Mungbean (Vigna radiata L. Wilczek) is a vital grain legume whose productivity is often constrained by limited phosphorus availability. Phosphorus deficiency hampers plant growth and alters root morphology, typically increasing the root-to-shoot ratio. Enhancing phosphorus-use efficiency (PUE) is therefore essential for improving mungbean performance under nutrient-limited conditions. To explore the genetic basis of PUE, transcriptome sequencing was performed on leaves, stems, and roots of two contrasting genotypes: Pusa 1333 (phosphorus-efficient) and PS 16 (phosphorus-inefficient). RNA-seq analysis revealed 833 upregulated and 1081 downregulated genes across tissues. In roots, 137 genes were upregulated and 477 downregulated; in stems, 365 were upregulated and 294 downregulated; and in leaves, 331 were upregulated and 310 downregulated. Gene Ontology (GO) classification indicated enrichment in biological processes related to growth, metabolism, and cellular organization. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that differentially expressed genes (DEGs) were predominantly associated with metabolic activity and secondary metabolite biosynthesis. Notably, eight DEGs were consistently expressed across all three tissues. Among these, Vradi05g03810 (polygalacturonase) interacted with cell wall–related genes, suggesting a role in modulating cell wall dynamics under phosphorus stress. Vradi05g03870 (RD22-like glycosyl transferase) interacted with stress-responsive proteins, indicating involvement in dehydration tolerance. These findings provide novel insights into the molecular regulation of PUE in mungbean and highlight candidate genes for breeding programs aimed at improving phosphorus efficiency and resilience under nutrient-limited conditions.

Mungbean (Vigna radiata) is a short-duration legume crop, and identifying suitable housekeeping genes is crucial for accurate gene expression analysis under both normal and stress conditions, as they enable proper normalization of target gene expression data. To address this gap, we carried out a comparative analysis of ten housekeeping genes to determine the most appropriate reference genes for assessing target gene expression. Among the ten genes, EF-1 ALPHA, CYP1, and GAPDH showed the most stable expression across different tissues compared with the other reference genes. All three genes exhibited lower Ct values and better stability scores across different algorithms than the other genes tested. Further, the suitability of these selected reference genes was validated by examining the expression of PHENYLALANINE AMMONIA LYASE (PAL), CINNAMOYL ALCOHOL DEHYDROGENASE (CAD), IRREGULAR XYLEM 14 (IRX14), and RIBULOSE-1,5-BISPHOSPHATE CARBOXYLASE/OXYGENASE (RuBisCO) in three different mungbean genotypes and tissue types. We further tested the expression of RuBisCO in seedlings under salt and osmotic stress and found that its expression was lower than that in control plants, which correlated with the seedling phenotype. Moreover, we validated the efficacy of these reference genes by examining the expression of selected target genes and found that all were upregulated in mungbean yellow mosaic virus-infected leaves compared with non-infected leaves. Based on these findings, we propose that EF-1 ALPHA is the best reference gene, followed by CYP1 and GAPDH, for analyzing gene expression across different tissues and developmental stages under both normal and stress conditions in mungbean.

Mungbean (Vigna radiata L.) is one of the most important yet genomically under-researched leguminous food crop. Its productivity is low due to the complex nature of yield realization, which is regulated by various yield-contributing traits. Thus, understanding the genetic basis of these traits is essential for developing an ideal genotype with high yield. In this study, mapping of quantitative trait loci (QTLs) was conducted for six yield-contributing traits using a recombinant inbred line (RIL) population (166 number), developed by crossing two contrasting genotypes (Pusa Baisakhi × PMR-1). The genotyping-by-sequencing (GBS) of RILs was used to construct the genetic map using 1347 single nucleotide polymorphism (SNPs). The QTL mapping using multiple interval mapping (MIM) and composite interval mapping (CIM) has identified 17 yield contributing QTLs of which, 4 for number of leaves/plant (NL), 3 for plant height (PH), 2 for SPAD value, 3 for 100 seed weight (SW), 2 for number of pods/plant (NP), and 3 for total grain yield (GY). The Logarithm of Odds (LOD) scores for these QTLs ranged from ~ 3-9, while phenotypic variance explained (PVE) ranged from ~ 9-24%. Several candidate genes with mRNA expression and protein-altering mutations were identified as having a direct role in key processes like growth (LOC106756212, LOC106776425, LOC106777991), flowering (LOC106777903, LOC106768860), metabolism (LOC106757749, LOC106758189), etc. The candidate genes are validated through digital gene expression analysis. In addition, Insertion-Deletion (InDel) markers were also developed for the identified QTLs which hold broad applications for the improvement of yield-related traits in mungbean.

Quantitative trait loci (QTL) mapping is used for the precise localization of genomic regions regulating various traits in plants. Two major QTLs regulating Soil Plant Analysis Development (SPAD) value ( qSPAD-7-1 ) and trichome density ( qTric-7-2 ) in mungbean were identified using recombinant inbred line (RIL) populations (PMR-1×Pusa Baisakhi) on chromosome 7. Functional analysis of QTL region identified 35 candidate genes for SPAD value (16 No) and trichome (19 No) traits. The candidate genes regulating trichome density on the dorsal leaf surface of the mungbean include VRADI07G24840, VRADI07G17780 , and VRADI07G15650, which encodes for ZFP6, TFs bHLH DNA-binding superfamily protein, and MYB102, respectively. Also, candidate genes having vital roles in chlorophyll biosynthesis are VRADIO7G29860, VRADIO7G29450 , and VRADIO7G28520 , which encodes for s-adenosyl-L-methionine, FTSHI1 protein, and CRS2-associated factor, respectively. The findings unfolded the opportunity for the development of customized genotypes having high SPAD value and high trichome density having a possible role in yield and mungbean yellow vein mosaic India virus (MYMIV) resistance in mungbean.

Background Lignin and xylan are important determinants of cell wall structure and lignocellulosic biomass digestibility. Genetic manipulations that individually modify either lignin or xylan structure improve polysaccharide digestibility. However, the effects of their simultaneous modifications have not been explored in a similar context. Here, both individual and combinatorial modification in xylan and lignin was studied by analysing the effect on plant cell wall properties, biotic stress responses and integrity sensing. Results Arabidopsis plant co-harbouring mutation in FERULATE 5-HYDROXYLASE (F5H) and overexpressing Aspergillus niger acetyl xylan esterase (35S:AnAXE1) were generated and displayed normal growth attributes with intact xylem architecture. This fah1-2/35S:AnAXE1 cross was named as hyper G lignin and hypoacetylated (HrGHypAc) line. The HrGHypAc plants showed increased crystalline cellulose content with enhanced digestibility after chemical and enzymatic pre-treatment. Moreover, both parents and HrGHypAc without and after pre-treating with glucuronyl esterase and alpha glucuronidase exhibited an increase in xylose release after xylanase digestion as compared to wild type. The de-pectinated fraction in HrGHypAc displayed elevated levels of xylan and cellulose. Furthermore, the transcriptomic analysis revealed differential expression in cell wall biosynthetic, transcription factors and wall-associated kinases genes implying the role of lignin and xylan modification on cellular regulatory processes. Conclusions Simultaneous modification in xylan and lignin enhances cellulose content with improved saccharification efficiency. These modifications loosen cell wall complexity and hence resulted in enhanced xylose and xylobiose release with or without pretreatment after xylanase digestion in both parent and HrGHypAc. This study also revealed that the disruption of xylan and lignin structure is possible without compromising either growth and development or defense responses against Pseudomonas syringae infection.

Background Dissection of complex plant cell wall structures demands a sensitive and quantitative method. FTIR is used regularly as a screening method to identify specific linkages in cell walls. However, quantification and assigning spectral bands to particular cell wall components is still a major challenge, specifically in crop species. In this study, we addressed these challenges using ATR-FTIR spectroscopy as it is a high throughput, cost-effective and non-destructive approach to understand plant cell wall composition. This method was validated by analysing different varieties of mungbean which is one of the most important legume crop grown widely in Asia. Results Using standards and extraction of a specific component of cell wall components, we assigned 1050-1060 cm -1 and 1390-1420 cm -1 wavenumbers that can be widely used to quantify cellulose and lignin, respectively, in Arabidopsis, Populus , rice and mungbean. Also, using KBr as a diluent, we established a method which can relatively quantify the cellulose and lignin composition among different tissue types of the above species. We further used this method to quantify cellulose and lignin in field-grown mungbean genotypes. The ATR-FTIR-based study revealed the cellulose content variation ranges from 27.9% to 52.37%, and the lignin content variation ranges from 13.77% to 31.6% in mungbean genotypes. Conclusion Cell wall composition in different mungbean genotypes was determined by the developed FT-IR-based method, which was cross-validated using canonical wet-chemistry methods. Overall, our data suggested that ATR-FTIR can be used for the relative quantification of lignin and cellulose in different plant species. This method can be used for rapid screening of cell wall composition in large number of germplasms of different crops including mungbean.

The seed size and shape in lentil (Lens culinaris Medik.) are important quality traits as these influences the milled grain yield, cooking time, and market class of the grains. Linkage analysis was done for seed size in a RIL (F5:6) population derived by crossing L830 (20.9 g/1000 seeds) with L4602 (42.13 g/1000 seeds) which consisted of 188 lines (15.0 to 40.5 g/1000 seeds). Parental polymorphism survey using 394 SSRs identified 31 polymorphic primers, which were used for the bulked segregant analysis (BSA). Marker PBALC449 differentiated the parents and small seed size bulk only, whereas large seeded bulk or the individual plants constituting the large-seeded bulk could not be differentiated. Single plant analysis identified only six recombinant and 13 heterozygotes, of 93 small-seeded RILs (<24.0 g/1000 seed). This clearly showed that the small seed size trait is very strongly regulated by the locus near PBLAC449; whereas, large seed size trait seems governed by more than one locus. The PCR amplified products from the PBLAC449 marker (149bp from L4602 and 131bp from L830) were cloned, sequenced and BLAST searched using the lentil reference genome and was found amplified from chromosome 03. Afterward, the nearby region on chromosome 3 was searched, and a few candidate genes like ubiquitin carboxyl-terminal hydrolase, E3 ubiquitin ligase, TIFY-like protein, and hexosyltransferase having a role in seed size determination were identified. Validation study in another RIL mapping population which is differing for seed size, showed a number of SNPs and InDels among these genes when studied using whole genome resequencing (WGRS) approach. Biochemical parameters like cellulose, lignin, and xylose content showed no significant differences between parents and the extreme RILs, at maturity. Various seed morphological traits like area, length, width, compactness, volume, perimeter, etc., when measured using VideometerLab 4.0 showed significant differences for the parents and RILs. The results have ultimately helped in better understanding the region regulating the seed size trait in genomically less explored crops like lentils.

Plant cell wall-derived oligosaccharides, i.e., damage-associated molecular patterns (DAMPs), could be generated after pathogen attack or during normal plant development, perceived by cell wall receptors, and can alter immunity and cell wall composition. Therefore, we hypothesised that xylo-oligosaccharides (XOS) could act as an elicitor and trigger immune responses. To test this, we treated Arabidopsis with xylobiose (XB) and investigated different parameters. XB-treatment significantly triggered the generation of reactive oxygen species (ROS), activated MAPK protein phosphorylation, and induced callose deposition. The combination of XB (DAMP) and flg22 a microbe-associated molecular pattern (MAMP) further enhanced ROS response and gene expression of PTI marker genes. RNA sequencing analysis revealed that more genes were differentially regulated after 30 min compared to 24 h XB-treated leaves, which correlated with ROS response. Increased xylosidase activity and soluble xylose level after 30 min and 3 h of XB-treatment were observed which might have weakened the DAMP response. However, an increase in total cell wall sugar and a decrease in uronic acid level was observed at both 30 min and 24 h. Additionally, arabinose, rhamnose, and xylose levels were increased in 30 min, and glucose was increased in 24 h compared to mock-treated leaves. The level of jasmonic acid, abscisic acid, auxin, and cytokinin were also affected after XB treatment. Overall, our data revealed that the shortest XOS can act as a DAMP, which triggers the PTI response and alters cell wall composition and hormone level. Xylobiose treatment in Arabidopsis rosette leaf induces plant-triggered immunity (PTI) responses, alters cell wall polysaccharide composition and influences growth hormone levels.

Microgreens have been used for raw consumption and are generally viewed as healthy food. This study aimed to optimize the yield parameters, shelf life, sensory evaluation and characterization of total aerobic bacteria (TAB), yeast and mold (Y&M), Escherichia coli, Salmonella spp., and Listeria spp. incidence in mungbean (Vigna radiata (L.) Wilczek), lentil (Lens culinaris Medikus subsp. culinaris), and Indian mustard (Brassica juncea (L.) Czern & Coss.) microgreens. In mungbean and lentil, seeding-density of three seed/cm(2), while in Indian mustard, eight seed/cm(2) were recorded as optimum. The optimal time to harvest mungbean, Indian mustard, and lentil microgreens were found as 7(th), 8(th), and 9(th) day after sowing, respectively. Interestingly, seed size was found highly correlated with the overall yield in both mungbeans (r(2) = .73) and lentils (r(2) = .78), whereas no such relationship has been recorded for Indian mustard microgreens. The target pathogenic bacteria such as Salmonella spp. and Listeria spp. were not detected; while TAB, Y&M, Shigella spp., and E. coli were recorded well within the limit to cause any human illness in the studied microgreens. Washing with double distilled water for two minutes has shown some reduction in the overall microbial load of these microgreens. The results provided evidence that microgreens if grown and stored properly, are generally safe for human consumption. This is the first study from India on the safety of mungbean, lentils, and Indian mustard microgreens.

Market class, cooking time, quality, and milled grain yield are largely influenced by the seed size and shape of the lentil (Lens culinaris Medik.); thus, they are considered to be important quality traits. To unfold the pathways regulating seed size in lentils, a transcriptomic approach was performed using large-seeded (L4602) and small-seeded (L830) genotypes. The study has generated nearly 375 million high-quality reads, of which 98.70% were properly aligned to the reference genome. Among biological replicates, very high similarity in fragments per kilobase of exon per million mapped fragments values (R > 0.9) showed the consistency of RNA-seq results. Various differentially expressed genes associated mainly with the hormone signaling and cell division pathways, transcription factors, kinases, etc. were identified as having a role in cell expansion and seed growth. A total of 106,996 unigenes were used for differential expression (DE) analysis. String analysis identified various modules having certain key proteins like Ser/Thr protein kinase, seed storage protein, DNA-binding protein, microtubule-associated protein, etc. In addition, some growth and cell division–related micro-RNAs like miR3457 (cell wall formation), miR1440 (cell proliferation and cell cycles), and miR1533 (biosynthesis of plant hormones) were identified as having a role in seed size determination. Using RNA-seq data, 5254 EST-SSR primers were generated as a source for future studies aiming for the identification of linked markers. In silico validation using Genevestigator® was done for the Ser/Thr protein kinase, ethylene response factor, and Myb transcription factor genes. It is of interest that the xyloglucan endotransglucosylase gene was found differentially regulated, suggesting their role during seed development; however, at maturity, no significant differences were recorded for various cell wall parameters including cellulose, lignin, and xylose content. This is the first report on lentils that has unfolded the key seed size regulating pathways and unveiled a theoretical way for the development of lentil genotypes having customized seed sizes.

Mungbean (Vignaradiata L. Wilczek) is an early maturing legume grown predominantly in Asia for its protein-rich seeds. P deficiency can lead to several physiological disorders which ultimately result in a low grain yield in mungbean. The genetic dissection of PUpE (Puptake efficiency) and PUtE (P utilization efficiency) traits are essential for breeding mungbean varieties with a high P uptake and utilization efficiency. The study involves an association mapping panel consisting of 120 mungbean genotypes which were phenotyped for total dry weight, P concentration, total P uptake, and P utilization efficiency under low P (LP) and normal P (NP) conditions in a hydroponic system. A genotyping-by-sequencing (GBS) based genome-wide association study (GWAS) approach was employed to dissect the complexity of PUpE and PUtE traits at the genetic level in mungbean. This has identified 116 SNPs in 61 protein-coding genes and of these, 16 have been found to enhance phosphorous uptake and utilization efficiency in mungbeans. We identified six genes with a high expression (VRADI01G04370, VRADI05G20860, VRADI06G12490, VRADI08G20910, VRADI08G00070 and VRADI09G09030) in root, shoot apical meristem and leaf, indicating their role in the regulation of P uptake and utilization efficiency in mungbean. The SNPs present in three genes have also been validated using a Sanger sequencing approach.

Lentil grains with high nutritional value qualify as a promising candidate for alleviation of micronutrient malnutrition in South Asia and North Africa. Genetic variation for micronutrient concentration in germplasm is prerequisite for biofortification of this crop. In the present study, ninety-six lentil genotypes consisting of Indian (released varieties, advanced breeding lines and germplasm lines) and Mediterranean (germplasm lines and landraces) lines were evaluated for grain iron (Fe) and zinc (Zn) concentrations and the stability of these traits was studied across three different locations in India. The pooled analysis of variance revealed significant genotype, environment and genotype by environment interaction (GEI) mean squares for both the micronutrients. Stability analysis employing the AMMI model elucidated the first two interaction principal components as significant and cumulatively explained 100% of GEI variation. The first two components explained 55.9% and 44.1% of the GEI sum of squares for grain iron and 50.8% and 49.2% for grain zinc concentration, respectively. No correlation between grain iron and zinc concentration was observed. Among 96 lines, genotypes IG 49, P 16214, ILL 147 and P 2118 were found to be relatively stable, having higher mean iron and zinc concentrations with low modified AMMI stability value (MASV), modified AMMI stability index (MASI) and genotype selection index (GSI). The identified promising genotypes (high Fe: P 16214, IG 115, P 2127 and IC 560812 and high Zn: P 8115, P 3234, LL 461 and IC 560812) can be utilized for studying the genetics of grain Fe and Zn concentration by developing mapping populations and for biofortification of Indian lentil.

Mungbean (Vigna radiata L. Wilczek) is an annual grain legume crop affected by low availability of phosphorus. Phosphorus deficiency mainly affects the growth and development of plants along with changes in root morphology and increase in root-to-shoot ratio. Deciphering the genetic basis of phosphorus use efficiency (PUE) traits can benefit our understanding of mungbean tolerance to low-phosphorus condition. To address this issue, 144 diverse mungbean genotypes were evaluated for 12 PUE traits under hydroponics with optimum- and low-phosphorus levels. The broad sense heritability of traits ranged from 0.63 to 0.92 and 0.58 to 0.92 under optimum- and low-phosphorus conditions, respectively. This study, reports for the first time such a large number of genome wide Single nucleotide polymorphisms (SNPs) (76,160) in mungbean. Further, genome wide association study was conducted using 55,634 SNPs obtained by genotyping-by-sequencing method. The results indicated that total 136 SNPs shared by both GLM and MLM models were associated with tested PUE traits under different phosphorus regimes. We have identified SNPs with highest p value (-log(10)(p)) for some traits like, TLA and RDW with p value (-log(10)(p)) of more than 6.0 at LP/OP and OP condition. We have identified nine SNPs (three for TLA and six for RDW trait) which was found to be present in chromosomes 8, 4, and 7. One SNP present in Vradi07g06230 gene contains zinc finger CCCH domain. In total, 71 protein coding genes were identified, of which 13 genes were found to be putative candidate genes controlling PUE by regulating nutrient uptake and root architectural development pathways in mungbean. Moreover, we identified three potential candidate genes VRADI11G08340, VRADI01G05520, and VRADI04G10750 with missense SNPs in coding sequence region, which results in significant variation in protein structure at tertiary level. The identified SNPs and candidate genes provide the essential information for genetic studies and marker-assisted breeding program for improving low-phosphorus tolerance in mungbean.

Post Arabidopsis thaliana , 55 genomes comprising 49 different plant species have been decoded by use of clone-by-clone, whole genome shotgun and next-generation sequencing approaches. The structural outcomes of these sequenced genomes shed light on their genomic constitution, particularly the way genes, transposable elements and genetic markers are orga-nized within the genomes. The functional outcomes provide a brief account of specific phenotypic trait characteristics of crop genomes by digging deep into the genetic make-up of transcription factors, regulato-ry elements and gene families governing multiple ag-ronomic traits in these crop plants. The comparative and evolutionary outcomes deduce the genetic basis of biological diversity and basic process of ge-nome evolution by analysing the syntenic relationships among genes and genomes/chromosomes of the se-quenced crop plants. Therefore, a revisit to published genome sequence landmarks in 30 major cultivated food crops constituting major groups (cereals, leg-umes, vegetables, fruits, oil seeds and fibres) would significantly assist us to gain a detailed insight into their genome organization and dissect the structural, functional, comparative and evolutionary intricacies for identifying species- and lineage-specific genes con-trolling multiple characteristics in crop plants. The es-sential inputs obtained will be helpful in devising efficient strategies to develop high-yielding climate-ready crop varieties through translational genomics.

Identification of potential genes/alleles governing complex seed-protein content (SPC) is essential in marker-assisted breeding for quality trait improvement of chickpea. Henceforth, the present study utilized an integrated genomics-assisted breeding strategy encompassing trait association analysis, selective genotyping in traditional bi-parental mapping population and differential expression profiling for the first-time to understand the complex genetic architecture of quantitative SPC trait in chickpea. For GWAS (genome-wide association study), high-throughput genotyping information of 16376 genome-based SNPs (single nucleotide polymorphism) discovered from a structured population of 336 sequenced desi and kabuli accessions [with 150-200 kb LD (linkage disequilibrium) decay] was utilized. This led to identification of seven most effective genomic loci (genes) associated [10-20% with 41% combined PVE (phenotypic variation explained)] with SPC trait in chickpea. Regardless of the diverse desi and kabuli genetic backgrounds, a comparable level of association potential of the identified seven genomic loci with SPC trait was observed. Five SPC-associated genes were validated successfully in parental accessions and homozygous individuals of an intra-specific desi RIL (recombinant inbred line) mapping population (ICC 12299 × ICC 4958) by selective genotyping. The seed-specific expression, including differential up-regulation (>four fold) of six SPC-associated genes particularly in accessions, parents and homozygous individuals of the aforementioned mapping population with a high level of contrasting SPC (21-22%) was evident. Collectively, the integrated genomic approach delineated diverse naturally occurring novel functional SNP allelic variants in six potential candidate genes regulating SPC trait in chickpea. Of these, a non-synonymous SNP allele-carrying zinc finger transcription factor gene exhibiting strong association with SPC trait was found to be the most promising in chickpea. The informative functionally relevant molecular tags scaled-down essentially have potential to accelerate marker-assisted genetic improvement by developing nutritionally rich chickpea cultivars with enhanced SPC.

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.

The present study exploited integrated genomics-assisted breeding strategy for genetic dissection of complex branch number quantitative trait in chickpea. Candidate gene-based association analysis in a branch number association panel was performed by utilizing the genotyping data of 401 SNP allelic variants mined from 27 known cloned branch number gene orthologs of chickpea. The genome-wide association study (GWAS) integrating both genome-wide GBS- (4556 SNPs) and candidate gene-based genotyping information of 4957 SNPs in a structured population of 60 sequenced desi and kabuli accessions (with 350-400 kb LD decay), detected 11 significant genomic loci (genes) associated (41% combined PVE) with branch number in chickpea. Of these, seven branch number-associated genes were further validated successfully in two inter (ICC 4958 × ICC 17160)- and intra (ICC 12299 × ICC 8261)-specific mapping populations. The axillary meristem and shoot apical meristem-specific expression, including differential up- and down-regulation (4-5 fold) of the validated seven branch number-associated genes especially in high branch number as compared to the low branch number-containing parental accessions and homozygous individuals of two aforesaid mapping populations was apparent. Collectively, this combinatorial genomic approach delineated diverse naturally occurring novel functional SNP allelic variants in seven potential known/candidate genes [PIN1 (PIN-FORMED protein 1), TB1 (teosinte branched 1), BA1/LAX1 (BARREN STALK1/LIKE AUXIN1), GRAS8 (gibberellic acid insensitive/GAI, Repressor of ga13/RGA and Scarecrow8/SCR8), ERF (ethylene-responsive element-binding factor), MAX2 (more axillary growth 2) and lipase] governing chickpea branch number. The useful information generated from this study have potential to expedite marker-assisted genetic enhancement by developing high-yielding cultivars with more number of productive (pods and seeds) branches in chickpea.

The SNP-based high-resolution QTL mapping mapped eight major genomic regions harbouring robust QTLs governing seed-Fe and Zn concentrations (39.4% combined phenotypic variation explained/PVE) on six chromosomes of an intra-specific high-density genetic linkage map (1.56 cM map-density). 24620 SNPs discovered from genome-wide GBS (genotyping-by-sequencing) and 13 known cloned Fe and Zn contents-related chickpea gene-orthologs were genotyped in a structured population of 92 sequenced desi and kabuli accessions. The large-scale 16591 SNP genotyping- and phenotyping-based GWAS (genome-wide association study) identified 16 genomic loci/genes associated (29% combined PVE) with seed-Fe and Zn concentrations. Of these, 11 trait-associated SNPs in the genes linked tightly with eight QTLs were validated by QTL mapping. The seed-specific expression, including pronounced differential-regulation of 16 trait-associated genes particularly in accessions/mapping individuals with contrasting level of seed-Fe and Zn contents was apparent. Collectively, the aforementioned rapid integrated genomic strategy led to delineate novel functional non-synonymous and regulatory SNP allelic-variants from 16 known/candidate genes, including three strong trait-associated genes (encoding late embryogenesis abundant and yellow stripe-like 1 protein, and vacuolar protein sorting-associated protein) and eight major QTLs regulating seed-Fe and Zn concentrations in chickpea. These essential inputs thus have potential to be deployed in marker-assisted genetic enhancement for developing nutritionally-rich iron/zinc-biofortified chickpea cultivars.

We discovered 26785 and 16573 high-quality SNPs differentiating two parental genotypes of a RIL mapping population using reference desi and kabuli genome-based GBS assay. Of these, 3625 and 2177 SNPs have been integrated into eight desi and kabuli chromosomes, respectively in order to construct ultra-high density (0.20–0.37 cM) intra-specific chickpea genetic linkage maps. One of these constructed high-resolution genetic map has potential to identify 33 major genomic regions harbouring 35 robust QTLs (PVE: 17.9–39.7%) associated with three agronomic traits, which were mapped within <1 cM mean marker intervals on desi chromosomes. The extended LD (linkage disequilibrium) decay (~15 cM) in chromosomes of genetic maps have encouraged us to use a rapid integrated approach (comparative QTL mapping, QTL-region specific haplotype/LD-based trait association analysis, expression profiling and gene haplotype-based association mapping) rather than a traditional QTL map-based cloning method to narrow-down one major seed weight (SW) robust QTL region. It delineated favourable natural allelic variants and superior haplotype-containing one seed-specific candidate embryo defective gene regulating SW in chickpea. The ultra-high-resolution genetic maps, QTLs/genes and alleles/haplotypes-related genomic information generated and integrated strategy for rapid QTL/gene identification developed have potential to expedite genomics-assisted breeding applications in crop plants, including chickpea for their genetic enhancement.