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Grass pea ( Lathyrus sativus L.) is a protein-rich legume widely cultivated in drought-prone areas of Asia and Africa. Despite its resilience and nutritional value, Lathyrus suffers from limited genetic variability and the persistent problem of β-ODAP toxicity, which restricts consumption and warrants focused breeding initiatives. Developing high-yielding, low-ODAP varieties is critical for food safety and agricultural productivity. The present study employed gamma irradiation (250, 300, 350 Gy) to induce mutagenesis in seeds of cultivar NLK-73. Through successive generational selection (up to M₄), 29 promising mutants were evaluated in a randomized block design. Phenotypic and yield attributes were measured, along with ODAP quantification using spectrophotometry. Data analysis included ANOVA, estimation of genetic parameters, heritability, and genetic advance. Significant genetic variability was observed among M₄ mutants for all evaluated traits. The analysis of variance indicated highly significant differences ( p  < 0.01) among genotypes for days to flowering, maturity, plant height, branches/plant, pods/plant, 100-seed weight, seed yield, and ODAP content. High heritability (> 60%) and substantial genetic advance were found for key traits such as branches and pods per plant, suggesting additive genetic action. Ten mutants (notably NLM-12, NLM-20, NLM-23) surpassed checks in seed yield (23–24.5 g/plant vs. 13.9 g/plant) with proportionately lower ODAP content, marking them as candidates for breeding programs and further evaluation. Gamma ray mutagenesis effectively broadened variability in Lathyrus sativus , enabling selection of superior M₄ mutants with enhanced yield and reduced ODAP content. The results suggest the feasibility of developing safer, high-yielding grass pea cultivars, warranting further validation. Adoption of mutation breeding should continue for rapid improvement of grass pea, focusing on reducing β-ODAP to trace levels while maximizing germplasm diversity and yield. Multi-location field trials are recommended to confirm stability of desirable traits. Molecular characterization and marker-assisted selection to expedite breeding for low-ODAP, high-protein lines is warranted. Exploration of alternative mutagens and advanced genomic tools will facilitate precise genetic improvement.

Background The MADS-box gene family possesses significant potential to improve crop production under harsh conditions by regulating growth, development, and the expression of floral organs. The grass pea (Lathyrus sativus), a crop grown predominantly in arid and semi-arid regions, could benefit greatly from the functions of MADS-box genes, which are not yet well characterized in this promising plant. Results In this study, a comprehensive analysis of all MADS-box genes in grass pea was performed at both the genomic and transcriptomic levels. A total of 46 genes were identified and classified based on their MADS-box domains. A comparative phylogenetic analysis with apple, Arabidopsis, and rice categorized the grass pea genes into 31 type I genes (M , M , M ) and 15 type II genes (MIKCc, MIKC*). Annotation analysis revealed variations in the intron-exon structures of the genes, with most type I genes being intronless. Ten distinct conserved motifs were identified across the genes. Structural analysis revealed the presence of MEF2-like and SRF-TF domains in the grass pea proteins. Protein-protein interaction analysis revealed extensive interactions among type II MADS-box genes, while enrichment analysis showed their involvement in various aspects of plant life, particularly floral organ development. Examination of the cis-elements in the promoter regions of the genes revealed up to 76 potential cis-elements, which were categorized into four groups based on their putative role in transcriptional regulation. RNA-seq was used to profile gene expression under different conditions to gain insights into their potential functional significance. Quantitative PCR (qPCR) analysis validated the expression levels of eight selected genes (LSMADS_D1, LSMADS_R5, LSMADS_R7, LSMADS_R9, LSMADS_D11, LSMADS_D13, LSMADS_R13, and LSMADS_D29) under salt stress conditions and confirmed their involvement in stress responses. Conclusion This study represents the first genome-wide exploration of the MADS-box gene family in grass pea. Our results provide valuable insights that could improve our understanding of the plant’s genomics, contribute to strengthening its resilience to challenging conditions, and help position it as an important crop in arid regions.

Background Tribe Fabeae comprises about 380 legume species, including some of the most ancient and important crops like lentil, pea, and broad bean. Breeding efforts in legume crops rely on a detailed knowledge of closest wild relatives and geographic origin. Relationships within the tribe, however, are incompletely known and previous molecular results conflicted with the traditional morphology-based classification. Here we analyse the systematics, biogeography, and character evolution in the tribe based on plastid and nuclear DNA sequences. Results Phylogenetic analyses including c. 70% of the species in the tribe show that the genera Vicia and Lathyrus in their current circumscription are not monophyletic: Pisum and Vavilovia are nested in Lathyrus , the genus Lens is nested in Vicia . A small, well-supported clade including Vicia hirsuta , V. sylvatica , and some Mediterranean endemics, is the sister group to all remaining species in the tribe. Fabeae originated in the East Mediterranean region in the Miocene (23–16 million years ago (Ma)) and spread at least 39 times into Eurasia, seven times to the Americas, twice to tropical Africa and four times to Macaronesia. Broad bean ( V. faba ) and its sister V. paucijuga originated in Asia and might be sister to V. oroboides . Lentil ( Lens culinaris ssp. culinaris ) is of Mediterranean origin and together with eight very close relatives forms a clade that is nested in the core Vicia , where it evolved c. 14 Ma. The Pisum clade is nested in Lathyrus in a grade with the Mediterranean L. gloeosperma , L. neurolobus , and L. nissolia . The extinct Azorean endemic V. dennesiana belongs in section Cracca and is nested among Mediterranean species. According to our ancestral character state reconstruction results, ancestors of Fabeae had a basic chromosome number of 2n=14, an annual life form, and evenly hairy, dorsiventrally compressed styles. Conclusions Fabeae evolved in the Eastern Mediterranean in the middle Miocene and spread from there across Eurasia, into Tropical Africa, and at least seven times to the Americas. The middle-Atlantic islands were colonized four times but apparently did not serve as stepping-stones for Atlantic crossings. Long-distance dispersal events are relatively common in Fabeae (seven per ten million years). Current generic and infrageneric circumscriptions in Fabeae do not reflect monophyletic groups and should be revised. Suggestions for generic level delimitation are offered.

Grass pea (Lathyrus sativus L.) is an important legume crop grown mainly in South Asia and Sub-Saharan Africa. This underutilized legume can withstand harsh environmental conditions including drought and flooding. During drought-induced famines, this protein-rich legume serves as a food source for poor farmers when other crops fail under harsh environmental conditions; however, its use is limited because of the presence of an endogenous neurotoxic nonprotein amino acid β-N-oxalyl-l-α,β-diaminopropionic acid (β-ODAP). Long-term consumption of Lathyrus and β-ODAP is linked to lathyrism, which is a degenerative motor neuron syndrome. Pharmacological studies indicate that nutritional deficiencies in methionine and cysteine may aggravate the neurotoxicity of β-ODAP. The biosynthetic pathway leading to the production of β-ODAP is poorly understood, but is linked to sulfur metabolism. To date, only a limited number of studies have been conducted in grass pea on the sulfur assimilatory enzymes and how these enzymes regulate the biosynthesis of β-ODAP. Here, we review the current knowledge on the role of sulfur metabolism in grass pea and its contribution to β-ODAP biosynthesis. Unraveling the fundamental steps and regulation of β-ODAP biosynthesis in grass pea will be vital for the development of improved varieties of this underutilized legume.

Grass pea (Lathyrus sativus L.) is a rich source of protein cultivated as an insurance crop in Ethiopia, Eritrea, India, Bangladesh, and Nepal. Its resilience to both drought and flooding makes it a promising crop for ensuring food security in a changing climate. The lack of genetic resources and the crop’s association with the disease neurolathyrism have limited the cultivation of grass pea. Here, we present an annotated, long read-based assembly of the 6.5 Gbp L. sativus genome. Using this genome sequence, we have elucidated the biosynthetic pathway leading to the formation of the neurotoxin, β-L-oxalyl-2,3-diaminopropionic acid (β-L-ODAP). The final reaction of the pathway depends on an interaction between L. sativus acyl-activating enzyme 3 (LsAAE3) and a BAHD-acyltransferase (LsBOS) that form a metabolon activated by CoA to produce β-L-ODAP. This provides valuable insight into the best approaches for developing varieties which produce substantially less toxin.

Key message Established an Agrobacterium-mediated hairy root transformation system for gene function analysis in Lathyrus sativus . Arabidopsis mutant complementation and genome editing in Lathyrus confirmed role of LsOCS in the oxalate metabolism. Grass pea ( Lathyrus sativus ) is a resilient legume cultivated for its protein-rich seeds and fodder. However, the presence of a naturally occurring neurotoxin, β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP), which causes neurolathyrism, limits its extensive cultivation. This paper reports the in-planta characterization of oxalyl-CoA synthetase (OCS), an enzyme involved in oxalate metabolism and important in the oxalylating step leading to β-ODAP production in Lathyrus . For this, we used complementation experiments in an Arabidopsis OCS mutant. The LsOCS -complemented lines showed oxalate content similar to wild-type levels, and the analysis of seeds by field emission scanning electron microscope (FESEM) showed that the LsOCS -complemented lines were rescued from seed-coat defects found in the mutant seeds. We used genome editing of LsOCS in Lathyrus hairy roots to further characterize LsOCS function. The mutations in LsOCS resulted in the accumulation of oxalate in the hairy roots of Lathyrus , as observed in Arabidopsis mutants, but did not affect the ODAP levels. The hairy root genome editing system could serve as a rapid tool for functional studies of Lathyrus genes and optimizing the agronomic traits.

Grass pea ( Lathyrus sativus L.) stands out as an excellent choice for sustainable agriculture, thanks to its favorable agronomic characteristics, including a robust root system that penetrates deeply into the soil and its resilience against various biotic and abiotic stressors. In this study, dry-matter yield and seed yield of 16 grass pea genotypes were evaluated in rain-fed conditions at “Gachsaran”, “Mehran”, “Kuhdasht”, and “Shirvan-Chardavol” locations in Iran for three consecutive years. The experimental field trials were carried out using a randomized complete block design, and each experimental setup was replicated three times. The descriptive statistics showed a mean value of 4.030 (ton/ha) and 1.530 (ton/ha), with phenotypic coefficients of 54.77 and 61.56 for dry-matter yield and seed yield, respectively. The projection of geographical, climatic, and edaphic variables into yield measurements depicted remarkable divergence among the four studied environments. Elevation exerts a greater influence on both dry matter and seed yields in the Mehran location as compared to other environments. The climatic factors of rainfall and relative humidity played an important role in “Gachsaran” and “Shirvan-Chardavol”, respectively. While for seed yield, the temperature-related attributes were more significant in the “Mehran” location. Low broad-sense heritability was observed, and the R 2 for genotype-environment interaction showed the existence of GEI for dry-matter yield (0.126) and seed yield (0.223). Both AMMI1 and AMMI2 could recognize unstable genotypes from other ones, and both AMMI’s identified genotypes G10 and G3 as high-yielding and stable genotypes. BLUP-based stability indices revealed G10 and G13 as superior genotypes for seed yield and dry-matter yield, respectively. Three and two mega-environments were identified using a GGE biplot for dry-matter yield and seed yield. For dry-matter-identified mega-environments, the G1, G13, and G2, and for seed-yield-recognized mega-environments, the G10 and G15 can be introduced. “Mehran” and “Gachsaran” out of the studied locations possessed diverse distributions considering dry-matter yield and seed yield and for further GE interaction studies, it is better to establish adaptability trials in these locations. The study concludes that for the promotion of sustainable agriculture in rain-fed regions, taking into account the influence of environmental factors, cultivation of the identified grass pea genotypes holds promise.

Rain-fed regions have a low quantity of rainfall with an asymmetric distribution. Therefore, by promoting plants like Lathyrus sativus L., as a legume adapted to unfavorable environments, genotypes with high fodder capacity under such conditions would assist food security worldwide. Here, 16 grass pea genotypes were examined in four rain-fed regions during 2016–2017, 2017–2018, and 2018–2019. Dry fodder yield (DY), plant height (PH), days to flowering (DF), and wet fodder yield (WY) were recorded across 12 test environments. Regarding MLM analysis of variance, LRT ENV and LRT ENV×GEN were significant for all studied traits. Phenotypic variance ranged between 1.42 (DY) to 86.9 (PH). Results showed the possibility of grass pea improvement through selection regarding calculated accuracy of selection (> 0.5). PLS regression emphasized the significant role of rainfall during December, January, February, March and April on DY and WY of grass pea. The DY of 16 genotypes across environments varied between 3.4 t/ha (G12 and G16) to 4.6 t/ha (G11). The WY also varied between 16.9 t/ha (G12) and 22.0 t/ha (G8). AMMI analysis revealed G2, and G6 and BLUP-based indices showed G8, and G11 as climate-resilient genotypes with stable DY and WY in rain-fed regions. In this study, WAASB×DY and WAASB×WY plots with equal weights of 50/50 for stability and performance showed G2, G6 as stable genotypes with high DY and WY values. Simultaneous selection based on overall recorded traits using MTSI index addressed G9 > G2 as promising genotypes. Although the polygon view of genotype by yield*trait depicted G1 and G11 as promising grass pea genotypes but G2, and G9 also had positive intermediate superiority indexes without any weakness considering studied traits. It is concluded WAASB×yield > AMMI > BLUP in terms of comprehensiveness in yield stability analysis of grass pea. Also, superiority index as complementary statistics could be incorporated into simultaneous multi-trait stability approaches for achieving exact selection. The identified grass pea genotypes have promising potential in rain-fed regions and could be good candidates for commercial production.

Summary Soya bean (Glycine max) and grass pea (Lathyrus sativus) seeds are important sources of dietary proteins; however, they also contain antinutritional metabolite oxalic acid (OA). Excess dietary intake of OA leads to nephrolithiasis due to the formation of calcium oxalate crystals in kidneys. Besides, OA is also a known precursor of β‐N‐oxalyl‐L‐α,β‐diaminopropionic acid (β‐ODAP), a neurotoxin found in grass pea. Here, we report the reduction in OA level in soya bean (up to 73%) and grass pea (up to 75%) seeds by constitutive and/or seed‐specific expression of an oxalate‐degrading enzyme, oxalate decarboxylase (FvOXDC) of Flammulina velutipes. In addition, β‐ODAP level of grass pea seeds was also reduced up to 73%. Reduced OA content was interrelated with the associated increase in seeds micronutrients such as calcium, iron and zinc. Moreover, constitutive expression of FvOXDC led to improved tolerance to the fungal pathogen Sclerotinia sclerotiorum that requires OA during host colonization. Importantly, FvOXDC‐expressing soya bean and grass pea plants were similar to the wild type with respect to the morphology and photosynthetic rates, and seed protein pool remained unaltered as revealed by the comparative proteomic analysis. Taken together, these results demonstrated improved seed quality and tolerance to the fungal pathogen in two important legume crops, by the expression of an oxalate‐degrading enzyme.

Background: There is an abundance of the neuroactive β-N-oxalyl-L-α,β-diaminopropionic acid (β-ODAP) in grass pea (Lathyrus sativus), pea (Pisum sativum), and several Chinese traditional herbs such as Panax notoginseng. It is well known for its dose- and context-dependent effects on its toxicological characteristics (inducing neurodegenerative neurolathyrism upon excessive consumption) or for its pharmacological effects (including neuroprotection and wound healing). Therefore, reducing β-ODAP levels improves the safety profile of β-ODAP-containing species for utilization, whereas increasing them facilitates their isolation and purification. LsBAHD3 acyltransferase, named after the first letter of BEAT benzylalcohol O-acetyltransferase (BEAT), anthocyanin O-hydroxycinnamoyltransferase (AHCT), anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT), and deacetylvindoline 4-Oacetyltransferase (DAT), was proven to be β-ODAP synthetase. Methods: In this report, the interaction of miR172f with LsBAHD3 was investigated through bioinformatic analysis and transient co-expression assays in Nicotiana benthamiana. Functions of miR172f in β-ODAP biosynthesis were also investigated through knockdown in the hairy roots of L. sativus and via transcriptomic analysis. Results: The results suggest that the knockdown of miR172f in hairy roots of L. sativus increased β-ODAP content via targets to LsBAHD3. In this process, protein ubiquitination, cysteine and methionine metabolism, enzyme regulator activity, and so on were associated with β-ODAP biosynthesis. Conclusions: These results identify miR172f as a novel regulator of β-ODAP biosynthesis through targeting of LsBAHD3, offering new insight into the gene expression of β-ODAP synthetase and the genetic network governing β-ODAP biosynthesis in L. sativus.