The 5′ Noncoding Region of Grapevine Chrome Mosaic Nepovirus RNA-2 Triggers a Necrotic Response on Three Nicotiana spp

The 5′ noncoding region (NCR) of grapevine chrome mosaic nepovirus (GCMV) was cloned in a viral vector derived from potato virus X (PVX). The recombinant virus obtained was inoculated to Nicotiana benthamiana, N. clevelandii, and N. tabacum plants. Infected plants developed necrotic symptoms in place of the vein clearing and mosaic typically observed after inoculation with PVX. Northern (RNA) blot analysis showed that the replication of PVX was not specifically altered by the presence of the GCMV 5′ NCR. Inoculation of recombinant PVX harboring deleted forms of the GCMV 5′ NCR showed that the three stem-loop structures at the 3′ end of the 5′ NCR (nucleotides 153 to 206) are dispensable for the induction of necrosis. Further deletion analysis indicated that neither the 5′-most 70 nucleotides of the 5′ NCR nor the downstream region (nucleotides 71 to 217) alone is able to induce the necrotic symptoms. In the presence of both the sequence encoding the GCMV coat protein and the GCMV 3′ NCR, the GCMV 5′ NCR failed to induce necrosis in the PVX background. The mechanisms by which the expression of the 5′ NCR might modify PVX symptoms are discussed.

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The ORF1 Products of Tombusviruses Play a Crucial Role in Lethal Necrosis of Virus-Infected Plants

Journal of Virology ◽

10.1128/jvi.74.23.10873-10881.2000 ◽

2000 ◽

Vol 74 (23) ◽

pp. 10873-10881 ◽

Cited By ~ 35

Author(s):

József Burgyán ◽

Csaba Hornyik ◽

György Szittya ◽

Dániel Silhavy ◽

György Bisztray

Keyword(s):

Transgenic Plants ◽

Viral Replication ◽

Potato Virus ◽

Nicotiana Benthamiana ◽

Crucial Role ◽

Viral Vector ◽

Potato Virus X ◽

Wild Type ◽

Systemic Necrosis

ABSTRACT Hybrids of cymbidium ringspot (CymRSV) and carnation Italian ringspot (CIRV) tombusviruses were used to identify viral symptom determinants responsible for the generalized necrosis in tombusvirus-infected plants. Surprisingly, symptoms of Nicotiana benthamiana infected with CymRSV/CIRV hybrids were distinctly different. It was demonstrated that not all chimeras expressing wild-type (wt) levels of p19 protein caused systemic necrosis as both parents CymRSV and CIRV did. We showed here that hybrids containing chimeric ORF1 were not able to induce lethal necrosis even if the viral replication of these constructs was not altered significantly. However, if a wt p33 (product of ORF1) of CymRSV was provided intrans in transgenic plants expressing p33 and its readthrough product p92, the lethal necrosis characteristic to tombusvirus infection was restored. In addition, the expression of p33 by a potato virus X viral vector in N. benthamiana caused severe chlorosis and occasionally necrosis, indicating the importance of p33 in wt symptoms of tombusviruses. Thus, our results provide evidence that elicitation of the necrotic phenotype requires the presence of the wt p33 in addition to the p19 protein of tombusviruses.

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Corrigendum to “Nicotiana benthamiana protein, NPCIP1, interacting with Potato virus X coat protein plays a role as susceptible factor for viral infection” [Virology 386 (2009) 257–269]

Virology ◽

10.1016/j.virol.2009.06.019 ◽

2009 ◽

Vol 391 (1) ◽

pp. 149-150

Author(s):

Mi-Ri Park ◽

Sang-Ho Park ◽

Sang-Yun Cho ◽

Kook-Hyung Kim

Keyword(s):

Coat Protein ◽

Viral Infection ◽

Potato Virus ◽

Nicotiana Benthamiana ◽

Potato Virus X

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NSvc4 Encoded by Rice Stripe Virus Targets Host Chloroplasts to Suppress Chloroplast-Mediated Defense

Viruses ◽

10.3390/v14010036 ◽

2021 ◽

Vol 14 (1) ◽

pp. 36

Author(s):

Zongdi Li ◽

Chenyang Li ◽

Shuai Fu ◽

Yu Liu ◽

Yi Xu ◽

...

Keyword(s):

Nicotiana Benthamiana ◽

Actin Filaments ◽

Movement Protein ◽

Infectious Clone ◽

Rice Stripe Virus ◽

Potato Virus X ◽

Host Factors ◽

Ros Production ◽

Rsv Infection ◽

Detailed Function

Our previous research found that NSvc4, the movement protein of rice stripe virus (RSV), could localize to the actin filaments, endoplasmic reticulum, plasmodesmata, and chloroplast, but the roles of NSvc4 played in the chloroplast were opaque. Here, we confirm the accumulation of NSvc4 in the chloroplasts and the N-terminal 1–73 amino acids of NSvc4 are sufficient to localize to chloroplasts. We provide evidence to show that chloroplast-localized NSvc4 can impair the chloroplast-mediated immunity. Expressing NSvc4 in Nicotiana benthamiana leaves results in the decreased expression of defense-related genes NbPR1, NbPR2, and NbWRKY12 and the inhibition of chloroplast-derived ROS production. In addition, generation of an infectious clone of potato virus X (PVX) carrying NSvc4 facilitates PVX infection in N. benthamiana plants. Moreover, we identify two chloroplast-related host factors, named NbGAPDH-A and NbPsbQ1, both of which can interact with NSvc4. Knockdown of NbGAPDH-A or NbPsbQ1 can both promote RSV infection. Our results decipher a detailed function of NSvc4 in the chloroplast.

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Identification of a Movement Protein of the Tenuivirus Rice Stripe Virus

Journal of Virology ◽

10.1128/jvi.01696-08 ◽

2008 ◽

Vol 82 (24) ◽

pp. 12304-12311 ◽

Cited By ~ 93

Author(s):

Ruyi Xiong ◽

Jianxiang Wu ◽

Yijun Zhou ◽

Xueping Zhou

Keyword(s):

Nicotiana Benthamiana ◽

Fluorescent Protein ◽

Severe Disease ◽

Rice Stripe Virus ◽

Potato Virus X ◽

Infected Leaf ◽

Biolistic Bombardment ◽

Movement Proteins ◽

Green Fluorescent

ABSTRACT Rice stripe virus (RSV) is the type member of the genus Tenuivirus. RSV has four single-stranded RNAs and causes severe disease in rice fields in different parts of China. To date, no reports have described how RSV spreads within host plants or the viral and/or host factor(s) required for tenuivirus movement. We investigated functions of six RSV-encoded proteins using trans-complementation experiments and biolistic bombardment. We demonstrate that NSvc4, encoded by RSV RNA4, supports the intercellular trafficking of a movement-deficient Potato virus X in Nicotiana benthamiana leaves. We also determined that upon biolistic bombardment or agroinfiltration, NSvc4:enhanced green fluorescent protein (eGFP) fusion proteins localize predominantly near or within the walls of onion and tobacco epidermal cells. In addition, the NSvc4:eGFP fusion protein can move from initially bombarded cells to neighboring cells in Nicotiana benthamiana leaves. Immunocytochemistry using tissue sections from RSV-infected rice leaves and an RSV NSvc4-specific antibody showed that the NSvc4 protein accumulated in walls of RSV-infected leaf cells. Gel retardation assays revealed that the NSvc4 protein interacts with single-stranded RNA in vitro, a common feature of many reported plant viral movement proteins (MPs). RSV NSvc4 failed to interact with the RSV nucleocapsid protein using yeast two-hybrid assays. Taken together, our data indicate that RSV NSvc4 is likely an MP of the virus. This is the first report describing a tenuivirus MP.

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EF-G–induced ribosome sliding along the noncoding mRNA

Science Advances ◽

10.1126/sciadv.aaw9049 ◽

2019 ◽

Vol 5 (6) ◽

pp. eaaw9049 ◽

Cited By ~ 5

Author(s):

M. Klimova ◽

T. Senyushkina ◽

E. Samatova ◽

B. Z. Peng ◽

M. Pearson ◽

...

Keyword(s):

Messenger Rna ◽

Elongation Factor ◽

Transfer Rna ◽

Noncoding Region ◽

Forward Movement ◽

Stem Loop ◽

A Site ◽

Hydrolysis Of ◽

Reading Frames ◽

Factor G

Translational bypassing is a recoding event during which ribosomes slide over a noncoding region of the messenger RNA (mRNA) to synthesize one protein from two discontinuous reading frames. Structures in the mRNA orchestrate forward movement of the ribosome, but what causes ribosomes to start sliding remains unclear. Here, we show that elongation factor G (EF-G) triggers ribosome take-off by a pseudotranslocation event using a small mRNA stem-loop as an A-site transfer RNA mimic and requires hydrolysis of about two molecules of guanosine 5′-triphosphate per nucleotide of the noncoding gap. Bypassing ribosomes adopt a hyper-rotated conformation, also observed with ribosomes stalled by the SecM sequence, suggesting common ribosome dynamics during translation stalling. Our results demonstrate a new function of EF-G in promoting ribosome sliding along the mRNA, in contrast to codon-wise ribosome movement during canonical translation, and suggest a mechanism by which ribosomes could traverse untranslated parts of mRNAs.

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Interplay between potato virus X and RNA granules in Nicotiana benthamiana

Virus Research ◽

10.1016/j.virusres.2019.197823 ◽

2020 ◽

Vol 276 ◽

pp. 197823

Author(s):

Gabriel Robles-Luna ◽

Nicolás Furman ◽

María Florencia Barbarich ◽

Nicolás Carlotto ◽

Alejandra Attorresi ◽

...

Keyword(s):

Potato Virus ◽

Nicotiana Benthamiana ◽

Potato Virus X ◽

Rna Granules

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Heterologous expression of biologically active Mambalgin-1 peptide as a new potential anticancer, using a PVX-based viral vector in Nicotiana benthamiana

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-020-01838-x ◽

2020 ◽

Vol 142 (2) ◽

pp. 241-251 ◽

Cited By ~ 1

Author(s):

Ghaffar Khezri ◽

Bahram Baghban Kohneh Rouz ◽

Hamideh Ofoghi ◽

Seyed Javad Davarpanah

Keyword(s):

Heterologous Expression ◽

Nicotiana Benthamiana ◽

Viral Vector ◽

Biologically Active

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Phenotypes and Functional Effects Caused by Various Viral RNA Silencing Suppressors in Transgenic Nicotiana benthamiana and N. tabacum

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-21-2-0178 ◽

2008 ◽

Vol 21 (2) ◽

pp. 178-187 ◽

Cited By ~ 50

Author(s):

Shahid Aslam Siddiqui ◽

Cecilia Sarmiento ◽

Erkki Truve ◽

Harry Lehto ◽

Kirsi Lehto

Keyword(s):

Mosaic Virus ◽

Potato Virus ◽

Nicotiana Benthamiana ◽

Rna Silencing ◽

Fluorescent Protein ◽

Potato Virus X ◽

Mottle Virus ◽

African Cassava Mosaic Virus ◽

Rice Yellow Mottle Virus ◽

Green Fluorescent

RNA silencing suppressor genes derived from six virus genera were transformed into Nicotiana benthamiana and N. tabacum plants. These suppressors were P1 of Rice yellow mottle virus (RYMV), P1 of Cocksfoot mottle virus, P19 of Tomato bushy stunt virus, P25 of Potato virus X, HcPro of Potato virus Y (strain N), 2b of Cucumber mosaic virus (strain Kin), and AC2 of African cassava mosaic virus (ACMV). HcPro caused the most severe phenotypes in both Nicotiana spp. AC2 also produced severe effects in N. tabacum but a much milder phenotype in N. benthamiana, although both HcPro and AC2 affected the leaf tissues of the two Nicotiana spp. in similar ways, causing hyperplasia and hypoplasia, respectively. P1-RYMV caused high lethality in the N. benthamiana plants but only mild effects in the N. tabacum plants. Phenotypic alterations produced by the other transgenes were minor in both species. Interestingly, the suppressors had very different effects on crucifer-infecting Tobamovirus (crTMV) infections. AC2 enhanced both spread and brightness of the crTMV-green fluorescent protein (GFP) lesions, whereas 2b and both P1 suppressors enhanced spread but not brightness of these lesions. P19 promoted spread of the infection into new foci within the infiltrated leaf, whereas HcPro and P25 suppressed the spread of crTMV-GFP lesions.

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