Institutional Publications

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Subject: search.filters.subject.reactive oxygen species

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    OsNUOR enhances disease susceptibility by interfering with reactive oxygen species homeostasis and ferroptosis-like cell death
    (Oxford University Press, 2026)
    Necrotrophic fungal pathogens such as Rhizoctonia solani, the causal agent of rice (Oryza sativa) sheath blight disease, enhance reactive oxygen species (ROS) production to induce necrosis in infected tissues. Here, we present evidence that the host alternative NADH:ubiquinone oxidoreductase (OsNUOR) facilitates R. solani infection by promoting an oxidative-stress-enriched environment and inducing iron-dependent ferroptosis-like cell death. OsNUOR overexpression (OE) lines exhibit enhanced disease susceptibility, whereas knock-out (KO) lines developed through genome editing demonstrate increased resistance. Infected OE lines have enhanced accumulation of ROS, lipid peroxides, and ferric ions (Fe3+); a significant reduction in antioxidative enzyme (including glutathione peroxidase) activity; and depletion of glutathione levels. In KO lines, the redox status of infected tissues is maintained, and the antioxidative defense is activated. Our data suggest that upregulation of OsNUOR induces mitochondrial ROS accumulation and modulates redox signalling, leading to Fe3+ accumulation and lipid peroxidation that promote necrosis in rice. KO lines are compromised in these processes and therefore exhibit disease resistance. We demonstrate that treatment with ferroptosis inhibitors prevents necrotic lesions, whereas ferroptosis inducers enhance disease severity. Overall, our study reveals the importance of ferroptosis-like cell death in promoting necrosis during R. solani infection in rice.
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    CBL1/9-CIPK6 complex negatively regulates respiratory burst oxidase homolog D in Arabidopsis thaliana
    (John Wiley & Sons, 2026) Vishwakarma, Niraj Kumar; Yadav, Shalini; Sardar, Atish; Choudhary, Megha; Chattopadhyay, Debasis
    Plant innate immune response is a well-balanced process with positive and negative regulations for the plants to survive. Calcium signaling is essential for pathogen-associated molecular pattern (PAMP)-driven respiratory burst oxidase homolog D (RBOHD)-mediated reactive oxygen species (ROS) burst. We show that calcium sensors calcineurin B like protein 1 (CBL1) and CBL9 and their interacting protein kinase CIPK6 negatively regulate RBOHD activity and immune response in Arabidopsis thaliana. Arabidopsis mutant cbl1cbl9, like cipk6, exhibited enhanced resistance and ROS production when infected with the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). CBL1 and CBL9 enhanced kinase activity of CIPK6. CBL1/9-CIPK6 module interacts with RBOHD at the plasma membrane. CIPK6 along with CBL1 reduces RBOHD activity in planta. CIPK6 phosphorylates the N-terminal cytoplasmic domain of RBOHD at a non-conserved (S33) and a conserved (S39) serine residue. While S39 phosphorylation increased RBOHD activity, S33 phosphorylation drastically reduced it and superseded the effect of S39 phosphorylation. We propose a model that CIPK6 phosphorylates RBOHD at S33 to suppress its activity to balance ROS generation in post-PTI situation in Arabidopsis. Our study reports a direct mechanism of negative regulation of ROS production and plant immune response by a calcium-signaling module in Arabidopsis thaliana.
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    Cellular responses in the pigeonpea wild relative Cajanus platycarpus to Helicoverpa armigera herbivory: The role of methionine sulfoxide reductase B1 (CpMSRB1) in enhanced defense
    (American Phytopathological Society, 2025) Rathinam, Maniraj; Dokka, Narasimham; Senthil, Kameshwaran; Mahawar, Shivangi; Tyagi, Shaily; Rengarajan, Dineshkumar; Vijayaraghavareddy, Preethi; Iyyappan, Yuvaraj; YB, Basavaraj; Reddy, Sandeep; T, Vinutha; G, Rama Prashat; Sinha, Subodh Kumar; Dash, Prasanta K.; Sreeman, Sheshshayee; Majee, Manoj; Sreevathsa, Rohini
    Understanding key cellular mechanisms leading to improved defense against various stressors is essential for cultivating robust nutritious crops capable of flourishing in diverse environments. We present an in-depth characterization of the defense response in the pigeonpea wild relative Cajanus platycarpus to herbivory by pod borer Helicoverpa armigera. To fight the attacking pest, C. platycarpus strategically activated non-enzymatic reactive oxygen species (ROS) scavengers and unleashed methionine sulfoxide reductases to safeguard the integrity of methionine residues. We unveiled for the first time physical interaction between CpMSRB1 and chorismate mutase (CpCM1.1), a pivotal player in the phenylpropanoid pathway. This association fueled the synthesis of phenylpropanoids and enhanced ROS scavenging crucial for repelling herbivores. Repairing CpCM1.1 also boosted salicylic acid production, coordinating defense signaling with jasmonic acid. Additionally, heterologous expression of CpMSRB1 in tomato improved defense against herbivory by enhanced ROS scavenging and polyphenol production. This study demonstrates the role of CpMSRB1 in protecting a major enzyme in the shikimate pathway, reinforcing defense against H. armigera.
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    Serine hydroxymethyltransferase6 is involved in growth and resistance against pathogens via ethylene and lignin production in Arabidopsis
    (John Wiley & Sons, 2024) Singh, Pooja; Kumari, Aprajita; Khaladhar, Vemula Chandra; Singh, Namrata; Pathak, Pradeep Kumar; Kumar, Vinod; Kumar, Ritika Jantu; Jain, Priyanka; Thakur, Jitendra K.; Fernie, Alisdair R.; Bauwe, Hermann; Raghavendra, A.S.; Gupta, Kapuganti Jagadis
    Photorespiratory serine hydroxymethyltransferases (SHMTs) are important enzymes of cellular one-carbon metabolism. In this study, we investigated the potential role of SHMT6 in Arabidopsis thaliana. We found that SHMT6 is localized in the nucleus and expressed in different tissues during development. Interestingly SHMT6 is inducible in response to avirulent, virulent Pseudomonas syringae and to Fusarium oxysporum infection. Overexpression of SHMT6 leads to larger flowers, siliques, seeds, roots, and consequently an enhanced overall biomass. This enhanced growth was accompanied by increased stomatal conductance and photosynthetic capacity as well as ATP, protein, and chlorophyll levels. By contrast, a shmt6 knockout mutant displayed reduced growth. When challenged with Pseudomonas syringae pv tomato (Pst) DC3000 expressing AvrRpm1, SHMT6 overexpression lines displayed a clear hypersensitive response which was characterized by enhanced electrolyte leakage and reduced bacterial growth. In response to virulent Pst DC3000, the shmt6 mutant developed severe disease symptoms and becomes very susceptible, whereas SHMT6 overexpression lines showed enhanced resistance with increased expression of defense pathway associated genes. In response to Fusarium oxysporum, overexpression lines showed a reduction in symptoms. Moreover, SHMT6 overexpression lead to enhanced production of ethylene and lignin, which are important components of the defense response. Collectively, our data revealed that SHMT6 plays an important role in development and defense against pathogens.
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    Nitric oxide regulation of plant metabolism
    (Elsevier B.V., 2022) Gupta, Kapuganti Jagadis; Kaladhar, Vemula Chandra; Fitzpatrick, Teresa B; Fernie, Alisdair R; Møller, Ian Max; Loake, Gary J
    Nitric oxide (NO) has emerged as an important signal molecule in plants, having myriad roles in plant development. In addition, NO also orchestrates both biotic and abiotic stress responses, during which intensive cellular metabolic reprogramming occurs. Integral to these response is the location of NO biosynthetic and scavenging pathways in diverse cellular compartments, enabling plants to effectively organize signal transduction pathways. NO regulates plant metabolism and in turn, metabolic pathways reciprocally regulate NO accumulation and function. Thus, these diverse cellular processes are inextricably linked. This review addresses the numerous redox pathways, located in the various subcellular compartments, which produce NO, in addition to the mechanisms underpinning NO scavenging. We focus on how this molecular dance is integrated into the metabolic state of the cell. Within this context, a reciprocal relationship between NO accumulation and metabolite production is often apparent. We additionally showcase cellular pathways including those associated with nitrate reduction that provide evidence for this integration of NO function and metabolism. Finally, we discuss the potential importance of the biochemical reactions governing NO levels in determining plant responses to a changing environment.
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    Isolation and measurement of respiration and structural studies of purified mitochondria from heterotrophic plant tissues
    (John Wiley & Sons, 2021) Pandey, Sonika; Kumari, Aprajita; Singh, Pooja; Gupta, Kapuganti Jagadis
    Mitochondria are the power houses of eukaryotic cells. These organelles contain various oxidoreductase complexes. Electron transfer from different reducing equivalents channeled via these complexes drives proton translocation across the inner mitochondrial membrane, leading to ATP generation. Plant mitochondria contain alternative NAD(P)H dehydrogenases, alternative oxidase, and uncoupling protein, and TCA cycle enzymes are located in their matrix. Apart from ATP production, mitochondria are also involved in synthesis of vitamins and cofactors and participate in fatty acid, nucleotide, photorespiratory, and antioxidant metabolism. Recent emerging evidence suggests that mitochondria play a role in redox signaling and generation of reactive oxygen and nitrogen species. For mitochondrial studies, it is essential to isolate physiologically active mitochondria with good structural integrity. In this article, we explain a detailed procedure for isolation of mitochondria from various heterotrophic tissues, such as germinating chickpea seeds, potato tubers, and cauliflower florets. This procedure requires discontinuous Percoll gradient centrifugation and can give a good yield of mitochondria, in the range of 4 to 8 mg per 50 g tissue with active respiratory capacity. After MitoTracker staining, isolated mitochondria can be visualized by using a confocal microscope. The structure of mitochondria can be monitored by scanning electron microscopy.
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    Phytoglobin-NO cycle and AOX pathway play a role in anaerobic germination and growth of deepwater rice
    (John Wiley & Sons, 2021) Kumari, Aprajita; Singh, Pooja; Kaladhar, Vemula Chandra; Manbir; Paul, Debarati; Pathak, Pradeep Kumar; Gupta, Kapuganti Jagadis
    An important and interesting feature of rice is that it can germinate under anoxic conditions. Though several biochemical adaptive mechanisms play an important role in the anaerobic germination of rice but the role of phytoglobin-nitric oxide cycle and alternative oxidase pathway is not known, therefore in this study we investigated the role of these pathways in anaerobic germination. Under anoxic conditions, deepwater rice germinated much higher and rapidly than aerobic condition and the anaerobic germination and growth were much higher in the presence of nitrite. The addition of nitrite stimulated NR activity and NO production. Important components of phytoglobin-NO cycle such as methaemoglobin reductase activity, expression of Phytoglobin1, NIA1 were elevated under anaerobic conditions in the presence of nitrite. The operation of phytoglobin-NO cycle also enhanced anaerobic ATP generation, LDH, ADH activities and in parallel ethylene levels were also enhanced. Interestingly nitrite suppressed the ROS production and lipid peroxidation. The reduction of ROS was accompanied by enhanced expression of mitochondrial alternative oxidase protein and its capacity. Application of AOX inhibitor SHAM inhibited the anoxic growth mediated by nitrite. In addition, nitrite improved the submergence tolerance of seedlings. Our study revealed that nitrite driven phytoglobin-NO cycle and AOX are crucial players in anaerobic germination and growth of deepwater rice.
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    The uncoupling of respiration in plant mitochondria: keeping reactive oxygen and nitrogen species under control
    (Oxford University Press, 2021) Popov, Vasily N.; Syromyatnikov, Mikhail Y.; Fernie, Alisdair R.; Chakraborty, Subhra; Gupta, Kapuganti Jagadis; Igamberdiev, Abir U.
    Plant mitochondrial respiration involves the operation of various alternative pathways. These pathways participate, both directly and indirectly, in the maintenance of mitochondrial functions though they do not contribute to energy production, being uncoupled from the generation of an electrochemical gradient across the mitochondrial membrane and thus from ATP production. Recent findings suggest that uncoupled respiration is involved in reactive oxygen species (ROS) and nitric oxide (NO) scavenging, regulation, and homeostasis. Here we discuss specific roles and possible functions of uncoupled mitochondrial respiration in ROS and NO metabolism. The mechanisms of expression and regulation of the NDA-, NDB- and NDC-type non-coupled NADH and NADPH dehydrogenases, the alternative oxidase (AOX), and the uncoupling protein (UCP) are examined in relation to their involvement in the establishment of the stable far-from-equilibrium state of plant metabolism. The role of uncoupled respiration in controlling the levels of ROS and NO as well as inducing signaling events is considered. Secondary functions of uncoupled respiration include its role in protection from stress factors and roles in biosynthesis and catabolism. It is concluded that uncoupled mitochondrial respiration plays an important role in providing rapid adaptation of plants to changing environmental factors via regulation of ROS and NO.
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    Nitric oxide accelerates germination via the regulation of respiration in chickpea
    (Oxford University Press, 2019) Pandey, Sonika; Kumari, Aprajita; Shree, Manu; Kumar, Vinod; Singh, Pooja; Bharadwaj, Chellapilla; Loake, Gary J.; Parida, Swarup K.; Masakapalli, Shyam Kumar; Gupta, Kapuganti Jagadis
    Seed germination is crucial for the plant life cycle. We investigated the role of nitric oxide (NO) in two chickpea varieties that differ in germination capacity: Kabuli, which has a low rate of germination and germinates slowly, and Desi, which shows improved germination properties. Desi produced more NO than Kabuli and had lower respiratory rates. As a result of the high respiration rates, Kabuli had higher levels of reactive oxygen species (ROS). Treatment with the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) reduced respiration in Kabuli and decreased ROS levels, resulting in accelerated germination rates. These findings suggest that NO plays a key role in the germination of Kabuli. SNAP increased the levels of transcripts encoding enzymes involved in carbohydrate metabolism and the cell cycle. Moreover, the levels of amino acids and organic acids were increased in Kabuli as a result of SNAP treatment. 1 H-nuclear magnetic resonance analysis revealed that Kabuli has a higher capacity for glucose oxidation than Desi. An observed SNAP-induced increase in 13C incorporation into soluble alanine may result from enhanced oxidation of exogenous [13C]glucose via glycolysis and the pentose phosphate pathway. A homozygous hybrid that originated from a recombinant inbred line population of a cross between Desi and Kabuli germinated faster and had increased NO levels and a reduced accumulation of ROS compared with Kabuli. Taken together, these findings demonstrate the importance of NO in chickpea germination via the control of respiration and ROS accumulation.
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    Reactive oxygen species, nitric oxide production and antioxidant gene expression during development of aerenchyma formation in wheat
    (Taylor & Francis Group, 2018) Wany, Aakanksha; Gupta, Kapuganti Jagadis
    In response to hypoxia, plant roots produce very high levels of nitric oxide. Recently, it was demonstrated that NO and ethylene both are essential for development of aerenchyma in wheat roots under hypoxia. Increased NO under hypoxia correlated with induction of NADPH oxidase gene expression, ROS production and lipid peroxidation in cortical cells. Tyrosine nitration was prominent in cells developing aerenchyma suggesting that NO and ROS play a key role in development of aerenchyma. However, the role of antioxidant genes during development of aerenchyma is not known, therefore, we checked gene expression of various antioxidants such as SOD1, AOX1A, APX and MnSOD at different time points after hypoxia treatment and found that expression of these genes elevated in 2 h but downregulated in 24 h where development of aerenchyma is prominent. Further, we found that plants growing under ammonium nutrition displayed delayed aerenchyma development. Taken together, new insights presented in this short communication highlighted additional regulatory role of antioxidants gene expression during aerenchyma development.