Publications of NIPGR Scientists

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Author: search.filters.author.Chattopadhyay, DebasisSubject: search.filters.subject.root

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    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, Debasis
    Cytokinin 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.
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    The MicroRNA397b-LACCASE2 module regulates root lignification under water and phosphate deficiency
    (American Society of Plant Biologists, 2020) Khandal, Hitaishi; Singh, Amar Pal; Chattopadhyay, Debasis
    Deficiency of water and phosphate induce lignin deposition in roots. LACCASEs, a family of cell wall-localized multi-copper oxidases, are involved in lignin biosynthesis. We demonstrate here that LACCASE2 (LAC2) acts as a negative regulator of lignin deposition in root vascular tissues during water deficit. An Arabidopsis T-DNA insertion mutant of LAC2 displayed a short primary root and high lignin deposition in root vascular tissues. However, restoration of LAC2 expression rescued these phenotypes. LAC2 expression was significantly downregulated under water deficit and post-transcriptionally regulated by microRNA397b (miR397b) in roots under normal and water deficit conditions. Downregulation of miR397b activity increased LAC2 expression and root length, and decreased lignin content in root vasculature. Similarly, phosphate (Pi) deficiency inversely affected miR397b and LAC2 expression. Lignin deposition in the root elongation zone under Pi-limited conditions was dependent on LAC2 expression. Localized iron accumulation and callose deposition in the root elongation zone under Pi-deficiency increased with LAC2-dependent lignification, suggesting a direct relationship between these processes. Our study reveals a regulatory role for the miR397b-LAC2 module in root lignification during water- and phosphate deficiency.
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    CBL-interacting protein kinase 25 contributes to root meristem development
    (Oxford University Press, 2019) Meena, Mukesh Kumar; Vishwakarma, Niraj Kumar; Tripathi, Vineeta; Chattopadhyay, Debasis
    Coordination of auxin and cytokinin activities determines root meristem size during post-embryonic development. Calcineurin B-like proteins (CBLs) and their interacting protein kinases (CIPKs) constitute signaling modules that relay calcium signals. Here we report that CIPK25 is involved in regulating the root meristem size. Arabidopsis plants lacking CIPK25 expression displayed a short root phenotype and a slower root growth rate with less meristem cells. This phenotype was rescued by restoration of CIPK25 expression. CIPK25 interacted with CBL4 and -5, and displayed a strong gene expression in the flower and root except in the cell proliferation domain in the root apical meristem. Its expression in root was positively and negatively regulated by auxin and cytokinin, respectively. cipk25 T-DNA insertion line was compromised in auxin transport and auxin-responsive promoter activity. cipk25 mutant line showed altered expression of auxin efflux carriers and an Aux/IAA family gene SHY2. Decreased PIN1 and PIN2 expression in cipk25 mutant line was completely restored when combined with SHY2 loss-of-function mutation resulting in recovery of root growth. SHY2 and PIN1 expression was partially regulated by cytokinin even in absence of CIPK25, suggesting existence of CIPK25-independent cytokinin signaling pathway(s) as well. Our results suggested a role of CIPK25 in root meristem 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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    Role of CIPK6 in root growth and auxin transport
    (Landes Bioscience, 2009) Tripathi, Vineeta; Syed, Nazia; Laxmi, Ashverya; Chattopadhyay, Debasis
    In our recent publication,1 we have shown that a T-DNA insertion in Arabidopsis CIPK6 gene encoding a CBL-interacting protein kinase caused reduction in expression of the gene and emergence of lateral roots. The change in phenotype in the mutant line was likely due to reduction in shoot-to-root acro- petal and the root tip basipetal auxin transport. Here we report identification of a homozygous knockout line of AtCIPK6 (atcipk6) with no detectable expression of the gene in normal growth condition. The knockout line exhibited considerable decrease in growth rate of the taproot as well as in emergence of lateral roots. The mutant line also showed reduction in the root tip basipetal and shoot-to-root acropetal auxin transport. Relative rate of auxin transport and the root phenotype of the atcipk6 closely matched with those of pgp4-1, an Arabidopsis line mutated in PGP4. This gene encodes an ABC integral membrane transporter, which functions in polar auxin transport. These observations strengthen our earlier proposal that CIPK6 is probably involved in polar auxin transport and indicate that it may function through the PGP4 auxin transporter.
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    CIPK6, a CBL-interacting protein kinase is required for development and salt tolerance in plant
    (Wiley-Blackwell, 2009) Tripathi, Vineeta; Parasuraman, Boominathan; Laxmi, Ashverya; Chattopadhyay, Debasis
    Calcineurin B-like proteins (CBL) and CBL-interacting protein kinases (CIPK) mediate plant responses to a variety of external stresses. Here we report that Arabidopsis CIPK6 is also required for the growth and development of plants. Phenotype of tobacco plants ectopically expressing a homologous gene (CaCIPK6) from the leguminous plant chickpea (Cicer arietinum) indicated its functional conservation. A lesion in AtCIPK6 significantly reduced shoot-to-root and root basipetal auxin transport, and the plants exhibited developmental defects such as fused cotyledons, swollen hypocotyls and compromised lateral root formation, in conjunction with reduced expression of a number of genes involved in auxin transport and abiotic stress response. The Arabidopsis mutant was more sensitive to salt stress compared to wild-type, while overexpression of a constitutively active mutant of CaCIPK6 promoted salt tolerance in transgenic tobacco. Furthermore, tobacco seedlings expressing the constitutively active mutant of CaCIPK6 showed a developed root system, increased basipetal auxin transport and hypersensitivity to auxin. Our results provide evidence for involvement of a CIPK in auxin transport and consequently in root development, as well as in the salt-stress response, by regulating the expression of genes.