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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.

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.

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.

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.

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.) 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.

• Background and Aims A test was made of the hypothesis that papilionate legume flowers filter pollinators according to their ability to exert strength to open flowers to access rewards. In addition, interactions with pollen vectors were expected to explain the structural complexity of the architecture of these flowers since operative flower strength may be determined by a combination of morphological traits which form part of an intrafloral functional module. • Methods Six papilionate species were studied: Collaea argentina, Desmodium uncinatum, Galactia latisiliqua, Lathyrus odoratus, Spartium junceum and Tipuana tipu. Measurements were made of the strength needed to open keels and the strength that pollinators were capable of exerting. Morphological traits of all petals were also measured to determine which of them could be either mutually correlated or correlated with operative strength and moment of strength and participated in a functional module. • Key Results It was observed that pollinators were capable in all cases of exerting forces higher and often several times higher than that needed to access floral rewards, and no association could be detected between floral operative strength and strength exerted by the corresponding pollinators. On the other hand, strong and significant correlations were found among morphometric traits and, of these, with operative strength and moment. This was particularly evident among traits of the keel and the wings, presumably involved in the functioning of the floral moveable mechanism. • Conclusions Though visitors are often many times stronger than the operative strength of the flowers they pollinate, exceptionally weak bees such as Apis mellifera cannot open the strongest flowers. On the other hand, strong correlations among certain petal morphometric traits (particularly between the keel and wings) give support to the idea that an intrafloral module is associated with the functioning of the mechanism of these legume flowers. In addition, the highly significant correlations found across petals support the view of functional phenotypic integration transcending the ontogenetic organization of flower structure.

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/Objectives: The increased frequency of extreme weather events related to climate change, including the occurrence of extreme temperatures, severely affects crop yields, impairing global food security. Heat stress resulting from temperatures above 30 °C is associated with poor germination performance and stand establishment. The combination of climate-resilient crop genotypes and tailored seed priming treatments might represent a reliable strategy to overcome such drawbacks. This work explores the potential of hydropriming as a tool to mitigate the heat-stress-mediated impact on germination performance in orphan legumes. Methods: For each tested species (Lathyrus sativus L., Pisum sativum var. arvense and Trigonella foenum-graecum L.), two accessions were investigated. Germination tests were performed at 25 °C, 30 °C, 35 °C and 40 °C to assess the heat stress tolerance threshold. Hydropriming was then applied and germination tests were performed at 40 °C to test the impact of the treatment on the seeds’ ability to cope with heat stress. An alkaline comet assay and Quantitative Real Time-Polymerase Chain Reaction were performed on embryos excised from primed and control seeds. Results: Phenotyping at the germination and seedling development stage highlighted the accession-specific beneficial impact of hydropriming under heat stress conditions. In L. sativus seeds, the alkaline comet assay revealed the dynamics of heat stress-induced DNA damage accumulation, as well as the repair patterns promoted by hydropriming. The expression patterns of genes involved in DNA repair and antioxidant response were consistently responsive to the hydropriming and heat wave conditions in L. sativus accessions.