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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.