Institutional Publications

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Subject: search.filters.subject.reactive oxygen species (ROS)

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    Flavonols affect the interrelated glucosinolate and camalexin biosynthetic pathways in Arabidopsis thaliana
    (Oxford University Press, 2024) Naik, Jogindra; Tyagi, Shivi; Rajput, Ruchika; Kumar, Pawan; Pucker, Boas; Bisht, Naveen C.; Misra, Prashant; Stracke, Ralf; Pandey, Ashutosh
    Flavonols are structurally and functionally diverse biomolecules involved in plant biotic and abiotic stress tolerance, pollen development, and inhibition of auxin transport. Despite the ubiquitous nature and multifunctionality of flavonols in land plants, their effects on global gene expression and signaling pathways are unclear. To explore the roles of flavonol metabolites in signaling, we performed comparative transcriptome and targeted metabolite profiling of seedlings from the flavonol-deficient Arabidopsis (Arabidopsis thaliana) loss-of-function mutant flavonol synthase1 (fls1) with and without exogenous supplementation of flavonol derivatives (kaempferol, quercetin, and rutin). Our RNA-seq results indicated that flavanols modulate various biological and metabolic pathways, with significant alteration in camalexin and aliphatic glucosinolate synthesis. Flavonols negatively regulated camalexin biosynthesis but appeared to promote the accumulation of aliphatic glucosinolates via transcription factor–mediated upregulation of biosynthesis genes. Interestingly, upstream amino acid biosynthesis genes involved in methionine and tryptophan synthesis were altered under flavonol deficiency and exogenous supplementation. Quercetin treatment significantly upregulated aliphatic glucosinolate biosynthesis genes compared to kaempferol and rutin. In addition, expression and metabolite analysis of the transparent testa7 mutant, which lacks hydroxylated flavonol derivatives, clarified the role of quercetin in the glucosinolate biosynthesis pathway. This study elucidates the molecular mechanisms by which flavonols interfere with signaling pathways, their molecular targets, and the multiple biological activities of flavonols in plants.
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    Revisiting the role of MAPK signalling pathway in plants and its manipulation for crop improvement
    (John Wiley & Sons, 2023) Manna, Mrinalini; Rengasamy, Balakrishnan; Sinha, Alok Krishna
    The mitogen-activated protein kinase (MAPK) pathway is an important signalling event associated with every aspect of plant growth, development, yield, abiotic and biotic stress adaptation. Being a central metabolic pathway, it is a vital target for manipulation for crop improvement. In this review, we have summarised recent advancements in understanding involvement of MAPK signalling in modulating abiotic and biotic stress tolerance, architecture and yield of plants. MAPK signalling cross talks with reactive oxygen species (ROS) and abscisic acid (ABA) signalling events in bringing about abiotic stress adaptation in plants. The intricate involvement of MAPK pathway with plant's pathogen defence ability has also been identified. Further, recent research findings point towards participation of MAPK signalling in shaping plant architecture and yield. These make MAPK pathway an important target for crop improvement and we discuss here various strategies to tweak MAPK signalling components for designing future crops with improved physiology and phenotypes.
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    Methionine sulfoxide reductase B5 plays a key role in preserving seed vigor and longevity in rice (Oryza sativa)
    (John Wiley & Sons, 2022) Hazra, Abhijit; Varshney, Vishal; Verma, Pooja; Kamble, Nitin Uttam; Ghosh, Shraboni; Achary, Rakesh Kumar; Gautam, Shikha; Majee, Manoj
    Oxidation of methionine leads to the formation of methionine S-sulfoxide and methionine R-sulfoxide, which can be reverted by two types of Methionine Sulfoxide Reductase (MSR), MSRA and MSRB, respectively. Despite the role of MSR enzymes being elucidated in various physiological processes, regulation and implication of MSR in seeds remained poorly explored. In this study, through molecular, biochemical, and genetic studies using seed-specific overexpression and RNAi lines of OsMSRB5 in Oryza sativa, we demonstrate the role of OsMSRB5 in maintaining seed vigor and longevity. We show that age-induced reduced vigor and viability of seeds is correlated with reduced MSR activity and increased methionine sulfoxide (MetSO) formation. OsMSRB5 expression increases during seed maturation and predominantly localizes in the embryo. Further analyses on transgenic lines reveal the role of OsMSRB5 in modulating reactive oxygen species (ROS) homeostasis to preserve seed vigor and longevity. We show that ascorbate peroxidase (APX) and PROTEIN L-ISOASPARTYL METHYLTRANSFERASE (PIMT) undergo MetSO modification in seeds that affect their functional competence. OsMSRB5 physically interacts with these proteins and reverts this modification to facilitate their functions and preserve seed vigor and longevity of seeds. Our results thus illustrate the role of OsMSRB5 in preserving seed vigor and longevity by modulating ROS homeostasis in seeds.
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    A conserved NAG motif is critical to the catalytic activity of galactinol synthase, a key regulatory enzyme of RFO biosynthesis
    (Portland Press, 2021) Salvi, Prafull; Kumar, Bhavnesh; Kamble, Nitin Uttam; Hazra, Abhijit; Majee, Manoj
    Galactinol synthase (GolS) catalyzes the key regulatory step in the biosynthesis of Raffinose Family Oligosaccharides (RFOs). Even though the physiological role and regulation of this enzyme has been well studied, little is known about active site amino acids and the structure-function relationship with substrates of this enzyme. In the present study, we investigate the active site amino acid and structure function relationship for this enzyme. Using a combination of three-dimensional homology modelling, molecular docking along with a series of deletion, site directed mutagenesis followed by in vitro biochemical and in vivo functional analysis; we have studied active site amino acids and their interaction with the substrate of chickpea and Arabidopsis GolS enzyme. Our study reveals that the GolS protein possesses GT8 family-specific several conserved motifs in which NAG motif plays a crucial role in substrate binding and catalytic activity of this enzyme. Deletion of entire NAG motif or deletion or the substitution (with alanine) of any residues of this motif results in complete loss of catalytic activity in in vitro condition. Furthermore, disruption of NAG motif of CaGolS1 enzyme disrupts it’s in vivo cellular function in yeast as well as in planta. Together, our study offers a new insight into the active site amino acids and their substrate interaction for the catalytic activity of GolS enzyme. We demonstrate that NAG motif plays a vital role in substrate binding for the catalytic activity of galactinol synthase that affects overall RFO synthesis.
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    Surviving the odds: from perception to survival of fungal phytopathogens under host-generated oxidative burst
    (Elsevier B.V., 2021) Singh, Yeshveer; Nair, Athira Mohandas; Verma, Praveen K.
    Fungal phytopathogens pose a serious threat to global crop production. Only a handful of strategies are available to combat these fungal infections, and the increasing incidence of fungicide resistance is making the situation worse. Hence, the molecular understanding of plant–fungus interactions remains a primary focus of plant pathology. One of the hallmarks of host–pathogen interactions is the overproduction of reactive oxygen species (ROS) as a plant defense mechanism, collectively termed the oxidative burst. In general, high accumulation of ROS restricts the growth of pathogenic organisms by causing localized cell death around the site of infection. To survive the oxidative burst and achieve successful host colonization, fungal phytopathogens employ intricate mechanisms for ROS perception, ROS neutralization, and protection from ROS-mediated damage. Together, these countermeasures maintain the physiological redox homeostasis that is essential for cell viability. In addition to intracellular antioxidant systems, phytopathogenic fungi also deploy interesting effector-mediated mechanisms for extracellular ROS modulation. This aspect of plant–pathogen interactions is significantly under-studied and provides enormous scope for future research. These adaptive responses, broadly categorized into ‘‘escape’’ and ‘‘exploitation’’ mechanisms, are poorly understood. In this review, we discuss the oxidative stress response of filamentous fungi, their perception signaling, and recent insights that provide a comprehensive understanding of the distinct survival mechanisms of fungal pathogens in response to the host-generated oxidative burst.
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    Regulating the regulator: nitric oxide control of post-translational modifications
    (John Wiley & Sons, Inc., 2020) Gupta, Kapuganti Jagadis; Kolbert, Zsuzsanna; Durner, Jorg; Lindermayr, Christian; Corpas, Francisco J.; Brouquisse, Renaud; Barroso, Juan B.; Saima, Umbreen; Palma, José M; Hancock, John T.; Petrivalsky, Marek; Wendehenne, David; Loake, Gary J.
    Nitric oxide (NO) is perfectly suited for duties as a redox signalling molecule. A key route for NO bioactivity occurs via protein S‐nitrosation, the addition of a NO moiety to a protein cysteine (Cys) thiol (‐SH) to form a S‐nitrosothiol (SNO). This process is thought to underpin a myriad of cellular processes in plants linked to development, environmental responses and immune function. Here we collate emerging evidence showing that NO bioactivity regulates a growing number of diverse post‐translational modifications (PTMs) including SUMOylation, phosphorylation, persulfidation and acetylation. We provide examples of how NO orchestrates these processes to mediate plant adaptation to a variety of cellular cues.
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    Arabidopsis Protein L-Isoaspartyl Methyltransferase repairs isoaspartyl damage to antioxidant enzymes and increases heat and oxidative stress tolerance
    (American Society for Biochemistry and Molecular Biology, 2020) Ghosh, Shraboni; Kamble, Nitin Uttam; Verma, Pooja; Salvi, Prafull; Petla, Bhanu Prakash; Roy, Shweta; Rao, Venkateswara; Hazra, Abhijit; Varshney, Vishal; Kaur, Harmeet; Majee, Manoj
    Stressful environments accelerate the formation of isoaspartyl (isoAsp) residues in proteins, which detrimentally affect protein structure and function. The enzyme Protein L-Isoaspartyl Methyltransferase (PIMT) repairs other proteins by reverting deleterious isoAsp residues to functional aspartyl residues. PIMT function previously has been elucidated in seeds, but its role in plant survival under stress conditions remains undefined. Herein, we used molecular, biochemical, and genetic approaches, including protein overexpression and knockdown experiments, in Arabidopsis to investigate the role of PIMTs in plant growth and survival during heat and oxidative stresses. We demonstrate that these stresses increase isoAsp accumulation in plant proteins, that PIMT activity is essential for restricting isoAsp accumulation, and that both PIMT1 and PIMT2 play an important role in this restriction and Arabidopsis growth and survival. Moreover, we show that PIMT improves stress tolerance by facilitating efficient reactive oxygen species (ROS) scavenging and thereby protecting the functionality of antioxidant enzymes from isoAsp-mediated damage during stress. Specifically, biochemical and MS/MS analyses revealed that antioxidant enzymes acquire deleterious isoAsp residues during stress, which adversely affect their catalytic activities, and that PIMT repairs the isoAsp residues and thereby restores antioxidant enzyme function. Collectively, our results suggest that the PIMT-mediated protein repair system is an integral part of the stress tolerance mechanism in plants, in which PIMTs protect antioxidant enzymes that maintain proper ROS homeostasis against isoAsp-mediated damage in stressful environments.
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    Nitrate, NO and ROS signaling in stem cell homeostasis
    (Cell Press, 2018) Wany, Aakanksha; Foyer, Christine H.; Gupta, Kapuganti Jagadis
    Shoot and root growth is facilitated by stem cells in the shoot and root apical meristems (SAM and RAM). Recent reports have demonstrated a close link between nitrogen nutrition, nitric oxide (NO), and reactive oxygen species (ROS) in the regulation of SAM and RAM functions in response to nitrogen availability.