Induction of cell death by tospoviral protein NSs and the motif critical for cell death does not control RNA silencing suppression activity

Singh, Ajeet; Permar, Vipin; Jain, Rakesh; Goswami, Suneha; Kumar, Rajeev; Canto, Tomás; Palukaitis, Peter; Praveen, Shelly

doi:10.1016/j.virol.2017.05.003

Cited by 4 publications

(10 citation statements)

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“…However, the efficacy of full-length NSs is not known. NSs protein of GBNV was shown as a pathogenic factor as it induced necrosis symptom and plant cell death ( Goswami et al, 2012 ; Singh et al, 2017 ). In this study, NSs being a symptom determinant, the dsRNA derived from the full-length gene was highly effective in protecting from GBNV infection in cowpea as well as in N. benthamiana plants.…”

Section: Discussionmentioning

confidence: 99%

“…The NSs protein of GBNV has been shown as a bifunctional enzyme having both ATPase and phosphatase activities ( Lokesh et al, 2010 ). Subsequently, the NSs protein of GBNV was found to be the symptom determinant, suppressor of RNA silencing, and inducer of plant cell death ( Goswami et al, 2012 ; Singh et al, 2017 ). GBNV infects a wide variety of important crops such as cowpea, mungbean, peanut, potato, tomato, soybean, and urdbean.…”

Section: Introductionmentioning

confidence: 99%

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Direct Foliar Application of dsRNA Derived From the Full-Length Gene of NSs of Groundnut Bud Necrosis Virus Limits Virus Accumulation and Symptom Expression

et al. 2021

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Groundnut bud necrosis virus (GBNV) is the most significant member of the genus Orthotospovirus occurring in the Indian subcontinent. There is hardly any effective measure to prevent GBNV in crop plants. In order to develop GBNV infection prevention procedure, we examined the effect of the direct foliar application of double-stranded RNA (dsRNA) derived from the full-length NSs gene (1,320 nucleotides) of GBNV. The bacterially expressed dsRNA to the non-structural (dsNSs) gene of GBNV was purified and delivered to plants as an aqueous suspension containing 0.01% Celite for evaluating its efficacy in preventing GBNV infection in systemic host, Nicotiana benthamiana as well as in local lesion and systemic host, cowpea cv. Pusa Komal (Vigna unguiculata). The dsNSs application and challenge-inoculation were conducted in three different combinations, where plants were challenge-inoculated with GBNV a day after, immediately, and a day before the application of dsNSs. N. benthamiana plants, which were not treated with dsRNA showed severe systemic wilting and death by 9–16 days post-inoculation (dpi). The non-treated cowpea plants exhibited many chlorotic and necrotic lesions on the cotyledonary leaves followed by systemic necrosis and death of the plants by 14–16 dpi. The dsNSs treated plants in all the combinations showed significant reduction of disease severity index in both N. benthamiana and cowpea. The treatment combination where the GBNV inoculation was conducted immediately after the dsNSs treatment was found to be the most effective treatment in preventing symptom expression. The viral RNA analysis by real time PCR also showed 20 and 12.5 fold reduction of GBNV in cowpea and N. benthamiana, respectively. Our results suggest that the foliar application of dsRNA derived from the full-length NSs gene of GBNV through Celite is successful in delivering long dsRNA leading to effective prevention of GBNV infection.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Direct Foliar Application of dsRNA Derived From the Full-Length Gene of NSs of Groundnut Bud Necrosis Virus Limits Virus Accumulation and Symptom Expression

et al. 2021

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“…The activities of different antioxidant enzymes (CAT, SOD, GR and APX) were determined at room temperature. CAT activity was performed according to [31] with some minor changes. The CAT activity reaction mixture was composed of 0.05 M phosphate buffer (pH 7.5), 0.020 M hydrogen peroxide and 30 µl enzyme extract.…”

Section: Enzyme Extraction and Activity Assaysmentioning

confidence: 99%

“…The inhibition of photochemical reduction of NBT (nitroblue tetrazolium) during the presence of ribo avin was used to assess SOD's activity, following the methodology outlined by [31]. The reaction mixture was composed of 0.05 M phosphate buffer (pH 7.8), 75 µM NBT, 13 mM methionine, 2 µM ribo avin, 0.1 mM EDTA, and 100 µl of enzyme extract, making a nal volume of 1.5 ml.…”

Section: Enzyme Extraction and Activity Assaysmentioning

confidence: 99%

Supplementation of mineral nutrients limits the viral RNA accumulation and systemic spread in Vigna unguiculata infected with Groundnut bud necrosis virus

Singh,

Narwal,

Mishra

et al. 2023

Preprint

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Groundnut bud necrosis virus (GBNV), a tripartite ambisense RNA genome containing plant virus fall under the family Bunyaviridae and genus Tospovirus. It infects majorly the crops belonging to the leguminosae and solanaceae families, causing characteristic symptoms such as chlorotic and necrotic spots on the leaves. Previously we reported that GBNV infection cycle induces the necrotic cell death at inoculated site and early senescence at the systemic site in the GBNV inoculated plants. In the current study we tried to understand the effect of macro and micro-nutrient supplementation on the viral disease development during GBNV infection in cowpea plant. We found that supplementation of macro-nutrients P (Phosphorus) and K (Potassium), and micro-nutrient Zn (Zinc) reduced the viral accumulation and inhibits the viral spread to systemic site. Whereas the supplementation of micro-nutrient Mn (Manganese) did not show any effect on disease development. Infected plants showed the chlorotic spots at inoculated site, those turned to necrotic as the infection progressed. Occurrence of oxidative stress was judged by the increased concentration of H2O2 (ROS) both at inoculated and systemic sites. ROS concentration was lower in the plants supplemented with nutrients P, K and Zn than the Mn supplemented and non-supplemented plants. We also we conducted a comparative analysis of antioxidants enzymes activities and examined oxidative stress responsive micro RNAs (miRNAs) along with their corresponding target transcripts in cowpea plants infected with GBNV at both inoculated and systemic sites. Antioxidants enzymes activities such as catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and ascorbate peroxidase (APX), were more induced in the plants supplemented with nutrients P, K and Zn than the supplemented with Mn and non-supplemented plants during viral infection. We also found the up-regulation of miR319 controlling metacaspase, responsible for initiation of cell death and down-regulation of miR398 and mir482 controlling COX-5b (Cytochrome C oxidase subunit 5b) and NBS-LRR genes, respectively in the plants supplemented with nutrients P, K and Zn than the supplemented with Mn and non-supplemented plants. Production of stress signals in plant induced with supplementation of macro and micro-nutrients with reduction in viral symptoms both at inoculation and systemic sites are also discussed.

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“…Plants infiltrated with empty vector (pBinAR) did not show visible lesions. NSs protein of groundnut bud necrosis virus (GBNV), previously reported to induce cell death, was used as a positive control (Singh et al, 2017). Intriguingly, Rβ-βC1/35S also caused HR-like necrosis.…”

Section: Overexpression Of βV1 Induces Hypersensitive Responsementioning

confidence: 99%

Functional characterization of a new ORF βV1 encoded by radish leaf curl betasatellite

et al. 2022

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Whitefly-transmitted begomoviruses infect and damage a wide range of food, feed, and fiber crops worldwide. Some of these viruses are associated with betasatellite molecules that are known to enhance viral pathogenesis. In this study, we investigated the function of a novel βV1 protein encoded by radish leaf curl betasatellite (RaLCB) by overexpressing the protein using potato virus X (PVX)-based virus vector in Nicotiana benthamiana. βV1 protein induced lesions on leaves, suggestive of hypersensitive response (HR), indicating cell death. The HR reaction induced by βV1 protein was accompanied by an increased accumulation of reactive oxygen species (ROS), free radicals, and HR-related transcripts. Subcellular localization through confocal microscopy revealed that βV1 protein localizes to the cellular periphery. βV1 was also found to interact with replication enhancer protein (AC3) of helper virus in the nucleus. The current findings suggest that βV1 functions as a protein elicitor and a pathogenicity determinant.

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Induction of cell death by tospoviral protein NSs and the motif critical for cell death does not control RNA silencing suppression activity

Cited by 4 publications

References 49 publications

Direct Foliar Application of dsRNA Derived From the Full-Length Gene of NSs of Groundnut Bud Necrosis Virus Limits Virus Accumulation and Symptom Expression

Direct Foliar Application of dsRNA Derived From the Full-Length Gene of NSs of Groundnut Bud Necrosis Virus Limits Virus Accumulation and Symptom Expression

Supplementation of mineral nutrients limits the viral RNA accumulation and systemic spread in Vigna unguiculata infected with Groundnut bud necrosis virus

Functional characterization of a new ORF βV1 encoded by radish leaf curl betasatellite

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