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Interaction between rice stripe virus Disease- Specific Protein and Host PsbP Enhances virus Symptoms

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Molecular Plant 2014年第4期7卷 691-708页

作者： Lingfang Kong Jianxiang Wu Lina Lu Yi Xu Xueping Zhou State Key Laboratory of rice Biology Institute of Biotechnology Zhejiang University Hangzhou 310058 People's Republic of China

rice stripe virus （RSV） causes severe diseases in Oryza sativa （rice） in many Eastern Asian countries. Diseasespecific protein （SP） of RSV is a non-structural protein and its accumulation level in rice plant was shown to determine the severity of RSV symptoms. Here, we present evidence that expression of RSV SP alone in rice or Nicotiana benthamiana did not produce visible symptoms. Expression of SP in these two plants, however, enhanced RSV- or Potato virus X （PVX）- induced symptoms. Through yeast two-hybrid screening, GST pull-down, and bimolecular fluorescence complementation assays, we demonstrated that RSV SP interacted with PsbP, a 23-kDa oxygen-evolving complex protein, in both rice and N. benthamiana. Furthermore, our investigation showed that silencing of PsbP expression in both plants increased disease symptom severity and virus accumulation. Confocal microscopy using N, benthamiana protoplast showed that PsbP accu- mulated predominantly in chloroplast in wild-type N. benthamiana cells. In the presence of RSV SP, most PsbP was recruited into cytoplasm of the assayed cells. In addition, accumulation of SP during RSV infection resulted in alterations of chloroplast structure and function. Our findings shed light on the molecular mechanism underlying RSV disease symptom development.

关键词： rice stripe virus disease-specific protein interaction PsbP symptom.

Laodelphax striatellus Atg8 facilitates rice stripe virus infection in an autophagy-independent manner

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Insect Science 2021年第2期28卷 315-329页

作者： Yuan-Ling Yu Meng-Ting Zhang Yan Huo Ji-Liang Tang Qing Liu Xiao-Ying Chen Rong-Xiang Fang Li-Li Zhang State Key Laboratory of Plant Genomics Institute of MicrobiologyChinese Academy of ScienceBeijingChina CAS Center for Excellence in Biotic Interactions University of the Chinese Academy of SciencesBeijingChina State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources Guangxi UniversityNanningChina National Plant Gene Research Center BeijingChina

rice stripe virus(RSV)is the causative agent of rice stripe disease and is completely dependent on insect vectors for its plant-to-plant *** striatellus is the major insect vector for *** this study,we explored the interactions be-tween RSV infection and *** autophagy,a potential intrinsic antiviral mechanism in *** found that *** autophagic activity did not affect RSV infection;however,the autophagy related-8(Atg8)gene significantly enhanced virus ***-ing RSV initial infection within the *** midgut,silencing of Atg8 expression significantly decreased the phosphorylation of c-Jun N-terminal kinase(p-JNK);however,when RSV infection is absent,silencing of Atg8 did not alter p-JNK *** results indicated that Atg8 might activate the JNK machinery by allowing more virus infection into *** further revealed that Atg8-deficiency significantly decreased RSV accumu-lation on the surface of the insect midgut epithelial cells,suggesting a receptor trafficking function of the y-aminobutyric acid receptor-associated protein *** the RSV ovary entry as a model,in which vitellogenin receptor(V gR)mediates RSV cell entry,we clarified that Atg8-deficiency decreased the abundance of V gR localizing on the cytomem-brane and disturbed the attachment of RSV in the germarium ***,these results revealed an autophagy-independent function of *** Atg8 that enhances RSV initial infection by increasing virus attachment on the infection sites.

关键词： Atg8 insect autophagy Laodelphax striatellus receptor trafficking rice stripe virus viral initial infection

Genetic variability and evolution of rice stripe virus

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2013年第10期14卷 875-885页

作者： Ling-zhe HUANG Li-xia RAO Xue-ping ZHOU Jian-xiang WU State Key Laboratory of rice Biology Institute of Biotechnology Zhejiang University

rice stripe virus(RSV) is the type member of the genus *** is known to have four segmented,single-stranded RNA molecules and causes rice stripe disease in the rice fields of China,Japan,and *** on the complete genomic sequences of the determined 6 RSV isolates(from Yunnan,Jiangsu,Zhejiang,and Liaoning Provinces,China) and 27 other RSV isolates(from Yunnan,Jiangsu,Anhui,Henan,and Shandong Provinces of China,also Japan and Korea) downloaded from GenBank,we provided a genotyping profile of RSV field isolates and described the population structure of *** RSV isolates,except isolate CX,could be divided into two subtypes,one including 6 isolates from Yunnan Province,and the other including 26 isolates from different parts of China,Japan,and Korea,which were referred to as subtype II and subtype I,*** amino acid distances between subtypes range from 0.053 to *** isolates in Yunnan Province were genetically differentiated from other parts of China,Japan,and Korea and showed infrequent gene *** RSV populations collected from other parts of China,Japan,and Korea were only composed of subtype I and showed very low genetic *** speculated that isolate CX may be the result of recombination of isolates from two *** potential recombination events were detected in RNA4 of isolate CX.

关键词： rice stripe virus Genetic variability Genetic evolution

Surface Display of rice stripe virus NSvc2 and Analysis of Its Membrane Fusion Activity

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Virologica Sinica 2012年第2期27卷 100-108页

作者： Shu-ling Zhao Xue-juan Dai Jian-sheng Liang Chang-yong Liang College of Bioscience and Biotechnology Yangzhou UniversityYangzhou 225009China

rice stripe virus (RSV) infects rice and is transmitted in a propagative manner by the small brown planthopper. How RSV enters an insect cell to initiate the infection cycle is poorly understood. Sequence analysis revealed that the RSV NSvc2 protein was similar to the membrane glycoproteins of several members in the family Bunyaviridae and might induce cell membrane fusion. To conveniently study the membrane fusion activity of NSvc2, we constructed cell surface display vectors for expressing Nsvc2 on the insect cell surface as the membrane glycoproteins of the enveloped viruses. Our results showed that NSvc2 was successfully expressed and displayed on the surface of insect Sf9 cells. When induced by low pH, the membrane fusion was not observed in the cells that expressed NSvc2. Additionally, the membrane fusion was also not detected when co-expressing Nsvc2 and the viral capsid protein on insect cell surface. Thus, RSV NSvc2 is probably different from the phlebovirus counterparts, which could suggest different functions. RSV might enter insect cells other than by fusion with plasma or endosome membrane.

关键词： rice stripe virus NSvc2 Surface display Membrane fusion

Development of a colloidal gold-based immunochromatographic strip for rapid detection of rice stripe virus

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2019年第4期20卷 343-354页

作者： De-qing HUANG Rui CHEN Ya-qin WANG Jian HONG Xue-ping ZHOU Jian-xiang WU State Key Laboratory of rice Biology Institute of Biotechnology Zhejiang University State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural Sciences

rice stripe virus(RSV) causes dramatic losses in rice production worldwide. In this study, two monoclonal antibodies(MAbs) 16E6 and 11 C1 against RSV and a colloidal gold-based immunochromatographic strip were developed for specific, sensitive, and rapid detection of RSV in rice plant and planthopper samples. The MAb 16E6 was conjugated with colloidal gold and the MAb 11C1 was coated on the test line of the nitrocellulose membrane of the test strip. The specificity of the test strip was confirmed by a positive reaction to RSV-infected rice plants and small brown planthopper(SBPH), and negative reactions to five other rice viruses, healthy rice plants, four other vectors of five rice viruses, and non-viruliferous SBPH. Sensitivity analyses showed that the test strip could detect the virus in RSV-infected rice plant tissue crude extracts diluted to 1:20 480(w/v, g/mL), and in individual viruliferous SBPH homogenate diluted to 1:2560(individual SPBH/μL). The validity of the developed strip was further confirmed by tests using field-collected rice and SBPH samples. This newly developed test strip is a low-cost, fast, and easy-to-use tool for on-site detection of RSV infection during field epidemiological studies and paddy field surveys, and thus can benefit decision-making for RSV management in the field.

关键词： Monoclonal antibody Colloidal gold-based immunochromatographic strip rice stripe virus rice Small brown planthopper

rice stripe virus NS2 Protein Interacts with SGS3 Protein of rice

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Rice Stripe Virus NS2 Protein Interacts with SGS3 Protein of...

Screening of rice stripe virus(RSV)-resistant Germplasm from Landraces of rice(Oryza sativa L.)

中国植物病理学会第九届全国会员代表大会暨2010年学术年会

作者： XIAO Dong-lai,WEI Tai-yun,WU Zu-jian~*,XIE Lian-hui (Key Laboratry of Plant Virology of Fujian Province,Fuzhou 350002,Fujian)

Yeast two-hybrid assay was used to screen the rice cDNA library using rice stripe virus(RSV) NS2 protein as a bait.A novel rice protein interacting with RSV NS2 was obtained, which was homologous to Arabidopsis thaliana SGS3(AtSGS3) protein and was designated as *** full-length OsSGS3 gene was amplified by RT-PCR and cloned into yeast expression vector *** two-hybrid assay indicated that NS2 could interact with full-length ***3 possessed the three domains typical of AtSGS3:zinc finger,XS and coiled *** OsSGS3 deletion mutants:OsSGS3-l,which contained the znic finger domain, OsSGS3-2,which contained the XS domain,and OsSGS3-3,which contained the coiled coil domain were *** two-hybrid assay indicated that NS2 could interact with full-length OsSGS3,OsSGS3-l,OsSGS3-2 and *** localization studies indicated that OsSGS3-YFP accumulated in cytoplasmic foci that perfectly resembled those of AtSGS3 and NS2-CFP accumulated in cell wall and nuclei in Nicotiana benthamiana *** confirmed the interaction between OsSGS3 and NS2 by using co-IP,BiFC,and *** results of colocalization revealed that a considerable proportion of OsSGS3 was redistributed to nucleus by *** speculated that the redistribution of OsSGS3 by NS2 might disrupt the normal function of ***3 has been demonstrated to be involved in small RNA *** explore the possible biological implications of the interaction of RSV NS2 with OsSGS3,we first detected the expression patterns of 5 auxin responsive factors(ARFs) that had been domonstrated to be potential targets of trans acting *** results indicated that the overall accumulation levels of the 5 genes increased in RSV infected rice.

关键词： rice stripe virus NS2 yeast two hybrid Bimolecular fluorescence complementation(BiFC)

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Agricultural Science & Technology 2014年第5期15卷 809-813页

作者：虞利俊范方军杨杰王军朱金燕李文奇唐玉邦徐磊仲维功江苏省农业科学院农业设施与装备研究所江苏南京210014 江苏省农业科学院粮食作物研究所江苏南京210014

[Objective] The aim of this study was to screen rice strip virus （RSV）-resistant landraces. [Method] The resistance of 119 rice landraces to rice stripe virus was identified in field spontaneously infected with smal plant-hopper. [Result] There were 55 landraces resistant to rice strip disease in 56 indica rice landraces, but on-ly two resistant to rice strip disease in 63 japonica rice landraces. [Conclusion] The results revealed that there were abundant rice landscapes resistant to RSV in Chi-na, and these varieties can be used to develop more genes resistant to RSV.

关键词： rice landrace rice stripe virus Resistance identification

rice St#pe virus Interferes with S-acylation of Remorin and Induces Its Autophagic Degradation to Facilitate virus Infection

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Molecular Plant 2018年第2期11卷 269-287页

作者： Shuai Fu Yi Xu Chenyang Li Yi Li dianxiang Wu Xueping Zhou State Key Laboratory of rice Biology Institute of Biotechnology Zhejiang University Hangzhou 310058 China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100193 China State Key Laboratory of Protein and Plant Gene Research College of Life Sciences Peking University Beijing 100871 China Section of Plant Pathology and Plant-Microbe Biology School of Integrated Plant Science Cornell University Ithaca NY USA

Remorins are plant-specific membrane-associated proteins and were proposed to play crucial roles in plant-pathogen interactions. However, little is known about how pathogens counter remorin-mediated host responses. In this study, by quantitative whole-proteome analysis we found that the remorin protein （NbREM1） is downregulated early in rice stripe virus （RSV） infection. We further discovered that the turn- over of NbREM1 is regulated by S-acylation modification and its degradation is mediated mainly through the autophagy pathway. Interestingly, RSV can interfere with the S-acylation of NbREM1, which is required to negatively regulate RSV infection by restricting virus cell-to-cell trafficking. The disruption of NbREM1 S-acylation affects its targeting to the plasma membrane microdomain, and the resulting accumulation of non-targeted NbREM1 is subjected to autophagic degradation, causing downregulation of NbREMI. Moreover, we found that RSV-encoded movement protein, NSvc4, alone can interfere with NbREM1 S-acylation through binding with the C-terminal domain of NbREM1 the S-acylation of OsREM1.4, the homologous remorin of NbREM1, and thus remorin-mediated defense against RSV in rice, the original host of RSV, indicating that downregulation of the remorin protein level by interfering with its S-acylation is a common strategy adopted by RSV to overcome remorin-mediated inhibition of virus movement.

关键词： remorins S-acylation autophagy rice stripe virus movement

Dimeric artificial microRNAs mediate highly efficient RSV and RBSDV resistance in transgenic rice plants

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Dimeric artificial microRNAs mediate highly efficient RSV an...