Publications of NIPGR Scientists

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Start date: 2020Subject: search.filters.subject.Legumes

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    Isolation of cytoskeleton and cytoskeleton-bound polysome, and identification of cytoskeletal proteins from a grain legume
    (Springer Nature Publishing AG, 2026) Kumar, Sunil; Chakraborty, Sohela; Chakraborty, Subhra; Chakraborty, Niranjan
    The plant cytoskeleton is an essential component of cellular architecture, enabling various critical metabolic processes, including cell division, differentiation, expansion and intracellular transport. It consists primarily of three distinct filamentous structures: microtubules, microfilaments and intermediate filaments. These structures are not static; they undergo continuous remodeling in response to environmental signals and developmental cues, which allow plants to adapt to changing conditions. The microtubules and actin filaments have previously been successfully isolated from various plant tissues, contributing to our understanding of their functions. Among the diverse plant families, legumes (Fabaceae) stand out as the third largest, encompassing approximately 20,000 species. They hold significant agricultural importance, ranking second to cereals in global crop production. To fully grasp the developmental and adaptive processes in legumes, it is essential to identify and understand their regulatory components. This chapter focuses on the isolation of cytoskeletal proteins from chickpea, a prominent grain legume, facilitating biochemical and proteomic analyses that may uncover new insights into the functioning of the cytoskeleton in legumes.
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    Identification of tRNA-derived fragments in legumes
    (Springer Nature Publishing AG, 2026) Arora, Simran; Aftab, Sahrish; Shree, Tanu; Kumar, Shailesh
    The tRNA-derived noncoding RNAs (tncRNAs) belong to the novel class of noncoding RNAs, acting as important components of genome regulatory circuits. In planta, the mechanism of generation and function of tncRNAs is not fully elucidated. Production of important leguminous plants like chickpea, Medicago and soybean is majorly hampered due to different biotic and abiotic stresses. Identification and characterization of tncRNAs in legumes may open a new paradigm for molecular biologists to make novel tools for the improved varieties of legumes for sustainable agriculture. The first step in the study of tncRNAs is to identify and annotate them in small RNA sequencing datasets. Here, we have demonstrated the tncRNA Toolkit for the identification and annotation of tncRNAs in a small RNA sequencing dataset of the important legume crop chickpea.
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    Genome editing in legumes: Current status
    (Springer Nature, 2025) Sharma, Aman; Chattopadhya, Debasis
    Legume crops are indispensable for global food security due to their high protein content and phytonutraceutical properties. Despite their importance, legume crops suffer from poor crop yields due to limited areas of cultivation, various biotic and abiotic stress factors, as well as poor agriculture infrastructure. The increasing global population as well as aggravating climate change has further worsened the situation. The recent revolution in genome sequencing and annotation has opened the gateway for legume crop improvement through advanced molecular breeding tools such as genomic selection, genetic engineering, and genome editing. Gene editing by CRISPR-Cas9 and its derivative systems is a powerful tool to make specific genetic changes at precise positions in the genome facilitating rapid crop improvement through precision breeding. In this chapter, an overview of the basic machinery of CRISPR-Cas9 and allied gene editing systems is provided followed by a comprehensive outline of the execution of a basic CRISPR-Cas9 gene editing experiment including various tools and techniques for construct designing and result analysis. The application of CRISPR-Cas9-mediated gene editing in several legume species is also discussed. We have also discussed the major constraints and bottlenecks faced during the practical implementation of genome editing techniques in legume crops.
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    CaLAP1 and CaLAP2 orchestrate anthocyanin biosynthesis in the seed coat of Cicer arietinum
    (Springer Nature Publishing AG, 2024) Singh, Samar; Pal, Lalita; Rajput, Ruchika; Chhatwal, Himani; Singh, Nidhi; Chattopadhyay, Debasis; Pandey, Ashutosh
    The seed coat color is a major economic trait in leguminous crop chickpea (Cicer arietinum). Anthocyanins and proanthocyanidins (PAs) are two classes of flavonoids that mainly contribute to the flower, seed coat and color of Desi chickpea cultivars. Throughout the land plant lineage, the accumulation of anthocyanins and PAs is regulated by MYB and bHLH transcription factors (TFs), which form an MBW (MYB, bHLH, and WD40) complex. Here, we report two R2R3-MYB TFs in chickpea belonging to the anthocyanin-specific subgroup-6, CaLAP1 (Legume Anthocyanin Production 1), and CaLAP2 (Legume Anthocyanin Production 2), which are mainly expressed in the flowers and developmental stages of the seeds. CaLAP1 and CaLAP2 interact with TT8-like CabHLH1 and WD40, forming the MBW complex, and bind to the promoter sequences of anthocyanin- and PA biosynthetic genes CaCHS6, CaDFR2, CaANS, and CaANR, leading to anthocyanins and PA accumulation in the seed coat of chickpea. Moreover, these CaLAPs partially complement the anthocyanin-deficient phenotype in the Arabidopsis thaliana sextuple mutant seedlings. Overexpression of CaLAPs in chickpea resulted in significantly higher expression of anthocyanin and PA biosynthetic genes leading to a darker seed coat color with higher accumulation of anthocyanin and PA. Our findings show that CaLAPs positively modulate anthocyanin and PA content in seed coats, which might influence plant development and resistance to various biotic and abiotic stresses.
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    Chickpea (Cicer arietinum) PHO1 family members function redundantly in Pi transport and root nodulation
    (Elsevier B.V., 2024) Mani, Balaji; Maurya, Kanika; Kohli, Pawandeep Singh; Giri, Jitender
    Phosphorus (P), a macronutrient, plays key roles in plant growth, development, and yield. Phosphate (Pi) transporters (PHTs) and PHOSPHATE1 (PHO1) are central to Pi acquisition and distribution. Potentially, PHO1 is also involved in signal transduction under low P. The current study was designed to identify and functionally characterize the PHO1 gene family in chickpea (CaPHO1s). Five CaPHO1 genes were identified through a comprehensive genome-wide search. Phylogenetically, CaPHO1s formed two clades, and protein sequence analyses confirmed the presence of conserved domains. CaPHO1s are expressed in different plant organs including root nodules and are induced by Pi-limiting conditions. Functional complementation of atpho1 mutant with three CaPHO1 members, CaPHO1, CaPHO1;like, and CaPHO1;H1, independently demonstrated their role in root to shoot Pi transport, and their redundant functions. To further validate this, we raised independent RNA-interference (RNAi) lines of CaPHO1, CaPHO1;like, and CaPHO1;H1 along with triple mutant line in chickpea. While single gene RNAi lines behaved just like WT, triple knock-down RNAi lines (capho1/like/h1) showed reduced shoot growth and shoot Pi content. Lastly, we showed that CaPHO1s are involved in root nodule development and Pi content. Our findings suggest that CaPHO1 members function redundantly in root to shoot Pi export and root nodule development in chickpea.
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    Dissecting chickpea genomic loci associated with the root penetration responsive traits in compacted soil
    (Springer Nature Publishing AG, 2024) Donde, Ravindra; Kohli, Pawandeep Singh; Pandey, Mandavi; Sirohi, Ujjwal; Singh, Bhagat; Giri, Jitender
    Soil compaction is a major concern for modern agriculture, as it constrains plant root growth, leading to reduced resource acquisition. Phenotypic variation for root system architecture (RSA) traits in compacted soils is present for various crops; however, studies on genetic associations with these traits are lacking. Therefore, we investigated RSA traits in diferent soil compaction levels and identifed signifcant genomic associations in chickpea. We conducted a Genome-Wide Association Study (GWAS) of 210 chickpea accessions for 13 RSA traits under three bulk densities (BD) (1.1BD, 1.6BD, and 1.8BD). Soil compaction decreases root exploration by reducing 12 RSA traits, except average diameter (AD). Further, AD is negatively correlated with lateral root traits, and this correlation increases in 1.8BD, suggesting the negative efect of AD on lateral root traits. Interestingly, we identifed probable candidate genes such as GLP3 and LRX for lateral root traits and CRF1-like for total length (TL) in 1.6BD soil. In heavy soil compaction, DGK2 is associated with lateral root traits. Reduction in laterals during soil compaction is mainly due to delayed seedling establishment, thus making lateral root number a critical trait. Interestingly, we also found a higher contribution of the GxE component of the number of root tips (Tips) to the total variation than the other lateral traits. We also identifed a pectin esterase, PPE8B, associated with Tips in high soil compaction and a signifcantly associated SNP with the relative change in Tips depicting a trade-of between Tips and AD. Identifed genes and loci would help develop soil-compaction-resistant chickpea varieties.
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    Abiotic stress impact on the interaction between Macrophomina phaseolina and crop plants
    (Springer Nature Publishing AG, 2024) Ranjan, Shubhashish; Mirchandani, Rishabh; Senthil-Kumar, Muthappa
    Macrophomina phaseolina (Tassi.) Goid is an emerging pathogen that causes diseases like dry root rot and charcoal rot in more than 100 plant families. Abiotic stresses such as drought, salinity, and heat exacerbate this fungal effect and predispose crops to pathogen attacks. Importantly, these combined stresses lead to significant crop yield losses under field conditions. In this, we review the interaction between the devastating pathogen M. phaseolina and several abiotic stresses that are more likely to occur in scenarios of climate change. Drought, heat, and salinity are the major stresses that interact with M. phaseolina in the field. We discuss several field studies, unique physiological and molecular responses, and their mechanisms of control in response to combined stress. The net effect of these interactions depends on a multitude of factors; thus, these interactions modify the impact of biotic stresses on plants by altering their susceptibility. The aim of this review is to provide an overview of what is currently known about M. phaseolina and abiotic stress interactions, as well as several other edaphic factors that interact with plants. We briefly discuss the role of drought, salinity, heat stress, and edaphic factors (such as pH, N, P, K, etc.) that influence pathogen infection in plants. Furthermore, we discuss possible management strategies to combat crop loss due to combined stress. Thus, we suggest the future aspect of combined stress breeding, along with the use of multi-omics techniques and genome editing approaches, to develop cultivars that exhibit stability in a combined stress environment.
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    Genome-wide identification, characterization, and expression profiling of 14-3-3 genes in legumes
    (Springer Nature Publishing AG, 2022) Chakraborty, Srija; Soundararajan, Prabhakaran; Kumar, Shailesh
    In plants, a large multigene family encodes 14-3-3 proteins, which are commonly found in eukaryotes. They are involved in plant development and environmental stress regulation. The current study aims to identify and characterize the 14-3-3 gene family in four important legumes, viz. Cicer arietinum, Cajanus cajan, Vigna radiata and Arachis hypogaea. The 14-3-3 proteins were clustered into ε and non-ε groups based on phylogenetic analysis, which was further confrmed by gene structure analysis, and motif composition analysis. Our study suggests that segmental duplication events played a pivotal role in the evolution and expansion of this gene family. Evidence of both stress-responsive, and hormone-responsive cis-regulatory elements in the promoter region of the 14-3-3 genes points to the possibility that they might be involved in the interplay between hormone and stress signalling pathways. The expression profling using RNA-seq data sets showed a variation in expression level in most 14-3-3 genes under multiple stress conditions, and in diferent developmental stages. Hence, this study afrms the participation of the 14-3-3 genes in multiple stress regulation, and in growth and development of the legumes. The results obtained from this study provide crucial information for improved understanding, and stress response analysis of the 14-3-3 gene family in C. arietinum, C. cajan, V. radiata and A. hypogaea.
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    BURP domain-containing genes in legumes: genome-wide identification, structure, and expression analysis under stresses and development
    (Springer Nature Publishing AG, 2022) Chitkara, Pragya; Poddar, Nikita; Singh, Amarjeet; Kumar, Shailesh
    BURP domain-containing proteins are a plant-specific protein family which play an important role in plant metabolism and development. These proteins have also been involved in various abiotic and biotic stress responses. In this study, genome-wide identification and characterization of BURP domain protein encoding gene family is performed in four important legumes, Phaseolus vulgaris, Cicer arietinum, Cajanus cajan, and Vigna radiata. BURP genes were distributed randomly across chromosomes in all four legume plants. The phylogenetic analysis classified all BURP proteins into five major subfamilies, namely, USP-like, RD22-like, BNM2-like, PG1β-like, and BURPV. Our findings revealed that BURP gene family descended from common ancestors with segmental gene duplication events playing a critical role in their evolution and expansion in legumes. The intron–exon and conserved protein motifs analysis revealed that BURP genes are structurally conserved in legumes. The promoter analysis revealed the presence of hormone, and stress-responsive cis-regulatory elements in BURP promoters, implying that BURP functions in both hormone and abiotic stress signaling. Global expression analysis revealed that several BURP genes in all four legumes express differentially during plant development, and under biotic and abiotic stresses. This indicates crucial role of BURP proteins in regulating the development of legumes and adaptation to different abiotic/biotic stresses. This study will provide the starter for cloning and detail functional investigation of BURP proteins in legume crops.
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    Quantitative phosphoproteomic analysis of legume using TiO2-based enrichment coupled with isobaric labeling
    (Springer Nature Publishing AG, 2020) Barua, Pragya; Lande, Nilesh Vikram; Kumar, Sunil; Chakraborty, Subhra; Chakraborty, Niranjan
    Phosphorylation of proteins is the most dynamic protein modification, and its analysis aids in determining the functional and regulatory principles of important cellular pathways. The legumes constitute the third largest family of higher plants, Fabaceae, comprising about 20,000 species and are second to cereals in agricultural importance on the basis of global production. Therefore, an understanding of the developmental and adaptive processes of legumes demands identification of their regulatory components. The most crucial signature of the legume family is the symbiotic nitrogen fixation, which makes this fascinating and interesting to investigate phosphorylation events. The research on protein phosphorylation in legumes has been focused primarily on two model species, Medicago truncatula and Lotus japonicus. The development of reciprocal research in other species, particularly the crops, is lagging behind which has limited its beneficial uses in agricultural productivity. In this chapter, we outline the titanium dioxide-based enrichment of phosphopeptides for nuclear proteome analysis of a grain legume, chickpea.