利用小RNA深度测序技术鉴定上海地区‘红美人’柑橘病毒种类

Identification of the viruses from Hongmeiren citrus in Shanghai by small RNA sequencing

为明确上海地区‘红美人’柑橘的病毒种类,2020年4月在崇明、金山、奉贤、嘉定、闵行和浦东采集疑似病毒感染的叶片样品11份,采用小RNA深度测序技术结合RT-PCR对不同来源的混合样品进行病毒种类鉴定。结果表明,样品中含有柑橘衰退病毒Citrus tristeza virus(CTV)、柑橘黄化脉明病毒Citrus yellow vein clearing virus(CYVCV)、柑橘树皮裂纹类病毒Citrus bark cracking viroid(CBCVd)和柑橘类病毒ⅤCitrus viroid V(CVd-Ⅴ)。同时发现,上海地区‘红美人’柑橘病毒复合侵染现象普遍。基于CTV和CYVCV的CP全长基因及CBCVd和CVd-Ⅴ全基因组序列系统进化分析结果表明:除了CYVCV的分离物与地理来源具有明显的相关性外,其余病毒及类病毒均没有严格的地域相关性。研究结果为开展‘红美人’柑橘种苗病毒检测及病毒病的防治奠定基础。

To identify the viruses and viroids from‘Hongmeiren’citrus in Shanghai,11 citrus leaf samples with the virus-like symptoms were collected from the districts of Chongming,Jinshan,Fengxian,Jiading,Minhang and Pudong in April,2020.The samples were analyzed by small RNA deep sequencing.The results showed that Citrus tristeza virus(CTV),Citrus yellow vein clearing virus(CYVCV),Citrus bark cracking viroid(CBCVd)and Citrus viroid V(CVd-Ⅴ)were detected in the samples by small RNA sequencing,which were confirmed by RT-PCR.The mixed infection of viruses/viroids was common in‘Hongmeiren’citrus.Phylogenetic analysis was conducted based on the full-length CP gene sequences of CTV and CYVCV,and the full-length genome sequences of CBCVd and CVd-Ⅴ,respectively.The results revealed that only CYVCV showed an obvious correlation with geographical origin.The study laid a foundation for the development of viral diagnosis and prevention in‘Hongmeiren’citrus seedlings.