辽宁省番茄上番茄斑驳花叶病毒的分子鉴定  

Molecular identification of tomato mottle mosaic virus in tomatoes from Liaoning province

作　　者：涂丽琴 吴淑华[1] 高丹娜 吉颖 程兆榜[1] 周益军[1] 朱月林[2] 刘勇[3] 季英华[1] TU Li-qin;WU Shu-hua;GAO Dan-na;JI Ying;CHENG Zhao-bang;ZHOU Yi-jun;ZHU Yue-lin;LIU Yong;JI Ying-hua(Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China;Institute of Plant Protection,Hunan Academy of Agricultural Sciences,Changsha 4110125,China)

机构地区：[1]江苏省农业科学院植物保护研究所,江苏南京210014 [2]南京农业大学园艺学院,江苏南京210095 [3]湖南省农业科学院植物保护研究所,湖南长沙410125

出　　处：《江苏农业学报》2020年第5期1126-1132,共7页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2018YFD0201208);国家自然科学基金项目(31572074、31770168);国家特色蔬菜产业技术体系项目(CARS-24-C-01);江苏省农业自主创新基金项目[CX(18)2005]

摘　　要：为明确2015年在辽宁省调查番茄病毒病时发现的病毒类型,对田间采集的番茄病样进行了分子检测。结果显示,在用烟草花叶病毒属通用检测引物进行RT-PCR扩增时22份样品中有6份扩增到800 bp左右的目的片段,扩增产物测序后BLAST分析结果表明它与已报道的番茄斑驳花叶病毒(Tomato mottle mosaic virus,ToMMV)(KF477193)同源性最高,为99.2%。为进一步确认,针对ToMMV编码的外壳蛋白(CP)设计了一对特异性引物,对样品进行RT-PCR检测,结果显示6份阳性样品都能扩增到目的条带,对其克隆测序后发现CP基因序列全长480 bp,编码1个相对分子质量约1.77×104的蛋白质,系统进化树表明该病毒与ToMMV的同源性最高,共同聚类于一个分支。这些结果表明在辽宁省番茄病株上检测到的病毒为ToMMV的1个分离物,这是在辽宁省番茄上发现番茄斑驳花叶病毒的首次报道。To clarify the types of viruses found in the process of investigation on tomato virus diseases from Liaoning province in 2015,the infected tomato samples collected in the fields were detected by molecular methods.The results showed that the target fragment of approximately 800 bp long was amplified from six samples of twenty-two samples in the process of RT-PCR amplification,using universal primers for Tobamovirus.The amplified products were sequenced and the results of BLAST analysis showed that,the sequence showed the highest homology with the reported tomato mottle mosaic virus(ToMMV)(KF477193),with a value of 99.2%.To make further confirmation of the above result,a pair of specific primers based on coat protein(CP)sequence coded by ToMMV were designed and used to do RT-PCR detection of the samples.The results showed that the target band could be amplified from six positive samples.The cloning and sequencing results of the target band revealed that the full length of CP gene was 480 bp,encoding a protein with a relative molecular weight of approximately 1.77×104.The results of the phylogenetic tree indicated that the virus sequence showed the highest homology with ToMMV and they all clustered into one branch.The above results indicate that the virus detected in infected tomatoes from Liaoning is an isolate of ToMMV,and this is the first report of ToMMV found in tomatoes from Liaoning province.

关 键 词：番茄斑驳花叶病毒 分子鉴定 番茄 

分 类 号：S436.412.1[农业科学—农业昆虫与害虫防治]