This document summarizes a doctoral seminar on the genetic improvement of Lathyrus sativus (grasspea) to reduce its neurotoxic amino acid content. It discusses the nutritional value and uses of Lathyrus, but notes it contains the neurotoxin beta-N-oxalyl-L-alpha,beta-diaminopropionic acid (ODAP), which can cause the motor neuron disease lathyrism. Various breeding approaches that have been used to develop low-ODAP varieties are described, including germplasm evaluation, hybridization, mutation breeding, marker-assisted selection, transgenic methods, and somaclonal variation. Case studies on somaclonal variation in Lathyrus are presented

1. Doctoral Seminar
on
Presented By
Zuge Sopan Shivaji
Ph.D. Scholar

2. Contents
Introduction
Nutritional Composition of Lathyrus
ODAP
Genetics of ODAP
Management and Removal of ODAP
Breeding Approaches
Case Study
Conclusion

3. Introduction
 Grasspea (Lathyrus sativus L.) is a potential valuable food crop.
 Native of South Europe and Western Asia region.
 Promising crop for adaptation under climate change.
 Hardy and penetrating root system
 Hardiest crop which can survive under drought, flood and
salinity.
 Fixes 108-125 kg N/ha.
 Globally, area under Lathyrus is 1.50 mha. with annual
production of 1.20 mt. India has 0.48 mha. area under Lathyrus
with 0.334 mt. production and 701 kg/ha. productivity (NFSM,
2016)
 Potential- 5 ton/ha. (Briggs et al. 1983)

4. State wise Area and Production of Lathyrus (%)
(NFSM, 2016)
Area (2013-15) Production (2013-15)

5. Nutritional Composition of Lathyrus Seed
On dry matter basis
Protein: 24-31% (Highest after Soybean)
Carbohydrate: 41-53.9%
Total dietary fibre: 2%
Fat:0.9-2%
Many minerals : Ca, Mg, Fe, Zn etc.
Essential amino acids
Low cholesterol
Polyphenols

6. Uses of Lathyrus

7. Merits are many then why ???
BOAA

8. β-N-oxalyl-L-α, β-diaminopropionic acid (β ODAP)
Structural analogue of the neurotransmitter glutamate.
Neurotoxin (non proteinaceous) responsible for the motor neuron
degeneration syndrome Neurolathyrism.
It causes due to continuous, excess (3-4 month) consumption of
lathyrus.
In past several epidemic occurred due to excess consumption of
this crop (eg. 1995-1997 drought during which 2000 people were
crippled in Ethiopia).
In India it is reported from state of Bengal, Madhya Pradesh,
Chhattisgarh, Uttar Pradesh and Maharashtra.

9. Neurotoxins and Their Mode of Action
Neurotoxins are substances that are poisonous or destructive to
nerve tissue.
Neurotoxins adversely affect function in both developing and
mature nervous tissue.
Chemical nature is proteinaceous and nonproteinaceous.
Common examples:
1. Lead
2. Ethanol (drinking alcohol),
3. Nitric oxide (NO)
4. Tetrodotoxin
5. ODAP

10. •
One stick stage Two sticks stage Crawler stage
Severity of Neurolathyrism

11. Genetics of ODAP
Polygenic trait with high rate of GxE interaction.
Gene action-
Non additive ((Dahiya and Jeswani, 1974), additive and
additive x additive effects for ODAP content (Pandey et
al., 2000).
Heritability(h2
) - 47-65%
Correlation of traits with ODAP
Days to maturity and seed yield- negative
Seed weight- positive, negative as well as no correlation.
Simultaneous selection for yield and low ODAP

12. Management and Removal of ODAP

13. 1. Germplasm evaluation
2. Varietal development
3. Mutation breeding
4. Development of markers for MAS
5. Transgenic development
6. Somaclonal variation
Breeding Approaches

14. Germplasm Evaluation
Large collection with high variability for ODAP is available.
ICARDA- largest collections of Lathyrus with more than 3300
accessions representing 45 species from 45 countries.
India has 2720 accessions in the national gene bank at New
Delhi and 2604 accessions as active collection at Indira Gandhi
Krishi Vishwavidyalaya (IGKV), Raipur.
Catalogue on characterisation and evaluation information on 63
traits of 1963 accessions of Lathyrus published by IGKV, Raipur
in 2008.
Bangladesh has in its collection 2078 local accessions.
Kumar et al., 2011

15. Evaluation of germplasm has been taken up sporadically for
different economic traits to identify useful donors for low
ODAP content and yield traits.
0.02-2.59%
None of the species is free from ODAP
A wide range of variability for crop duration, plant height, pods
per plant, seeds per pod ,seed weight, biomass, seed yield and
ODAP content.
Maintenance of genetic purity of germplasm with low ODAP
(up to 30% cross pollination).
Pusa 24
 0.2 % ODAP with 1655 kg/ ha. yield
 Maturity 125-130 days
Cont.

16. Accessions Evaluated for ODAP Content
Kumar et al., 2011

17. Varietal Development
ODAP content is a polygenic trait and is highly influenced by G,
E and G x E.
Germplasm with low ODAP has many undesirable agronomic
traits.
In order to combine low ODAP with high yield, appropriate
phenology and stress tolerance, breeding programs have been
undertaken.
 In India following varieties with low ODAP developed
Pusa 24 with 0.2% ODAP content
Prateek and Mahateora have been developed through
hybridization at IGKV, Raipur.

18.  Prateek (LS 82046 × A 60) has low ODAP (0.076%),
high yield (1.2–1.6 ton/ha.) and tolerant to powdery and
downy mildew.
 Mahateora (Ratan × JRL 2) has 0.074% ODAP with
1.5 ton/ha. yield and is suitable for rice fallows.
 Bangladesh- BARI Khesari-1 & 2 with 0.06 and 0.04%
ODAP were developed.
Cont.

19. Released Varieties Having Low ODAP Content
IGKV (2009)
Kumar et al., 2011
IGKV (2009)

20.  Mutation breeding aim to develop zero/low ODAP varieties.
 Recurrent irradiation and subsequent selection ODAP content.
 Gamma rays and Ethyl Methyl Sulphonate (EMS) used for
generating a wide spectrum of mutants with low ODAP
concentration.
 50 Kr gamma rays + 0.2% EMS > gamma rays > EMS (Nerkar,
1972)
 International
 Bina Khesari-1 gamma rays (250 Gy) in Bangladesh with
0.2% ODAP
 It widen the genetic base of cultivated germplasm by creation of
mutants with low ODAP, erect and determinate growth.
Mutation Breeding

21. Development of Markers for Marker Assisted
Selection (MAS)
Several DNA marker systems have been applied successfully
in Lathyrus diversity studies.
However, this success has not been translated into gene
discovery or development of trait-associated markers for
marker-assisted selection (MAS).
cDNA library is also available for development of markers.
There is an urgent need to develop a comprehensive genetic
map for Lathyrus.
Association mapping
QTL mapping

22. Regeneration protocol for development complete plants via
tissue culture is available.
Agrobacterium mediated transformation and particle
bombardment
Two terminal enzymes,
Oxalyl CoA synthetase
ODAP synthase (Sehgal et al., 1992)
Silencing of any of the two enzymes block ODAP synthesis.
 Anti-sense RNA technology
Effect of Methionine on ODAP
Transgenic Development

23. Molecular Cloning of Oxalate Decarboxylase
Gene
Oxalic acid is an essential starting substrate which is
involved in neurotoxin ODAP biosynthesis.
Oxalate decarboxylase synthesized by the basidiomycete
(Collybia velutipes) showed the single step breakdown
oxalic acid.
Gene for oxalate decarboxylase is already isolated and
sequenced.
More than 60% decrease in the level of total oxalate is
observed in transgenic Tobacco and Tomato.
This novel gene can be used to develop transgenic plants
with low ODAP content.
Kesarwni et al., 2000

24. Somaclonal Variation
Somaclonal variation is the variation seen in plants that have been
produced by plant tissue culture.
Chromosomal rearrangements are an important source of this
variation.
Characters affected include both qualitative and quantitative traits.
Mutations may be due varied nutrients, culture conditions and
mutagenic effects of metabolic products that accumulate in the
medium.
Ratan
Somaclon of Pusa 24
0.05 % ODAP
2530 kg/ ha. Yield potential.

25. Case Study
Four genotypes namely Nirmal (0.239), P24 (0.314), Nayagarh
local (0.438) and Dhenkanal local (0.402) were used for
development of somaclones.
Internodes were used as explants.
Chromosomal abnormalities and variation in one or more
morphological trait was observed.
Variation for ODAP was found in range of 0.045 to 0.531 in R1
generation.

26. Somaclones with Low ODAP Content Selected
from R2 Generation

27. CONCLUSION
Ban is not the solution, it requires research and budget
allocation.
Global attention is needed for genetic improvement.
Moreover, yield can be further increased with low ODAP in
Lathyrus.
Communication and collaborative research between medical
people, neuroscientists, geneticists and agronomists is necessary
for improvement of this crop.
GOI should take proper step to provide improved varieties to
farmer.
Eliminate the toxin from crop, not the crop from cultivation.

28. THANK YOU