广西番茄花叶病毒和番茄褪绿病毒的分子鉴定  

Molecular Identification of Tomato Mosaic Virus and Tomato Chlorosis Virus in Guangxi

作　　者：李战彪[1] 莫翠萍 陈锦清 秦碧霞[1] 崔丽贤[1] 谢慧婷[1] 蒋雅琴 汤亚飞[3] 蔡健和[1] LI Zhanbiao;MO Cuiping;CHEN Jinqing;QIN Bixia;CUI Lixian;XIE Huiting;JIANG Yaqin;TANG Yafei;CAI Jianhe(Plant Protection Research Institute,Guangxi Academy of Agricultural Sciences/Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China,Ministry of Agriculture and Rural Affairs/Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests,Nanning 530007;Vegetable Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007;Plant Protection Research Institute,Guangdong Academy of Agricultural Sciences/Guangdong Provincial Key Laboratory of High Technology for Plant Protection,Guangzhou 510640)

机构地区：[1]广西壮族自治区农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广西作物病虫害生物学重点实验室,南宁530007 [2]广西壮族自治区农业科学院蔬菜研究所,南宁530007 [3]广东省农业科学院植物保护研究所/广东省植物保护新技术重点实验室,广州510640

出　　处：《中国农学通报》2023年第28期92-98,共7页Chinese Agricultural Science Bulletin

基　　金：广西自然科学基金项目“番茄环纹斑点病毒(TZSV)NSs蛋白的互作寄主因子筛选、鉴定及功能解析”(2020GXNSFAA259003);广西农业科学院基本科研业务专项项目“与番茄环纹斑点病毒(TZSV)NSs蛋白互作的寄主因子筛选及功能鉴定”(桂农科2020YM92);广西农业科学院基本科研业务专项项目“广西重要粮食及特色经济作物病毒病防控关键技术研究”(桂农科2021YT071);广东省植物保护新技术重点实验室开放基金项目“粤桂辣椒病毒病病原的分子鉴定及优势病毒病理学特性研究”(植重2021-06)。

摘　　要：为明确引起广西番茄呈现褪绿斑驳和花叶症状的病毒病原,采集4个疑似病毒感染的番茄样品,利用小RNA深度测序、RT-PCR、序列分析、遗传进化树的构建等方法对样品进行鉴定及遗传进化分析。研究结果发现,小RNA数据中的41条contigs分别被注释为番茄花叶病毒(tomato mosaic virus,ToMV)和番茄褪绿病毒(tomato chlorosis virus,ToCV);RT-PCR证实了s RNA的结果,且所有样品均存在ToMV和To CV的复合侵染。通过拼接获得ToMV和ToCV RNA2的近全基因序列,ToMV近全基因序列为6383 bp,命名为ToMV-GX;ToCV RNA2的近全基因序列为8031 bp,命名为ToCV-GX。进化树分析发现,本研究获得的ToMV-GX与ToMV中国山西、内蒙古、呼和浩特3个分离物处于一个分支,说明其具有较近的亲缘关系;ToCV-GX与ToCV中国台湾、广东分离物处于一个大分支,说明具有较近的亲缘关系。上述结果证实,引起广西番茄呈现褪绿斑驳和花叶症状的病原是To MV和ToCV,本研究是ToMV和ToCV复合侵染广西番茄的首次报道。To clarify the pathogens of the tomato leaves exhibiting curling,mottle and mosaic symptoms in Guangxi,four samples suspected of virus infection were collected,and small RNA deep sequencing(sRNA),reverse transcription-polymerase chain reaction(RT-PCR),sequence analysis and phylogenetic trees construction were applied for the viruses detection and genetic and evolutionary analysis.The results showed that 41 contigs from sRNA were annotated as tomato mosaic virus(ToMV)and tomato chlorosis virus(ToCV),respectively.The results of sRNA were also confirmed by RT-PCR,and all the samples were mix-infected by ToMV and ToCV.Nearly complete genome sequences of ToMV and ToCV RNA2 Guangxi isolates were obtained by multiple DNA fragments splicing.The length of the nearly complete genome sequence of ToMV was 6383 bp,named as ToMV-GX.The nearly complete genome sequence of ToCV RNA2 was 8031 bp,named as ToCV-GX.Phylogenetic analysis showed that the ToMV-GX isolate clustered with the ToMV isolates from Shanxi,Inner Mongolia and Hohhot in a big branch,indicating their close evolutionary relationship.ToCV-GX isolates clustered with the ToCV isolates from Taiwan region and Guangdong in a big branch,indicating their close evolutionary relationship.The above results showed that ToMV and ToCV were the pathogens that infected tomato showing curling,mottle and mosaic symptoms,and this was the first report of ToMV and ToCV co-infected tomato in Guangxi.

关 键 词：番茄花叶病毒 番茄褪绿病毒 广西 小RNA深度测序 遗传进化分析 

分 类 号：S432.1[农业科学—植物病理学]