Quality Enhancement in Crops: Molecular

Approaches

Dr. Neelu Joshi

Assistant Professor
Department of Biotechnology and Bioinformatics
Padmashree Dr. D.Y. Patil University
Navi Mumbai

Quality Enhancement of
Crops
Quality- Degree of excellence
Nutritional quality
Organoleptic quality
Processing quality

Nutritional Quality
Proteins
Carbohydrate
Oil content
Vitamins
Minerals

Proteins
Classification of proteins on the basis of
solubility
Albumin fraction- water
Globulin-water
Prolamine- alcohol
Glutenin- dilute acid

Protein quality- amino acids content and

digestibility
Essential amino acidsNon essential amino acids

Deficient
cys

Cereals
Lys, tryp

Pulses
met,

Prolamine fraction
Crop
prolamine fraction
amino acid
Wheat
Maize
Barley
Sorghum
Rice
Pear millet

gliadin
zein
hordein
kafirin
oryzein
prolamine

limiting

lys, threo
lys, tryp
lys
lys, tryp
lys
lys

Aspartate pathway- The only

pathway for synthesis of essential amino
Key Enzymes
acids
Aspartate kinase (AK)bacterial-less sensitive to feedback
inhibition by lysine

DHDPS- Dihydrodipicolinate synthase(feedback insensitive)

Strategy for increased lysine in cereals
1. Expression of Corynebacterium DHDPS in canola
2. Expression of Corynebacterium DHDPS + E.coli AK in
soybean
3. Create sink for stable incorporation of lysine( gene for

lysine rich protein)

4.

Blocking lysine degradative pathway by cloning cDNA

encoding lysine degradative enzymes

Improvement of cystein in
pulses
Cystein biosynthesis (multiple codon)
Substitution of non-esssential amino acids

with cystein through:

a targeted mutagenesis
b codon engineering

Improvement of grain protein

quality
Wheat- Increased lysine content by transgenic approach
Rice-expression of human milk protein,expression of

lysine rich protein from soybean

Maize-lys rich mutants, QPM (Quality Protein Maize)
Pulse- increased lysine content in soybean and canola
Modification of gene for sulphur-poor protein for
enriching its nucleic acid sequence for S-amino acid:
-phaseolin from Phaseolus vulgaris
-vicilin from Vicia faba
Expression of synthetic protein (13% met) in sweet
potato

Starch:

synthesized and accumulates in plastids

called amyloplast

Amylose (unbranched)and amylopectin(branched)

Manipulation of the pathway leading to one or the

other form of starch

Amylose:amylopectin,; 30:70%, influence physico chem
properties of starch, its use in food production involve formation

of gel after heating the starch in water and cooling. Amylose tend
to aggregate and crystallize on cooling, whereas amylopectin gels
are more stable and desirable for food processing. Amylose rich
starch is a valuable feedstock for industrial purpose.

Rice: high amylose-hardiness; low-stickness; 22%ideal

GBSS(granule-bound starch synthase)-synthesis of amylose

chain; antisense inhibition in potato to get amylosefree starch

Starch branching enz(SBE A, SBE B)- antisense
inhibition to get high- amylose starch, useful for food
and industrial applications

Increased starch quantity

Starch biosynthesis regulated (allosteric)at a key

step catalysed by ADP-glucose pyrophosphorylase.

It is regulated by activator FbP and repressor AMP
and Pi.
Mutant ADP-GPP gene of E. coli
altered allosteric property, become less
dependent on FbP, less sensitive to inhibition by
AMP
gene (IglgC16)inserted and expressed in potato
35% more starch than control tubers
uniform frying, low oil consumption

Oil quality
Quantity and composition of fatty acids impart

unique physico-chemical properties to the oil and

decide their use
Capric Lauric Myristic palmitic stearic oleic linoleic linoleinic

erucic
c 10
18:3

c12
c22

c14

c16

c18

18:1

18:2

SFA- suitable for table purpose, solid

MUFA
PUFA-nutritionaly important but low shelf life, prone to

oxidation
Medium chain(c12, 14)-soaps and detergents

Metabolic engineering of
lipids
Improvement of existing lipid products
Engineering of novel lipid products

Genetic engineering of plant

oils
Goal

Purpose

Decrease linoleic acid in

soybean

Increased stability, less

tendency to spoil

Develop a temperate zone,

annual crop as source of
lauric acid

stabilizes supplies, lower

price soaps and detergents

Develop high erucic acid

plant oils-Brassica

Make industrial use of plant

oil economical, reduces
prices

Decreased levels of
unsaturated fatty acid in
Cocoa seeds

Make chocolates that

remain solid in warm
weather

Strategy to decrease
desaturation in PUFA oilseeds
Antisense suppression of stearoyl CoA

desaturase (which converts stearic to oleic),

results in decreased oleic and higher stearic
acid- oilseed rape.
-High stearic acid oil is used as cocoa butter
substitute.

Production of short chain fatty

acids
Oils of major oil crops-c16-c18; coconut and

palmoils are c8-c14

Lauric acid (c12)-soaps, detergents and
cosmetics
Synthesis of FA at a particular length is
terminated by the hydrolysis of acyl-ACP by
thioesterase.
Gene for acyl-ACP thioesterase from California
Bay tree cloned and introduced into oilseed
rape

Production of longer chain fatty

acids
Fatty acids longer than c18 used for industrial oil
Brassica -Erucic acid is good as industrial

oleochemical but nutritionaly unsuitable

conventional breeding has led to the development of:
high-erucic acid rape (HEAR)
low- erucic acid rape (LEAR)
Strategy- to overexpress the genes that encode
elongases
-To transfer enzyme activitiesthat preferentially
incorporate erucic acid into TAGs

Modification of degree of
saturation
Decreased levels of unsaturated fatty acid in
Cocoa seeds to make chocolates that remain
solid in warm weather.
High stearic acid oil in oilseed rape
Insertion of antisense construct of the
9desaturase gene resulted in decreased amount
of desaturase enzyme which in turn led to
decreased formation of oleic acid and a rise in the
proportion of stearic acid.
High oleic acid oil in oilseed rape
Antisense suppression of 12desaturase gene

Biofortification
I saw a few dying of hunger, of eating, more than
thousand

Hidden hunger-hunger despite eating

Micronutrient deficit food
Solution

-biofortification: development of micronutrientdense staple food crop

Provitamin A (-carotene)
Immature rice endosperm-GGDP_ -carotene
Requires 3 enzymes, absent in rice kernel

psy (phytoene synthase)-Daffodil

Lcy (lycopene beta cyclase)-Daffodil
Crt 1 (phytoene desaturase)-Erwinia uredovera
Golden Rice 1 (psy, crt 1 expressed in
endosperm)-Ingo potrykus
Golden Rice 2(psy of maize, crt 1) Syngenta
Group, 2005

Iron-rich Rice
Rice contain Phytate which entraps 95% of

dietary iron
Phytase-resist high temperature
Ferritin (iron storage protein)
Gene for cystein rich protein-for absorption of

iron