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中国农学通报

中国农学通报 ›› 2023, Vol. 39 ›› Issue (28): 92-98.doi: 10.11924/j.issn.1000-6850.casb2022-0787

• 植物保护·农药 • 上一篇    下一篇

广西番茄花叶病毒和番茄褪绿病毒的分子鉴定

李战彪1(), 莫翠萍1, 陈锦清1, 秦碧霞1, 崔丽贤1, 谢慧婷1, 蒋雅琴2, 汤亚飞3(), 蔡健和1()   

  1. 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
  • 收稿日期:2022-09-13 修回日期:2023-02-01 出版日期:2023-10-05 发布日期:2023-09-25
  • 通讯作者: 蔡健和，男，1964年出生，广西藤县人，研究员，硕士，主要研究方向植物病毒学。通信地址：530007 广西壮族自治区南宁市西乡塘区大学东路174号 广西农科院植保所，Tel：0771-3244461，E-mail：caijianhe@gxaas.net。 汤亚飞，女，1980年出生，湖南桃源人，研究员，博士，主要研究方向植物病毒学。通信地址：510640 广东省广州市金颖路7号 广东省农科院植保所，Tel：020-87597476，E-mail：yf.tang1314@163.com。
  • 作者简介:

    李战彪，男，1984年出生，河南封丘人，副研究员，博士，主要研究方向植物病毒学。通信地址：530007 广西壮族自治区南宁市西乡塘区大学东路174号 广西农科院植保所，Tel：0771-3240115，E-mail：。

  • 基金资助:
    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)

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

LI Zhanbiao1(), MO Cuiping1, CHEN Jinqing1, QIN Bixia1, CUI Lixian1, XIE Huiting1, JIANG Yaqin2, TANG Yafei3(), CAI Jianhe1()   

  1. 1 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
    2 Vegetable Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007
    3 Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences/ Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Guangzhou 510640
  • Received:2022-09-13 Revised:2023-02-01 Published-:2023-10-05 Online:2023-09-25

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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sRNA的结果，且所有样品均存在ToMV和ToCV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ToMV和ToCV，本研究是ToMV和ToCV复合侵染广西番茄的首次报道。

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

Abstract:

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 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.

Key words: tomato chlorosis virus, tomato mosaic virus, Guangxi, small RNA deep sequencing, genetic and evolutionary analysis