Publications of NIPGR Scientists
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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 The chickpea WIP2 gene underlying a major QTL contributes to lateral root development(Oxford University Press, 2024) Dwivedi, Vikas; Pal, Lalita; Singh, Shilpi; Singh, Nagendra Pratap; Parida, Swarup K.; Chattopadhyay, DebasisLateral root is a major component of root system architecture and lateral root count (LRC) positively contributes to yield under drought in chickpea. To understand the genetic regulation of LRC, a biparental mapping population derived from two chickpea accessions having contrasting LRC was genotyped by sequencing and phenotyped to map four major quantitative trait loci (QTLs) contributing to 13 to 32% of the LRC trait variation. A SNP tightly linked to the locus contributing to highest trait variation was located on the coding region of a gene (CaWIP2) orthologous to WIP2 gene of Arabidopsis thaliana. A polymorphic simple sequence repeat (SSR) in the CaWIP2 promoter showed differentiation between low vs. high LRC parents and mapping individuals suggesting its utility for marker-assisted selection. CaWIP2 promoter showed strong activities in chickpea apical root meristem and lateral root primordia. Expression of CaWIP2 under its native promoter in Arabidopsis wip2wip4wip5 mutant rescued its root-less phenotype to produce more lateral root than the wild type plants and led to formation of amyloplasts in the columella. CaWIP2 expression also induced expression of genes that regulate lateral root emergence. Our study identified a gene-based marker for LRC which will be useful to develop drought tolerant high-yielding chickpea.Item A superior gene allele involved in abscisic acid signaling enhances drought tolerance and yield in chickpea(Oxford University Press, 2023) Thakro, Virevol; Malik, Naveen; Basu, Udita; Srivastava, Rishi; Narnoliya, Laxmi; Daware, Anurag; Varshney, Nidhi; Mohanty, Jitendra K; Bajaj, Deepak; Dwivedi, Vikas; Tripathi, Shailesh; Jha, Uday Chand; Dixit, Girish Prasad; Singh, Ashok K; Tyagi, Akhilesh K.; Upadhyaya, Hari D; Parida, Swarup K.Identifying potential molecular tags for drought tolerance is essential for achieving higher crop productivity under drought stress. We employed an integrated genomics-assisted breeding and functional genomics strategy involving association mapping, fine mapping, map-based cloning, molecular haplotyping and transcript profiling in the introgression lines (ILs)- and near isogenic lines (NILs)-based association panel and mapping population of chickpea (Cicer arietinum). This combinatorial approach delineated a bHLH (basic helix-loop-helix) transcription factor, CabHLH10 (Cicer arietinum bHLH10) underlying a major QTL, along with its derived natural alleles/haplotypes governing yield traits under drought stress in chickpea. CabHLH10 binds to a cis-regulatory G-box promoter element to modulate the expression of RD22 (responsive to desiccation 22), a drought/ABA-responsive gene (via a trans-expression QTL), and two strong yield-enhancement photosynthetic efficiency (PE) genes. This, in turn, upregulates other downstream drought-responsive and abscisic acid signaling genes, as well as yield-enhancing PE genes, thus increasing plant adaptation to drought with reduced yield penalty. We showed that a superior allele of CabHLH10 introgressed into the NILs improved root and shoot biomass and PE, thereby enhancing yield and productivity during drought without compromising agronomic performance. Furthermore, overexpression of CabHLH10 in chickpea and Arabidopsis (Arabidopsis thaliana) conferred enhanced drought tolerance by improving root and shoot agro-morphological traits. These findings facilitate translational genomics for crop improvement and the development of genetically-tailored, climate-resilient, high-yielding chickpea cultivars.Item Biochemical analysis of anthocyanin and proanthocyanidin and their regulation in determining chickpea flower and seed coat colours(Oxford University Press, 2023) Pal, Lalita; Dwivedi, Vikas; Gupta, Santosh Kumar; Saxena, Samiksha; Pandey, Ashutosh; Chattopadhyay, DebasisFlower and seed coat colour are important agronomic traits in chickpea (Cicer arietinum L.). Cultivated chickpeas are of two types namely, desi (dark seeded, purple flower) and kabuli (light colour seeded, white flower). There has been limited information about the molecular mechanism underlying the colour variation of flower and seed coats in desi and kabuli chickpea. We profiled the anthocyanin and proanthocyanidin (PA) contents in chickpea flowers and seed coats. Tissue-specific silencing of two genes encoding a basic helix-loop-helix (CabHLH) protein and a tonoplast-localized multidrug and toxic compound extrusion (CaMATE1) transporter in a desi genotype resulted in the reduction in expressions of anthocyanin and PA biosynthetic genes and anthocyanin and PA contents in the flower and seed coat and produced flowers and seeds with kabuli characteristics. Transcriptional regulation of a subset of anthocyanin and PA biosynthetic genes by a natural CabHLH variant and transport assay of a natural CaMATE1 variant explained the association of these alleles with the kabuli phenotype. We carried out a detailed molecular characterization of these genes, and provided evidences that kabuli chickpea flower and seed colour phenotype can be derived by manipulation of single genes in a desi chickpea background.Item Root-specific expression of chickpea cytokinin oxidase/dehydrogenase 6 leads to enhanced root growth, drought tolerance and yield without compromising nodulation(John Wiley & Sons, 2020) Khandal, Hitaishi; Gupta, Santosh Kumar; Dwivedi, Vikas; Mandal, Drishti; Sharma, Nilesh Kumar; Vishwakarma, Niraj Kumar; Pal, Lalita; Choudhary, Megha; Francis, Aleena; Malakar, Paheli; Singh, Nagendra Pratap; Sharma, Kapil; Sinharoy, Senjuti; Singh, Narendra Pratap; Sharma, Rameshwar; Chattopadhyay, DebasisCytokinin group of phytohormones regulate root elongation and branching during post‐embryonic development. Cytokinin degrading enzymes cytokinin oxidases/dehydrogenases (CKXs) have been deployed to investigate biological activities of cytokinin and to engineer root growth. We expressed chickpea cytokinin oxidase 6 (CaCKX6) under the control of a chickpea root‐specific promoter of CaWRKY31 in Arabidopsis thaliana and chickpea having determinate and indeterminate growth patterns, respectively, to study the effect of cytokinin depletion on root growth and drought tolerance. Root‐specific expression of CaCKX6 led to a significant increase in lateral root number and root biomass in Arabidopsis and chickpea without any penalty to vegetative and reproductive growth of shoot. Transgenic chickpea lines showed increased CKX activity in root. Soil‐grown advanced chickpea transgenic lines exhibited higher root‐to‐shoot biomass ratio and enhanced long‐term drought tolerance. These chickpea lines were not compromised in root nodulation and nitrogen fixation. The seed yield in some lines was up to 25% higher with no penalty in protein content. Transgenic chickpea seeds possessed higher levels of zinc, iron, potassium and copper. Our results demonstrated the potential of cytokinin level manipulation in increasing lateral root number and root biomass for agronomic trait improvement in an edible legume crop with indeterminate growth habit.Item A repeat length variation in myo-inositol monophosphatase gene contributes to seed size trait in chickpea(Nature Publishing Group, 2017) Dwivedi, Vikas; Parida, Swarup K.; Chattopadhyay, DebasisChickpea (Cicer arietinum L.) is the third most important food legume crop. Seed size is the most economically important trait for chickpea. To understand the genetic regulation of seed size in chickpea, the present study established a three-way association of CT repeat length variation of a simple sequence repeat (SSR) in myo-inositol monophosphatase gene (CaIMP) with seed weight and phytic acid content by large scale validation and genotyping in a set of genetically diverse germplasm accessions and two reciprocal intra-specific mapping populations. Germplasms and mapping individuals with CT repeat-length expansion in the 5' untranslated region of CaIMP exhibited a pronounced increase in CaIMP protein level, enzymatic activity, seed-phytate content and seed weight. A chickpea transient expression system demonstrated this repeat-length variation influenced the translation of CaIMP mRNA, apparently by facilitating translation initiation. Our analyses proposed that the SSR marker derived from 5' UTR of a CaIMP gene is a promising candidate for selection of seed size/weight for agronomic trait improvement of chickpea.Item Investigation of genes encoding calcineurin B-like protein family in legumes and their expression analyses in chickpea (Cicer arietinum L.)(PLOS, 2015) Meena, Mukesh K.; Ghawana, Sanjay; Sardar, Atish; Dwivedi, Vikas; Khandal, Hitaishi; Roy, Riti; Chattopadhyay, DebasisCalcium ion (Ca2+) is a ubiquitous second messenger that transmits various internal and external signals including stresses and, therefore, is important for plants' response process. Calcineurin B-like proteins (CBLs) are one of the plant calcium sensors, which sense and convey the changes in cytosolic Ca2+-concentration for response process. A search in four leguminous plant (soybean, Medicago truncatula, common bean and chickpea) genomes identified 9 to 15 genes in each species that encode CBL proteins. Sequence analyses of CBL peptides and coding sequences (CDS) suggested that there are nine original CBL genes in these legumes and some of them were multiplied during whole genome or local gene duplication. Coding sequences of chickpea CBL genes (CaCBL) were cloned from their cDNAs and sequenced, and their annotations in the genome assemblies were corrected accordingly. Analyses of protein sequences and gene structures of CBL family in plant kingdom indicated its diverse origin but showed a remarkable conservation in overall protein structure with appearance of complex gene structure in the course of evolution. Expression of CaCBL genes in different tissues and in response to different stress and hormone treatment were studied. Most of the CaCBL genes exhibited high expression in flowers. Expression profile of CaCBL genes in response to different abiotic stresses and hormones related to development and stresses (ABA, auxin, cytokinin, SA and JA) at different time intervals suggests their diverse roles in development and plant defence in addition to abiotic stress tolerance. These data not only contribute to a better understanding of the complex regulation of chickpea CBL gene family, but also provide valuable information for further research in chickpea functional genomics.Item Expression of chickpea CIPK25 enhances root growth and tolerance to dehydration and salt stress in transgenic tobacco(Frontiers Media S.A., 2015) Meena, Mukesh K.; Ghawana, Sanjay; Dwivedi, Vikas; Roy, Ansuman; Chattopadhyay, DebasisCalcium signaling plays an important role in adaptation and developmental processes in plants and animals. A class of calcium sensors, known as Calcineurin B-like (CBL) proteins sense specific temporal changes in cytosolic Ca(2+) concentration and regulate activities of a group of ser/thr protein kinases called CBL-interacting protein kinases (CIPKs). Although a number of CIPKs have been shown to play crucial roles in the regulation of stress signaling, no study on the function of CIPK25 or its orthologs has been reported so far. In the present study, an ortholog of Arabidopsis CIPK25 was cloned from chickpea (Cicer arietinum). CaCIPK25 gene expression in chickpea increased upon salt, dehydration, and different hormonal treatments. CaCIPK25 gene showed differential tissue-specific expression. 5'-upstream activation sequence (5'-UAS) of the gene and its different truncated versions were fused to a reporter gene and studied in Arabidopsis to identify promoter regions directing its tissue-specific expression. Replacement of a conserved threonine residue with an aspartic acid at its catalytic site increased the kinase activity of CaCIPK25 by 2.5-fold. Transgenic tobacco plants overexpressing full-length and the high active versions of CaCIPK25 displayed a differential germination period and longer root length in comparison to the control plants. Expression of CaCIPK25 and its high active form differentially increased salt and water-deficit tolerance demonstrated by improved growth and reduced leaf chlorosis suggesting that the kinase activity of CaCIPK25 was required for these functions. Expressions of the abiotic stress marker genes were enhanced in the CaCIPK25-expressing tobacco plants. Our results suggested that CaCIPK25 functions in root development and abiotic stress tolerance.