Publications of NIPGR Scientists

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    Comparative investigation on chemical and green synthesized titanium dioxide nanoparticles against chromium (VI) stress eliciting differential physiological, biochemical, and cellular attributes in Helianthus annuus L.
    (Elsevier B.V., 2024) Kumar, Dharmendra; Mariyam, Safoora; Gupta, Kapuganti Jagadis; Thiruvengadam, Muthu; Ghodake, Gajanan Sampatrao; Xing, Baoshan; Seth, Chandra Shekhar
    Nanotechnology is a new scientific area that promotes unique concepts to comprehend the optimal mechanics of nanoparticles (NPs) in plants under heavy metal stress. The present investigation focuses on effects of synthetic and green synthesized titanium dioxide nanoparticles (TiO2 NPs and gTiO2 NPs) against Cr(VI). Green TiO2 NPs have been produced from plant leaf extract (Ricinus communis L.). Synthesis was confirmed employing an array of optical spectroscopic and electron microscopic techniques. Chromium strongly accelerated H2O2 and MDA productions by 227 % and 266 % at highest chromium concentration (60 mg/kg of soil), respectively, and also caused DNA damage, and decline in photosynthesis. Additionally, anomalies were observed in stomatal cells with gradual increment in chromium concentrations. Conversely, foliar applications of TiO2 NPs and gTiO2 NPs considerably mitigated chromium stress. Sunflower plants treated with modest amounts of green TiO2 NPs had significantly better growth index compared to chemically synthesized ones. Principal component analysis highlighted the variations among photosynthetic attributes, oxidative stress markers, and antioxidant defense systems. Notably, gTiO2 supplementation to the Cr(VI) strained plants minimized PC3 production which is a rare report so far. Conclusively, gTiO2 NPs have been identified to be promising nano-based nutrition resource for farming applications.
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    Heterologous overexpression of PDH45 gene of pea provides tolerance against sheath blight disease and drought stress in rice
    (Elsevier B.V., 2022) Sahoo, Ranjan Kumar; Chandan, Ravindra Kumar; Swain, Durga Madhab; Tuteja, Narendra; Jha, Gopaljee
    Biotic and abiotic stress tolerant crops are required for sustainable agriculture as well as ensuring global food security. In a previous study, we have reported that heterologous overexpression of pea DNA helicase (PDH45), a DEAD-box family member protein, provides salinity stress tolerance in rice. The improved management of photosynthetic machinery and scavenging of reactive oxygen species (ROS) are associated with PDH45 mediated salinity stress tolerance. However, the role of PDH45 in biotic and other abiotic stress (drought) tolerance remains unexplored. In the present study, we have generated marker-free transgenic IR64 rice lines that overexpress PDH45 under the CaMV35S promoter. The transgenic rice lines exhibited a significant level of tolerance against sheath blight disease, caused by Rhizoctonia solani, a polyphagous necrotrophic fungal pathogen. The defense as well as antioxidant responsive marker genes were significantly upregulated in the PDH45 overexpressing (OE) rice lines, upon pathogen infection. Moreover, the OE lines exhibited tolerance to drought stress and various antioxidant as well as drought responsive marker genes were significantly upregulated in them, upon drought stress. Overall, the current study emphasizes that heterologous overexpression of PDH45 provides abiotic as well as biotic stress tolerance in rice. Tolerance against drought as well as sheath blight disease by overexpression of a single gene (PDH45) signifies the practical implication of the present study. Moreover, considering the conserved nature of the gene in different plant species, we anticipate that PDH45 can be gainfully deployed to impart tolerance against multiple stresses in agriculturally important crops.
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    Genomic dissection of ROS detoxifying enzyme encoding genes for their role in antioxidative defense mechanism against Tomato leaf curl New Delhi virus infection in tomato
    (Elsevier B.V., 2021) Sharma, Namisha; Muthamilarasan, Mehanathan; Dulani, Priya; Prasad, Manoj
    In the present study, genes encoding for six major classes of enzymatic antioxidants, namely superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), Peroxidase (Prx) and glutathione S-transferase (GST) are identified in tomato. Their expression was studied in tomato cultivars contrastingly tolerant to ToLCNDV during virus infection and different hormone treatments. Significant upregulation of SlGR3, SlPrx25, SlPrx75, SlPrx95, SlGST44, and SlGST96 was observed in the tolerant cultivar during disease infection. Virus-induced gene silencing of SlGR3 in the tolerant cultivar conferred disease susceptibility to the knock-out line, and higher accumulation (~80%) of viral DNA was observed in the tolerant cultivar. Further, subcellular localization of SlGR3 showed its presence in cytoplasm, and its enzymatic activity was found to be increased (~65%) during ToLCNDV infection in the tolerant cultivar. Knock-out lines showed ~3- and 3.5-fold reduction in GR activity, which altogether underlines that SlGR3 is vital component of the defense mechanism against ToLCNDV infection.
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    Host gamma aminobutyric acid metabolic pathway is involved in resistance against Rhizoctonia solani
    (American Phytopathological Society, 2021) Rani, Mamta; Jha, Gopaljee
    Rhizoctonia solani is a highly destructive necrotrophic fungal pathogen having a diverse host range, including rice and tomato. Previously R. solani infection in rice has been found to cause large-scale readjustment in host primary metabolism and accumulation of various stress associated metabolites such as gamma aminobutyric acid (GABA). In this study, we report upregulation of GABA shunt genes during pathogenesis of R. solani in rice as well as tomato. The exogenous application of GABA provided partial resistance against R. solani infection in both the hosts. Further, using virus induced gene silencing (VIGS) approach, we knocked down the expression of some of the tomato genes involved in GABA biosynthesis (glutamate decarboxylase; GAD) and GABA catabolism (GABA-transaminase; GABA-T and succinic semialdehyde dehydrogenase; SSADH) to study their role in host defense against R. solani infection. The silencing of each of these genes was found to enhance disease susceptibility in tomato. Overall the results from gene expression analysis, exogenous chemical treatment and gene silencing studies suggest that GABA pathway plays a positive role in plant resistance against necrotrophic fungal pathogen R. solani.
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    Ectopic overexpression of cytosolic ascorbate peroxidase gene (Apx1) improves salinity stress tolerance in Brassica juncea by strengthening antioxidative defense mechanism
    (Springer Nature, 2020) Saxena, Saurabh C.; Salvi, Prafull; Kamble, Nitin Uttam; Joshi, Pankaj K.; Majee, Manoj; Arora, Sandeep
    Salinity stress is considered to be a key constrain that reduces the crop productivity by impairing plant growth and development. During salt stress condition, an underlying mechanism for reduction in crop yield is increase in ROS level that can potentially harm cellular macromolecules, leading to disruption of essential physiological and biochemical processes. Plants possess a complex antioxidative defense machinery for scavenging these ROS. Ascorbate peroxidase (APX, E.C. 1.11.1.11), is a crucial antioxidant enzyme involved in Ascorbate–Glutathione pathway that primarily detoxifies the negative impact of H2O2 in cell. The efficient scavenging of H2O2 is a prerequisite for enhanced tolerance to salinity stress. Here, we have inspected whether over-expression of APX could provide protection against salinity stress. Cytosolic ascorbate peroxidase (Apx1) gene, isolated from Arabidopsis thaliana, was chosen as the candidate gene for strengthening the antioxidative defense system of Brassica juncea. Physiological parameters were employed to analyze the growth status of transgenic plants. Leaf disc assay was done to evaluate the salinity stress tolerance potential of transgenic plants, using several physiological and biochemical parameters. Under salinity stress, the transgenic plants performed well as compared to their non-transgenic counterparts; as revealed through greater proline accumulation, increased chlorophyll stability index, lower chlorophyll a/b ratio, and higher antioxidative enzyme activities. Further, the lower H2O2 levels were well correlated with lesser membrane damage as measured through MDA content. Collectively, our results clearly depicted that ectopic overexpression of AtApx1 gene was able to confer salinity stress tolerance by strengthening the antioxidative defense system in B. juncea.
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    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 Jagadis
    Nitrate, 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.
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    Morpho-physiological and transcriptome profiling reveal novel zinc deficiency-responsive genes in rice
    (Springer, 2017) Bandyopadhyay, Tirthankar; Mehra, Poonam; Hairat, Suboot; Giri, Jitender
    Intensive farming has depleted the soil zinc (Zn) availability resulting in decreased crop productivity. Here, we attempt to understand the Zn deficiency response in rice through temporal transcriptome analysis. For this, rice seedlings were raised under Zn-deficient conditions up to 4 weeks followed by Zn re-supply for 3 days. Zn-deficient plants developed characteristic deficiency symptoms such as leaf bronzing, decrease in biomass, total chlorophyll, PSII efficiency, decreased carbonic anhydrase activity and increased ROS production. Interestingly, severe alterations in root system architecture were also observed. Comprehensive transcriptome analyses of rice seedlings were carried out after 2 (DEF2W) and 4 weeks (DEF4W) of Zn deficiency with respect to transcriptome profiles of corresponding Zn sufficient conditions (SUF2W, SUF4W). Additionally, to detect the potential Zn-responsive genes, transcriptome profile of Zn-recovered seedlings was compared with DEF4W. All differentially expressed Zn-responsive genes were categorized into early and late Zn deficiency response, and a set of 77 genes, induced and repressed on Zn deficiency and re-supply, respectively, was identified. These genes could be used as low Zn-responsive marker genes. Further, genes involved in membrane transport, phytosiderophore activity and organic acid biosynthesis showed high differential expression. Additionally, the present study unravelled several genes putatively associated with alterations in root system architecture under Zn deficiency and provides novel insights into the interpretation of morpho-physiological, biochemical and molecular regulation of zinc deficiency responses in rice.
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    Arsenic stress activates MAP kinase in rice roots and leaves
    (Elsevier, 2011) Rao, Kudupudi Prabhakara; Vani, Gubbala; Kumar, Kundan; Wankhede, Dhammaprakash Pandhari; Misra, Mohit; Gupta, Meetu; Sinha, Alok Krishna
    The toxic metalloid arsenite has become a potential threat to rice growing regions leading to serious contamination in food chain. In the present study effect of different physiological concentration of arsenite that is toxic and triggers the molecular events were evaluated in rice seedlings. Along with severe effect on the growth of rice seedling, production of reactive oxygen species (ROS) and nitric oxide (NO) in arsenite treated rice roots was also observed. Activation of a 42kDa mitogen activated protein kinase (MAPK/MPK) by arsenite was observed in rice leaves and 42 and 44kDa in roots in dose dependent manner. The activated MAPK could be immunoprecipitated with anti-phospho-tyrosine antibody, 4G10. The kinetic of MAPK activation by arsenite was found to be dose dependent. Transcript analysis of MAPK family and immunokinase assay in arsenite treated rice seedling revealed significant level of induction in OsMPK3 transcripts in leaves and OsMPK3, OsMPK4 transcripts in roots. Among MAPK kinase (MKKs) gene family, OsMKK4 transcripts were found to be induced in arsenite treated rice leaves and roots. In-silico homology modeling and docking analysis supported OsMPK3-OsMKK4 interaction. The data indicates that arsenite stress is transduced through MAPK signaling cascade in rice.