Transcriptome-wide association mapping provides insights into the genetic basis and candidate genes governing flowering, maturity and seed weight in rice bean (Vigna umbellata).

BACKGROUND: Rice bean (Vigna umbellata), an underrated legume, adapts to diverse climatic conditions with the potential to support food and nutritional security worldwide. It is used as a vegetable, minor food crop and a fodder crop, being a rich source of proteins, minerals, and essential fatty acids. However, little effort has been made to decipher the genetic and molecular basis of various useful traits in this crop. Therefore, we considered three economically important traits i.e., flowering, maturity and seed weight of rice bean and identified the associated candidate genes employing an associative transcriptomics approach on 100 diverse genotypes out of 1800 evaluated rice bean accessions from the Indian National Genebank. RESULTS: The transcriptomics-based genotyping of one-hundred diverse rice bean cultivars followed by pre-processing of genotypic data resulted in 49,271 filtered markers. The STRUCTURE, PCA and Neighbor-Joining clustering of 100 genotypes revealed three putative sub-populations. The marker-trait association analysis involving various genome-wide association study (GWAS) models revealed significant association of 82 markers on 48 transcripts for flowering, 26 markers on 22 transcripts for maturity and 22 markers on 21 transcripts for seed weight. The transcript annotation provided information on the putative candidate genes for the considered traits. The candidate genes identified for flowering include HSC80, P-II PsbX, phospholipid-transporting-ATPase-9, pectin-acetylesterase-8 and E3-ubiquitin-protein-ligase-RHG1A. Further, the WRKY1 and DEAD-box-RH27 were found to be associated with seed weight. Furthermore, the associations of PIF3 and pentatricopeptide-repeat-containing-gene with maturity and seed weight, and aldo-keto-reductase with flowering and maturity were revealed. CONCLUSION: This study offers insights into the genetic basis of key agronomic traits in rice bean, including flowering, maturity, and seed weight. The identified markers and associated candidate genes provide valuable resources for future exploration and targeted breeding, aiming to enhance the agronomic performance of rice bean cultivars. Notably, this research represents the first transcriptome-wide association study in pulse crop, uncovering the candidate genes for agronomically useful traits.

Assessment of diversity in anti-nutrient profile, resistant starch, minerals and carbohydrate components in different ricebean (Vigna umbellata) accessions.

Ricebean accessions (n = 38) cultivated in India were evaluated for their comprehensive nutrient, anti-nutrients and mineral composition. Protein and total dietary fibre ranged between 23.23 and 27.33 and 12.27 to 16.69 g/100 g, respectively. Among the oligosaccharides, verbascose was not detected, however, raffinose and stachyose ranged between 47 and 186 and 117 to 5765 mg/100 g, respectively. Among the free sugars, sucrose was found dominating (up to 370 mg/100 g). Resistant starch (4.13 to 8.62 %), iron (3.49 to 7.46 mg/100 g), zinc (1.90 to 3.72 mg/100 g) and selenium (0.28 to 4.48 µg/100 g) varied significantly (p < 0.05) among ricebean samples. Phytic acid, saponin, trypsin inhibitor and oxalate analysed in ricebean accessions ranged between 303 and 760 mg/100 g, 19 to 46 mg/g, 309 to 1076 mg/100 g and 219 to 431 mg/100 g, respectively. Multivariate analysis using hierarchical clustering analysis (HCA), and principal component analysis (PCA) was employed to decipher the diversity of nutrients and anti-nutrients across the ricebean accessions. Based on HCA, dendrogram-1 (nutrients) and dendrogram-2 (minerals, anti-nutrients) were produced, having four clusters in each. In the dendrogram-1 and 2, the largest cluster had (n = 21) and (n = 15) accessions, respectively. The PCA analyse the uncorrelated set of variables (principal components) and it condenses a large set of data variables. Based on the eigenvalue >1, a total of eight PCs were formed contributing total variance of 78.8 %. The factor loading contribution in the PC1 and PC2 were from iron, fructose, glucose, raffinose and total dietary fibre, selenium (Se) and protein, respectively.

Rice bean-adzuki bean multitrait near infrared reflectance spectroscopy prediction model: a rapid mining tool for trait-specific germplasm.

In the present era of climate change, underutilized crops such as rice beans and adzuki beans are gaining prominence to ensure food security due to their inherent potential to withstand extreme conditions and high nutritional value. These legumes are bestowed with higher nutritional attributes such as protein, fiber, vitamins, and minerals than other major legumes of the Vigna family. With the typical nutrient evaluation methods being expensive and time-consuming, non-invasive techniques such as near infrared reflectance spectroscopy (NIRS) combined with chemometrics have emerged as a better alternative. The present study aims to develop a combined NIRS prediction model for rice bean and adzuki bean flour samples to estimate total starch, protein, fat, sugars, phytate, dietary fiber, anthocyanin, minerals, and RGB value. We chose 20 morphometrically diverse accessions in each crop, of which fifteen were selected as the training set and five for validation of the NIRS prediction model. Each trait required a unique combination of derivatives, gaps, smoothening, and scatter correction techniques. The best-fit models were selected based on high RSQ and RPD values. High RSQ values of >0.9 were achieved for most of the studied parameters, indicating high-accuracy models except for minerals, fat, and phenol, which obtained RSQ <0.6 for the validation set. The generated models would facilitate the rapid nutritional exploitation of underutilized pulses such as adzuki and rice beans, showcasing their considerable potential to be functional foods for health promotion.

Evaluation and identification of advanced lentil interspecific derivatives resulted in the development of early maturing, high yielding, and disease-resistant cultivars under Indian agro-ecological conditions.

The genetic base revealed by pedigree records of the majority of released cultivars appears to be narrow in major pulse crops, including lentils, because of the frequent use of the same parents and their derivatives in crop improvement programs. Therefore, corrective measures are needed to widen the genetic base by involving the genetic resources of a distinct gene pool. In this direction, rigorous efforts were made to introgress wild Lens taxa, L. culinaris ssp. orientalis, and L. ervoides into the backgrounds of cultivated varieties. Subsequently, genetic materials were advanced through the single seed descent method of breeding along with a rapid generation advancement (normal and off-season) approach. Two F10:11 interspecific derivatives of lentils were evaluated in augmented block design at two locations, viz. International Centre for Agricultural Research in Dry Areas (ICARDA) and Sher-e-Kashmir University of Agricultural Sciences and Technology (SKUAST), India. The analysis of variance showed remarkable variability for all target characters at both locations. The heritability estimates were high, and correlation analysis exhibited a significant association between the majority of traits assessed at ICARDA and SKUAST, India. Further, SKUAST identified the most promising lines as "Jammu Lentil 144" and "Jammu Lentil 71." These derivatives were further validated separately for their agronomic potential and resistance against major biotic stresses. The results revealed that Jammu Lentil 144 and Jammu Lentil 71 produced 16.65 and 9.40% more seed yield than local and national checks, including earliness, by 25 and 15 days, respectively. These promising interspecific derivatives were also found to be resistant to fusarium wilt, root rot, pod borer, and aphid infestations. The standard agronomy of these cultivars has also been assessed consecutively for 2 years at SKUAST. Overall, the pre-breeding efforts have resulted in the development of early maturing, high-yielding, and disease-resistant lentil cultivars for the Jammu region of India.

Identification of promising chickpea interspecific derivatives for agro-morphological and major biotic traits.

The wild Cicer species is well-known for having climate-resilient and productivity-enhancing traits of interest. Therefore, wide hybridization could be used as a realistic strategy for introgressing prospective traits from wild species into the cultivated gene pool. The present study was, thus, undertaken to evaluate F7 chickpea interspecific derivatives derived from Cicer reticulatum Ladiz. and C. echinospermum P. H. Davis wild annual Cicer species. As a result, a set of six interspecific crosses were advanced using the single seed descent (SSD) method of breeding. The F7 generation of these crosses was assessed in two diverse agro-ecological regions of India. The data revealed a wide range of variation with respect to seed yield and its important component traits, which resulted in the identification of the most promising derivatives carrying desirable characters as indicated by range, mean, and coefficient of variation. Further, fruitful heterosis was also estimated as promising selection criteria for identifying superior lines for earliness and high seed yield, including resistance against prevailing stresses (ascochyta blight, botrytis gray mold, dry root rot, and fusarium wilt). The superior derivatives carrying putative characters could be recommended for further breeding and selection of genetic materials for developing suitable genotypes.

Agro-Morphological Characterization and Nutritional Profiling of Traditional Himalayan Crop Landraces for Their Promotion Toward Mainstream Agriculture.

The northwest Indian Himalayas are often regarded as a biological hotspot for the presence of rich agro-biodiversity harboring locally adapted traditional crop landraces facing utter neglect owing to modern agricultural systems promoting high-yielding varieties. Addressing this challenge requires extricating the potential of such cultivars in terms of agro-morphological and nutritional attributes. In this study, 29 traditional crop landraces of maize (11), paddy (07), finger millet (03), buckwheat (05), and naked barley (03) were characterized and evaluated for target traits of interest. In maize, Chitkanu emerged as an early maturing landrace (107 days) with high concentrations of zinc (Zn), iron (Fe), and potassium (K), and Safed makki showed the highest 100-seed weight (28.20 g). Similarly, Bamkua dhan exhibited high concentrations of K and phosphorus (P), and Lamgudi dhan showed a high protein content (14.86 g/100 g) among paddy landraces. Ogla-I and Phapra-I showed high contents of protein (14.80 g/100 g) and flavonoids (20.50 mg/g) among buckwheat landraces, respectively, followed by Nei-I, which exhibited the highest protein content (15.66 g/100 g) among naked barley landraces. Most of the target traits varied significantly (p < 0.05) among evaluated samples, except those associated with finger millet landraces. The grouping pattern obtained by principal component analysis (PCA) and multidimensional scaling (MDS) was congruent with the geographical relationship among the crop landraces. This study led to the identification of elite crop landraces having useful variations that could be exploited in plant breeding programs and biofortification strategies for future crop improvement. Our endeavor would aid in conserving the depleting Himalayan agro-biodiversity and promoting versatile traditional crops toward mainstream agriculture vis-à-vis future nutritional security.

Transcriptome Analysis Reveals Key Pathways and Candidate Genes Controlling Seed Development and Size in Ricebean (Vigna umbellata).

Ricebean (Vigna umbellata) is a lesser known pulse with well-recognized potential. Recently, it has emerged as a legume with endowed nutritional potential because of high concentration of quality protein and other vital nutrients in its seeds. However, the genes and pathways involved in regulating seed development and size are not understood in this crop. In our study, we analyzed the transcriptome of two genotypes with contrasting grain size (IC426787: large seeded and IC552985: small seeded) at two different time points, namely, 5 and 10 days post-anthesis (DPA). The bold seeded genotype across the time points (B5_B10) revealed 6,928 differentially expressed genes (DEGs), whereas the small seeded genotype across the time point (S5_S10) contributed to 14,544 DEGs. We have also identified several candidate genes for seed development-related traits like seed size and 100-seed weight. On the basis of similarity search and domain analysis, some candidate genes (PHO1, cytokinin dehydrogenase, A-type cytokinin, and ARR response negative regulator) related to 100-seed weight and seed size showed downregulation in the small seeded genotype. The MapMan and KEGG analysis confirmed that auxin and cytokinin pathways varied in both the contrasting genotypes and can therefore be the regulators of the seed size and other seed development-related traits in ricebeans. A total of 51 genes encoding SCF TIR1/AFB , Aux/IAA, ARFs, E3 ubiquitin transferase enzyme, and 26S proteasome showing distinct expression dynamics in bold and small genotypes were also identified. We have also validated randomly selected SSR markers in eight accessions of the Vigna species (V. umbellata: 6; Vigna radiata: 1; and Vigna mungo: 1). Cross-species transferability pattern of ricebean-derived SSR markers was higher in V. radiata (73.08%) than V. mungo (50%). To the best of our knowledge, this is the first transcriptomic study conducted in this crop to understand the molecular basis of any trait. It would provide us a comprehensive understanding of the complex transcriptome dynamics during the seed development and gene regulatory mechanism of the seed size determination in ricebeans.

Identification of Allelic Variation in Drought Responsive Dehydrin Gene Based on Sequence Similarity in Chickpea (Cicer arietinum L.).

Chickpea (Cicer arietinum L.) is an economically important food legume grown in arid and semi-arid regions of the world. Chickpea is cultivated mainly in the rainfed, residual moisture, and restricted irrigation condition. The crop is always prone to drought stress which is resulting in flower drop, unfilled pods, and is a major yield reducer in many parts of the world. The present study elucidates the association between candidate gene and morpho-physiological traits for the screening of drought tolerance in chickpea. Abiotic stress-responsive gene Dehydrin (DHN) was identified in some of the chickpea genotypes based on the sequence similarity approach to play a major role in drought tolerance. Analysis of variance revealed a significant effect of drought on relative water content, membrane stability index, plant height, and yield traits. The genotypes Pusa1103, Pusa362, and ICC4958 were found most promising genotypes for drought tolerance as they maintained the higher value of osmotic regulations and yield characters. The results were further supported by a sequence similarity approach for the dehydrin gene when analyzed for the presence of single nucleotide polymorphisms (SNPs) and indels. Homozygous indels and single nucleotide polymorphisms were found after the sequencing in some of the selected genotypes.

Evaluation and identification of wild lentil accessions for enhancing genetic gains of cultivated varieties.

Domesticated lentil has a relatively narrow genetic base globally and most released varieties are susceptible to severe biotic and abiotic stresses. The crop wild relatives could provide new traits of interest for tailoring novel germplasm and cultivated lentil improvement. The primary objective of this study was to evaluate wild lentil accessions for identification of economically viable agro-morphological traits and resistance against major biotic stresses. The study has revealed substantial variations in seed yield and its important component characters. Further, the diversity analysis of wild accessions showed two major clusters which were bifurcated into sub-clusters, thereby suggesting their wider genetic divergence. However, principal component analysis exhibited that seed yield plant-1, number of seeds plant-1, number of pods plant-1, harvest index and biological yield plant-1 contributed significantly to the total genetic variation assessed in wild lentil taxa. Moreover, some of the wild accessions collected from Syria and Turkey regions showed resistance against more than one disease indicating rich diversity of lentil genetic resources. The identification of most promising genotypes carrying resistance against major biotic stresses could be utilized in the cultivated or susceptible varieties of lentil for enhancing genetic gains. The study has also identified some trait specific accessions, which could also be taken into the consideration while planning distant hybridization in lentil.

Refinement of Draft Genome Assemblies of Pigeonpea (Cajanus cajan).

Genome assembly of short reads from large plant genomes remains a challenge in computational biology despite major developments in next generation sequencing. Of late several draft assemblies have been reported in sequenced plant genomes. The reported draft genome assemblies of Cajanus cajan have different levels of genome completeness, a large number of repeats, gaps, and segmental duplications. Draft assemblies with portions of genome missing are shorter than the referenced original genome. These assemblies come with low map accuracy affecting further functional annotation and the prediction of gene components as desired by crop researchers. Genome coverage, i.e., the number of sequenced raw reads mapped onto a certain location of the genome is an important quality indicator of completeness and assembly quality in draft assemblies. The present work aimed to improve the coverage in reported de novo sequenced draft genomes (GCA_000340665.1 and GCA_000230855.2) of pigeonpea, a legume widely cultivated in India. The two recently sequenced assemblies, A1 and A2 comprised 72% and 75% of the estimated coverage of the genome, respectively. We employed an assembly reconciliation approach to compare the draft assemblies and merge them, filling the gaps by employing an algorithm size sorting mate-pair library to generate a high quality and near complete assembly with enhanced contiguity. The majority of gaps present within scaffolds were filled with right-sized mate-pair reads. The improved assembly reduced the number of gaps than those reported in draft assemblies resulting in an improved genome coverage of 82.4%. Map accuracy of the improved assembly was evaluated using various quality metrics and for the presence of specific trait-related functional genes. Employed pair-end and mate-pair local libraries helped us to reduce gaps, repeats, and other sequence errors resulting in lengthier scaffolds compared to the two draft assemblies. We reported the prediction of putative host resistance genes against Fusarium wilt disease by their performance and evaluated them both in wet laboratory and field phenotypic conditions.

Transgressive segregations for agronomic improvement using interspecific crosses between C. arietinum L. x C. reticulatum Ladiz. and C. arietinum L. x C. echinospermum Davis species.

The wild species of chickpea have tremendous potential for enhancing genetic gains of cultigen and have resistant accessions against major biotic and abiotic stresses. In the present study, two wild annual accessions, one each of C. reticulatum Ladiz. (ILWC 229) and C. echinospermum Davis (ILWC 246) were assessed for their agro-morphological features and hybridized with different cultivated varieties (BGD 72, PBG 5, ICKG 96029, Pusa 372 and JG 11) of chickpea. Fertile F1 plants were developed as revealed by their normal meiotic chromosomal configuration including high pollen stainability percentage and seed set. The effect of genetic and non-genetic factors on crossability performance with respect to pod and seed set was also evident under two growing conditions of North-Western Indian Himalayas. The segregation analysis using F2 phenotypic ratio of some distinct morphological (plant growth habit, stem pigmentation at seedling stage and testa texture) characters indicated their monogenic inheritance pattern. The study would also be useful to chickpea breeders to identify true to type interspecific plants. Further, the F1, F2 and F3 generations of all seven crosses along with parents were evaluated under natural field condition to determine the extent of variability created into the cultivated background of chickpea. There was a wide range of variation in F3 population against cold stress, suggesting selection of tolerant recombinant lines at an early stage. We also studied fruitful heterosis (%) as a useful approach, instead of residual heterosis to identify better performing transgressive segregants. The values of most of the interspecific crosses for important traits assessed in F2 and F3 generations were higher than that of better parent, suggesting isolation of inbred vigour for pod numbers and earliness. The results indicated that wild Cicer annual accessions of C. reticulatum and C. echinospermum species can be exploited after proper screening for traits of interest for diversification of cultivated gene pool and subsequent use in chickpea improvement.

Bio-fortification potential of global wild annual lentil core collection.

Lentil, generally known as poor man's' meat due to its high protein value is also a good source of dietary fiber, antioxidants and vitamins along with fast cooking characteristics. It could be used globally as a staple food crop to eradicate hidden hunger, if this nutritionally rich crop is further enriched with essential minerals. This requires identification of essential mineral rich germplasm. So, in the present study, a core set of 96 wild accessions extracted from 405 global wild annual collections comprising different species was analyzed to determine its bio-fortification potential. Impressive variation (mg/100 g) was observed for different minerals including Na (30-318), K (138.29-1578), P (37.50-593.75), Ca (4.74-188.75), Mg (15-159), Fe (2.82-14.12), Zn (1.29-12.62), Cu (0.5-7.12), Mn (1.22-9.99), Mo (1.02-11.89), Ni (0.16-3.49), Pb (0.01-0.58), Cd (0-0.03), Co (0-0.63) and As (0-0.02). Hierarchical clustering revealed high intra- and inter-specific variability. Further, correlation study showed positive significant association among minerals and between minerals including agro-morphological traits. Accessions representation from Turkey and Syria had maximum variability for different minerals. Diversity analysis exhibited wide geographical variations across gene-pool in core set. Potential use of the identified trait-specific genetic resources could be initial genetic material, for genetic base broadening and biofortification of cultivated lentil.

Comparative Agronomic Performance and Reaction to Fusarium wilt of Lens culinaris × L. orientalis and L. culinaris × L. ervoides derivatives.

The development of transgressive phenotype in the segregating populations has been speculated to contribute to niche divergence of hybrid lineages, which occurs most frequently at larger genetic distances. Wild Lens species are considered to be more resistant against major biotic and abiotic stresses than that of the cultivated species. In the present study, we assessed the comparative agronomic performance of lentil (Lens culinaris subsp. culinaris) inter-sub-specific (L. culinaris subsp. orientalis) and interspecific (L. ervoides) derivatives, also discussed its probable basis of occurrence. The F3, F4, and F5 inter sub-specific and interspecific populations of ILL8006 × ILWL62 and ILL10829 × ILWL30, respectively revealed a substantial range of variation for majority of agro-morphological traits as reflected by the range, mean and coefficient of variation. A high level of fruitful heterosis was also observed in F3 and F4 progeny for important traits of interest. Phenotypic coefficient of variation (PCV) was higher in magnitude than genotypic coefficient of variation (GCV) in all generations for several quantitative characters. The results showed high heritability estimates for majority of traits in conjunction with low to high genetic advance in F3 and F4 generations. Further, F5 progeny of ILL10829 × ILWL30, manifested resistant disease reaction for fifteen recombinant inbred lines (RILs) against (Fusarium oxysporum f. sp. lentis (Vasd. Srin.) Gord.). The multilocation agronomic evaluation of both crosses showed better results for earliness, desirable seed yield and Fusarium wilt resistance under two agro-ecological regions of north-western India. These better performing recombinants of ILL8006 × ILWL62 and ILL10829 × ILWL30 can be advanced for further genetic improvement and developing high yielding disease resistant cultivars of lentil.

A High-Resolution InDel (Insertion-Deletion) Markers-Anchored Consensus Genetic Map Identifies Major QTLs Governing Pod Number and Seed Yield in Chickpea.

Development and large-scale genotyping of user-friendly informative genome/gene-derived InDel markers in natural and mapping populations is vital for accelerating genomics-assisted breeding applications of chickpea with minimal resource expenses. The present investigation employed a high-throughput whole genome next-generation resequencing strategy in low and high pod number parental accessions and homozygous individuals constituting the bulks from each of two inter-specific mapping populations [(Pusa 1103 × ILWC 46) and (Pusa 256 × ILWC 46)] to develop non-erroneous InDel markers at a genome-wide scale. Comparing these high-quality genomic sequences, 82,360 InDel markers with reference to kabuli genome and 13,891 InDel markers exhibiting differentiation between low and high pod number parental accessions and bulks of aforementioned mapping populations were developed. These informative markers were structurally and functionally annotated in diverse coding and non-coding sequence components of genome/genes of kabuli chickpea. The functional significance of regulatory and coding (frameshift and large-effect mutations) InDel markers for establishing marker-trait linkages through association/genetic mapping was apparent. The markers detected a greater amplification (97%) and intra-specific polymorphic potential (58-87%) among a diverse panel of cultivated desi, kabuli, and wild accessions even by using a simpler cost-efficient agarose gel-based assay implicating their utility in large-scale genetic analysis especially in domesticated chickpea with narrow genetic base. Two high-density inter-specific genetic linkage maps generated using aforesaid mapping populations were integrated to construct a consensus 1479 InDel markers-anchored high-resolution (inter-marker distance: 0.66 cM) genetic map for efficient molecular mapping of major QTLs governing pod number and seed yield per plant in chickpea. Utilizing these high-density genetic maps as anchors, three major genomic regions harboring each of pod number and seed yield robust QTLs (15-28% phenotypic variation explained) were identified on chromosomes 2, 4, and 6. The integration of genetic and physical maps at these QTLs mapped on chromosomes scaled-down the long major QTL intervals into high-resolution short pod number and seed yield robust QTL physical intervals (0.89-2.94 Mb) which were essentially got validated in multiple genetic backgrounds of two chickpea mapping populations. The genome-wide InDel markers including natural allelic variants and genomic loci/genes delineated at major six especially in one colocalized novel congruent robust pod number and seed yield robust QTLs mapped on a high-density consensus genetic map were found most promising in chickpea. These functionally relevant molecular tags can drive marker-assisted genetic enhancement to develop high-yielding cultivars with increased seed/pod number and yield in chickpea.

Transcriptome landscape of perennial wild Cicer microphyllum uncovers functionally relevant molecular tags regulating agronomic traits in chickpea.

The RNA-sequencing followed by de-novo transcriptome assembly identified 11621 genes differentially xpressed in roots vs. shoots of a wild perennial Cicer microphyllum. Comparative analysis of transcriptomes between microphyllum and cultivated desi cv. ICC4958 detected 12772 including 3242 root- and 1639 shoot-specific microphyllum genes with 85% expression validation success rate. Transcriptional reprogramming of microphyllum root-specific genes implicates their possible role in regulating differential natural adaptive characteristics between wild and cultivated chickpea. The transcript-derived 5698 including 282 in-silico polymorphic SSR and 127038 SNP markers annotated at a genome-wide scale exhibited high amplification and polymorphic potential among cultivated (desi and kabuli) and wild accessions suggesting their utility in chickpea genomics-assisted breeding applications. The functional significance of markers was assessed based on their localization in non-synonymous coding and regulatory regions of microphyllum root-specific genes differentially expressed predominantly in ICC 4958 roots under drought stress. A high-density 490 genic SSR- and SNP markers-anchored genetic linkage map identified six major QTLs regulating drought tolerance-related traits, yield per plant and harvest-index in chickpea. The integration of high-resolution QTL mapping with comparative transcriptome profiling delineated five microphyllum root-specific genes with non-synonymous and regulatory SNPs governing drought-responsive yield traits. Multiple potential key regulators and functionally relevant molecular tags delineated can drive translational research and drought tolerance-mediated chickpea genetic enhancement.

mQTL-seq delineates functionally relevant candidate gene harbouring a major QTL regulating pod number in chickpea.

The present study used a whole-genome, NGS resequencing-based mQTL-seq (multiple QTL-seq) strategy in two inter-specific mapping populations (Pusa 1103 × ILWC 46 and Pusa 256 × ILWC 46) to scan the major genomic region(s) underlying QTL(s) governing pod number trait in chickpea. Essentially, the whole-genome resequencing of low and high pod number-containing parental accessions and homozygous individuals (constituting bulks) from each of these two mapping populations discovered >8 million high-quality homozygous SNPs with respect to the reference kabuli chickpea. The functional significance of the physically mapped SNPs was apparent from the identified 2,264 non-synonymous and 23,550 regulatory SNPs, with 8-10% of these SNPs-carrying genes corresponding to transcription factors and disease resistance-related proteins. The utilization of these mined SNPs in Δ (SNP index)-led QTL-seq analysis and their correlation between two mapping populations based on mQTL-seq, narrowed down two (Caq(a)PN4.1: 867.8 kb and Caq(a)PN4.2: 1.8 Mb) major genomic regions harbouring robust pod number QTLs into the high-resolution short QTL intervals (Caq(b)PN4.1: 637.5 kb and Caq(b)PN4.2: 1.28 Mb) on chickpea chromosome 4. The integration of mQTL-seq-derived one novel robust QTL with QTL region-specific association analysis delineated the regulatory (C/T) and coding (C/A) SNPs-containing one pentatricopeptide repeat (PPR) gene at a major QTL region regulating pod number in chickpea. This target gene exhibited anther, mature pollen and pod-specific expression, including pronounced higher up-regulated (∼3.5-folds) transcript expression in high pod number-containing parental accessions and homozygous individuals of two mapping populations especially during pollen and pod development. The proposed mQTL-seq-driven combinatorial strategy has profound efficacy in rapid genome-wide scanning of potential candidate gene(s) underlying trait-associated high-resolution robust QTL(s), thereby expediting genomics-assisted breeding and genetic enhancement of crop plants, including chickpea.

Genome-wide high-throughput SNP discovery and genotyping for understanding natural (functional) allelic diversity and domestication patterns in wild chickpea.

We identified 82489 high-quality genome-wide SNPs from 93 wild and cultivated Cicer accessions through integrated reference genome- and de novo-based GBS assays. High intra- and inter-specific polymorphic potential (66-85%) and broader natural allelic diversity (6-64%) detected by genome-wide SNPs among accessions signify their efficacy for monitoring introgression and transferring target trait-regulating genomic (gene) regions/allelic variants from wild to cultivated Cicer gene pools for genetic improvement. The population-specific assignment of wild Cicer accessions pertaining to the primary gene pool are more influenced by geographical origin/phenotypic characteristics than species/gene-pools of origination. The functional significance of allelic variants (non-synonymous and regulatory SNPs) scanned from transcription factors and stress-responsive genes in differentiating wild accessions (with potential known sources of yield-contributing and stress tolerance traits) from cultivated desi and kabuli accessions, fine-mapping/map-based cloning of QTLs and determination of LD patterns across wild and cultivated gene-pools are suitably elucidated. The correlation between phenotypic (agromorphological traits) and molecular diversity-based admixed domestication patterns within six structured populations of wild and cultivated accessions via genome-wide SNPs was apparent. This suggests utility of whole genome SNPs as a potential resource for identifying naturally selected trait-regulating genomic targets/functional allelic variants adaptive to diverse agroclimatic regions for genetic enhancement of cultivated gene-pools.

An integrated genomic approach for rapid delineation of candidate genes regulating agro-morphological traits in chickpea.

The identification and fine mapping of robust quantitative trait loci (QTLs)/genes governing important agro-morphological traits in chickpea still lacks systematic efforts at a genome-wide scale involving wild Cicer accessions. In this context, an 834 simple sequence repeat and single-nucleotide polymorphism marker-based high-density genetic linkage map between cultivated and wild parental accessions (Cicer arietinum desi cv. ICC 4958 and Cicer reticulatum wild cv. ICC 17160) was constructed. This inter-specific genetic map comprising eight linkage groups spanned a map length of 949.4 cM with an average inter-marker distance of 1.14 cM. Eleven novel major genomic regions harbouring 15 robust QTLs (15.6-39.8% R(2) at 4.2-15.7 logarithm of odds) associated with four agro-morphological traits (100-seed weight, pod and branch number/plant and plant hairiness) were identified and mapped on chickpea chromosomes. Most of these QTLs showed positive additive gene effects with effective allelic contribution from ICC 4958, particularly for increasing seed weight (SW) and pod and branch number. One robust SW-influencing major QTL region (qSW4.2) has been narrowed down by combining QTL mapping with high-resolution QTL region-specific association analysis, differential expression profiling and gene haplotype-based association/LD mapping. This enabled to delineate a strong SW-regulating ABI3VP1 transcription factor (TF) gene at trait-specific QTL interval and consequently identified favourable natural allelic variants and superior high seed weight-specific haplotypes in the upstream regulatory region of this gene showing increased transcript expression during seed development. The genes (TFs) harbouring diverse trait-regulating QTLs, once validated and fine-mapped by our developed rapid integrated genomic approach and through gene/QTL map-based cloning, can be utilized as potential candidates for marker-assisted genetic enhancement of chickpea.

Global wild annual Lens collection: a potential resource for lentil genetic base broadening and yield enhancement.

Crop wild relatives (CWRs) are invaluable gene sources for various traits of interest, yet these potential resources are themselves increasingly threatened by the impact of climate change as well as other anthropogenic and socio-economic factors. The prime goal of our research was to cover all aspects of wild Lens genetic resource management like species characterization, agro-morphological evaluation, diversity assessment, and development of representative sets for its enhanced utilization in lentil base broadening and yield improvement initiatives. We characterized and evaluated extensively, the global wild annual Lens taxa, originating from twenty seven counties under two agro-climatic conditions of India consecutively for three cropping seasons. Results on various qualitative and quantitative characters including two foliar diseases showed wide variations for almost all yield attributing traits including multiple disease resistance in the wild species, L. nigricans and L. ervoides accessions. The core set developed from the entire Lens taxa had maximum representation from Turkey and Syria, indicating rich diversity in accessions originating from these regions. Diversity analysis also indicated wide geographical variations across genepool as was reflected in the core set. Potential use of core set, as an initial starting material, for genetic base broadening of cultivated lentil was also suggested.