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Hongyan Chen - One of the best experts on this subject based on the ideXlab platform.
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Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector.
Frontiers in Microbiology, 2019Co-Authors: Haitao Wang, Juan Wang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan Chen, Xiao-feng ZhangAbstract:Abstract: Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their Insect Vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix cincticeps). Infection route of RYSV in the internal organs of its Insect Vector after ingestion of virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of intestinal regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as a alternative means of rapid viral dissemination in an Insect Vector.
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Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector
Frontiers Media S.A., 2019Co-Authors: Haitao Wang, Juan Wang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan ChenAbstract:Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their Insect Vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix cincticeps). The infection route of RYSV in the internal organs of its Insect Vector after ingestion of the virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of midgut regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS, then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as an alternative means of rapid viral dissemination in an Insect Vector
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Development of Continuous Cell Culture of Brown Planthopper To Trace the Early Infection Process of Oryzaviruses in Insect Vector
2016Co-Authors: Hongyan Chen, Qianzhuo Mao, Qifei Liu, Limin Zheng, Dongsheng JiaAbstract:Rice ragged stunt virus (RRSV), an oryzavirus in the family Reoviridae, is transmitted by the brown planthopper,Nilaparvata lugens, in a persistent-propagative manner. Here, we established a continuous cell line of brown planthopper to investigate the mechanism underlying the formation of the viroplasm, the putative site for viral replication and assembly, during infection of RRSV in its Insect Vector cells. Within 24 h of viral infection of cultured cells, the viroplasm had formed and contained the viral nonstructural proteins Pns6 and Pns10, known to be constituents of viroplasm. Core capsid protein P3, core particles, and newly synthesized viral RNAs were accumulated inside the viroplasm, while outer capsid protein P8 and virions were accumulated at the periphery of the viroplasm, confirming that the viroplasm induced by RRSV infection was the site for viral replication and assembly. Pns10 formed viroplasm-like inclusions in the absence of viral infection, suggesting that the viroplasmmatrix was largely composed of Pns10. Pns6 was recruited in the viroplasm by direct interaction with Pns10. Core capsid protein P3 was recruited to the viroplasm through specific association with Pns6. Knockdown of Pns6 and Pns10 expression using RNA inter-ference inhibited viroplasm formation, virion assembly, viral protein expression, and viral double-stranded RNA synthesis. Thus, the present study shows that both Pns6 and Pns10 of RRSV play important roles in the early stages of viral life cycle in its Insect Vector cells, by recruiting or retaining components necessary for viral replication and assembly. IMPORTANC
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assembly of viroplasms by viral nonstructural protein pns9 is essential for persistent infection of rice gall dwarf virus in its Insect Vector
Virus Research, 2015Co-Authors: Limin Zheng, Hongyan ChenAbstract:Abstract Rice gall dwarf virus (RGDV), a plant reovirus, is transmitted by leafhopper Vector Recilia dorsalis in a persistent-propagative manner. In a sequential study of RGDV infection of its Insect Vector, the virus initially infected the filter chamber epithelium, then directly crossed the basal lamina into the visceral muscles, from where it spread throughout the entire midgut and hindgut. Finally, RGDV spread into the salivary glands. During RGDV infection of the continuous cultured cells of R. dorsalis , viroplasm that was mainly comprised of viral nonstructural protein Pns9 was formed and acted as the site of viral replication and assembly of progeny virions. Knockdown of Pns9 expression in cultured Insect Vector cells using synthesized dsRNAs from the Pns9 gene strongly inhibited viroplasm formation and viral infection. The microinjection of dsRNAs from the Pns9 gene strongly abolished viroplasm formation in the initially infected filter chamber epithelium and prevented viral spread into leafhopper visceral muscles. These results indicated that the assembly of viroplasms was essential for the persistent infection and spread of RGDV in its Insect Vector.
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Virus-Induced Tubule: a Vehicle for Rapid Spread of Virions through Basal Lamina from Midgut Epithelium in the Insect Vector
Journal of Virology, 2014Co-Authors: Hongyan Chen, Aiming Wang, Haitao WangAbstract:The plant reoviruses, plant rhabdoviruses, tospoviruses, and tenuiviruses are transmitted by Insect Vectors in a persistent propagative manner. These viruses induce the formation of viral inclusions to facilitate viral propagation in Insect Vectors. The intestines of Insect Vectors are formed by epithelial cells that lie on the noncellular basal lamina surrounded by visceral muscle tissue. Here, we demonstrate that a recently identified plant reovirus, southern rice black-streaked dwarf virus (SRBSDV), exploits virus-containing tubules composed of virus-encoded nonstructural protein P7-1 to directly cross the basal lamina from the initially infected epithelium toward visceral muscle tissues in the intestine of its Vector, the white-backed planthopper (Sogatella furcifera). Furthermore, such tubules spread along visceral muscle tissues through a direct interaction of P7-1 and actin. The destruction of tubule assembly by RNA interference with synthesized double-stranded RNA targeting the P7-1 gene inhibited viral spread in the Insect Vector in vitro and in vivo. All these results show for the first time that a virus employs virus-induced tubule as a vehicle for viral spread from the initially infected midgut epithelium through the basal lamina, facilitating the rapid dissemination of virus from the intestine of the Insect Vector. IMPORTANCE Numerous plant viruses are transmitted in a persistent manner by sap-sucking Insects, including thrips, aphids, planthoppers, and leafhoppers. These viruses, ingested by the Insects, establish their primary infection in the intestinal epithelium of the Insect Vector. Subsequently, the invading virus manages to transverse the basal lamina, a noncellular layer lining the intestine, a barrier that may theoretically hinder viral spread. The mechanism by which plant viruses cross the basal lamina is unknown. Here, we report that a plant virus has evolved to exploit virus-induced tubules to pass through the basal lamina from the initially infected midgut epithelium of the Insect Vector, thus revealing the previously undescribed pathway adapted by the virus for rapid dissemination of virions from the intestine of the Insect Vector.
Haitao Wang - One of the best experts on this subject based on the ideXlab platform.
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Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector.
Frontiers in Microbiology, 2019Co-Authors: Haitao Wang, Juan Wang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan Chen, Xiao-feng ZhangAbstract:Abstract: Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their Insect Vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix cincticeps). Infection route of RYSV in the internal organs of its Insect Vector after ingestion of virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of intestinal regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as a alternative means of rapid viral dissemination in an Insect Vector.
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Rice Yellow Stunt Nucleorhabdovirus Matrix Protein Mediates Viral Axonal Transport in the Central Nervous System of Its Insect Vector
Frontiers Media S.A., 2019Co-Authors: Haitao Wang, Juan Wang, Qian Zhang, Tianbao Zeng, Yuemin Zheng, Hongyan ChenAbstract:Persistently transmitted plant viruses encounter multiple membrane and tissue barriers in the process of completing their infection routes within their Insect Vectors. Some of these viruses have been reported to overcome the elaborate barriers of the central nervous system (CNS) to travel through the nervous tissues, but the specific mechanisms of this process remain unknown. Here, we report the axonal transport mechanism of rice yellow stunt virus (RYSV), a nucleorhabdovirus, in the CNS of the green rice leafhopper (Nephotettix cincticeps). The infection route of RYSV in the internal organs of its Insect Vector after ingestion of the virus was investigated by immunofluorescence microscopy. RYSV was first detected in the epithelial cells of midgut regions, from where it proceeded to the nervous system, and finally into the salivary glands. We then utilized immunofluorescence and electron microscopy to investigate the distribution of RYSV particles within the leafhopper CNS, demonstrating that non-enveloped viral particles distributed along the microtubule-based neurofilaments in the axon cytoplasm following the direct interaction of leafhopper α-tubulin with the RYSV M protein. Tubulin inhibitors inhibited the dissemination of RYSV to the CNS, then into the salivary glands in leafhoppers. We therefore describe a mechanism of plant virus transport through CNS axons as an alternative means of rapid viral dissemination in an Insect Vector
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Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in Insect Vector cells.
Virology Journal, 2018Co-Authors: Haitao Wang, Juan Wang, Zhijun Fu, Xiao-feng ZhangAbstract:Background In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of Insect Vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in Insect Vector is also unclear.
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development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus rice yellow stunt virus in Insect Vector cells
Virology Journal, 2018Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng ZhangAbstract:In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of Insect Vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in Insect Vector is also unclear. In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected Insect cells. Early during RYSV infection in the Insect Vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV.
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Development of leafhopper cell culture to trace the early infection process of a nucleorhabdovirus, rice yellow stunt virus, in Insect Vector cells
BMC, 2018Co-Authors: Haitao Wang, Juan Wang, Taiyun Wei, Yunjie Xie, Xiao-feng ZhangAbstract:Abstract Background In China, the rice pathogen Rice yellow stunt virus (RYSV), a member of the genus Nucleorhabdovirus in the family Rhabdoviridae, was a severe threat to rice production during the1960s and1970s. Fundamental aspects of the biology of this virus such as protein localization and formation of the RYSV viroplasm during infection of Insect Vector cells are largely unexplored. The specific role(s) of the structural proteins nucleoprotein (N) and phosphoprotein (P) in the assembly of the viroplasm during RYSV infection in Insect Vector is also unclear. Methods In present study, we used continuous leafhopper cell culture, immunocytochemical techniques, and transmission electron microscopy to investigate the subcellular distributions of N and P during RYSV infection. Both GST pull-down assay and yeast two-hybrid assay were used to assess the in vitro interaction of N and P. The dsRNA interference assay was performed to study the functional roles of N and P in the assembly of RYSV viroplasm. Results Here we demonstrated that N and P colocalized in the nucleus of RYSV-infected Nephotettix cincticeps cell and formed viroplasm-like structures (VpLSs). The transiently expressed N and P are sufficient to form VpLSs in the Sf9 cells. In addition, the interactions of N/P, N/N and P/P were confirmed in vitro. More interestingly, the accumulation of RYSV was significantly reduced when the transcription of N gene or P gene was knocked down by dsRNA treatment. Conclusions In summary, our results suggest that N and P are the main viral factors responsible for the formation of viroplasm in RYSV-infected Insect cells. Early during RYSV infection in the Insect Vector, N and P interacted with each other in the nucleus to form viroplasm-like structures, which are essential for the infection of RYSV
Sekman Wong - One of the best experts on this subject based on the ideXlab platform.
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itraq based quantitative proteomics suggests mitophagy involvement after rice black streaked dwarf virus acquisition in Insect Vector small brown planthopper laodelphax striatellus fallen
Journal of Proteomics, 2021Co-Authors: Haoqiu Liu, Prem Prakash Das, Jianhua Zhang, Man Wang, Qingsong Lin, Yijun Zhou, Sekman WongAbstract:Abstract Plant viruses trigger numerous responses in their Insect Vectors. Using iTRAQ-based quantitative proteomics analysis, early responses of the Insect Vector, the small brown planthopper (Laodelphax striatellus Fallen, SBPH), after acquiring Rice black-streaked dwarf virus (RBSDV) at 3 days and 5 days post first access to diseased plants (padp) were revealed. A total of 582 differentially abundant proteins (DAPs) in SBPH with a fold change >1.500 or Biological significance RBSDV causes severe yield loss in rice plants. RBSDV is transmitted efficiently only through SBPH. It is important to understand how RBSDV infects SBPH in a persistent, circulative and propagative manner. However, there has been no study on the interaction between RBSDV and SBPH at the early acquisition stage using a proteomics approach. In this study, we combined iTRAQ technique and LC-MS/MS to analyze the Vector proteomics at both the initial and latent infection stages after RBSDV acquisition and verified the results by RT-qPCR. Our results revealed that significantly low DAPs were involved in various pathways, including biosynthesis of secondary metabolites, ribosomes, carbon metabolism, biosynthesis of amino acids and TCA cycle. Further clustering of the DAPs revealed significant changes in SBPH mitochondria, including decreased proteins in mitochondrial ribosomes and electron transport chain complex I, II and V. On the other hand, there was a high abundance of Parkin, suggesting the occurrence of mitochondria damage and subsequent Parkin-mediated mitophagy for clearance of impaired mitochondria. Moreover, the decreased level of PMPCB in terms of gene expression and protein abundance suggested decreased PINK1 turnover, promoting Parkin/PINK1-mediated mitophagy. Further analysis on autophagy/mitophagy-related gene transcription level indicated up-regulation of OPTN, p62/SQSTM1, TAX1BP1 and TBK1, promoting selective autophagy in SBPH after RBSDV acquisition. These findings provided new insights into the effects of RBSDV on SBPH after early acquisition by selective degradation of mitochondria, especially on reprogramming of energy metabolism and decreased mitochondria biogenesis, to prevent apoptosis and prolong the life span of SBPH post virus acquisition.
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itraq based quantitative proteomics suggests mitophagy involvement after rice black streaked dwarf virus acquisition in Insect Vector small brown planthopper laodelphax striatellus fallen
Journal of Proteomics, 2021Co-Authors: Haoqiu Liu, Prem Prakash Das, Jianhua Zhang, Man Wang, Qingsong Lin, Yijun Zhou, Sekman WongAbstract:Plant viruses trigger numerous responses in their Insect Vectors. Using iTRAQ-based quantitative proteomics analysis, early responses of the Insect Vector, the small brown planthopper (Laodelphax striatellus Fallen, SBPH), after acquiring Rice black-streaked dwarf virus (RBSDV) at 3 days and 5 days post first access to diseased plants (padp) were revealed. A total of 582 differentially abundant proteins (DAPs) in SBPH with a fold change >1.500 or <0.667 (p-value < 0.05) were identified. The proteomic analysis in SBPH at 3 days padp revealed 106 highly abundant proteins and 193 of low abundance, while 5 days padp revealed 214 highly abundant proteins and 182 of low abundance. Among them, 51 highly abundant proteins and 42 of low abundance were shown consistently at both 3 days and 5 days padp. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis mapping and Gene Ontology (GO) term classification suggested impairment of mitochondria in SBPH after RBSDV acquisition, and the 77 out of 582 differentially abundant SBPH proteins analyzed by the STRING program revealed the interaction network of the mitochondrial DAPs, showing an overall down-regulation of mitochondrial proteins including the electron transport chain proteins and mitochondrial ribosome proteins. The high abundance of Parkin at 5 days padp suggests that activation of mitophagy induced degradation of mitochondria occurred. Further verification of autophagy/mitophagy-related genes by reverse-transcription quantitative RT-PCR (RT-qPCR) in SBPH after RBSDV acquisition showed up-regulation of the autophagy receptors Optineurin (OPTN), Sequestosome-1 (SQSTM1, also known as p62) and Tax1-binding protein 1 (TAX1BP1) which targets ubiquitinated damaged mitochondria during mitophagy. The phosphorylation of the three autophagy receptors may be up-regulated through an increase of transcription level TRAF-associated NFκB activator (TANK)-binding kinase 1 (TBK1). As a result, an overall reduction in the abundance of mitochondrial proteins was observed and the selective autophagic degradation was up-regulated through increased transcription level of OPTN, p62/SQSTM1, TAX1BP1 and TBK1. Therefore, acquisition of RBSDV associated with up-regulated autophagy and selective mitochondrial degradation in SBPH suggest prevention of mitochondrial-mediated apoptosis and extension of the Vector life span. BIOLOGICAL SIGNIFICANCE: RBSDV causes severe yield loss in rice plants. RBSDV is transmitted efficiently only through SBPH. It is important to understand how RBSDV infects SBPH in a persistent, circulative and propagative manner. However, there has been no study on the interaction between RBSDV and SBPH at the early acquisition stage using a proteomics approach. In this study, we combined iTRAQ technique and LC-MS/MS to analyze the Vector proteomics at both the initial and latent infection stages after RBSDV acquisition and verified the results by RT-qPCR. Our results revealed that significantly low DAPs were involved in various pathways, including biosynthesis of secondary metabolites, ribosomes, carbon metabolism, biosynthesis of amino acids and TCA cycle. Further clustering of the DAPs revealed significant changes in SBPH mitochondria, including decreased proteins in mitochondrial ribosomes and electron transport chain complex I, II and V. On the other hand, there was a high abundance of Parkin, suggesting the occurrence of mitochondria damage and subsequent Parkin-mediated mitophagy for clearance of impaired mitochondria. Moreover, the decreased level of PMPCB in terms of gene expression and protein abundance suggested decreased PINK1 turnover, promoting Parkin/PINK1-mediated mitophagy. Further analysis on autophagy/mitophagy-related gene transcription level indicated up-regulation of OPTN, p62/SQSTM1, TAX1BP1 and TBK1, promoting selective autophagy in SBPH after RBSDV acquisition. These findings provided new insights into the effects of RBSDV on SBPH after early acquisition by selective degradation of mitochondria, especially on reprogramming of energy metabolism and decreased mitochondria biogenesis, to prevent apoptosis and prolong the life span of SBPH post virus acquisition.
Toshihiro Omura - One of the best experts on this subject based on the ideXlab platform.
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tubular structure induced by a plant virus facilitates viral spread in its Vector Insect
PLOS Pathogens, 2012Co-Authors: Qian Chen, Toshihiro Omura, Qianzhuo Mao, Lianhui Xie, Qifei Liu, Hongyan Chen, Takumi Shimizu, Tamaki Ueharaichiki, Taiyun WeiAbstract:Rice dwarf virus (RDV) replicates in and is transmitted by a leafhopper Vector in a persistent-propagative manner. Previous cytopathologic and genetic data revealed that tubular structures, constructed by the nonstructural viral protein Pns10, contain viral particles and are directly involved in the intercellular spread of RDV among cultured leafhopper cells. Here, we demonstrated that RDV exploited these virus-containing tubules to move along actin-based microvilli of the epithelial cells and muscle fibers of visceral muscle tissues in the alimentary canal, facilitating the spread of virus in the body of its Insect Vector leafhoppers. In cultured leafhopper cells, the knockdown of Pns10 expression due to RNA interference (RNAi) induced by synthesized dsRNA from Pns10 gene strongly inhibited tubule formation and prevented the spread of virus among Insect Vector cells. RNAi induced after ingestion of dsRNA from Pns10 gene strongly inhibited formation of tubules, preventing intercellular spread and transmission of the virus by the leafhopper. All these results, for the first time, show that a persistent-propagative virus exploits virus-containing tubules composed of a nonstructural viral protein to traffic along actin-based cellular protrusions, facilitating the intercellular spread of the virus in the Vector Insect. The RNAi strategy and the Insect Vector cell culture provide useful tools to investigate the molecular mechanisms enabling efficient transmission of persistent-propagative plant viruses by Vector Insects.
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Assembly of the viroplasm by viral non-structural protein Pns10 is essential for persistent infection of rice ragged stunt virus in its Insect Vector.
Journal of General Virology, 2012Co-Authors: Dongsheng Jia, Lianhui Xie, Nianmei Guo, Fusamichi Akita, Toshihiro OmuraAbstract:Rice ragged stunt virus (RRSV), an oryzavirus, is transmitted by brown planthopper in a persistent propagative manner. In this study, sequential infection of RRSV in the internal organs of its Insect Vector after ingestion of virus was investigated by immunofluorescence microscopy. RRSV was first detected in the epithelial cells of the midgut, from where it proceeded to the visceral muscles surrounding the midgut, then throughout the visceral muscles of the midgut and hindgut, and finally into the salivary glands. Viroplasms, the sites of virus replication and assembly of progeny virions, were formed in the midgut epithelium, visceral muscles and salivary glands of infected Insects and contained the non-structural protein Pns10 of RRSV, which appeared to be the major constituent of the viroplasms. Viroplasm-like structures formed in non-host Insect cells following expression of Pns10 in a baculovirus system, suggesting that the viroplasms observed in RRSV-infected cells were composed basically of Pns10. RNA interference induced by ingestion of dsRNA from the Pns10 gene of RRSV strongly inhibited such viroplasm formation, preventing efficient virus infection and spread in its Insect Vectors. These results show that Pns10 of RRSV is essential for viroplasm formation and virus replication in the Vector Insect.
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Sequential infection of Rice dwarf virus in the internal organs of its Insect Vector after ingestion of virus.
Virus research, 2011Co-Authors: Qian Chen, Toshihiro Omura, Tamaki Uehara-ichiki, Taiyun WeiAbstract:Confocal microscopy revealed that Rice dwarf virus (RDV) initially accumulated in epithelial cells of the filter chamber of leafhopper Vector Nephotettix cincticeps 2 days after acquisition access feeding on diseased plants. Subsequently, RDV accumulation progressed to the anterior midgut, and then spread to the nervous system before infection of other organs. Furthermore, RDV accumulation progressed to the visceral muscles surrounding the anterior midgut. Later, RDV accumulation was detected in other parts of the alimentary canal, salivary glands and the follicular cells of the ovarioles in viruliferous Insect Vector. Our results suggest that RDV may use the muscle or neural tissues for viral dissemination from the infected Vector's midgut into other tissues.
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Entry of Rice Dwarf Virus into Cultured Cells of Its Insect Vector Involves Clathrin-Mediated Endocytosis
Journal of Virology, 2007Co-Authors: Tamaki Ichiki-uehara, Hiroyuki Hibino, Toshihiro OmuraAbstract:Electron microscopy revealed that the entry of Rice dwarf virus (RDV) into Insect Vector cells involved endocytosis via coated pits. The treatment of cells with drugs that block receptor-mediated or clathrin-mediated endocytosis significantly reduced RDV infectivity. However, the drug that blocks caveola-mediated endocytosis had a negligible effect on such infection. Infection was also inhibited when cells had been pretreated with bafilomycin A1, which interferes with acidification of endosomes. Moreover, immunofluorescence staining indicated that the virus is internalized into early endosomes. Together, our data indicate that RDV enters Insect Vector cells through receptor-mediated, clathrin-dependent endocytosis and is sequestered in early endosomes.
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The P2 Protein of Rice Dwarf Phytoreovirus Is Required for Adsorption of the Virus to Cells of the Insect Vector
Journal of Virology, 1998Co-Authors: Toshihiro Omura, Akira Kikuchi, Boxiong Zhong, Masato Wada, Masatoshi Tomaru, Wakako Maruyama, Yasuo Watanabe, Ikuo KimuraAbstract:Intact particles of rice dwarf phytoreovirus adsorbed to and entered monolayer-cultured cells of the Insect Vector Nephotettix cincticeps and multiplied within the cells. Particles that lacked the P2 protein neither attached to nor infected such cells. Furthermore, P2-free particles obtained from a transmission-competent isolate of the virus were unable to infect Insect Vectors that had been allowed to feed on these virus particles through a membrane. However, when such virus particles were injected into Insects via a glass capillary tube they successfully infected the Insects, which became able to transmit the virus. These results support the hypothesis that, while P2-free particles can neither interact with nor infect cells in the intestinal tract of the Insect Vector, they do retain the ability to infect such cells when physically introduced into the hemolymph by injection.
Qian Chen - One of the best experts on this subject based on the ideXlab platform.
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Autophagy pathway induced by a plant virus facilitates viral spread and transmission by its Insect Vector.
PLOS Pathogens, 2017Co-Authors: Yong Chen, Manman Li, Qian Chen, Wei WuAbstract:Many viral pathogens are persistently transmitted by Insect Vectors and cause agricultural or health problems. Generally, an Insect Vector can use autophagy as an intrinsic antiviral defense mechanism against viral infection. Whether viruses can evolve to exploit autophagy to promote their transmission by Insect Vectors is still unknown. Here, we show that the autophagic process is triggered by the persistent replication of a plant reovirus, rice gall dwarf virus (RGDV) in cultured leafhopper Vector cells and in intact Insects, as demonstrated by the appearance of obvious virus-containing double-membrane autophagosomes, conversion of ATG8-I to ATG8-II and increased level of autophagic flux. Such virus-containing autophagosomes seem able to mediate nonlytic viral release from cultured cells or facilitate viral spread in the leafhopper intestine. Applying the autophagy inhibitor 3-methyladenine or silencing the expression of Atg5 significantly decrease viral spread in vitro and in vivo, whereas applying the autophagy inducer rapamycin or silencing the expression of Torc1 facilitate such viral spread. Furthermore, we find that activation of autophagy facilitates efficient viral transmission, whereas inhibiting autophagy blocks viral transmission by its Insect Vector. Together, these results indicate a plant virus can induce the formation of autophagosomes for carrying virions, thus facilitating viral spread and transmission by its Insect Vector. We believe that such a role for virus-induced autophagy is common for Vector-borne persistent viruses during their transmission by Insect Vectors.
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Tubules of plant reoviruses exploit tropomodulin to regulate actin-based tubule motility in Insect Vector.
Scientific Reports, 2017Co-Authors: Qian Chen, Linghua Zhang, Yanshuang ZhangAbstract:: Plant reoviruses are known to exploit virion-packaging tubules formed by virus-encoding non-structural proteins for viral spread in Insect Vectors. Tubules are propelled by actin-based tubule motility (ABTM) to overcome membrane or tissue barriers in Insect Vectors. To further understand which Insect factors mediate ABTM, we utilized yeast two-hybrid and bimolecular fluorescence complementation assays to test interactions between tubule protein Pns10 of rice dwarf virus (RDV), a plant reovirus, and proteins of its Insect Vector, the leafhopper Nephotettix cincticeps. Tropomodulin (Tmod), vitellogenin, and lipophorin precursor of N. cincticep displayed positive and strong interaction with Pns10, and actin-associated protein Tmod interacted with Pns10 in pull-down assay and the co-immunoprecipitation system. Further, we determined Pns10 tubules associated with Tmod in cultured cells and midgut of N. cincticep. The expression dynamic of Tmod was consistent with that of Pns10 and the fluctuation of RDV accumulation. Knockdown of Tmod inhibited the Pns10 expression and viral accumulation, thus decreasing the viruliferous rates of leafhopper. These results suggested that Tmod was involved in viral spread by directly interacting with Pns10 tubules, finally promoting RDV infection. This study provided direct evidence of plant reoviruses utilizing an actin-associated protein to manipulate ABTM in Insect Vectors, thus facilitating viral spread.
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Small interfering RNA pathway modulates persistent infection of a plant virus in its Insect Vector
Scientific Reports, 2016Co-Authors: Haitao Wang, Qian ChenAbstract:Plant reoviruses, rhabdoviruses, tospoviruses, and tenuiviruses are transmitted by Insect Vectors in a persistent-propagative manner. How such persistent infection of plant viruses in Insect Vectors is established and maintained remains poorly understood. In this study, we used rice gall dwarf virus (RGDV), a plant reovirus, and its main Vector leafhopper Recilia dorsalis as a virus–Insect system to determine how the small interference (siRNA) pathway modulates persistent infection of a plant virus in its Insect Vector. We showed that a conserved siRNA antiviral response was triggered by the persistent replication of RGDV in cultured leafhopper cells and in intact Insects, by appearance of virus-specific siRNAs, primarily 21-nt long, and the increased expression of siRNA pathway core components Dicer-2 and Argonaute-2. Silencing of Dicer-2 using RNA interference strongly suppressed production of virus-specific siRNAs, promoted viral accumulation, and caused cytopathological changes in vitro and in vivo. When the viral accumulation level rose above a certain threshold of viral genome copy (1.32 × 1014 copies/μg Insect RNA), the infection of the leafhopper by RGDV was lethal rather than persistent. Taken together, our results revealed a new finding that the siRNA pathway in Insect Vector can modulate persistent infection of plant viruses.
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nonstructural protein pns4 of rice dwarf virus is essential for viral infection in its Insect Vector
Virology Journal, 2015Co-Authors: Qian Chen, Linghua ZhangAbstract:Background Rice dwarf virus (RDV), a plant reovirus, is mainly transmitted by the green rice leafhopper, Nephotettix cincticeps, in a persistent-propagative manner. Plant reoviruses are thought to replicate and assemble within cytoplasmic structures called viroplasms. Nonstructural protein Pns4 of RDV, a phosphoprotein, is localized around the viroplasm matrix and forms minitubules in Insect Vector cells. However, the functional role of Pns4 minitubules during viral infection in Insect Vector is still unknown yet.
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nonstructural protein pns12 of rice dwarf virus is a principal regulator for viral replication and infection in its Insect Vector
Virus Research, 2015Co-Authors: Qian ChenAbstract:Abstract Plant reoviruses are thought to replicate and assemble within cytoplasmic structures called viroplasms. The molecular mechanisms underling the formation of the viroplasm during infection of rice dwarf virus (RDV), a plant reovirus, in its leafhopper Vector cells remain poorly understood. Viral nonstructural protein Pns12 forms viroplasm-like inclusions in the absence of viral infection, suggesting that the viroplasm matrix is basically composed of Pns12. Here, we demonstrated that core capsid protein P3 and nonstructural protein Pns11 were recruited in the viroplasm by direct interaction with Pns12, whereas nonstructural protein Pns6 was recruited through interaction with Pns11. The introduction of dsRNA from Pns12 gene into cultured Insect Vector cells or intact Insect strongly inhibited such viroplasm formation, preventing efficient viral spread in the leafhopper in vitro and in vivo . Thus, nonstructural protein Pns12 of RDV is a principal regulator for viral replication and infection in its Insect Vector.