地黄花叶病毒河南分离物检测鉴定及其MP序列分析  

Detection and identification of Rehmannia glutinosa mosaic virus Henan isolate and its MP sequence analysis

作　　者：郭枭 冯陈尉 庄新建 丁诗文 董卓倬 贺振[1] 张坤[1] GUO Xiao;FENGCHEN Wei;ZHUANG Xinjian;DING Shiwen;DONG Zhuozhuo;HE Zhen;ZHANG Kun(College of Horticulture and Plant Protection,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学园艺与植物保护学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2022年第3期102-107,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(31801699);江苏省自然科学基金青年基金项目(BK20180904);广东省植物保护新技术重点实验室开放课题(植重2021-07);中央本级重大增减支项目(2060302-1904-11)。

摘　　要：2019年夏,在河南温县中草药园内发现种植的地黄呈现疑似病毒侵染所表现出的褪绿、花叶等症状,采集该样品,用Trizol法提取地黄样品总RNA,分离其siRNA,并进行文库构建和siRNA高通量测序。将测序结果经接头去除、拼接组装后(Velvet Software 0.7.31,k=17),与NCBI数据库中序列进行核酸比对分析,发现该样品中存在着地黄花叶病毒(Rehmannia mosaic virus,ReMV)和瓜类褪绿黄化病毒(cucurbit chlorotic yellows virus,CCYV)的部分片段。通过RT-PCR确认该样品存在ReMV的侵染,使用商业化的ELISA试剂盒进一步确认该样品被ReMV侵染过。进一步克隆ReMV河南分离物的MP序列,并结合NCBI数据库中发布的该病毒MP序列,构建了基于MP序列的ReMV河南分离物系统发育树。该研究丰富了ReMV群体的传播范围及其基因组遗传信息,为后续ReMV的深入研究奠定了基础。In the summer of 2019,the Rehmannia glutinosa leaves exhibited chlorosis and mosaic symptoms that suspected of virus infection in the Chinese herbal garden located in County Wen,Henan Province.The symptomatic samples were collected and performed total RNAs extraction by Trizol reagent.The siRNA was isolated and used for siRNA library construction.Reads obtained by high-through sequencing were further trimmed and assembled by software(Velvet Software 0.7.31,k=17),then performed blastN in NCBI database.The results showed that the sample contained partial fragments from Rehmannia mosaic virus(ReMV)and cucurbit chlorotic yellows virus(CCYV).RT-PCR and commercial ELISA kit confirmed the infection of ReMV in the collected sample.The present study cloned the movement protein gene(MP)sequence of ReMV Henan isolate,and constructed a phylogenetic tree of ReMV Henan isolate based on the cloned and existing MP sequence in NCBI.The results of this study enrich the host range and the genetic information of the ReMV,and provide a basis for in-depth research on ReMV in the future.

关 键 词：地黄 地黄花叶病毒 检测方法 基因克隆 序列分析 

分 类 号：S435.672[农业科学—农业昆虫与害虫防治]