Legumes have been part of the human diet since the early ages of agriculture. Legumes are consumed in many forms: seedling and young leaves are eaten in salads, fresh immature pods and seeds provide a green vegetable, and dry seeds are cooked in various dishes. Legume seeds provide an exceptionally varied nutrient profile, including proteins, fibres, vitamins and minerals. Breeding for nutritional quality entails an improvement primarily in protein quantity and quality which are of paramount significance. PROBLEMS AND PROSPECTS OF BREEDING FOR NUTRITIONAL QUALITY • Negative correlation between yield and protein content. • Negative correlation between protein and sulphur containing amino acids • Lack of proper field screening technique.

1. - D H AN U J A N
2 0 1 9 5 0 8 0 0 5
I M . S C . , G P B
Breeding for nutritional quality in
pulses

2. PULSES
 Legumes have been part of the human diet since the early ages
of agriculture.
 Legumes are consumed in many forms: seedling and young
leaves are eaten in salads, fresh immature pods and seeds
provide a green vegetable, and dry seeds are cooked in various
dishes.
 Legume seeds provide an exceptionally varied nutrient profile,
including proteins, fibres, vitamins and minerals

3. QUALITY
 The food value of legumes seeds is high. The calorie value per unit
weight is comparable to cereals.
 Additionally the protein content is double besides being fair source of
minerals and vitamins. Legume protein is cheaper.
 In grain legumes there is a deficiency in the sulphur containing amino
acid content of legume seeds.
 Grain legume seeds have reasonable quantities of thiamin and
nicotinic acid as well as the nutritionally important minerals, calcium
and iron but they contain little retinol, riboflavin and ascorbic acid.
They also contain a variety of anti nutritional factors.

4.  Breeding for nutritional quality entails an improvement primarily
in protein quantity and quality which are of paramount
significance.
 The quality of protein depends on its amino acid profile and net
protein utilization (NPU), i.e. biological value.
 In legumes the primary aim is to correct the amino acid balance,
i.e., to elevate the content of limiting amino acids, rather than
increasing the protein content per se.

5. PROTEIN
The protein in pulses
ranged from 14.9-
34.6%.
The most proteins are
located in cotyledons
and embryonic axis and
seed coat contains very
little protein.
 Protein of pulses is of two types : metabolic
protein (enzymatic and structural) and storage
protein.
 Cotyledons - 93% of methionine and tryptophan of
whole seed; Embryo – 2.5% protein.
 The composition of amino acids in seed is
influenced by phosphorous, molybdenum and
phosphorous level in soil. The application of
sulphur containing fertilizer enhances cystine
content of chickpea protein.

6. BIOLOGICAL VALUES
 Wide variation and range from 32-78%.
 Low biological value of pulses is due to low methionine content.
 The pulse protein has poor digestibility. The low biological value and
digestibility of pulse protein is attributed to presence of protease inhibitors
and anti nutritional factors.
 The availability and utilization of protein are influenced by the amino acid
composition, digestibility of proteins and presence of anti nutritional factors.

7. CARBOHYDRATES
The total carbohydrate of pulses ranges from 53.3-68%.
Carbohydrates include mono and oligosaccharides, starch and other
polysaccharides. Starch is most abundant carbohydrate and varies
from 31.5-53.6%. Ingestion of large of large quantity of pulses is known
to cause flatulence in human beings.
Members of raffinose
family of sugars are
not digested by man
because human
mucosa lacks the
hydrolytic enzyme α-
1, 6-galactosidase.

8. OLIGO
SACCHARIDES
Raffinose,
Stachyose,
Verbascose and
Ajugose -
predominate in most
pulses accounting
for 31.1-76% of
Soluble sugars.
 Verbascose is major oligosaccharide in
mungbean, urdbean and pigeonpea.
 Stachyose is major oligosaccharide in
chickpea, lentil and cowpea.
 Ajugose is found in peas. Raffinose is
moderate in all pulses.

9. CRUDE FIBER
 Crude fiber consists of cellulose, hemicelluloses, lignin, pectic
and cutin substance. Pulses contain good amount of crude
fiber (1.2-13.5%).
Cellulose is major
component in
chickpea and peas
whereas
hemicelluloses is
important for
pigeonpea,
urdbean and lentil

10. LIPIDS
 Lipids include free fatty acids,
mono, di and triacyl glycerol,
phospholipids, sterols, sterol
esters, glycolipids, lipoproteins.
 Total lipid content of pulses
range from 1.0 – 4.99%.
 Total lipid content varies with
variety, origin, location and
climate.
 Greengram – Potassium
 Bengalgram - Fibre, protein,
potassium, Vit C
 Moth beans –Ca, K, Fe, Cu,
Na, Zn
 Cowpea - Vit A ,folic Acid, Zn,
P
MINERALS

11. PROBLEMS AND PROSPECTS OF
BREEDING FOR NUTRITIONAL QUALITY
Negative correlation between yield and
protein content.
Negative correlation between protein and
sulphur containing amino acids
Lack of proper field screening technique.

12. NEGATIVE CORRELATION BETWEEN YIELD AND
PROTEIN CAN BE ALTERED BY BREEDING
 A concurrent choice of both yield and protein will largely mitigate the situation.
 Variation for both the traits coexists between crop species, and between
varieties within a crop.
 Availability of mutant genes responsible for a higher amount of protein content
in grains (such as opaque-2 in maize) could further enlarge the spectrum of
genetic variation.
 With such a range of variability, making inter-family selection for high yield,
followed by intra-family selection for high protein content from selected
productive families, would thus offset the ill – effects of the negative
correlation between yield and protein.

13. NEGATIVE CORRELATION BETWEEN PROTEIN
QUALITY AND QUANTITY CAN BE MANIPULATED
 In legumes the negative correlations between protein content
and methionine and cystein content can be modified as have
been done in case of cereals.
 This apart, as in cereals, several mutants higher in protein
content and better in amino acid profile have been also
identified in legumes.
 Appropriate genetic manipulation of these mutants may bring
about an appreciable improvement in both protein quantity
and quality.

14. BETTER SCREENING TECHNIQUES ARE AVAILABLE
 Many procedures for a chemical analysis of the protein and amino
acid profile have been developed.
 Require double screening – consuming process.
 These techniques involve both rapid chemical analysis of protein
and amino acid profile of grains in the lab, and improved field
techniques including the use of genetic markers associated with
high protein content.
 Suitable methods for screening for protein content are dye binding
coupled with automated N determination or by using IR reflectance.

15. BROAD SENSE HERITABILITY
S. No Crop Broad sense
Heritability for
protein content
Reference
1 White lupin 83% Green et al .(1977)
2 Soybean 40- 80% Johnson et al., 1955)
3 Chickpea 75% Sandhu et al.( 1968)
4 Dry beans 30-64% Leleji et al.(, 1972)
5 Fababean 54% Bond(1977)
6 Beans 49.3% (Broad sense)
& 20 %(narrow
sense)
Dickson and Hackler,
1973
7 Kabuli
Chickpea
High Singh et al (1986)
9 Fababean High Robertson et al (1986

16. ENVIRONMENTAL INFLUENCE
 Seed protein content in grain legumes is strongly influenced by the
environment.
 The protein content was positively associated with the sum of temperature
from sowing to maturity and with the temperature during flowering and
beginning of seed filling, while it was negatively associated with July
precipitations.
 All environmental factors that impact nitrogen nutrition, such as drought
stress, soil compacting, root diseases and pests may also influence seed
protein content through their impact on nitrogen availability.
 Despite this, seed protein content heritability is generally moderate to high

17. GENETIC CONTROL OF PROTEIN CONTENT
• Mungbean - High governed by dominant genes; mostly non-
additive variance, Biparental mating system and modified diallel
selective mating system.
• Chickpea - Predominance of additive effects for protein content
and tryptophan content but for sulphur content non-additive
effects
• Soybean - Additive > dominance gene action

18. GENETIC VARIABILITY FOR QUALITY TRAITS IN
GRAIN LEGUMES
 Sufficient variation exists for the main components in legume
seeds since
 oil concentrations vary from 1 to >40 %
 carbohydrate from 2-3 to 60 %
 protein from about 15 to 50 % of the dry weight.
 Manipulation of the proportion of these components is difficult
because of complex biochemical machinery about which very
little is known, e.g. what is the biological regulation whereby
soya beans are oil seeds while broad beans mainly store starch.

19. CORRELATION BETWEEN PROTEIN CONTENT
AND GRAIN YIELD
 The reported negative correlation seems to be
small enough to allow selection of plants high in
protein without reducing yield.
 Under this situation selection for higher protein
content should be performed only among lines
yielding at least at the same level as the original
populations; the production of proteins per
hectare must be taken as a selection criterion.
 The alternative possibility is to select for
increased yield while trying to maintain the
protein level constants.

20. RECURRENT SELECTION
• Recurrent selection was also used to increase seed protein
content and seed yield in soybean.
• Recurrent mass selection for 2 cycles, based on a desired grain
index, used for simultaneous increase of seed yield and seed
protein percentage, could increase seed protein content from
21.9 to 24.6 %.
• Selection on the basis of single plant protein content in
segregating generations resulted in increase of protein content
with little change in yield

21. SELECTION STRATEGY FOR IMPROVEMENT OF
PROTEIN CONTENT
 Depends on factors such as the crop species involved, the
breeding system of crop species, the selection objectives, the
influence of the environmental factors.
 Traits which are governed by major genes and have high
heritability estimates should be selected in early segregating
generations e.g., seed colour and seed texture.
 Traits which are governed by polygenes and have low
heritability estimates should be selected in F4 and F5.

22.  For the trait like protein content between family selection
should be practiced in later generations among the superior
families already selected for higher yield.
 When the negative correlation between protein content and
grain yield is low, selection should be made first for high yield
and then within high yielding families, plants having the
highest percentage protein should be identified for further
selection and inter-mating.
Contd…

23. POSSIBILITY OF IMPROVEMENT IN AMINO ACID
COMPOSITION THROUGH SELECTION
 The main difficulties for a direct selection are due to the low variability
for each amino acid, the relationships among the amino acids and the
negative correlations between the protein content and the sulphur
amino acid level.
 A possibility is there for increasing the sulphur amino acid content
without decreasing the protein content. Lines with both high protein
and sulphur amino acid contents were obtained in Vicia faba, and in
other species.
 Particular attention in the selection work with legumes should,
however, be given to the tryptophan content, which is the third limiting

24. RELATION OF GRAIN PROTEIN CONTENT AND
AMINO ACID CONTENT
 In Phaseolus vulgaris seed protein content was positively
correlated with leucine content (Kosson, 1989).
 Genotypes with high protein content and high lysine content
were developed in soybean (Tymchuk et al., 1990).
 Similarly mutants with high protein, lysine and seed yiled were
identified in Vigna mungo and Vigna radiata.

25. PROTEIN CONTENT AND OIL
 Highly negative correlations between protein and oil are well
documented in soybean and varieties.
 Negative correlations reported between starch and protein
content
 Negative correlations have been reported between seed size
and protein content in pigeonpea but some promising lines
with high protein content and large seed size have been
obtained at ICRISAT suggesting the possibility of
improvement of protein content and yield contributing traits

26. POTENTIAL LEVERS FOR PROTEIN CONTENT
IMPROVEMENT
 Seed protein content is the relative accumulation of
proteins and dry matter in the seeds depends on the sink
and source strength.
 Seed protein increased dramatically when the
source/sink ratio increased.

27. IMPROVING SEED SINK STRENGTH
 Functional interactions exist among the different seed
constituents: for example, the disruption of the r gene
abolishes starch synthesis in pea seeds, leading to a wrinkled
seed phenotype.
 Elevated sucrose content impacted the accumulation of
storage protein families
 By knocking down the accumulation of one of the
constituents, the percentage of the others will increase.
However, this may have a detrimental effect on seed yield.

28. IMPROVING NITROGEN SUPPLY TO THE
SEED
 Pea mutants with absence of N2 fixation activity produce
lower seed yield and protein content, which can be
alleviated by adequate mineral fertilization, whereas an
autoregulation mutant of pea displaying a
supernodulating phenotype has a reduced shoot biomass
and seed yield, associated with higher seed protein
content

29. Grain legume seeds
bring in the diet
carbohydrates (lipids,
starches, fibres) and
minor seed
compounds which
will influence
positively or
negatively protein
bio-availability by
impacting digestibility
or acceptability
Factors Nutrient Major dietary source
Promoters
1. Prebiotics: inulin and
fructans
Fe, Zn, Ca Lentils, chicory, garlic
2. Beta-carotene Fe, Zn Lentil, pea, chickpea, green and orange
vegetables
3. Selenium I Lentil, pea, chickpea, sea food
4. Organic acids: ascorbic
acid
Fe, Zn Lentils, fresh fruits and vegetables
5. Amino acids Fe, Zn Animal meat
Inhibitors
1. Phytic acid Fe, Zn, Ca All legumes, cereals
2. Fiber Fe, Zn All legumes, cereals
3. Haemagglutinins Fe, Zn Most legumes, wheat
4. Phenolics Fe, Zn All legumes
5. Heavy metals Zn Contaminated legumes and leafy vegetables
FACTORS PRESENT IN PULSES THAT PROMOTE OR
INHIBITS MICRONUTRIENTS BIOAVAILABILITY

30. TRYPSIN INHIBITORS:
 Trypsin inhibitors are present in most grain legume seeds. High inhibiting
activities found in soybean seeds, reduced by processing. Null mutants
identified in soybean allows for production of low trypsin inhibitor cultivars.
 In pea, large genetic variability is available for the activity of Bowman-Birk
trypsin/chymotrypsin inhibitor proteins (TIA).
 The polymorphism in coding and promoter sequences of genes at Tri
locus accounts for most of the variation in TIA and this allowed to initiate
MAS.
 However, if low TIA activity is a benefit in pig or poultry feed digestibility,
recent data suggest that high contents of trypsin inhibitors in foods should
be positive, since a reduction of HT29 colon cancer cells.

31. LECTINS
 Most grain legumes cotyledons contain lectins
 In plants, lectins are very diverse and are involved in plant
defense or symbiosis with Rhizobia.
 Some natural variability exists for lectin hemagluglutinin
activity.
 However, the low content and toxicity of lectins together with
the complexity of lectin roles did not allow for the definition of
a breeding target for this trait.

32. ALPHA-GALACTOSIDES
 Major alpha-galactosides in grain legume seeds are raffinose,
stachyose and verbascose.
 Highly probable prebiotic properties which may be of interest
against colorectal Cancer.
 Even if some genetic variation exists, genetic tool to monitor
these contents have never been worked out, due to
competition with easy cooking or technological treatments
such as soaking.

33. TANNINS AND
FLAVONOID
COMPOUNDS
Flavonoids are
phenolic
compounds
involved in
determination of
seed coat
colours and in
the tanning
power on
proteins. They
bind to proteins
and reduce
digestibility.
 The diverse colours of common beans were
suggested to be important sources of dietary
antioxidants.
 In pea and faba bean, a single gene mutation
has a pleiotropic effect eliminating tannins from
seed coat and determining a white flower trait.
They increase protein digestibility in pigs or
poultry by 10 % when compared to tannin-
containing lines.
 The health benefit of proanthocyanidins may
deserve some attention.

34. PHYTIC ACID
 It is commonly found in cereal and legume seeds and its anti-
nutritional effect is associated with mineral-complexing
(especially Zn, Ca and Fe) and inactivation of digestive
enzymes.
 Phytic acid may have protective effects such as a decrease of
the risk of iron-mediated colon cancer and lowering serum
cholesterol and triglycerides.
 In common bean, 5 QTL were identified that controlled total
and net seed phytate content.

36. ACCEPTABILITY OF LEGUME SEED PROTEINS IN
FOOD
 Lipoxygenase activity can cause unpleasant tastes and aromas
when reacting with seed lipids.
 In soybean and pea, null mutants were found for 3 and 2 LOX
genes respectively. Their molecular characterization has well
progressed and offers possibilities of breeding for lipoxygenase-free
varieties.
 Saponins contribute to the bitterness of peas as well as that of
soybean.
 Saponins have been studied for their positive hypo-

37. BIOFORTIFICATION
 Biofortification is a method by which the nutritional value
of crops can be enhanced with the help of breeding,
transgenic techniques, or agronomic practices.
 Biofortification is one of the feasible way to reduce
malnutrition problem among underserved and
malnourished rural people in more cost efficient manner.

38. BIOAVAILABILITY OF PROTEIN
 Legumes contain many antinutrients which need to be minimized to
improve the bioavailability of micronutrients.
 It was found that inter-specific breeding of mungbean (methionine 0.17
g/kg) with black gram (high methionine 1.8–2.0 g/kg) significantly
enhanced the quality of protein in mungbean. The hybrid contains γ-
glutamyl-S-methyl-cysteine and γ-glutamyl-methionine, found in mung
bean and black gram respectively.
 Transgenic approaches for production of sulfur amino acid rich crop of
narbon bean (Vicia narbonensis), lupin (Lupinus angustifolius), forage
alfalfa has been studied.

39. BIOAVAILABILITY OF IRON (FE) AND ZINC
(ZN)
 Bioavailability of iron and zinc ranges from 5-15% and 18–34%
respectively.
 Presence of phytate in pulses and legumes are responsible for the low
bioavailability of iron and zinc.
 Phytic acid form complex with Ca, Mg, Cu, Fe and reduces its solubility.
 Concentration of phytic acid and zinc is found more in higher temperature
regime (8.8 mg/g and 69 mg/kg, respectively) comparative to lower
temperature regime (6.7 mg/g and 61 mg/kg, respectively) and the same
trend is found with Fe also (116 vs. 113 mg/kg). Thus phytic acid
concentration decreases when seeds expose to low temperature.

40. IRON
BIOAVAILABILITY
OF FIELD PEAS
Biofortification doesn’t
only focus on
increasing compounds.
Two approaches: One is
to increase the iron
content, and the other is
to make the iron more
bioavailable
“If you can double the
bioavailability, that’s like
adding twice as much
iron,”
Decreasing phytate ,
increase bioavailability
of iron.
 The approach for increasing iron bioavailability in field
peas is not focused on existing levels of iron in the
seeds. Instead, researchers decrease the levels of
phytate.
 Warkentin and colleagues have developed several lines
of low-phytate peas, derived from a high-performing
variety called CDC Bronco.
 Compared bioavailability in relation to seed coat color
for low- and normal-phytate peas. Darker field pea
seeds often have higher levels of polyphenols, which
inhibit iron absorption. The iron concentration of whole
seeds was significantly greater for the darkest seeds,
but the bioavailability of iron was significantly lower in
dark seeds compared with non-pigmented varieties.

41. BIOFORTIFIED SOYABEAN
 Biofortification of soybean sprout with a solution of ZnSO4 (10 or 20 μg/ml) has
significantly enhanced the quantity of zinc and also had good bio accessibility
(Zou, Tao, et al.,).
 Biofortification of soybean is done with different concentration of strontium ion
(0.5mM–3.0mM).
 At concentrations up to 1.5mM, strontium stimulated plant growth by 19.42% FW
(14.70% DW) and 22.62% FW (22.66% DW) for the shoots and roots,
respectively. Although concentrations above 2mM were showed toxic impacts.
 In vitro studies showed greater impact of strontium salts in treating osteoporosis
related problem and absence of toxicity in animals and humans.

42. BIOFORTIFIED BEAN
 CIAT has developed a biofortified bean (Phaseolus vulgaris L.)
through breeding of crops containing iron up to 0.1 g/kg.
 Ascorbic acid increases the absorption of iron (Fe) present in plant
origin foods by forming Fe(III) complexes and reducing amount of
Fe 3+ to more soluble and bioavailable Fe 2+.
 Therefore ascorbate level in plant foods can be increased by using
rDNA technology which would help to reduce the negative impact of
phytate and polyphenlos in staple foods on bioavailability of Fe and
also make these foods as essential source of significant nutrient
and vitamin C.

43. TRANSGENIC SOYBEAN
 The soybean has been targeted to increase provitamin A (beta-carotene),
a monounsaturated ω-9 fatty acid (oleic acid) and seed protein contents
by expressing bacterial PSY gene .
 The cysteine content of soybean seeds has been increased through over
expression of the sulfur assimilatory enzyme, O-acetylserine
sulfhydrylase.
 Increased methionine and cysteine by overexpressing the maize zein
protein.
 Antisense RNA technology has been used to reduce the amount of linoleic
acid and palmitic acid and increase the amount of oleic acid by inhibition

44.  Soybean is low in isoflavone which is associated with benefits
such as decreased risk of heart disease, reduced menopausal
symptoms, and reduced risk of some hormone-related cancers.
 Isoflavone content has been enhanced in soybean seeds by the
combination of maize C1 and R transcription factor-driven gene
activation and suppression of a competing pathway
 Importance of ω-3 fatty acid content in soy-bean is evident from
the fact that a large number of cultivars with improved oleic,
linoleic, have been released by private companies.
Contd…

45.  Common bean methionine
content has been increased by
the express ion of methionine-
rich storage albumin from
Brazil nut
 Its methionine content has
been increased by the
expression of sunflower seed
albumin gene.
Transgenic Common
Beans (Phaseolus
vulgaris)
Transgenic Lupines
(Lupinus angustifolius)

46. LENTIL BREEDING
 There is a positive correlation of iron and zinc synthesis with protein
synthesis, therefore lentil varieties with higher iron, zinc, and protein
content can be developed together (ICARDA, HarvestPlus).
 High iron and zinc lentil varieties, five in Bangladesh (Barimasur-4,
Barimasur-5, Barimasur-6, Barimasur-7, and Barimasur-8), seven in
Nepal (ILL 7723, Khajurah-1, Khajurah-2, Shital, Sisir Shekhar, Simal),
two in India (L4704, Pusa Vaibhav), one in Ethiopia (Alemaya), and two in
Syria (Idlib-2, Idlib-3) has been released by ICARDA, HarvestPlus
biofortification program till date.
 Lentil varieties have been screened for variation in Se content.

47. COW PEA BREEDING
 Cow pea which is also known as poor man meat, rich in protein
content has been biofortified for iron content by means of
breeding methods.
 Pant Lobia-1 (2008), Pant Lobia-2 (2010), Pant Lobia-3 (2013),
and Pant Lobia-4 (2014) varieties with increased iron content
have been released by GB Pant University, Pantnagar, India in
collaboration to HarvestPlus.

48. BEAN
BREEDING
Studies till date
suggest that the iron
content of the
common bean
(P.vulgaris) could be
increased by 60–
80%, while zinc
content would be
more modest,
perhaps around
50%.
 High heritability has been observed in iron and
zinc content in common bean.
 HarvestPlus is working in this direction and
promoting iron biofortified beans in several
developing countries. They have released 10 Fe-
biofortified common bean varieties in Rwanda
(RWR 2245, RWR 2154, MAC 42 MAC 44, CAB 2, RWV 1129,
RWV 3006, RWV 3316, RWV 3317, RWV 2887).
 Also released ten biofortified iron bean varieties
in the Democratic Republic of Congo, i.e., COD
MLB 001, COD MLB 032, HM 21-7, RWR 2245, PVA 1438,
COD MLV 059, VCB 81013, Nain de Kyondo, Cuarentino,
Namulenga.

49. REFERENCES
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non-conventional interventions in crop improvement, 1st Edition, M D Publishers
New Delhi,
 Burstin, J., Gallardo, K., Mir, R. R., Varshney, R. K., and Duc, G. (2011).
“Improving protein content and nutrition quality,” in Biology and Breeding of Food
Legumes, eds A. Pratap and J. Kumar (Wallingford, CT:CAB International), 314–
328.
doi: 10.1079/9781845937669.0314
 Smartt, J., Winfield, P.J. & Williams, D. (1975). A strategy for the improvement of
protein quality in pulses by breeding. Euphytica 24, 447–451.
 Kumar S, Pandey G. (2020 ) Biofortification of pulses and legumes to enhance
nutrition. Heliyon. doi:10.1016/j.heliyon.2020.e03682
 Monika G, Sharma Natasha, Sharma Saloni, Kapoor Payal, Kumar Aman,
Chunduri Venkatesh, Arora Priya. (2018). Biofortified Crops Generated by
Breeding, Agronomy, and Transgenic Approaches Are Improving Lives of Millions
of People around the World, Frontiers in Nutrition, Vol 5, doi:

50. THANK
YOU