Publications of NIPGR Scientists
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Item Phosphate deficiency inducible OsGDPD5 affects root growth by regulating sugar-auxin crosstalk(John Wiley & Sons, 2025) Verma, Lokesh; Pandey, Mandavi; Bhatia, Chitra; Mehra, Poonam; Singh, Bhagat; Giri, JitenderGlycerophosphodiester phosphodiesterases (GDPDs) enzymes are known to be involved in phospholipids degradation pathways, where glycerophosphodiesters are hydrolyzed to glycerol-3-phosphate (G3P) and corresponding alcohol. In plants, GDPDs are involved in phosphate deficiency adaptive responses and have been shown to impact root length, but the precise mechanism remains unclear. This study focuses on the rice GDPD5 gene and its role in regulating primary root growth. Our research demonstrates that OsGDPD5 encodes a functional GDPD enzyme and could hydrolyze glycerophosphocholine and glycerophosphorylethanolamine. At transcriptional levels, OsGDPD5 is preferentially expressed in the root tip and regulated by transcription factor OsPHR2. We have used CRISPR/Cas9 to generate OsGDPD5 knock-out lines, allowing us to explore its role in root growth. Our findings show that osgdpd5 mutants had a shorter primary root, which could be restored to a normal level by the exogenous application of sugar or G3P. Further, knocking out OsGDPD5 alters endogenous levels of G3P and sugars, affecting auxin biosynthesis in the root and, ultimately, primary root growth. In this manner, OsGDPD5 has a crucial role in regulating physiological processes, specifically sugar and auxin signaling, which are known to be involved in root growth regulation in rice. Our research thus unraveled a link between rice phosphate deficiency-responsive lipid remodeling and root growth via sugar-hormone signaling.Item Lignin deposition in chickpea root xylem under drought(Taylor & Francis Group, 2020) Sharma, Nilesh Kumar; Gupta, Santosh KumarIn our recent publication, we have shown that a member of the Laccase family, LACCASE2 (LAC2) acted as a negative regulator of lignin deposition in the root xylem tissue of Arabidopsis thaliana. LAC2 messenger RNA (mRNA) level was post-transcriptionally regulated by microRNA 397b, which showed increased expression under water and phosphate deficiency, resulting in the downregulation of LAC2 expression In this report, we have investigated root growth and lignin deposition in an economically important legume crop chickpea (Cicer arietinum L.) in response to natural drought in soil-grown condition. In contrast to the growth retardation of Arabidopsis root in mannitol-supplemented medium, chickpea root showed an increase in length in low soil moisture condition. Lignin estimation in the primary root showed an increase in lignin content, which was substantiated by staining of root xylem. Drought treatment enhanced the expression of four out of six LAC genes tested, while the expression of two was downregulated. Our preliminary study indicateed a molecular mechanism of lignin deposition in chickpea root xylem during drought.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 The MicroRNA397b-LACCASE2 module regulates root lignification under water and phosphate deficiency(American Society of Plant Biologists, 2020) Khandal, Hitaishi; Singh, Amar Pal; Chattopadhyay, DebasisDeficiency 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.Item A novel glycerophosphodiester phosphodiesterase improves phosphate deficiency tolerance in rice(John Wiley & Sons, 2019) Mehra, Poonam; Pandey, Bipin K.; Verma, Lokesh; Giri, JitenderSoil phosphate (Pi) deficiency is major constraint for rice cultivation world‐wide. Cellular membranes account for one‐third of cellular P (Phosphorus) in the form of phospholipids. Therefore, remobilization of Pi from membrane phospholipids under Pi deficiency can be an important strategy to improve PUE (Phosphorus Use Efficiency). GDPDs (Glycerophosphodiester phosphodiesterases) hydrolyse intermediate product of phospholipid catabolism, glycerophosphodiesters to glycerol‐3‐phosphate (G3P); a precursor for P and non P‐lipid biosynthesis. Here, we show that OsGDPD2 is a Pi deficiency responsive gene which is transcriptionally regulated by OsPHR2. In silico analysis of active site residues and enzymatic assays confirmed phosphodiesterase activity of OsGDPD2. All overexpression lines showed higher GDPD activity, Pi content, root growth and biomass accumulation as compared to wild‐type. Conversely, silencing of OsGDPD2 led to decreased GDPD activity and Pi content. Notably, most of the P‐containing metabolites and fatty acids were elevated in transgenic lines. Further, quantitative analysis of polar lipids revealed higher accumulation of several classes of phospholipids and galactolipids in overexpression lines indicating a potential role of OsGDPD2 in de novo glycerolipid biosynthesis. Thus, present study provides insights into novel physiological roles of OsGDPD2 in low Pi acclimation in rice.Item CBL-interacting protein kinase 25 contributes to root meristem development(Oxford University Press, 2019) Meena, Mukesh Kumar; Vishwakarma, Niraj Kumar; Tripathi, Vineeta; Chattopadhyay, DebasisCoordination 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.Item Striking the right chord: Signaling enigma during root gravitropism(Frontiers Media S.A., 2017) Singh, Manjul; Gupta, Aditi; Laxmi, AshveryaPlants being sessile can often be judged as passive acceptors of their environment. However, plants are actually even more active in responding to the factors from their surroundings. Plants do not have eyes, ears or vestibular system like animals, still they "know" which way is up and which way is down? This is facilitated by receptor molecules within plant which perceive changes in internal and external conditions such as light, touch, obstacles; and initiate signaling pathways that enable the plant to react. Plant responses that involve a definite and specific movement are called "tropic" responses. Perhaps the best known and studied tropisms are phototropism, i.e., response to light, and geotropism, i.e., response to gravity. A robust root system is vital for plant growth as it can provide physical anchorage to soil as well as absorb water, nutrients and essential minerals from soil efficiently. Gravitropic responses of both primary as well as lateral root thus become critical for plant growth and development. The molecular mechanisms of root gravitropism has been delved intensively, however, the mechanism behind how the potential energy of gravity stimulus converts into a biochemical signal in vascular plants is still unknown, due to which gravity sensing in plants still remains one of the most fascinating questions in molecular biology. Communications within plants occur through phytohormones and other chemical substances produced in plants which have a developmental or physiological effect on growth. Here, we review current knowledge of various intrinsic signaling mechanisms that modulate root gravitropism in order to point out the questions and emerging developments in plant directional growth responses. We are also discussing the roles of sugar signals and their interaction with phytohormone machinery, specifically in context of root directional responses.Item JAZ repressors: Potential involvement in nutrients deficiency response in rice and chickpea(Frontiers Media S.A., 2015) Singh, Ajit P.; Pandey, Bipin K.; Deveshwar, Priyanka; Narnoliya, Laxmi; Parida, Swarup K.; Giri, JitenderJasmonates (JA) are well-known phytohormones which play important roles in plant development and defense against pathogens. Jasmonate ZIM domain (JAZ) proteins are plant-specific proteins and act as transcriptional repressors of JA-responsive genes. JA regulates both biotic and abiotic stress responses in plants; however, its role in nutrient deficiency responses is very elusive. Although, JA is well-known for root growth inhibition, little is known about behavior of JAZ genes in response to nutrient deficiencies, under which root architectural alteration is an important adaptation. Using protein sequence homology and a conserved-domains approach, here we identify 10 novel JAZ genes from the recently sequenced Chickpea genome, which is one of the most nutrient efficient crops. Both rice and chickpea JAZ genes express in tissue- and stimuli-specific manners. Many of which are preferentially expressed in root. Our analysis further showed differential expression of JAZ genes under macro (NPK) and micronutrients (Zn, Fe) deficiency in rice and chickpea roots. While both rice and chickpea JAZ genes showed a certain level of specificity toward type of nutrient deficiency, generally majority of them showed induction under K deficiency. Generally, JAZ genes showed an induction at early stages of stress and expression declined at later stages of macro-nutrient deficiency. Our results suggest that JAZ genes might play a role in early nutrient deficiency response both in monocot and dicot roots, and information generated here can be further used for understanding the possible roles of JA in root architectural alterations for nutrient deficiency adaptations.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.Item Role of CIPK6 in root growth and auxin transport(Landes Bioscience, 2009) Tripathi, Vineeta; Syed, Nazia; Laxmi, Ashverya; Chattopadhyay, DebasisIn 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.