit area has a direct bearing on cocoon harvest (Dandin et al.,

2003). More than 60 % production cost of silk is attributed to mulberry cultivation alone

(Yokoyama, 1962; Anonymous, 1997). Hence, cultivation of mulberry is the most crucial

factor in sericulture for the production of lustrous silk.

Mulberry (Morus spp.)

Mulberry (Morus spp.), the sole food plant of silkworm is a perennial, deep-rooted, fast

growing tree species and is widely adaptable to different agro climatic conditions. It produces

protein rich foliage and is believed to have had its origin on the lower slopes of the Himalayas.

The genus Morus belongs to the family Moraceae, having 68 species growing as tree

in wild and cultivated forms in different countries of the world (Sanjappa, 1989). But only five

species of Morus viz., M. indica, M. alba, M. bombycis, M. sinensis and M. multicaulis are

potentially important as food for the silkworm.

In India, mulberry is cultivated under a wide range of agro climatic and soil conditions

(Plate 1). Post 1990, the leaf yield level of mulberry has increased from 30 - 35 tonnes/ha/yr
and reached the present exorbitant level of 50 - 60 tonnes. This increase has been brought

about by the development and release of new high-yielding varieties, and has resulted in

higher cocoon production and crop stability (Dandin et al., 2003).

Further, mulberry is perhaps one of the very rare tree species, which can serve all the

important requirements of mankind namely food, medicine, fodder, fibre, fuel etc., and is

known a wonder plant (Chowdary et al., 2009a; Chowdary and Mala, 2009b). The mulberry

leaf plays a major role (38.2 %) among the factors contributing for a successful silkworm

cocoon crop to obtain good quality of raw silk (Miyashita, 1986).

Various research institute of Central Silk Board in the country has evolved high yielding

mulberry varieties suitable for the different agro climatic conditions of the country. Further, it is

very well established the requirement of higher doe of fertilizers for maximizing the production

of quality mulberry leaves to the tune of 50 – 60 tonnes/ha/yr. This in turn has increased

cocoon productivity and reduced the leaf-cocoon ratio in southern states of India (Dandin et

al., 2003).

For increased qualitative mulberry leaf production various organic fertilizers (farm yard

manure, green manures & oil cakes), chemical fertilizers (nitrogenous, phosphatic and

potassium), biofertilisers (nitrogen fixers &, phosphate solubulizers) and plant growth

promoters (salicylic acid, seriboost, triacontanol, zinc sulphate, ferrous sulphate) have been

recommended during mulberry cultivation. Simultaneously, bioformulations of different

antagonistic microbes (Pseudomonas fluorescens, Trichoderma harzianum and T. viride)

have also been recommended for crop protection especially for soilborne diseases (Dandin et

al., 2003; Sharma et al., 2003).

The present project work has been carried out with the following objective:
  To evaluate the morpho physiolgoical variations in some of the elite mulberry

genotypes.

MATERIALS, METHODS & RESULTS

I. General procedures of cleaning glassware

Glassware such as beakers, conical and round bottom flasks, test tubes, watch

glasses, pipettes, etc. were thoroughly washed in detergent solution (0.1 %) after rinsing with

potassium dichromate (K2Cr2O7). Then this glassware were washed with several rounds of

clean water, was kept on a draining rack for a few hours. Finally the drying of the glassware

was done in an oven at 110 – 120°C for 1h. Further, sterilization of glassware was done by

keeping them in a hot air oven at 160°C for 1h.

II. Morphological parameters
The plant growth parameters were recorded on 65th day after pruning of experimental

plot maintained under irrigated conditions as per the recommended package of practices

(Dandin et al., 2003). Data on plant growth parameters (5 plants randomly selected) includes

average plant height (cm), number of branches/plant, number of leaves/plant, average leaf

yield/plant, moisture content of leaf were estimated as follows.

(a). Average plant height (cm)

The length of each shoot of the plant was measured from the plant’s crown (20 cm

above the ground level 0) to the tip of each plant and the average plant length was expressed

in cm.

(b). Number of branches/plant

The total numbers of branches/plant were counter and tabulated.

(c). Number of leaves/plant

The total numbers of leaves/plant were counter and tabulated.

(d). Average leaf yield/plant

The leaf yield/plant assessed by harvesting all the leaves from plants and fresh weight

was taken. From the fresh weight the leaf yield/plant was calculated by using the following

formula:

Total fresh weight of leaves from five plants (g)

Leaf yield/plant (g) =
5

(e). Determination of leaf moisture content

Fresh mulberry leaves from the longest shoot of the plant of each genotype were

collected from ten randomly selected plants in pre-weighed polythene bags and closed with
rubber bands to avoid loss of moisture from the leaves. Immediately after transportation to the

laboratory, the samples were weighed for fresh weight along with pre-weighed polythene

bags. The fresh weight of leaf samples was then determined by subtracting the weight of the

polythene bags. The leaf samples were then dried in a hot air oven at 65 – 70°C temperature

till a constant weight was arrived. Three replications were maintained. The percentage of leaf

moisture was then calculated by using the following formula on fresh weight basis

(Anonymous, 1990).

Fresh weight of leaf – Dry weight of leaf

Leaf moisture (%) = × 100
Fresh weight of leaf

Table1: Growth and yield parameters of elite mulberry genotypes under

irrigated conditions
III. Physiological parameters
Measurement of leaf water potential in different
SL NO VARIETIS HEIHGT NUMBER NUMBE LEAF MOISTURE
OF THE OF R OF YIELD / %
PLANT BRANCHE LEAVES/ PLANT
(cm) S /PLANT PLANT
1 V1 126.7 16 247 0.650 75.5
2 S 1635 120.2 13 176 0.350 70.7
3 MYSORE 103.5 18 254 0.293 60.5
LOCAL
4 RC 1 102.5 15 182 0.385 70.2

5 RC 2 108.3 17 189 0.450 75.3

6 S 13 106.9 17 173 0.350 70.5
7 S 34 105.4 18 165 0.363 60.8

Mulberry varieties

Aim: To measure the leaf water potential energy of water\unit mass of water in a biological

system.

Principle In a thermodynamic sense it is an indicator of energy stands of water in system.

Water from soil absorbed by mulberry roots and conducted up warded against gravitational

pull to the apex of the plant. There by disturbing the water to different organs only due to

gradients in water potential. Water potential is a drying force for SPAC< soil plant

atmospheric condition>. Water potential is indicated on “ψ. Water potential depends on y

component potentials under the same atmospheric pressure and temperature which are

pressure potential; osmotic potential or solute potential; gravitation potential and metric

potential shown as below ψw= ψg+ψop+ψmp+ψpp

Gravitational potential: The position of water in the system.

Osmotic potential: Dissolved solutes in water.

Metric potential: Metric forces, which bind water in the system i.e., soil or leaf Tissue.

Pressure potential: Hydrostatic pressure or trigger pressure in the system.

Overall water potential is a tool to axes water stress and plant metabolism. Water potential

controls the movement of water. Water potential is measured in terms of pressure units like

bars, MPa and Kilo Pascal’s

10 Bars =1 MPa

10 Mpa= 1 KPa

Water potential in the biological system is always a negative

(-), With reference to the pure water the water potential is “o” Hence, measurement of leaf

water potential in mulberry is important to understand biomass production.

Materials:

 Index leaves of different mulberry varieties (V-1, S-36, S-34, Mysore Local, S-13, RC-1

and RC-2).

 Blade

 Water potential cups (4 cm diameter)

Instruments: Dew point potential meter (Model wp-4) Decagon devices. Inc.

This is the fastest instrument for measuring water potential giving reading directly in

MPa. It measures water potential of both leaf as well as the soil from 0to 60 MPa. With

accuracy of MPa. WP-4 uses the chilled mirror dew point technique to measure the water

potential of the sample. Instrument contains led indicator light on left side and functional keys

on right side. Sample drawer and LCD (liquid crystals display). It is a portable instrument.
Procedure:

Index mulberry leaves of different verities were harvested and kept on Plastic covers and

brought to the laboratory. After wiping the surface with tissue paper to remove dust circular

disc of 4cm diameter were cut with blade by the use of sample cups from 3 sides of each leaf

and 3 replication were maintained. Leaf disc were kept in for temperature equilibrium on the

instrument for 5-10 mins, by keeping inside cups. Later on the lids are removed and cups

were put into the drawer and the water potential measurement were taken. The water

potential of leaf disc samples were measured by relating the sample water potential reading

to the vapour pressure of air in equilibrium. WP-4 measures water potential by equilibrating

the liquid phase of water with vapour stage of water in the closed chamber with the help of

sensor block, a dew point sensor, and temperature sensor and infrared thermometer. From

these measurement the vapour pressure of the air is computed at dewpiont temperature in

the water potential of samples in equilibrium and water potential readings of samples

displaying on LCD.

Result: The measurement of leaf water potential of different mulberry samples, which are

noted on mega, Pascal’s (Mpa) are below in the table.

Inference: It is inferred that Mysore local is having highest water potential followed by S-13,

S-36, V-1, RC-2, S-34 varieties.

Table2: Water potential in different elite mulberry genotypes

Varieties Tempe Avg PP Avg MPa Avg

rature
V-1 31.0 31.1 40.83 4.84 -6.65 6.64

31.1 40.85 -6.90

31.2 .0.83 -6.54

S-36 1.5 31.53 4.78 4.87 -5.78 -6.67

1.5 4.94 -8.45
1.6 4.90 -5.78

S-34 31.3 3 4.75 4.78 -5.48 -6.10

1.66
31.5 4.95 -8.66
31.3 4.63 -4.16

Mysore 31.7 3 4.93 4.93 -8.27 -8.15

local 1.7
31.7 4.94 -8.37
31.7 4.91 -7.83

S-13 31.8 3 4.86 4.91 -7.02 -7.83

1.9
31.9 4.93 -8.13
32.0 4.94 -8.34
 RC-1 32.1 3 4.72 4.74 -5.05 -5.34
2.1
32.1 4.75 -5.06
32.1 4.76 -5.39
RC-2 32.1 3 4.80 4.82 -6.09 -6.44
2.03
32.1 4.82 -6.36
31.9 4.85 -6.86

b) Measurement of leaf relative water content and

water saturation deficit in different mulberry verities

Aim: To determine the leaf relative water content and water saturation deficit in different
mulberry varieties.

Principle: Water is the important input for mulberry growth and crop productivity. The entire

plant water relation depends upon the contents of water on the plant system .All

physiobiochemicall activities depend on the plant water status. Water status in the plant can

be measured in terms of leaf moisture content and moisture retention capacity and relative

water content (RWC). Relative water content is defined as water or moisture content of

mulberry leafs in relation to absorption and retention capacity of the plants for physiological

efficiency. RWC is an intergraded measure of plant water status in composition to turgidity.

Relative water content is a physiological tool to understand plant water status, which directly

reflects the physiological mulberry. Water saturation deficiency is a indicator of percent water

reduction to its total turgidity. Cell turgidity reflects division and differentiates cells and growth

of the plants.

Materials:
Index leaves of different mulberry varieties (V-1, S-36, S-34, Mysore local, RC-1, RC-2)

 Sample Cup

 Petriplates

 Blade.

 Electronic weighing machine.

 Distilled water.

 Hot air oven.

Procedure:

Index leaves of different mulberry varieties are harvested and brought to the laboratory by

keeping in polythene cover. The leaves are wiped out with tissue paper to remove dust. With

the help samples cup 4 cm diameter 6 leaf discs was cut from each variety. Soon after this

the fresh weigh of the leaf discs are weighed in electronic beam balance and the leaf disc

were immersed in distilled water on petriplates under laboratory condition with ambient light

for 3hours. After, the leaf discs were taken out in the petriplates and water are wiped out by

using blotting paper. Then the turgid weights of leaf discs are recorded with the help of

electronic balance. Then the leaf disc are put in sample covers and kept in hot air oven for 24

hours to get constant dry weight for each sample variety. The oven dried (60 c) leaf disc was

measured for the dry weight of the sample.

Result: The leaf relative water content and water saturation deficit of different verities are as

follows.
Table 3: Relative water content water saturation deficit in different elite
mulberry genotypes

Sl. Varieties Fresh Turgid Dry weight

weight (gm) weight (gm) (gm)
No.

1 V1 1.395 1.686 0.496

2 S 13 1.270 1.485 0.416
3 S 34 1.365 1.536 0.404
4 S 36 1.209 1.363 0.384
5 Mysore 1.005 1.143 0.345
local

6 RC 1 1.223 1.339 0.393

7 RC 2 1.302 1.470 0.446

c) Determination of transpiration rate in mulberry

leaves
Aim: To determine the rate of transpiration in different mulberry varieties by gravimetric

method.

Principle: one of the most important process of plant function is uptake of water from the

soil .the mechanism for water uptake and transport with in the plant is transpiration, the loss

of water from aerial part of plant in the form of vapour. Transpiration plays several vital roles

in keeping the plant alive. For most the evaporation of water from leaf surface allows plants to
cool themselves. Without this ability the plants would quickly over heat and die on bright

sunny days. It also facilitates of nutrients uptake into the plants. Transpiration of water from

the plant creates gradients that drive movement in the soil towards the root.

Materials required:

 Twigs of different mulberry varieties with uniform number of leaves (V1, S 36, Mysore
local, RC 2, RC1, S 13, S 34).

 Conical flask

 Beaker and water

 Non absorbent cotton

 Aluminium foil

 Electronic balance

 Blade

Procedure: A 500ml beaker was taken with water in the lab and went to the field. In field the

longest branch of different mulberry were selected. Those branches were bent into the beaker

and the twig was cut under water with the help of a blade. Those twigs are taken and brought

to the laboratory and kept in water in the beaker. Then immediately the twigs were transfer

with uniform number of leaves into a conical flask with full of water. Then mouth of the conical

flask was plugged with non-absorbent cotton, and the sealed with alluminium foil by taking

care to avoid the evaporation. Next the entire setup was weighed in electronic balance and

recorded. It is the initial weight of this setup. Finally the entire setup was kept in open field

and bright sunlight and allowed it for 2hours. After exactly 2hours the setup was brought to

the lab and the final weight was taken.

Calculation: The transpiration rate is calculated in the different mulberry variety (V1, S 36,

Mysore local, RC1, RC2, S 13, S 34) by the following formula.

 Transpiration rate = (initial weight – final weight)/initial weight

TR %=(IW– FW)/ IW * 100

Result: By the above formula transpiration rate measured individually for 7 mulberry varieties.

Table4: Transpiration rate in elite mulberry genotypes

Sl. VARIETIES INITIAL WEIGHT FINAL WEIGHT

No. (gm) (gm)
1 V1 453.8 447.2
2 S 36 433.8 426.9
3 Mysore local 383 375.6
4 RC 2 383.4 375.8
5 RC 1 439.8 432.4
6 S 13 436.5 424.6
7 S 34 440.8 436.4
d) Study of phenomenon of osmosis in plant water
Relation

Aim: to study the phenomenon, of osmosis in plant water relations.

Objective: To understand the phenomenon of exosmosis and endosmosis using potato resin

osmoscopes.

Principle: Plant water status decides that crop growth in biomass production. Hence, study of

plant water relation is most important to understand physiology of crop productivity. Water

moves from its high potential to the lower one in the biological system by the phenomenon of

osmosis. Mass flow and diffusion of water between different plant cells takes place through a

semipermiable cell membrane from a higher concentration to lower concentration. Being a

universal solvent, water dissolves different solutes in the cell and hence decreasing in water

potential. Due to concentration gradient of different solutes water may be absorbed by the

cells, is called edosmosis, and if a cells loose water to the outside medium is known as
exosmosis. This phenomenon pertains in plant water relations, most specially for entry of

water into the mulberry roots in the soil media and translocation between different cells.

Materials required:

 Petriplates

 Distilled water

 Potato tubers

 Resins

 Blade

 1% sugar solution

 Blue ink

Procedure:

Endosmosis: A petriplates was taken and poured distilled water and then resins were kept in

it at five numbers. Then the setup was leave for 6 hours.

Observations: After 6 hours flaccid resins were swollen, due to the phenomenon of

endosmosis, i.e., the entry of distilled water into the resins to the turgid condition. Since the

dry resins contain less water (adhered to the cell contents), the water from the outside

medium enters to the resins and then it become turgid.

The water enters due to the phenomenon of osmosis by cell membrane.

Exosmosis: A potato tuber was taken; the skin was peeled off and cut to make a block. Than

it was scooped with a blade and made well. Than in the well-distilled water was added with a

drop of ink and the level of water was marked with a pin.

After that the entire setup was kept in 1% sugar solution on a petriplates. Next the setup was

leave for 6 hours.

Observation: after 6 hours it was observe that the water level of the potato well has come

down. Simultaneously, it was observed the volume of sugar solution has raised and attain

blue colour. Sugar solution contain low osmotic potential, then the distilled water, has high

osmotic potential.

e) Estimation of total nitrogen and crude protein content in

mulberry leaves

Aim; Estimation of leaf nitrogen was carried out by the Micro-Kjeldahal method

PRINCIPLE:

The sample is digested with concentrated H2SO4 in the presence of catalyst to convert

the nitrogen in protein or any other Organic material to ammonium sulphate. By steam

distillation of this salt in the presence of a strong alkali, ammonia is liberated and collected in

Boric acid solution as ammonium borate, which is estimated against a standard acid by

titration. On an average most proteins have 16% nitrogen in their composition. In other words,

1mg nitrogen equals 6.25mg of protein. Thus, by finding out the amount of ammonia formed,

from a known amount of sample, one can calculate the amount of protein present (1ml of 0.1

N Acid =1.401mg N).

REAGENTS;

 Concentrated H2SO4.

 Mercuric Oxide.

 Potassium sulphate
  Sodium Hydroxide – Sodium Thiosulphate solution; dissolve 600gms of NaoH
and 50 gms of Na2SO3.5H2O in water and make up to 1 lt.

 0.02N standard HCl or H2SO4.

 4% Boric Acid Solution; dissolve 4gms of H 3 BO3 in warm water and dilute
100ml.

Mixed indicator solution: mix two parts of 0.2% methyl red in ethanol with one part of

0.2 % methylene blue in ethanol (mix one part of 0.2% methyl red in ethanol with five parts of

o.2% bromocresol green in ethanol).

Procedure:

Estimation of leaf nitrogen was carried out by the Micro-Kjeldahal method, following the

basic principles laid out by Anonymous (1990). One hundred mg of leaf powder was placed in

a long necked micro-Kjeldahal flask and 10 ml of concentrated sulphuric acid was added to it.

Samples were digested in the micro digestion unit till the digested solution turned white in

colour. Later, 2 ml of 30 % hydrogen peroxide was added to it and digestion was continued till

the sample became clear. Two to three drops of methyl red and Bromocresol green mixed

indicators were added to 25 ml of Boric acid solution in a 250 ml conical flask and steam

distilled the digested samples in a Kjeldahal distillation unit. After distillation, the boric acid

solution was titrated against 0.1 N sulphuric acid solution till the green colour of the boric acid

changed to brick red. The total nitrogen content of the leaf samples was estimated by using

the following formula and then the crude protein content of the mulberry leaf was determined

multiplying the total nitrogen with the conversion factor of 6.25.

Calculation:

TV * N of avid* 0.014* vol of digested sample *100

Total Nitrogen (%) =
Weight of the sample (g) × Aliquot taken
 Crude protein (%) = Total Nitrogen × 6.25.

Reagents preparation: Catalyst mixture: Mix 20 g CuSO4 and 100 g K2SO4 finely by using

piton and mortar

Mixed indicator: Mix 0.1 g bromo cresol green with 0.07 g methyl red and dissolve in 100 ml

ethanol.

N HCl: N1V1 = N2V2;

500 X 0.1 = 11.3

500 X 0.1 = 4.42 ml concentrated HCl in 500 ml distilled water 11.3

In Kjeldhal flask fill NaOH 40 % solution (40 g in 100 ml)

Milli equivalent weight of nitrogen is 0.014

Table: 5. Total nitrogen and crude protein content in some elite mulberry
genotypes

Variety Total Nitrogen (%) Crude protein (%)

K2 1.96 12.25
V1 2.63 16.4
S 36 2.12 13.25
f) Estimation of available phosphorous.

Aim: To estimate the phosphorus in mulberry leaves

Principle:

Available phosphorus was estimated by the method of Olsen et al. (1954). In this

method the available phosphate reacts with ammonium molybdate, in an acid medium, to

form molybdophosphoric acid. The molybdo-phosphoric acid then reduces to a blue coloured

complex through reaction with stannous chloride. Absorbance readings were taken at 660

nm wavelength using a spectrophotometer. A standard curve was prepared from the

absorbance readings of different concentrations of potassium dihydrogen phosphate.

Procedure:

One gram of leaf was taken in a 250 ml conical flask. To this 20 ml of 0.5 N sodium

bicarbonate (pH 8.5) and a pinch of carbon black were added and continuously shaken for 30

minutes and then filtered through Whatman No.1 filter paper. Similarly, a blank was prepared

without using leaf. 5 ml of clean filtrate was placed in a 25 ml volumetric flask to which 5 ml of

1.5 % ammonium molybdate solution was added. After this, 10 ml of distilled water and 1 ml

of stannous chloride solution (10 g dissolved in 25 ml conc. HCl and then 0.5 ml of this diluted

to 66 ml with distilled water) were added and a final volume was made up of 25 ml, by adding

distilled water and allowing up to 30 minutes for colour development. This was read at 660 nm

wavelengths in a spectrophotometer. The phosphorus (P) content was calculated (kg/ha) by

comparing the spectrophotometer readings with the standard curve.

Calculation:
Available R × Total volume of extract × Volume made up × 106
P (kg/ha) = × 2.24
In leaf sample 106 × wt. of sample (g) × volume of aliquot taken

Where, R = µ g of P in the aliquot (obtained from the standard curve).

Table: 6. Available phosphorus content in some elite mulberry genotypes

Variety Available
phosphorus (%)

K2 0.22
V1 0.28
S 36 0.26

(f). Estimation of available potassium

Aim: To estimate the potassium in mulberry leaves

Principle:
 Potassium was extracted from the leaf with the help of saturated ammonium acetate

solution, by shaking, followed by filtration. The extract was determined by using a flame

photometer as described by Rao (1993). One gram of dry soil was placed in a 250 ml conical

flask; 20 ml of neutral ammonium acetate solution was added to it and shaken for 30 minutes.

Immediately after this, it was filtered through Whatman filter paper No.2. Potassium (K)

concentration in the extract was determined with the help of flame photometer. The reading

was compared with the standard curve to determine the amount of available potassium.

Reagent: 1.Potassium standard 100ppm of potassium dissolve 0.191g of KCL in some

volume of distill water and than make up the volume to one liter.

Materials: Flame photometer, pipettes, volumetric flask, etc.

Procedure:

Preparation of standard curve:

Take 0,1,2,3,4 and 5ml of 100ppm potassium solution in a separate 50ml volumetric flask.

Make up the volume to 50ml distill water and mix well. After adjusting the needle of flame

photometer to zero by feeding blank, adjust the needle to 100 by feeding maximum

concentrated k solution. Then feed the standard to record the flame photometer readings. Plot

flame photometer readings.

For plant sample:

1.Pipette out 5/10ml of digested sample into a 50ml volumetric flask and add

5/10ml of vanadomolybdate reagent

2. Make up the volume to 50ml and mix.

3.Read the colour intensity at 430nm after 30min.

 4.Compare the unknown sample absorbance with standard cure and calculate percent

of the sample

Calculation:

R × Volume of extract × 106

Available K (kg/ha) = × 2.24
106 × Wt. of the leaf sample taken (g)

Where, R = ppm or µ g of K (obtained from the standard curve).

Table: 7. Available potassium content in some elite mulberry genotypes

Variety Available potassium

(%)
K2 1.01
V1 1.26
S 36 1.11

Elite Mulberry genotypes

 V1

S 36

Mysore Local
RC 1

RC 2
S 13
 S 34

THEORY CUM DEMO ON

NUTRIENT DEFICIENCIES IN MULBERRY AND THEIR

MANAGEMENT

Mulberry (Morus spp.) is intensively cultivated for feeding of money spinning insect,

silkworm (Bombyx mori). Silkworm rearing under controlled environment for production of

lustrous silk is one of the major activities in India providing ample employment opportunities.

Hence maximization of quality leaf production is the vital aspect. Seventeen elements (9

macro and 8 micro nutrients) have been reported to be essential for the growth of mulberry

(Table 1). Plants take carbon, hydrogen and oxygen mostly from air and water, while the
other nutrients are taken from the soil especially in the form of ions. If the soils are deficient

with these nutrients, the plants show the deficiency symptoms. The deficiency symptoms can

be categorized in following groups.

Diagnosis of nutrient deficiencies in mulberry

• Chlorosis

• Necrosis or death of plant tissue

• Lack of new growth of terminal growth, resulting in resetting

• Accumulation of anthocynin and an appearance of reddish colour

• Stunting or reduced growth with either normal of dark green colour or yellowing.

Nutrient deficiency symptoms can be easily diagnosed in mulberry by visual observation

(Table 2 & Fig. 1)

Role of different macro and micro nutrients for qualitative mulberry leaf
production

Nitrogen
Nitrogen is the building block to form amino acids, proteins, DNA, RNA, amides,

amines, chlorophyll and structural constituents of cell wall etc. Hence it plays an important

role in mulberry foliage production.

Phosphorous
Phosphorous constitutes of enzymes, phospho proteins, phospho lipids and nucleic

acids. It promotes early maturity of leaf. It is a part of NADP, ADP, DNA and RNA, electron

transport in oxidation-reduction process. Translocation of sugar and starch etc.

Potassium
Potassium actively involves in maintenance of cell osmotic potential by regulating

through stomata. It also provides resistance to plant against lodging of pests/ diseases.

Besides these functions potassium involved in formation of ATP, starch and protein synthesis.

Calcium
Calcium as calcium pectate helps in cell elongation and cell division and also involves

in translocation of carbohydrates. It plays an important role in enzyme activation.

Magnesium
Magnesium constitutes as an essential molecule in chlorophyll. It acts as a co factor of

transphosphorylase, dehydrogenase and carboxylase enzymes. Associates in formation of

polypeptide chain from amino acids and also involves in formation of sugars.

Sulphur
Sulphur constitutes in cysteine and methionine formation specifically and vitamins and

hormone formation generally. It also involves in oxidation-reduction process.

Iron
Iron plays an important role in synthesis of chlorophyll, nitrogen fixation,

photosynthesis, electron transfer etc. Regulates respiratory enzymes, particularly cytochrome

enzyme.

Zinc
Zinc acts as an important component of number of enzymes, protein synthesis and

also regulates plant growth hormones i.e. auxins.

Manganese
Manganese helps for evolution of oxygen during photosynthesis and also a component

of number of enzymes. It is a structural component of certain metallo proteins. Activates

electron transport system.

Copper
Copper involved in several enzyme systems. It helps in formation of cell wall, electron

transport system and oxidation-reduction process.

Boron
Boron plays an important role in transport of sugars across the cell membrane, controls

formation of sugars and starch, transpiration, carbohydrate metabolism, amino acid

metabolism and protein synthesis.

Molybdenum
Molybdenum actively involved in Nitrogen cycle by formation of nitrate reductase

enzymes.

Chlorine
Chlorine involved in phosphorylation, photosynthesis and osmotic pressure regulation

by acting as an ion in counter balance to cat ions.

Silicon
Silicon helps in possessing rigidity of cell wall, acts as a protector and assists in

photosynthesis.

Sodium
Sodium helps in osmotic regulation and also fulfills the functions of potassium in some

cases.

Vanadium
Vanadium involved in oxidation-reduction process of cells.

Cobalt
Cobalt involved in nitrogen fixation process.
Conditions ideal for nutrient absorption

The ability of plants to absorb water and nutrients depends on permeability of root

surfaces, which in turn influenced by the metabolic activity of the roots. The metabolic activity

is increased by supplying oxygen to the roots and removing carbon dioxide. The ionic

absorption process of nutrients is most efficient under the following conditions, when

 The soil has a high concentration of nutrient ions.

 The soil is well aerated allowing oxygen to diffuse readily into the soil and carbon

dioxide diffuse out.

 The soil is sufficiently moist to permit ions in solution to contact large areas of root

surfaces moisture are the medium for transport.

 The leaves are exposed to adequate light

 If the nutrients are not made available to the plants as per the requirement some

symptoms may visible on the leaves.

Management of nutrient deficiencies

• It is advisable to use the inorganic fertilizers after the soil testing along with the organic

manures and biofertilizers.

• An emphasis should be laid down for incorporation of green manure in the soil so that

balanced nutrient management can be followed with out decreasing the leaf yield

producing potentiality in mulberry crop.

• Foliar application of Seriboost – a micro nutrient formulation. Mix 500 ml of Seriboost in

200 liters of water for 1 acre of garden. I spray – 23 to 25 days after pruning/ leaf

picking. II spray – 30 to 32 days after pruning / leaf picking.

Table 1. Essential elements and its available form to Mulberry

Elements Available form

A. Macronutrients
1. Sulfur So4
2. Phosphorous HPo4
3. Magnesium Mg2+
4. Calcium Ca2+
5. Potassium K+
6. Nitrogen No3, NH4
7. Oxygen O2, H2O
8. Carbon Co2
9. Hydrogen H2O
B. Micronutrients
1. Molybdinum Mo O-2
2. Copper Cu+4, Cu+2
3. Zinc Zn+2
4. Manganese Mn+2
5. Iron Fe2+, Fe3+
6. Boron Bo3, B4O7
7. Chlorine Cl
8. Nickel Ni
Table 2. Visual symptoms of nutrient deficiencies on Mulberry leaves

Element Deficiency Symptoms

On terminal leaves
Calcium Plants remain dark green, young bud leaves chlorotic,
finally death of terminal bud.
Boron Young leaves of the terminal buds with loose colour at the
base, death of the terminal bud.
On young leaves
Sulfur Light green leaf, veins still paler, no dead spot.
Iron Chlorosis, no spots, main vein typically green.
Copper Interveinal chlorosis, resetting and permanent wilting, leaf
detaches easily.
Manganese Leaf chlorotic, main and small vein dark green.
On old leaves
Nitrogen Dwarfing and abnormally light green plant leaf erect, light
green to yellow.
Phosphorous Dwarfing and abnormally dark green plant, leaf erect and
unusual narrow. In acute condition greenish brown to
black, bronzing on backside of leaf.
Potassium Chlorotic leaves, small dead spots or specks at the tips
and margins, rusty appearance, crimping and cupping of
margin and tips.
Magnesium Chlorosis starting from tips and margins, with no dead
spots, vein green, base necrosis in acute condition, leaf
detaches easily.
Zinc Leaf become narrow and small, lamina chlorotic, veins
green, dead spot develop all over the leaf including veins,
tips and margins.
Molybdenum Leaf lighter green, golden yellow to orange, dead spot all
over the area except veins, affected areas extrude
resinous gum through lower surfaces.

Fig. 1. Nutrient deficiency symptoms in mulberry

 Nitrogen deficiency Phosphorus deficiency

Potassium deficiency Calcium deficiency

Magnesium deficiency Zinc deficiency

 Sulfur deficiency Iron deficiency

Manganese deficiency Copper deficiency

f
i
c
i
 e
n
c
y
Application of Seriboost

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