Publications of NIPGR Scientists
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Item Differential regulation of nitric oxide mediated by phytoglobin1 plays a role in resistance during Botrytis cinerea infection in Arabidopsis thaliana(Springer Nature Publishing AG, 2026) Jaiswal, Rekha; Saini, Deepak; Swain, Jagannath; Gupta, Kapuganti JagadisBotrytis cinerea is a prominent necrotrophic pathogen responsible for gray mold disease, affecting a wide range of plant species, including economically vital crops such as tomatoes, grapes, strawberries, etc. Nitric oxide (NO) is considered as a crucial player in plant responses to biotic stress. NO homeostasis is regulated by phytoglobin (Pgb1), a potential scavenger of NO. However, the role of the Pgb1-NO cycle in regulating defense response against B. cinerea remains largely unknown. In the current study, we investigated the defense response of Arabidopsis thaliana against B. cinerea infection using antisense (Pgb1 AS) and overexpression (Pgb1 OE) lines, which produce differential levels of NO. The Pgb1 AS line accumulated higher NO levels and conferred resistance against B. cinerea infection, with reduced ROS levels, reduced cell death, and increased stomatal closure. Conversely, Pgb1 OE showed reduced NO levels accompanied by increased susceptibility. The elevated NO level in Pgb1 AS was associated with increased nitrate reductase (NR) activity and upregulation of NIA1 and NIA2 gene expression. Interestingly, ethylene-mediated defense pathway genes such as ERF1, ACS2, and ACS6 were upregulated while pathogen-related genes such as PR1, PR2, PR5, and NPR1 were downregulated in Pgb1 AS line. The elevated expression of ethylene genes corresponded with the higher ethylene levels in Pgb1 AS. Overall, our results confirmed the crucial role of phytoglobin-modulated NO in defense against B. cinerea infection by activating the ethylene-mediated defense pathway.Item The confluence of TOR signaling and plant growth regulators in development and stress responses(Oxford University Press, 2026) Saksena, Harshita B; Kumar, Mukesh; Samtani, Harsha; Sharma, Aishwarye; Rawat, Sanjay Singh; Awasthi, Prakhar; Botta, Halidev Krishna; Sandhya, Shital; Pande, Anjali; Naaz, Sheeba; Kushwah, Sunita; Shukla, Brihaspati N; Laxmi, AshveryaThe Target of Rapamycin (TOR) is an evolutionarily conserved protein kinase that serves as a crucial signaling hub, seamlessly integrating a wide range of internal and external signals to meticulously regulate cellular and organismal physiology. TOR is crucial in regulating the different phases of lifecycle in plants including embryogenesis, seed germination, meristem activation, root and leaf development, flowering and senescence. Beyond its central role in growth and development, emerging research has revealed its significant involvement in the response to environmental stresses. Even though plant growth regulators such as auxin, cytokinin (CK), brassinosteroid (BR), gibberellin (GA), abscisic acid (ABA), ethylene (ET), salicylic acid (SA), jasmonic acid (JA) and nitric oxide (NO) function as pivotal signaling molecules in modulating plant development and stress responses, how they coordinate with the energy status still remains obscure. Here we summarize the current findings on the dynamic interconnection between TOR and these discrete phytoregulators and their potential role in executing diverse biological processes in plants.Item Expanding roles for S-nitrosylation in the regulation of plant immunity(Elsevier B.V., 2023) Borrowman, Sam; Gupta, Kapuganti Jagadis; Loake, Gary J.Following pathogen recognition, plant cells produce a nitrosative burst resulting in a striking increase in nitric oxide (NO), altering the redox state of the cell, which subsequently helps orchestrate a plethora of immune responses. NO is a potent redox cue, efficiently relayed between proteins through its co-valent attachment to highly specific, powerfully reactive protein cysteine (Cys) thiols, resulting in formation of protein S-nitrosothiols (SNOs). This process, known as S-nitrosylation, can modulate the function of target proteins, enabling responsiveness to cellular redox changes. Key targets of S-nitrosylation control the production of reactive oxygen species (ROS), the transcription of immune-response genes, the triggering of the hypersensitive response (HR) and the establishment of systemic acquired resistance (SAR). Here, we bring together recent advances in the control of plant immunity by S-nitrosylation, furthering our appreciation of how changes in cellular redox status reprogramme plant immune function.Item The PHYTOGLOBIN-NO cycle regulates plant mycorrhizal symbiosis(Elsevier B.V., 2019) Kumari, Aprajita; Pathak, Pradeep Kumar; Loake, Gary J.; Gupta, Kapuganti JagadisThe production of the redox-active signaling molecule, NO, has long been associated with interactions between microbes and their host plants. Emerging evidence now suggests that specific NO signatures and cognate patterns of PHYTOGLOBIN1 (PHYTOGB1) expression, a key regulator of cellular NO homeostasis, may help determine either symbiosis or pathogenicity.Item Methods for measuring nitrate reductase, nitrite levels, and nitric oxide from plant tissues(Springer Nature Publishing AG, 2020) Wany, Aakanksha; Pathak, Pradeep Kumar; Gupta, Kapuganti JagadisNitrogen (N) is one of the most important nutrients which exist in both inorganic and organic forms. Plants assimilate inorganic form of N [nitrate (NO3−), nitrite (NO2−) or ammonium (NH4+)] and incorporate into amino acids. The metabolism of N involves a series of events such as sensing, uptake, and assimilation. The initial stage is sensing, triggered by nitrate or ammonium signals initiating signal transduction processes in N metabolism. The assimilation pathway initiates with NO3−/NH4+ transport to roots via specific high and low affinity (HATs and LATs) nitrate transporters or directly via ammonium transporters (AMTs). In cytosol the NO3− is reduced to NO2− by cytosolic nitrate reductase (NR) and the produced NO2− is further reduced to NH4+ by nitrite reductase (NiR) in plastids. NR has capability to reduce NO2− to nitric oxide (NO) under specific conditions such as hypoxia, low pH, and pathogen infection. The produced NO acts as a signal for wide range of processes such as plant growth development and stress. Here, we provide methods to measure NR activity, NO2− levels, and NO production in plant tissues.Item Using different forms of nitrogen to study hypersensitive response elicited by avirulent Pseudomonas syringae(Springer Nature Publishing AG, 2020) Singh, Namrata; Singh, Pooja; Pathak, Pradeep Kumar; Gupta, Kapuganti JagadisNitrate, ammonium, or a combination of both is the form of N available for nitrogen assimilation from soil by the plants. Nitrogen is an important and integral part of amino acids, nucleotides, and defense molecules. Hence it is very important to study the role of nitrate and ammonium nutrition in plant defense via hypersensitive response (HR). Shifting plants from ammonium nitrate Hoagland solution to nitrate Hoagland nutrition slightly enhances root length and leaf area. HR phenotype is different in nitrate and ammonium grown plants when challenged with avirulent Pseudomonas syringae DC3000 avrRpm1. HR is also associated with increased production of reactive oxygen species (ROS) and nitric oxide (NO). Hence to understand HR development it is essential to measure HR lesions, cell death, ROS, NO, and bacterial growth. Here we provide a stepwise protocol of various parameters to study HR in Arabidopsis in response to nitrate and ammonium nutrition.Item Measurement of nitrate reductase activity in tomato (Solanum lycopersicum L.) leaves under different conditions(Springer Nature Publishing AG, 2020) Bulle, Mallesham; Kishorekumar, Reddy; Pathak, Pradeep K.; Wany, Aakanksha; Gupta, Kapuganti JagadisNitrogen is one of the crucial macronutrients essential for plant growth, development, and survival under stress conditions. Depending on cellular requirement, plants can absorb nitrogen mainly in multiple forms such as nitrate (NO3−) or ammonium (NH4+) or combination of both via efficient and highly regulated transport systems in roots. In addition, nitrogen-fixing symbiotic bacteria can fix atmospheric nitrogen in to NH4+ via highly regulated complex enzyme system and supply to the roots in nodules of several species of leguminous plants. If NO3− is a primary source, it is transported from roots and then it is rapidly converted to nitrite (NO2−) by nitrate reductase (NR) (EC 1.6.6.1) which is a critical and very important enzyme for this conversion. This key reaction is mediated by transfer of two electrons from NAD(P)H to NO3−. This occurs via the three redox centers comprised of two prosthetic groups (FAD and heme) and a MoCo cofactor. NR activity is greatly influenced by factors such as developmental stage and various stress conditions such as hypoxia, salinity and pathogen infection etc. In addition, light/dark dynamics plays crucial role in modulating NR activity. NR activity can be easily detected by measuring the conversion of NO3− to NO2− under optimized conditions. Here, we describe a detailed protocol for measuring relative NR enzyme activity of tomato crude extracts. This protocol offers an efficient and straightforward procedure to compare the NR activity of various plants under different conditions.Item Nitric oxide and hydrogen sulfide crosstalk during heavy metal stress in plants(John Wiley & Sons, 2020) Shivaraj, Sheelavanta Matha; Vats, Sanskriti; Bhat, Javid Akhter; Dhakte, Priyanka; Goyal, Vinod; Khatri, Praveen; Kumawat, Surbhi; Singh, Akshay; Prasad, Manoj; Sonah, Humira; Sharma, Tilak Raj; Deshmukh, RupeshGases such as ethylene, hydrogen peroxide (H2O2), nitric oxide (NO), carbon monoxide (CO), and hydrogen sulfide (H2S) have been recognized as vital signaling molecules in plants and animals. Of these gasotransmitters, NO and H2S have recently gained momentum mainly because of their involvement in numerous cellular processes. It is therefore important to study their various attributes including their biosynthetic and signaling pathways. The present review provides an insight into various routes for the biosynthesis of NO and H2S as well as their signaling role in plant cells under different conditions, more particularly under heavy metal stress. Their beneficial roles in the plant's protection against abiotic and biotic stresses as well as their adverse effects have been addressed. This review describes how H2S and NO, being very small size molecules, can quickly pass through the cell membranes and triggers a multitude of responses to various factors, notably to various stress conditions like drought, heat, osmotic, heavy metal, and multiple biotic stresses. The versatile interactions between H2S and NO involved in the different molecular pathways have been discussed. In addition to the signaling role of H2S and NO, their direct role in post‐translational modifications is also considered. The information provided here will be helpful to better understand the multifaceted roles of H2S and NO in plants, particularly under stress conditions.Item A forty year journey: The generation and roles of NO in plants(Elsevier B.V., 2019) Kolbert, Zs; Barroso, J.B.; Brouquisse, R.; Corpas, F.J.; Gupta, Kapuganti Jagadis; Lindermayr, C.; Loake, G.J.; Palma, J.M.; Petřivalský, M.; Wendehenne, D.; Hancock, J.T.In this year there is the 40th anniversary of the first publication of plant nitric oxide (NO) emission by Lowell Klepper. In the decades since then numerous milestone discoveries have revealed that NO is a multifunctional molecule in plant cells regulating both plant development and stress responses. Apropos of the anniversary, these authors aim to review and discuss the developments of past concepts in plant NO research related to NO metabolism, NO signaling, NO's action in plant growth and in stress responses and NO's interactions with other reactive compounds. Despite the long-lasting research efforts and the accumulating experimental evidences numerous questions are still needed to be answered, thus future challenges and research directions have also been drawn up.Item Current approaches to measure nitric oxide in plants(Oxford University Press, 2019) Vishwakarma, Abhaypratap; Wany, Aakanksha; Pandey, Sonika; Bulle, Mallesham; Kumari, Aprajita; Kishorekumar, Reddy; Igamberdiev, Abir U.; Mur, Luis A.J.; Gupta, Kapuganti JagadisNitric oxide (NO), is now established as an important signalling molecule in plants where they influence growth, development and responses to stress. Despite of extensive research, the most appropriate methods to measure and localise these signalling radicals are debated and still needs investigation. Many confounding factors such as presence of other reactive intermediates, scavenging enzymes and compartmentation, influence how accurately each can be measured. Further, these signalling radicals have short half-lives ranging from seconds to minutes based on the cellular redox condition. Hence, it is necessary to use the sensitive and specific methods in order to understand the contribution of each signalling molecule to various biological processes. In this review, we will provide current state knowledge on NO measurement in plant samples, via various methods. We will also discuss advantages, limitations and wider applications of each method.