Browsing by Author "Panchal, Poonam"

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A citrate efflux transporter important for manganese distribution and phosphorus uptake in rice
(John Wiley & Sons, 2023) Panchal, Poonam; Bhatia, Chitra; Chen, Yi; Sharma, Meenakshi; Bhadouria, Jyoti; Verma, Lokesh; Maurya, Kanika; Miller, Anthony J.; Giri, Jitender
The plant citrate transporters, functional in mineral nutrient uptake and homeostasis, usually belong to the multidrug and toxic compound extrusion transporter family. We identified and functionally characterized a rice (Oryza sativa) citrate transporter, OsCT1, which differs from known plant citrate transporters and is structurally close to rice silicon transporters. Domain analysis depicted that OsCT1 carries a bacterial citrate-metal transporter domain, CitMHS. OsCT1 showed citrate efflux activity when expressed in Xenopus laevis oocytes and is localized to the cell plasma membrane. It is highly expressed in the shoot and reproductive tissues of rice, and its promoter activity was visible in cells surrounding the vasculature. The OsCT1 knockout (KO) lines showed a reduced citrate content in the shoots and the root exudates, whereas overexpression (OE) line showed higher citrate exudation from their roots. Further, the KO and OE lines showed variations in the manganese (Mn) distribution leading to changes in their agronomical traits. Under deficient conditions (Mn-sufficient conditions followed by 8 days of 0 μm MnCl2 · 4H2 O treatment), the supply of manganese towards the newer leaf was found to be obstructed in the KO line. There were no significant differences in phosphorus (P) distribution; however, P uptake was reduced in the KO and increased in OE lines at the vegetative stage. Further, experiments in Xenopus oocytes revealed that OsCT1 could efflux citrate with Mn. In this way, we provide insights into a mechanism of citrate-metal transport in plants and its role in mineral homeostasis, which remains conserved with their bacterial counterparts.
Monogalactosyl Diacylglycerol Synthase 3 affects phosphate utilization and acquisition in rice
(Oxford University Press, 2022) Verma, Lokesh; Bhadouria, Jyoti; Rupam, Bhunia Kumar; Singh, Shweta; Panchal, Poonam; Bhatia, Chitra; Eastmond, Peter J; Giri, Jitender
Galactolipids are essential for compensating for the loss of phospholipids by “membrane lipid remodeling” in plants under phosphorus (P) deficiency. Monogalactosyl diacylglycerol (MGDG) synthases catalyze the synthesis of MGDG which is further converted into digalactosyl diacylglycerol (DGDG), later replacing phospholipids in the extraplastidial membranes. However, the roles of these enzymes are not well explored in rice. In this study, the rice MGDG synthase gene, OsMGD3 was identified and functionally characterized. We showed that plant phosphate (Pi) status and transcription factor OsPHR2 are involved in the transcriptional regulation of OsMGD3. CRISPR/Cas9 knockout (KO) and overexpression (OE) lines of OsMGD3 were generated to explore its potential role in rice adaptation to Pi deficiency. Compared to WT, OsMGD3 KO lines displayed a reduction while OE lines showed an enhanced, Pi acquisition and utilization. Further, OsMGD3 showed a predominant role in roots, altering lateral root growth. Our comprehensive lipidomic analysis revealed the role of OsMGD3 in membrane lipid remodeling in addition to a role in regulating diacylglycerol and phosphatidic acid levels that affected the expression of Pi transporters. Our study highlights the role of OsMGD3 in affecting both internal P utilization and P acquisition in rice.
Organic acids: Versatile stress response roles in plants
(Oxford University Press, 2021) Panchal, Poonam; Miller, Anthony J; Giri, Jitender
Organic acids (OAs) are central to cellular metabolism. Many plant stress responses involve exudation of OAs at the root-soil interface that can improve soil mineral acquisition and toxic metal tolerance. Because of their simple structure, the Low Molecular Weight Organic Acids (LMWOAs) are widely studied. We discuss the conventional roles of OAs, along with some newly emerging roles in plant stress tolerance. OAs are more versatile in their role in plant stress tolerance and are efficient chelating agents when compared with other acids, such as amino acids. Root OA exudation is important in soil carbon sequestration. These functions are key processes combating climate change and helping with more sustainable food production. We briefly review the mechanisms behind enhanced biosynthesis, secretion and regulation of these activities under different stresses. Also, an outline of the transgenic approaches targeted towards the enhanced production and secretion of OAs is provided. A re-occurring theme of OAs in plant biology is their roles as either ‘acids’ modifying pH or ‘chelators’ binding metals or as ‘carbon sources’ for microbes. We argue that these multiple functions are key factors for understanding these molecules important roles in plant stress biology. Finally, we contemplate how the functions of OAs in plant stress responses can be made use of and what the important unanswered questions are.
Root-expressed rice PAP3b enhances secreted APase activity and helps utilize organic phosphate
(Oxford University Press, 2023) Bhadouria, Jyoti; Mehra, Poonam; Verma, Lokesh; Pazhamala, Lekha T; Rumi, Rumi; Panchal, Poonam; Sinha, Alok Krishna; Giri, Jitender
Phosphate (Pi) deficiency leads to the induction of purple acid phosphatases (PAPs) in plants, which dephosphorylates organic phosphorus complexes in the rhizosphere and intracellular compartments to release Pi. In this study, we demonstrate that OsPAP3b belongs to group III low molecular weight PAP, and is low Pi responsive, preferentially in roots. The expression of OsPAP3b is negatively regulated with Pi re-supply. Interestingly, OsPAP3b was found to be dual localized to the nucleus and secretome. Furthermore, OsPAP3b is transcriptionally regulated by OsPHR2 as substantiated by DNA-protein binding assay. Through in-vitro biochemical assays, we further demonstrate that OsPAP3b is a functional acid phosphatase with broad substrate specificity. Overexpression of OsPAP3b in rice led to increased secreted APase activity and improved mineralization of organic P sources, reflected in better growth of transgenics compared to wild type when grown on organic P as exogenous P substrate. Under Pi deprivation, OsPAP3b knockdown and knockout lines showed no significant changes in total P content and dry biomass. However, the expression of other phosphate starvation-induced (PSI) genes and the levels of metabolites were found to be altered in the overexpression and knockdown lines. In addition, in-vitro pull-down assay revealed multiple putative interacting proteins of OsPAP3b. Our data collectively suggest that OsPAP3b can aid in organic P utilization in rice. The APase isoforms behavior and nuclear localization indicate its additional role, possibly in stress signaling. Considering its important roles, OsPAP3b could be a potential target for improving low Pi adaptation in rice.
Soil carbon sequestration by root exudates
(Elsevier B.V., 2022) Panchal, Poonam; Preece, Catherine; Peñuelas, Josep; Giri, Jitender
Root exudates are well-known ‘labile’ sources of soil carbon that can prime microbial activity. Recent investigations suggest that the stability of labile carbon inputs in soil mostly depends upon the physical, chemical, and biological properties of the surroundings. Here, we propose that, in some ecosystems, such as forests and grasslands, root exudates can function as a source of soil organic carbon (SOC) that can be stabilized through various mechanisms leading to long-term sequestration. Increasing soil carbon sequestration is important for capturing atmospheric CO2 and combating climate change issues. Thus, there is an urgent need to preserve existing ecosystems and to adopt strategies such as afforestation, reforestation, and establishment of artificial grasslands to foster carbon sequestration through higher root exudate inputs in the soil.