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Molecular characterization and pathogenicity of an infectious clone of tomato leaf curl New Delhi virus isolate infecting Cucumis melo

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Stress Biology 2023年第1期3卷 546-554页

作者： Yuzhen Mei Lingmin Cai Yaqin Wang Fangfang Li Xiuling Yang Jinghua Yang Xueping Zhou State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou 310058ZhejiangChina State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China Laboratory of Germplasm Innovation and Molecular Breeding Institute of Vegetable ScienceZhejiang UniversityHangzhou 310058China Yazhou Bay Science and Technology City Hainan InstituteZhejiang UniversitySanya 572025China

Tomato leaf curl New Delhi virus(ToLCNDV)is a member of the genus Begomovirus,and causes devastating disease in the *** recent years,ToLCNDV was rapidly spreading in China and induces severe economic losses in *** this study,we sequenced and characterized the complete genome of ToLCNDV isolates from melon plants showing leaf curling and stunting symptoms in Jiangsu Province of *** constructed a full-length infectious cDNA clone of ToLCNDV,which could induce systemic infection with typical symptoms in Nicotiana benthamiana,Cit-rullus melo,and Citrullus lanatus plants through agrobacterium-mediated *** experimental evidence demonstrated that the virions produced in plants infected with the infectious clone of ToLCNDV are biologically active and *** also evaluated the resistance of commercial melon cultivars to ToLCNDV and found all testing melon cultivars were susceptible to ***,the reverse genetic system developed herein will facilitate further research on biological functions of proteins encoded by ToLCNDV and plant-ToLCNDV interactions,which might provide new insights into breeding resistance germplasm in crops.

关键词： Begomovirus Tomato leaf curl New Delhi virus infectious clone

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Characterization of a full-length infectious clone of bovine foamy virus 3026

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Virologica Sinica 2014年第2期29卷 94-102页

作者： Tiejun Bing Hong Yu Yue Li Lei Sun Juan Tan Yunqi Geng Wentao Qiao Key Laboratory of Molecular Microbiology and Biotechnology (Ministry of Education) and Key Laboratory of Microbial Functional Genomics (Tianjin) College of Life Sciences Nankai University Department of Molecular and Cellular Pharmacology the Vascular Biology Institute University of Miami School of Medicine National Laboratory of Biomacromolecules Center for Biological Imaging Institute of Biophysics Chinese Academy of Sciences

The biological features of most foamy viruses(FVs) are poorly understood, including bovine foamy virus(BFV). BFV strain 3026(BFV3026) was isolated from the peripheral blood mononuclear cells of an infected cow in Zhangjiakou, China. A full-length genomic clone of BFV3026 was obtained from BFV3026-infected cells, and it exhibited more than 99% amino acid(AA) homology to another BFV strain isolated in the USA. Upon transfection into fetal canine thymus cells, the full-length BFV3026 clone produced viral structural and auxiliary proteins, typical cytopathic effects, and virus particles. These results demonstrate that the full-length BFV3026 clone is fully infectious and can be used in further BFV3026 research.

关键词： bovine foamy virus infectious clone syncytium electron microscopy

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Construction Strategy and Affecting Factors of Positive-Strand RNA Virus infectious clone

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Journal of Northeast Agricultural University(English Edition) 2008年第4期15卷 80-87页

作者： SHEN Shichuan YUE Kuizhong College of Life Sciences Northeast Agricultural University Harbin 150030 China

Construction of infectious clones by full-length cDNA is basic and key for recovering RNA virus and is core of reverse *** this article,basic consideration and key technology were viewed and factors affecting infectivity of clones were also *** research advances were briefly introduced about positive-strand RNA viruses infectious ***,this article also reviewed the application of infectious clones.

关键词： positive-strand RNA virus infectious clone affecting factors application

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Molecular and biological characterization of melon-infecting squash leaf curl China virus in China

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Journal of Integrative Agriculture 2020年第2期19卷 570-577页

作者： WU Hui-jie LI Meng HONG Ni PENG Bin GU Qin-sheng Zhengzhou Fruit Research Institute Chinese Academy of Agricultural SciencesZhengzhou 450009P.R.China Huazhong Agricultural University Wuhan 430070P.R.China School of Food and Bioengineering Zhengzhou University of Light IndustryZhengzhou 450000P.R.China

It has been reported that squash leaf curl China virus(SLCCNV)infects some Cucurbitaceae crops except for melon(Cucumis melo L.).A new disease of melon exhibiting severe leaf curl and dwarfing was observed in Hainan Province of *** this study,the pathogen was identified as SLCCNV through biological and molecular *** isolate(SLCCNV-HN)possess a bipartite genome,DNA-A(HM566112.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062251.1)pumpkin and SLCCNV-Hn61(AM260205.1)squash isolates from China,whereas DNA-B(HM566113.1)with the highest nucleotide identity(99%)to SLCCNV-Hn(MF062252.1).Phylogenetic analyses based on the full-length SLCCNV-HN DNA-A and-B sequences indicated that SLCCNV-HN melon isolate is clustered with SLCCNV-Hn pumpkin,SLCCNV-Hn61 and SLCCNV-SY squash isolates from southern China,forming an independent *** clone of SLCCNV-HN was constructed and the melon plants were inoculated and the infection rate is 100%,the systemic symptoms in melon showed identical to those of melon plants infected in ***,melon plants transmission of this virus by Bemisia tabaci with a transmission rate of 95%(19/20)showed leaf curl and dwarf symptoms 15 days post transmission,thereby fulfilling Koch’s *** of genomic organization and phylogenetic trees indicated that SLCCNV-HN melon isolate belongs to the Begomovirus *** the best of our knowledge,this is the first characterization of meloninfecting SLCCNV through its genome,infectious clone and transmission.

关键词： melon squash leaf curl China virus(SLCCNV) genome infectious clone

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Complete genome sequence of two strawberry vein banding virus isolates from China

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BIOCELL 2021年第6期45卷 1695-1702页

作者： LEI JIANG SHUAI LI XIZI JIANG TONG JIANG School of Plant Protection Anhui Agricultural UniversityHefei230036China Anhui Province Key Laboratory of Integrated Pest Management on Crops Hefei230036China Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes Anhui Agricultural UniversityHefei230036China

It was rarely reported about strawberry vein banding virus(SVBV)genome sequence in China and most countries *** this work,we determined the complete genome sequences of two SVBV isolates in China,designated SVBV-AH and SVBV-BJ,that were obtained from naturally infected strawberry samples from Anhui province and Beijing city of China,*** complete genomes of SVBV-AH and SVBV-BJ were 7,862 nucleotides(nts)and 7,863 nts long,respectively,and both constituted with seven genes typical of the *** of complete nucleotide sequences showed that SVBV-AH and SVBV-BJ shared a significant nucleotide sequence identity of 97.7%of each other and had 85.7%and 86.0%sequence identity related to SVBV from the United States(SVBV-US),*** trees,based on the alignment of complete nucleotide sequences and amino acid sequences of Coat Protein(CP),both showed that SVBV-AH and SVBV-BJ clustered into one branch with all the other SVBV isolates,and other species of caulimoviruses clustered into another tree *** illustrated that all the SVBV isolates had an extremely high relationship but had a distant relationship with other species of *** further confirmed that SVBV-AH infectious clone could cause similar symptoms to SVBVinfected in strawberry under natural *** together,our study provided valuable information to elucidate the origin and dissemination of SVBV Chinese isolates,meanwhile providing the necessary vector for studying the gene functions of strawberry.

关键词： Strawberry vein banding virus Chinese isolate Complete genome Coat Protein(CP) Phylogenetic analysis infectious clone

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Infectivity of a genotype 4 hepatitis E virus cDNA clone by intrahepatic inoculation of laboratory rats

Infectivity of a genotype 4 hepatitis E virus cDNA clone by ...

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动物生态养殖、疫病防控、食品安全与人类健康——中国畜牧兽医学会家畜传染病学分会第八届全国会员代表大会暨第十五次学术研讨会

作者： Yumin Zhu Xiaoming Yu Yuanshu Zhang Yanyan Ni Fusheng Si Ruisong Yu Shijuan Dong Yaowei Huang Zhen Li Institute of Animal Science and Veterinary Medicine Shanghai Academy of Agricultural Sciences Shanghai Key Laboratory of Agricultural Genetics and Breeding College of Veterinary Medicine Nanjing Agricultural University Center for Molecular Medicine and infectious Diseases College of Veterinary Medicine Virginia Polytechnic Institute and State University

A cDNA clone of a genotype 4 swine hepatitis E virus (HEV) strain (SAAS-FX17), identified in Shanghai, has been constructed. Capped RNA transcripts were prepared in vitro and shown to be replication-competent in Huh7 cells. Sprague-Dawley (SD) rats administered the RNAtranscripts by intrahepatic inoculation developed active infections as evidenced by fecal virus shedding and sero-conversion to anti-HEV. The former was first detected between 23-30 days post-inoculation (dpi) and persisted until 45 dpi. Sera of rats inoculated with RNA transcripts became anti-HEV positive between 30 and 40 dpi, and reverted to anti-HEV negative at 52 dpi. Our data indicate for the first time that intrahepatic inoculation of rats with RNA transcripts of an HEV cDNAclone may serve as an alternative animal model for HEV research.

关键词： Hepatitis E virus (HEV) infectious clone genotype 4 rat animal model

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Development the GFP Expression Vector of Cucurbit chlorotic yellows virus

Development the GFP Expression Vector of Cucurbit chlorotic ...

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中国植物病理学会2017年学术年会

作者：施艳 Sun Xinyan Wei Ying Han Xiaoyu Chen Linlin Wang Zhenyue College of Plant Protection Henan Agricultural University

Cucurbit chlorotic yellows virus(CCYV),the recently found cucurbit-infecting crinivirus in the family Closteroviridae is among the largest single stranded positive-sense RNA *** bipartite RNA genome is comprised of 8607-nucleotide[nt](RNA1 and 8041-nt RNA2).Recent studies on CCYV were hampered by the lack of reverse genetic *** of full-length infectious cDNA clones will facilitate the investigation of viral determinants in virus replication and movement,as well as the interactions between viral proteins and host *** previous study developed two sets of full-length CCYV eDNA clones under the control of the T7 RNA polymerase and 35 S *** we constructed CCYV GFP expression vector using "add a gene" strategy according to the previous study on the closterovirus Citrus tristeza virus suggesting the "add a gene" strategy is better than ORF substitution and ORF fusion,and tested the GFP fluorescence on systemic leaves of cucumber ***-five days after agroinfiltration of pCCYVGFP together with pCBCCYVRNA1 and pCBP1/HC-Pro in Cucumis sativus,GFP fluorescence was observed using epifluorescence *** was worth noting that the GFP fluorescence was not only detectable in the leaf veins but also in the surrounding mesophyll tissues which is consistent with the previous study on the CCYV localization in leaf lamina using *** of the three different constructs,it was found that pCCYVGFPinoculated cucumber leaves showed the cell spread at 25 dpi,while the other two constructs were mainly in isolated ***,the fluorescence cell number of pCCYVGFP was significantly higher than pCCYVGFP.

关键词： CCYV infectious clone GFP expression

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