Publications of NIPGR Scientists

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    Genomic and expression analysis indicate the involvement of phospholipase C family in abiotic stress signaling in chickpea (Cicer arietinum)
    (Elsevier B.V., 2020) Sagar, Sushma; Biswas, Dipul Kumar; Singh, Amarjeet
    Phospholipase C proteins are phospholipid hydrolysing enzymes and crucial components of abiotic stress triggered lipid signaling in plants. PLCs are implicated in plant reaction to drought, salinity, and cold stress responses, however, characterization of the PLC family in the legume crop chickpea is missing. Here, we identify and describe nine PLC encoding genes in the chickpea genome. Phylogenetic analysis showed that the chickpea PLC family has evolved through a common path in dicots. Subcellular localization of fluorescence tagged proteins confirmed cytoplasmic and plasma membrane bound forms of PLCs in chickpea. The promoters of all the PLC genes are comprised of several hormone response related, development and abiotic stress related cis-regulatory elements. Expression analysis in five developmental stages (germination, seedling, vegetative, reproductive and senescence) showed significant expression of multiple PLCs in germination, vegetative and reproductive stages, suggesting their diverse role in various developmental processes. qRT-PCR expression analysis of the entire PLC gene family under drought, salt and cold stresses revealed that most PLC genes are differentially expressed in multiple abiotic stresses. These observations indicate the involvement of PLC gene family in abiotic stress signaling and responses in important legume crop. The present study opens new avenues for utilizing PLC- related information in biotechnological programs for abiotic stress tolerance and legume crop improvement.
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    Mediator subunit OsMED14_1 plays an important role in rice development
    (John Wiley & Sons, 2020) Malik, Naveen; Ranjan, Rajeev; Parida, Swarup K.; Agarwal, Pinky; Tyagi, Akhilesh K.
    Mediator, a multi‐subunit coactivator complex, regulates transcription in eukaryotes and is involved in diverse processes in Arabidopsis through its different subunits. Here, we have explored developmental aspects of one of the rice Mediator subunit gene OsMED14_1. We analyzed its expression pattern through RNA in‐situ hybridization and pOsMED14_1:GUS transgenics which showed its expression in roots, leaves, anthers and seeds prominently at younger stages, indicating possible involvement of this subunit in multiple aspects of rice development. To understand developmental roles of OsMED14_1 in rice, we generated and studied RNAi based knockdown rice plants which showed multiple effects including less height, narrower leaves and culms with reduced vasculature, lesser lateral root branching, defective microspore development, reduced panicle branching and seed set, and smaller seeds. Histological analyses showed that slender organs were caused by reduction in both cell number and cell size in OsMED14_1 knockdown plants. Flow cytometric analyses and expression analyses of cell‐cycle related genes revealed that defective cell‐cycle progression led to these defects. Expression analyses of auxin related genes and IAA immuno‐localization study indicated altered auxin level in these knockdown plants. Reduction of lateral root branching in knockdown plants was corrected by exogenous IAA supplement. OsMED14_1 physically interacts with transcription factors YABBY5, TDR and MADS29, possibly regulating auxin homeostasis and ultimately leading to lateral organ/leaf, microspore and seed development.
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    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, Debasis
    Calcium 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.
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    Genome-wide expressional and functional analysis of calcium transport elements during abiotic stress and development in rice
    (John Wiley & Sons, 2014) Singh, Amarjeet; Kanwar, Poonam; Yadav, Akhilesh K.; Mishra, Manali; Jha, Saroj K.; Baranwal, Vinay; Pandey, Amita; Kapoor, Sanjay; Tyagi, Akhilesh K.; Pandey, Girdhar K.
    Ca²⁺ homeostasis is required to maintain a delicate balance of cytosolic Ca²⁺ during normal and adverse growth conditions. Various Ca²⁺ transporters actively participate to maintain this delicate balance especially during abiotic stresses and developmental events in plants. In this study, we present a genome-wide account, detailing expression profiles, subcellular localization and functional analysis of rice Ca²⁺ transport elements. Exhaustive in silico data mining and analysis resulted in the identification of 81 Ca²⁺ transport element genes, which belong to various groups such as Ca²⁺-ATPases (pumps), exchangers, channels, glutamate receptor homologs and annexins. Phylogenetic analysis revealed that different Ca²⁺ transporters are evolutionarily conserved across different plant species. Comprehensive expression analysis by gene chip microarray and quantitative RT-PCR revealed that a substantial proportion of Ca²⁺ transporter genes were expressed differentially under abiotic stresses (salt, cold and drought) and reproductive developmental stages (panicle and seed) in rice. These findings suggest a possible role of rice Ca²⁺ transporters in abiotic stress and development triggered signaling pathways. Subcellular localization of Ca²⁺ transporters from different groups in Nicotiana benthamiana revealed their variable localization to different compartments, which could be their possible sites of action. Complementation of Ca²⁺ transport activity of K616 yeast mutant by Ca²⁺-ATPase OsACA7 and involvement in salt tolerance verified its functional behavior. This study will encourage detailed characterization of potential candidate Ca²⁺ transporters for their functional role in planta.
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    Comparative transcript profiling of TCP family genes provide insight into gene functions and diversification in rice and Arabidopsis
    (Academy Journals, 2010) Sharma, Rita; Kapoor, Meenu; Tyagi, Akhilesh K.; Kapoor, Sanjay
    Plant-specific TCP transcription factor family has been implicated in diverse aspects of growth and development. Rice and Arabidopsis genomes encode 26 and 24 TCP family genes, respectively. In this study, we have performed an inclusive analysis of their expression during 21 and 18 stages of development in rice and Arabidopsis, respectively. The assorted patterns of expression, exhibited by TCP family genes, provide an evidence for spatiotemporal regulation of their relative abundance throughout plant development. Further profiling of rice genes in three sub-stages of early panicle development revealed differential accumulation of nine genes during panicle initiation and organ development. QPCR-based expression profiling of selected rice genes, during four stages of anther, suggested their involvement in early anther development as well. Eleven genes of rice and seven of Arabidopsis were differentially expressed in response to three abiotic stress treatments viz., cold, dehydration and salt. In silico analysis of 5' regulatory regions of differentially expressed genes revealed the presence of previously characterized cis-regulatory elements. Duplications seem to have played major role in diversification of TCP family genes with 14 genes of rice and 10 of Arabidopsis lying on duplicated segments of the respective genomes. Most of the duplicated genes exhibited varied expression patterns. The knowledge obtained in this study will be useful for selection and assessment of the functions of individual genes using reverse genetics approaches.