Comprehensive characterization of protease inhibiting gene family, cis-regulatory elements, and protein interaction network in linseed and their expression upon bud fly infestation.

Linseed, also known as flax is an important oilseed crop with many potential uses in paint, textile, food and pharmaceutical industries. Susceptibility to bud fly (Dasyneura lini Barnes) infestation is a serious biotic concern leading to severe yield penalty in linseed. Protease inhibitors (PIs) are potential candidates that activate during the insect-pest attack and modulate the resistance. In the present study, we explored the PI candidates in the linseed genome and a total of 100 LuPI genes were identified and grouped into five distinct subgroups. The analysis of cis-acting elements revealed that almost all LuPI promoters contain several regulatory elementary related to growth and development, hormonal regulation and stress responses. Across the subfamilies of PIs, the specific domains are consistently found conserved in all protein sequences. The tissue-specific in-silico expression pattern via RNA-seq revealed that all the genes were regulated during different stress. The expression through qRT-PCR of 15 genes revealed the significant up-regulation of LuPI-24, LuPI-40, LuPI-49, LuPI-53, and LuPI-63 upon bud fly infestation in resistant genotype EC0099001 and resistant check variety Neela. This study establishes a foundation resource for comprehending the structural, functional, and evolutionary dimensions of protease inhibitors in linseed.

Investigating the impact of terminal heat stress on contrasting wheat cultivars: a comprehensive analysis of phenological, physiological, and biochemical traits.

Terminal heat stress has become one of the major threats due to global climate change which is significantly affecting the production and productivity of wheat crop. Therefore, it is necessary to identify key traits and genotypes to breed heat-tolerant wheat. The present study was undertaken with the objective of comparing the effects of heat stress (HSE) and extended heat stress (EHSE) on phenological-physio-biochemical traits of contrasting heat-tolerant and heat-susceptible genotypes during the reproductive phase. Phenological traits exhibited significant reduction under EHSE compared to HSE. Heat-tolerant genotypes maintained balanced phenological-physio-biochemical traits, while heat-sensitive genotypes showed significant reductions under both stress regimes. Among phenological traits, DM (R2 = 0.52) and BY (R2 = 0.44) have shown a positive effect on seed yield, indicating that biomass and crop duration contributed to the yield advantage under stress. During the grain filling stage, both the normalized difference vegetation index (NDVI) and chlorophyll (Chl) exhibited consistently positive impacts on grain yield under both HSE and EHSE conditions. This could be attributed to the enhanced photosynthesis resulting from delayed senescence and improved assimilate remobilization under terminal heat stress. The biochemical activity of superoxide dismutase (SOD), peroxidase (POX), and ascorbate peroxidase (APX) was induced in tolerant genotypes under HSE. The correlation of canopy temperature with phenological-physio-biochemical traits remained static under HSE and EHSE, suggesting CT as the best selection parameter for heat tolerance. The traits showing a positive association with yield and that are less affected under stress could be used for selecting tolerant genotypes under stress environments. These tolerant genotypes can be used to develop mapping populations to decipher the genes conferring tolerance as well as to study the molecular basis of tolerance.

Analysis of Auxin-Encoding Gene Family in Vigna radiata and It's Cross-Species Expression Modulating Waterlogging Tolerance in Wild Vigna umbellata.

Mungbean is known to be susceptible to waterlogging (WL) stress. Some of the wild species have the potential to tolerate this through various physiological and molecular mechanisms. Auxin Response Factor (ARF) and Auxin/Indole Acetic Acid (AUX/IAA), an early responsive gene family, has multiple functions in growth, development, and stress tolerance. Here, we report the first comprehensive analysis of the ARF and AUX/IAA gene family in mungbean. A total of 26 ARF and 19 AUX/IAA genes were identified from the mungbean genome. The ARF and AUX/IAA candidates were clearly grouped into two major clades. Further, the subgrouping within the major clades indicated the presence of significant diversity. The gene structure, motif analysis, and protein characterization provided the clue for further fundamental research. Out of the10 selected candidate genes, VrARF-5, VrARF-11, VrARF-25, and VrAUX/IAA-9 were found to significantly multiple-fold gene expression in the hypocotyl region of WL-tolerant wild relatives (PRR 2008-2) provides new insight into a role in the induction of lateral root formation under WL stress. The analysis provides an insight into the structural diversity of ARF and AUX/IAA genes in mungbean. These results increase our understanding of ARF and AUX/IAA genes and therefore offer robust information for functional investigations, which can be taken up in the future and will form a foundation for improving tolerance against waterlogging stress.

Morpho-Physiological and Biochemical Responses of Field Pea Genotypes under Terminal Heat Stress.

Field pea is one of the important short-duration cool season pulse crops which contributes significantly towards food and nutritional security. Two heat-susceptible (HS) and two heat-tolerant (HT) genotypes were selected from the previous study for further characterization. A significant variation was observed for morpho-physiological traits studied. Principal component analysis explained that first two principal components, i.e., PC1 and PC2 showed 76.5% of the total variance in optimal condition, whereas 91.2% of the total variance was covered by the first two PCs in heat stress environment. The seed yield per plant determined significant and positive association with superoxide dismutase and number of seeds per pod under optimal conditions, whereas under heat stress condition, it was positively associated with number of effective pods per plant, biological yield per plant, proline, pod length, number of seeds per pod, superoxide dismutase, and peroxidase. The significant reduction was noticed in the susceptible genotypes, whereas tolerant genotypes showed stable and non-significant reduction in chlorophyll content. Further, minimum cell damage and higher hydrogen peroxide production was noticed in the susceptible genotypes. In addition, the biochemical characterization of HS and HT genotypes revealed that the higher expression of peroxidase, superoxide dismutase, and catalase modulates the tolerant responses in HT genotypes. These genotypes were further used in developing heat-tolerant field pea genotypes.

Capturing agro-morphological variability for tolerance to terminal heat and combined heat-drought stress in landraces and elite cultivar collection of wheat.

Climate change has resulted in extreme temperature and drought around the globe, which poses serious threat to food security. Both heat and drought stress affects the production and productivity of wheat crop. The present study was undertaken to evaluate 34 landraces and elite cultivars of Triticum spp. for phenological and yield-related traits under optimum, heat, and combined heat-drought stress environments during 2020-2021 and 2021-2022. The pooled analysis of variance showed significant genotype × environment interaction, suggesting an influence of stress on trait expression. The trait performance of genotypes exhibited significant reduction under combined heat-drought stress as compared to optimum and heat stress environments. The maximum seed yield penalty was observed under combined heat-drought stress environment as compared to heat stress alone. Regression analysis indicated significant contribution of number of grains per spike towards stress tolerance. Based on Stress Tolerance Index (STI), genotypes Local-17, PDW 274, HI-8802, and HI-8713 were identified to be tolerant to both heat and combined heat and drought stress at Banda, whereas genotypes DBW 187, HI-8777, Raj 4120, and PDW 274 were tolerant at Jhansi location. The genotype PDW 274 showed stress tolerance under all treatments at both the locations. The genotypes PDW 233 and PDW 291 showed highest stress susceptibility index (SSI) across the environments. The number of grains per spike and test kernel weight were positively associated with seed yield across the environments and locations. The selected genotypes Local-17, HI 8802, and PDW 274 were identified to be the potential sources of heat and combined heat-drought tolerance, which may be utilized in hybridization to develop tolerant wheat genotypes and also for mapping of underlying genes/quantitative trait loci (QTLs).

Screening of Comprehensive Panel of Cultivated and Wild Vigna Species for Resistance to Pulse Beetle, Callosobruchus chinensis L.

Pulses are a key source of dietary proteins in human nutrition. Despite several efforts to increase the production, various constraints, such as biotic and abiotic factors, threaten pulse production by various means. Bruchids (Callosobruchus spp.) are the serious issue of concern, particularly in storage conditions. Understanding host-plant resistance at morphological, biochemical and molecular levels is the best way to minimize yield losses. The 117 mungbean (Vigna radiata L. Wilczek) genotypes, including endemic wild relatives, were screened for resistance against Callosobruchus chinensis; among them, two genotypes, PRR 2008-2 and PRR 2008-2-sel, which belong to V. umbellata (Thumb.), were identified as highly resistant. The expression of antioxidants in susceptible and resistant genotypes revealed that the activity of phenylalanine ammonia lyase (PAL) was upregulated in the highly resistant wild Vigna species and lower in the cultivated susceptible genotypes, along with other biomarkers. Further, the SCoT-based genotyping revealed SCoT-30 (200 bp), SCoT-31 (1200 bp) and SCoT-32 (300 bp) as unique amplicons, which might be useful for developing the novel ricebean-based SCAR markers to accelerate the molecular breeding programme.

Genome-Wide Analysis of Late Embryogenesis Abundant Protein Gene Family in Vigna Species and Expression of VrLEA Encoding Genes in Vigna glabrescens Reveal Its Role in Heat Tolerance.

Late embryogenesis abundant (LEA) proteins are identified in many crops for their response and role in adaptation to various abiotic stresses, such as drought, salinity, and temperature. The LEA genes have been studied systematically in several crops but not in Vigna crops. In this study, we reported the first comprehensive analysis of the LEA gene family in three legume species, namely, mung bean (Vigna radiata), adzuki bean (Vigna angularis), and cowpea (Vigna unguiculata), and the cross-species expression of VrLEA genes in a wild tetraploid species, Vigna glabrescens. A total of 201 LEA genes from three Vigna crops were identified harboring the LEA conserved motif. Among these 55, 64, and 82 LEA genes were identified in mung bean, adzuki bean, and cowpea genomes, respectively. These LEA genes were grouped into eight different classes. Our analysis revealed that the cowpea genome comprised all eight classes of LEA genes, whereas the LEA-6 class was absent in the mung bean genome. Similarly, LEA-5 and LEA-6 were absent in the adzuki bean genome. The analysis of LEA genes provides an insight into their structural and functional diversity in the Vigna genome. The genes, such as VrLEA-2, VrLEA-40, VrLEA-47, and VrLEA-55, were significantly upregulated in the heat-tolerant genotype under stress conditions indicating the basis of heat tolerance. The successful amplification and expression of VrLEA genes in V. glabrescens indicated the utility of the developed markers in mung bean improvement. The results of this study increase our understanding of LEA genes and provide robust candidate genes for future functional investigations and a basis for improving heat stress tolerance in Vigna crops.

Applications of Molecular Markers for Developing Abiotic-Stress-Resilient Oilseed Crops.

Globally, abiotic stresses, such as temperature (heat or cold), water (drought and flooding), and salinity, cause significant losses in crop production and have adverse effects on plant growth and development. A variety of DNA-based molecular markers, such as SSRs, RFLPs, AFLPs, SNPs, etc., have been used to screen germplasms for stress tolerance and the QTL mapping of stress-related genes. Such molecular-marker-assisted selection strategies can quicken the development of tolerant/resistant cultivars to withstand abiotic stresses. Oilseeds such as rapeseed, mustard, peanuts, soybeans, sunflower, safflower, sesame, flaxseed, and castor are the most important source of edible oil worldwide. Although oilseed crops are known for their capacity to withstand abiotic challenges, there is a significant difference between actual and potential yields due to the adaptation and tolerance to severe abiotic pressures. This review summarizes the applications of molecular markers to date to achieve abiotic stress tolerance in major oilseed crops. The molecular markers that have been reported for genetic diversity studies and the mapping and tagging of genes/QTLs for drought, heavy metal stress, salinity, flooding, cold and heat stress, and their application in the MAS are presented.

Genome-wide identification and characterization of Lectin receptor-like kinase (LecRLK) genes in mungbean (Vigna radiata L. Wilczek).

Lectins are a diverse group of proteins found throughout plant species. Numerous lectins are involved in many important processes such as organogenesis, defense mechanism, signaling, and stress response. Although the mungbean whole genome sequence has been published, distribution, diversification, and gene structure of lectin genes in mungbean are still unknown. A total of 73 putative lectin genes with kinase domain have been identified through BLAST and HMM profiling. Furthermore, these sequences could be classified into three families, such as G-type, L-type, and C-type VrLecRLKs. 59 out of 73 VrLecRLKs were distributed on to 11 chromosomes, whereas rest could not be anchored onto any specific chromosome. Gene structure analysis revealed a varying number of exons in 73 VrLecRLK genes. Gene ontology annotations were grouped into three categories like biological processes, cellular components and molecular functions, which were associated with signaling pathways, defense responses, transferase activity, binding activity, and kinase activity. The comprehensive and systematic studies of LecRLK genes family provides a reference and foundation for further functional analysis of VrLecRLK genes in mungbean.

Microsatellite-based association mapping for agronomic traits in mungbean (Vigna radiata L. Wilczek).

Mungbean (Vigna radiata L. Wilczek) is one of the most important warm season food legumes which contributes significantly towards nutritional security and environmental sustainability. Marker-trait association (MTA) for agronomic characters offer opportunities to deploy marker-assisted breeding for genetic amelioration of crops. This investigation was carried out with an objective to decipher population genetic structure of diverse Vigna accessions and detect microsatellite loci linked to major agronomic traits for mungbean improvement. The study was initiated with 290 diverse Vigna accessions including wild and cultivated accessions. A mungbean yellow mosaic India virus (MYMIV)-resistant association mapping panel was constructed to minimize the effect of yellow mosaic disease on crop performance. Among these, 117 accessions including 55 cultivated and 63 wild accessions were found highly resistant to MYMIV. After multi-environment phenotyping, a panel of 70 MYMIV-resistant mungbean accessions was subjected to analysis for assessing the population genetic structure as well as MTA for important agronomic traits. There was sufficient genetic variation among the 70-mungbean genotypes as depicted by 91 microsatellite markers. Population genetic structure analysis grouped the genotypes into five subpopulations. The locus GMES0162 (LG4) was strongly associated with days to first flowering, whereas loci CEDG 035 (LG8), DMB SSR001 (LG6), DMB SSR008 (LG4) and CEDG 168 (LG11) were associated with pod number. These marker-trait associations will be helpful in genetic improvement of mungbean through molecular breeding.

Association mapping for mungbean yellow mosaic India virus resistance in mungbean (Vigna radiata L. Wilczek).

The present study aimed to detect the marker-trait association of a selected diverse panel of 127 mungbean genotypes against mungbean yellow mosaic India virus (MYMIV). Virus-specific primers pairs viz., AC-abut/AV-abut and BC-abut/BV-abut confirmed the involvement of MYMIV in yellow mosaic disease development and the same was validated through restriction digestion analysis. 256 genome-wide microsatellite markers were screened on a test panel in which 93 polymorphic markers were used in association studies. Population structure analysis led to formation of six distinct subpopulations. 1097 alleles were detected among 127 test genotypes whereas number of alleles ranged 2-22 and PIC values ranged 0.27-0.92%, indicating ample amount of variation at genome level. 15 microsatellite markers were detected as associated with MYMIV resistance, among them three microsatellites explained 11-14% phenotypic variation. The specific regions close to CEDG293, DMB-SSR008 and DMB-SSR059 associated with MYMIV resistance were detected, located on linkage group 2, 4 and 9 and may prove useful in marker-assisted mungbean improvement programme for enhancing MYMIV resistance.