Publications of NIPGR Scientists
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Item Dissecting the genetic basis of seed-iron content in Chickpea using a combinatorial approach of QTL-Seq and molecular haplotyping(Springer Nature Publishing AG, 2026) Singh, Gourav; Chakraborty, Anirban; Singh, Sangeeta; Bhardwaj, Shubham; Parida, Swarup K.; Bhatia, SabhyataIn order to map the QTL(s) and genes regulating the complex seed-iron content (SFC) trait in chickpea, the quantitative trait locus (QTL)-seq approach was used. Whole genome re-sequencing of DNA bulks derived from a mapping population (ICC8261 × 1CC4958) contrasting for SFC led to the identification of three QTLs, [CaqFe4.1 (0.10 Mb), CaqFe4.2 (0.54 Mb) and CaqFe7.1 (0.83 Mb)] in chickpea. In-silico expression analysis of genes underlying the QTLs revealed their varied levels during stages of seed development. Moreover, estimation of Gʹ values of the SNPs identified in the QTL region revealed a SNP that generated synonymous variant of the MAIN-like-2 gene. Haplotype analysis of MAIN-like-2 in a diverse panel of chickpea germplasm varying for SFC further exemplified its haplotypes that displayed strong association to this trait. Homology-based protein interaction analysis coupled with quantitative-real time PCR based-expression analysis revealed several co-expressing co-chaperone and heat shock proteins including P23-1, HSP 90.5 and HSP90.6, having well established roles in seed development as protein components of MAIN-like-2 proteins in chickpea. The functional loci as well as the molecular signatures defined in this study have potential to expedite marker assisted breeding of iron-rich chickpea varieties.Item Deciphering the dynamics of enzymes associated with the synthesis of cryoprotectants during cold acclimation in contrasting chickpea genotypes(Springer Nature Publishing AG, 2025) Padhiar, Deeksha; Kaur, Sarbjeet; Rani, Anju; Jha, Uday Chand; Prasad, P. V. Vara; Sharma, Kamal Dev; Kumar, Sanjeev; Parida, Swarup K.; Siddique, Kadambot H. M.; Nayyar, HarshChickpea, a vital legume crop, is highly susceptible to cold stress, especially during its reproductive phase, resulting in significant flower and pod abortions and reduced seed yield. Our previous study demonstrated that cold acclimation is effective in enhancing cold tolerance but benefits only cold-tolerant (CT) genotypes, while cold-sensitive (CS) genotypes remain unaffected. In this extended study aimed at probing the detailed mechanisms of this differential response, we further examined the expression profiles of enzymes involved in the synthesis and breakdown of osmolytes (pyrroline-5-carboxylate synthase, proline dehydrogenase (PDH), betaine aldehyde dehydrogenase) and sugars (sucrose synthase, acid invertase, trehalose-6-phosphate synthase, trehalose-6-phosphate phosphatase, and trehalase activity), along with the expression of various antioxidants (superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase) in both CT and CS genotypes. Seeds of two contrasting chickpea genotypes, cold-tolerant ICC 17258 and cold-sensitive ICC 15567, were planted in pots during the first week of November in an outdoor field environment. After 40 days, the plants were transferred to walk-in growth chambers for cold acclimation at specific temperatures. Initially, the plants were exposed the plants to 25/18℃ (pre-acclimation stage; PAS) for 2 days, followed by a 21-day cold acclimation period with progressively decreasing temperatures over seven days for each cold acclimation stage (CAS): CAS1 (21/13℃), CAS2 (18/10℃), and CAS3 (15/8℃). Subsequently, the plants were subjected to cold stress at 13/7℃ for 15 days and then exposed to 30/23℃ (12 h day/night) until maturity. Our findings demonstrated that the expression of various enzymes involved in the synthesis of osmolytes and sugars in leaves, anthers, and ovules was significantly upregulated during the cold acclimation process in the CT chickpea genotypes but not in the CS genotypes. This enhanced metabolic activity, coupled with elevated levels of enzymatic antioxidants during the acclimation process, contributed to improved leaf water status, photosynthetic efficiency, and ultimately, superior reproductive performance (pollen germination, pollen viability, stigma receptivity, and ovule viability) under cold stress conditions compared to CS genotypes. The enhanced cold tolerance observed in the CT genotypes is likely attributable to their genetic predisposition and efficient stress defense mechanisms facilitated by the upregulated expression of cold-responsive enzymes.Item Exploring the synergistic effects of drought and heat stress on chickpea seed development: Insights into nutritional quality and seed yield(Elsevier B.V., 2024) Awasthi, Rashmi; Devi, Poonam; Jha, Uday Chand; Sharma, Kamal Dev; Roorkiwal, Manish; Kumar, Sanjeev; Pareek, Ashwani; Siddique, Kadambot H.M.; Prasad, PV Vara; Parida, Swarup K.; Nayyar, HarshGrowing chickpea (Cicer arietinum L.) faces significant challenges due to rising temperatures and drought stress, particularly during the reproductive and seed-filling phases. This study investigated the single and joint impacts of drought and heat stress on seed development, focusing on the responses of drought-tolerant (DT) and droughtsensitive (DS) chickpea genotypes. Initially raised in an outdoor environment (mean day and night temperature of 27 and 16±1 ◦C, respectively, light intensity of 1230–1440 µmol m− 2 s − 1 , relative humidity of 70/43 %) until seed filling (around 110–113 days after sowing) commenced. The plants were subsequently exposed to single or combined heat and drought stress under controlled conditions until maturity. Control pots were maintained at day and night temperature of 25 and 15 ◦C, respectively with 500 µmol m− 2 s − 1 light, 60–65 % RH, and regular irrigation, and drought-stressed pots were kept at 50 % field capacity under the same conditions of light and humidity. Heat stress in pots was gradually increased to 32(day)/20 ◦C (night) under regular irrigation, while combined stress pots experienced both drought (50 % field capacity) and heat stress conditions 32(day)/20 ◦C (night) under the same light and humidity conditions with irrigation. All stress treatments adversely affected cell membranes, photosynthesis, and water regulation, with more pronounced effects under combined stress. While heat stress increased stomatal conductance, drought and combined stress significantly reduced it. Seed filling rate and duration decreased under all stress conditions, especially combined stress. The stresses in combination severely reduced seed weight and pod numbers compared to individual stresses. Enzyme activities involved in starch and sucrose synthesis and hydrolysis substantially decreased under the combined stress. Seed composition elements (starch, storage proteins, sugars, fat, crude fiber, and ash) exhibited significant reductions across all stress treatments, particularly for the combined stress. Thus, under combined stresses, starch, proteins, and soulube sugars were markedly decreased to 13–20 %, 6.4–12.4 %, and 3–5 % in seeds, compared to 37–39 %, 21–24 %, and 6 % in control seeds. The DT genotype outperformed the DS genotype for all traits under individual and combined stress conditions. Principal component analysis revealed a complex interplay among various physiological responses (membrane damage, chlorophyll, chlorophyll fluorescence, relative leaf water content, and stomatal conductance), seed yield, and seed composition under the combined stress. This study highlighted that combined heat and drought stress severely impacted chickpea yield and nutritional traits, such as seed starch and protein content, compared to individual stresses underscoring the need to develop cultivars tolerant to this stress combination.Item Genome wide association studies for flowering time, shelling percentage, harvest index and related traits in chickpea (Cicer arietinum L.)(Springer Nature Publishing AG, 2024) Patil, N. Shivanagouda; Hegde, Venkatraman; Vinod, K. K.; Parida, Swarup K.; Jain, Pradeep KumarGrain yield is a product of biomass (BM) and harvest index (HI). HI, not BM, is the major constraint of chickpea productivity in the long season environments of northern Indian plains. This is the first genome wide association study (GWAS) of shelling percentage (Sh.%), HI and related traits in chickpea using genotyping by sequencing approach. Genotyping of 178 germplasm lines involving varieties, trait specific genotypes and advance breeding lines found 35,795 high-throughput single nucleotide polymorphisms (SNPs). The objectives of the study were to unravelling the genomic regions responsible for variation in flowering time, shelling percentage, HI and related traits in chickpea. The phenotypic analysis across three environments showed high variability and connections among HI and related traits. Population structure analysis revealed two sub-populations (k = 2). Linkage disequilibrium (LD) was extensive, and LD decay was relatively low. A total of 172 marker-trait associations (MTAs) were identified for HI and related traits using FarmCPU model, of which days to 50% flowering (DAF), HI and shelling percentage showed significant associations. The current study has identified consistent pleiotropic MTAs, SNC_021165.1_57891716 and SNC_021165.1_57917493, SNC_021164.1_6073633 and SNC_021165.1_33405826 for biomass, pod and seed yield/plant, pod and seed number/plant HI and Sh.%, respectively. All the MTAs observed for SYPL, PYPL and maximum number of MTAs for HI and BM were located on the same chromosome number 6. Further maximum number of MTAs for PPL and SN were distributed on chromosome 5. One of the important findings is that most of the MTAs for SYPL, PYPL, BM, and 100SW are distributed within 25 kb genomic region of chromosome 6. A total of 16 MTAs were retained for Insilco analysis (− log10(p) > 4.0) and searched for their candidate genes in a 100-kb flanking region against the reference genome. The maximum number of genes noticed for DAF (50), followed by Sh.% (48), HI (16), SYPL (13), BM (10), and PYPL (8). The transcripts for DAF, Arabinogalactan proteins and pentatricopeptide repeat were involved in flowering of Arabidopsis. For both PYPL and SYPL, MTA found linked to the genes coding for pentatricopeptide repeat, serine/threonine-protein kinase and E3 ubiquitin-protein ligase. These transcripts played role mainly in pollen maturation, fertilization and flowering time. Chaperone protein DnaJ linked to HI, and receptor-like protein kinase (RLK) for Sh.% were involved in improving yield in Arabidopsis and rice, respectively. The present study also validated one of the MTA linked to DAF which showed r2 value of 12.62%.Item Identification of a stable drought-tolerant high-yielding line for chickpea crop improvement(Springer Nature Publishing AG, 2025) Gupta, Santosh Kumar; Dwivedi, Vikas; Kute, Nandakumar Surendra; Francis, Philip; Parida, Swarup K.; Chattopadhyay, DebasisChickpea (Cicer arietinum L.) is grown in marginal land with low input and is, therefore, drought-prone. In order to develop a drought-tolerant line, a bi-parental recombinant inbred line (RIL) mapping population was generated by inter-crossing between two varieties JGK3 (ICCV 95334) and Himchana1 (ICCX-810800) having contrasting root traits. Ninety-two genetically diverse RILs of F8 generation were selected based on their total root length to root dry weight ratio (RL/DW). The leaf relative water content of these RILs under low soil moisture did not show any strong correlation with the RL/DW. Twenty RILs having high RL/DW were evaluated for seed yield in a field under rainfed condition without any supplementary irrigation. The best performing RIL, which performed better than the check varieties, was reevaluated for a further year under rainfed condition. The genotypic constitution of this superior low soil moisture tolerant individual RIL was determined by constructing its recombination map using genome-wide SNPs obtained through genotyping-by-sequencing. The RIL possesses the superior alleles of the genomic QTL region known to govern drought tolerance in chickpea. The phenotypic and genotypic characterization of RILs in our study identified a chickpea pre-breeding line that can be used as a genetic donor for developing drought-tolerant high-yielding chickpea varieties and our results provide an evidence that total root length to root dry weight ratio can be used as a quantitative trait for assessing drought tolerance.Item Uncovering DNA methylation landscapes to decipher evolutionary footprints of phenotypic diversity in chickpea(Oxford University Press, 2024) Daware, Anurag; Mohanty, Jitendra K.; Narnoliya, Laxmi; Singh, Akansha; Rathore, Deepanshi; Thakro, Virevol; Francis, Aleena; Singh, Nagendra Pratap; Francis, Philip; Tripathi, Shailesh; Chattopadhyay, Debasis; Parida, Swarup K.Genetic diversity and environmental factors are long believed to be the dominant contributor to phenotypic diversity in crop plants. However, it has been recently established that, besides genetic variation, epigenetic variation, especially variation in DNA methylation, plays a significant role in determining phenotypic diversity in crop plants. Therefore, assessing DNA methylation diversity in crop plants becomes vital, especially in the case of crops like chickpea, which has a narrow genetic base. Thus, in the present study, we employed whole-genome bisulfite sequencing to assess DNA methylation diversity in wild and cultivated (desi and kabuli) chickpea. This revealed extensive DNA methylation diversity in both wild and cultivated chickpea. Interestingly, the methylation diversity was found to be significantly higher than genetic diversity, suggesting its potential role in providing vital phenotypic diversity for the evolution and domestication of the Cicer gene pool. The phylogeny based on DNA methylation variation also indicates a potential complementary role of DNA methylation variation in addition to DNA sequence variation in shaping chickpea evolution. Besides, the study also identified diverse epi-alleles of many previously known genes of agronomic importance. The Cicer MethVarMap database developed in this study enables researchers to readily visualize methylation variation within the genes and genomic regions of their interest (http://223.31.159.7/cicer/public/). Therefore, epigenetic variation like DNA methylation variation can potentially explain the paradox of high phenotypic diversity despite the narrow genetic base in chickpea and can potentially be employed for crop improvement.Item Delineation of genes for a major QTL governing heat stress tolerance in chickpea(Springer Nature Publishing AG, 2024) Mohanty, Jitendra K.; Thakro, Virevol; Yadav, Antima; Nayyar, Harsh; Dixit, Girish P.; Agarwal, Pinky; Parida, Swarup K.; Jha, Uday ChandChickpea (Cicer arietinum) is a cool season grain legume experiencing severe yield loss during heat stress due to the intensifying climate changes and its associated gradual increase of mean temperature. Hence, understanding the genetic architecture regulating heat stress tolerance has emerged as an important trait to be addressed for enhancing yield and productivity of chickpea under heat stress. The present study is intended to identify the major genomic region(s) governing heat stress tolerance in chickpea. For this, an integrated genomics-assisted breeding strategy involving NGS-based high-resolution QTL-seq assay, QTL region-specifc association analysis and molecular haplotyping was deployed in a population of 206 mapping individuals and a diversity panel of 217 germplasm accessions of chickpea. This combinatorial strategy delineated a major 156.8 kb QTL genomic region, which was subsequently narrowed-down to a functional candidate gene CaHSFA5 and its natural alleles associated strongly with heat stress tolerance in chickpea. Superior natural alleles and haplotypes delineated from the CaHSFA5 gene have functional signifcance in regulating heat stress tolerance in chickpea. Histochemical staining, interaction studies along with diferential expression profling of CaHSFA5 and ROS scavenging genes suggest a cross talk between CaHSFA5 with ROS homeostasis pertaining to heat stress tolerance in chickpea. Heterologous gene expression followed by heat stress screening further validated the functional signifcance of CaHSFA5 for heat stress tolerance. The salient outcomes obtained here can have potential to accelerate multiple translational genomic analysis including marker-assisted breeding and gene editing in order to develop high-yielding heat stress tolerant chickpea varieties.Item eQTL-seq: a rapid genome-wide integrative genetical genomics strategy to dissect complex regulatory architecture of gene expression underlying quantitative trait variation in crop plants(Springer Nature Publishing AG, 2024) Mohanty, Jitendra K.; Jha, Uday Chand; Dixit, G. P.; Bharadwaj, Chellapilla; Parida, Swarup K.We developed a combinatorial next generation sequencing (NGS)-based high-throughput expression QTL-seq (eQTL-seq) strategy for rapid elucidation of complex genetic architecture and regulatory pathways of gene expression underlying quantitative trait variation in crop plants. This high-resolution genome-wide integrative genetical genomics strategy assists us to delineate functionally relevant molecular tags (QTLs/eQTLs, genes, master regulators/transcription factors, and alleles) regulating vital agronomic traits in order to expedite genomics-assisted breeding and crop improvement. As a proof-of-concept, the optimized eQTL-seq-led genetical genomics approach was employed in 192 mapping individuals of an intra-specific recombinant inbred line (RIL) population (desi accession ICC 4958 x kabuli accession ICC 12968) with contrasting seed weight trait. This led to identification and mapping of high-resolution major QTLs/eQTLs associated with seed weight based on genome resequence-derived genomic SNP-index (Delta(gSNP-index)), global transcriptome sequence-based expression SNP-index (Delta(eSNP-index)), and relative gene expression (RGE)-index (Delta(RGE-index)). The eQTL-seq scaled down the major seed weight QTLs/eQTLs into the potential candidate gene(s) governing cis- and trans-regulated expression to decipher the gene regulatory modules controlling complex quantitative seed weight trait by deriving global allele-specific gene expression polymorphism in chickpea. Henceforth, the efficacy of NGS-driven integrative eQTL-seq strategy as compared to traditional QTL mapping, fine-mapping (map-based cloning), and QTL-seq approaches that are widely adopted for delineation of candidate genes underlying major QTLs was evident in chickpea. This will further accelerate quantitative dissection of regulatory architecture by decoding the molecular genetic basis of gene expression variation controlling diverse complex phenotypic trait diversity in chickpea and other crop plants as well.Item The developmental dynamics in cool season legumes with focus on chickpea(Springer Nature Publishing AG, 2023) Basu, Udita; Parida, Swarup K.Chickpea is one of the most widely consumed grain legume world-wide. Advances in next-generation sequencing and genomics tools have led to genetic dissection and identification of potential candidate genes regulating agronomic traits in chickpea. However, the developmental particularities and its potential in reforming the yield and nutritional value remain largely unexplored. Studies in crops such as rice, maize, tomato and pea have highlighted the contribution of key regulator of developmental events in yield related traits. A comprehensive knowledge on the development aspects of a crop can pave way for new vistas to explore. Pea and Medicago are the close relatives of genus Cicer and the basic developmental events in these legumes are similar. However, there are some distinct developmental features in chickpea which hold potential for future crop improvement endeavours. The global chickpea germplasm encompasses wide range of diversities in terms of morphology at both vegetative and reproductive stages. There is an immediate need for understanding the genetic and molecular basis of this diversity and utilizing them for the yield contributing trait improvement. The review discusses some of the key developmental events which have potential in yield enhancement and the lessons which can be learnt from model legumes in this regard.Item Harnessing the hidden allelic diversity of wild Cicer to accelerate genomics-assisted chickpea crop improvement(Springer Nature Publishing AG, 2022) Mohanty, Jitendra Kumar; Jha, Uday Chand; Dixit, G.P.; Parida, Swarup K.Chickpea, commonly called Bengal gram or Garbanzo bean, faces a productivity crisis around the globe due to numerous biotic and abiotic stresses. The eroded genetic base of the cultivated Cicer gene pool is becoming a significant bottleneck in developing stress-resilient chickpea cultivars. In this scenario, the crop wild relatives (CWR) of chickpea, with the useful genomic wealth of their wild adaptation, give a ray of hope to improve the genetic background of the cultivated Cicer gene pool. To extrapolate these unearthed genomic diversities of wild, we require a thorough understanding of the pre-historic domestication episodes that are changing their shape with the expansion of the available scientific evidence. Keeping aforesaid in view, the current review article provides a glimpsed overview on several efforts done so far to reveal the mysterious origin and evolution of the Cicer gene pool, along with the constraints in their utilization for chickpea crop improvement. It encapsulates various stress-resilient CWR of chickpea and their use in several pre-breeding programs to develop numerous breeding populations for crop genetic enhancement. Further, this review will recapitulate the significant contributions of structural, functional and comparative genomics, pan-genomics and diverse genomics-assisted breeding strategy in dissecting the untapped trait-specific allelic/gene diversity and domestication pattern behind the CWR of chickpea, along with their potential and promises. We expect the newly explored genetic variations may be used in the breeding programs for re-wilding the cultigens’ genomic background to open a new avenue for genetic gain and crop improvement capacity of chickpea.
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