Publications of NIPGR Scientists

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Author: search.filters.author.Gupta, Kapuganti JagadisHas files: No

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    Expression analysis of ROS-related genes during the germination of chickpea (Cicer Arietinum L.) seeds
    (Springer Nature Publishing AG, 2026) Babuta, Priyanka; Samant, Sanjib Bal; Saini, Deepak; Gupta, Kapuganti Jagadis
    Seed germination is a critical physiological process that transforms a quiescent seed into a metabolically active seedling and is also a crucial factor in determining maximum crop production. This transition is influenced by various intrinsic and extrinsic factors. Interestingly, reactive oxygen species (ROS) plays an important role in breaking seed dormancy by oxidation of biomolecules, weakening of the testa and degradation of endosperm. Similarly, molecular internal oxygen is also considered vital for the transition of dormancy to seed germination. However, it is essential to establish a correlation between the internal oxygen and the generation of ROS during seed germination. This chapter details protocols for imaging internal oxygen concentrations using VisiSens and fluorescent detection of ROS using H2DCFDA in chickpea seeds, complemented by qPCR analysis of key ROS-related genes (RBOH, AOX 1, UCP 1, and NADH dehydrogenase). These findings from these methods help advance our understanding of the inverse relationship between molecular oxygen and ROS dynamics during seed germination.
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    Method for the measurement of ethylene during hypoxia in rice plants
    (Springer Nature Publishing AG, 2025) Jaiswal, Rekha; Gupta, Kapuganti Jagadis; Praveen, Afsana
    Ethylene is a versatile phytohormone that is involved in the regulation of both growth and development such as senescence, and also it can act as a signaling hormone during hypoxia. Ethylene acts alone or in interaction with different phytohormones and proteins to regulate numerous cellular processes. Accumulating evidence suggest that endogenous ethylene production and emission into atmosphere are modulated by various biotic and abiotic stresses. Since it is a gaseous hormone, a precise detection, particularly under low-oxygen (hypoxic) conditions, is important for understanding its role in regulatory processes and stress signaling pathways. Currently, measurement practices such as gas chromatography, electrochemical sensing, and optical sensing are widely employed to detect ethylene. These methods are distinct from each other in terms of sensitivity, time response, selectivity, and cost. However, each method has its own advantages and limitations. Gas chromatography (GC) is one of the best techniques that is applied for the separation and measurement of ethylene due to its volatile and supersensitive nature. In this chapter, we describe a detailed GC-based procedure specifically optimized for measuring ethylene levels during hypoxic stress application in (Oryza sativa) rice plants.
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    Method for the measurement of ethylene during pathogen infection in arabidopsis
    (Springer Nature Publishing AG, 2025) Jaiswal, Rekha; Gupta, Kapuganti Jagadis; Praveen, Afsana
    Ethylene is a gaseous phytohormone that plays an important role as a signaling molecule during pathogen attack, influencing disease resistance and defense responses in plants. A precise measurement of ethylene production upon pathogen challenge is essential to elucidate its role in plant–pathogen interactions. Gas chromatography (GC) is among the most accurate and sensitive techniques for detecting and quantifying ethylene emissions due to its selectivity and effectiveness with gaseous molecules. In this chapter, we provide a detailed procedure employing GC specifically adapted for measuring ethylene levels during pathogen infection (Botrytis cinerea) in Arabidopsis leaflets. Arabidopsis leaflets infected by the necrotrophic pathogen Botrytis cinerea exhibit increased ethylene emission, facilitating the activation of defense pathways and secondary metabolites such as camalexin. The present GC method captures ethylene dynamics at early infection stages, ensuring precise quantification critical for dissecting the molecular mechanisms of plant immunity.
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    Measurement of reactive oxygen species and nitric oxide from tomato plants in response to abiotic and biotic stresses
    (Springer Nature Publishing AG, 2024) Samant, Sanjib Bal; Manbir; Rekha; Swain, Jagannath; Singh, Pooja; Kumari, Aprajita; Gupta, Kapuganti Jagadis
    Nitric oxide (NO) is a free radical molecule that has been known to influence several cellular processes such as plant growth, development, and stress responses. NO together with reactive oxygen species (ROS) play a role in signaling process. Due to extremely low half-life of these radicals in cellular environment, it is often difficult to precisely monitor them. Each method has some advantages and disadvantages; hence, it is important to measure using multiple methods. To interpret the role of each signaling molecule in numerous biological processes, sensitive and focused methods must be used. In addition to this complexity, these Reactive Oxygen Species (ROS) and NO react with each other leads to nitro-oxidative stress in plants. Using tomato as a model system here, we demonstrate stepwise protocols for measurement of NO by chemiluminescence, DAF fluorescence, nitrosative stress by western blot, and ROS measurement by NBT and DAB under stress conditions such as osmotic stress and Botrytis infection. While describing methods, we also emphasized on benefits, drawbacks, and broader applications of these methods.
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    Metabolomic response to high light from pgrl1 and pgr5 mutants of Chlamydomonas reinhardtii
    (Springer Nature Publishing AG, 2023) Chouhan, Nisha; Marriboina, Sureshbabu; Kumari, Aprajita; Singh, Pooja; Yadav, Ranay Mohan; Gupta, Kapuganti Jagadis; Subramanyam, Rajagopal
    Chlamydomonas (C.) reinhardtii metabolomic changes in cyclic electron flow-dependent mutants are still unknown. Here, we used mass spectrometric analysis to monitor the changes in metabolite levels in wild-type, cyclic electron-deficient mutants pgrl1 and pgr5 grown under high-light stress. A total of 55 metabolites were detected using GC-MS analysis. High-light stress-induced selective anaplerotic amino acids in pgr5. In addition, pgr5 showed enhancement in carbohydrate, polyamine, and polyol metabolism by 2.5-fold under high light. In response to high light, pgr5 triggers an increase in several metabolites involved in regulating osmotic pressure. Among these metabolites are glycerol pathway compounds such as glycerol-3-phosphate and glyceryl-glycoside, which increase significantly by 1.55 and 3.07 times, respectively. In addition, pgr5 also enhanced proline and putrescine levels by 2.6- and 1.36-fold under high light. On the other hand, pgrl1-induced metabolites, such as alanine and serine, are crucial for photorespiration when subjected to high-light stress. We also observed a significant increase in levels of polyols and glycerol by 1.37- and 2.97-fold in pgrl1 under high-light stress. Both correlation network studies and KEGG pathway enrichment analysis revealed that metabolites related to several biological pathways, such as amino acid, carbohydrate, TCA cycle, and fatty acid metabolism, were positively correlated in pgrl1 and pgr5 under high-light stress conditions. The relative mRNA expression levels of genes related to the TCA cycle, including PDC3, ACH1, OGD2, OGD3, IDH3, and MDH4, were significantly upregulated in pgrl1 and pgr5 under HL. In pgr5, the MDH1 level was significantly increased, while ACS1, ACS3, IDH2, and IDH3 levels were reduced considerably in pgrl1 under high-light stress. The current study demonstrates both pgr5 and prgl1 showed a differential defense response to high-light stress at the primary metabolites and mRNA expression level, which can be added to the existing knowledge to explore molecular regulatory responses of prg5 and pgrl1 to high-light stress.
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    The functional role of nitric oxide in plant mitochondrial metabolism
    (Elsevier B.V., 2016) Gupta, Alok Kumar; Kumari, Aprajita; Mishra, Sonal; Wany, Aakanksha; Gupta, Kapuganti Jagadis
    In recent years, mitochondrial nitric oxide (NO) production has attracted increasing attention. Mitochondria generate NO using nitrite as a substrate. Cytochrome c oxidase and other components of the electron transport chain also contribute to NO generation. Accumulating evidence indicates that mitochondria are scavengers of NO. Furthermore, several genes encoding mitochondrial proteins, as well as mitochondrial proteins, are regulated by NO. In this chapter, we provided an overview of the mechanisms of NO generation and scavenging in mitochondria and of the NO-dependent regulation of proteins and genes encoding mitochondrial proteins. In addition, the functional roles of NO in mitochondrial metabolism, such as inhibition of aconitase, production of ATP and induction of alternative oxidase are presented.
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    Integrating classical and alternative respiratory pathway
    (John Wiley & Sons, 2015) Gupta, Kapuganti Jagadis; Neelwarne, Bhagyalakshmi; Mur, Luis A.J.
    Respiratory pathways are vital for plant carbon and energy metabolism, which is the main use of most assimilated carbohydrates. Most respiratory pathways are very well established, the prominent being glycolysis in cytosol and the tricarboxylic acid (TCA) cycle, which occurs in the matrix of mitochondria coupled with the electron transport chain (ETC) which functions along the inner mitochondrial membrane. This chapter integrates such alternative respiratory pathways with components of the classical oxidative-phosphorylative pathways. Mitochondrial electron transport generates ATP by using the reducing equivalents derived through the operation of the TCA-cycle. Currently most research on alternative electron transfer is focused on nonphosphorylating bypass mechanisms: a second oxidase – the alternative oxidase (AOX), an external NAD (P) H dehydrogenases in the first part of ETC, and also plant uncoupling mitochondrial proteins (PUCPs). Electron transfer flavoprotein (ETF) is an electron acceptor for at least nine mitochondrial matrix flavoprotein dehydrogenases.
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    Simultaneous isolation of root and leaf mitochondria from Arabidopsis
    (John Wiley & Sons, 2015) Gupta, Kapuganti Jagadis; Ewald, Ralph
    In all aerobic organisms mitochondria generate ATP via oxidative phosphorylation. For bulky tissues such as potato tubers and cauliflower and for larger crop plants such as tobacco, pea, soybean or etiolated seedlings it is possible to get a good yield of mitochondria. However, for tiny model plants like Arabidopsis it is very difficult to get sufficient quantities of mitochondria for various studies. Moreover, for comparative studies it is very important to isolate leaf and root mitochondria. Due to the lack of chlorophyll, root mitochondria isolation is often an easy task. Leaf mitochondria isolation has the advantage that higher amounts of tissues can be obtained from the plants in comparison to root material, but chlorophyll contamination can be a problem. This chapter describes how to isolate root mitochondria with sufficiently high yields, and how to obtain chlorophyll-free leaf mitochondria simultaneously.