菊花R病毒浙江湖州分离物的全基因组序列克隆及其进化和重组分析  

Genome-wide cloning,evolution,and recombination analyses of chrysanthemum virus R isolates from Huzhou city,Zhejiang,China

作　　者：李佳鹏 刘梦阳 吴潇银 叶增康 仲雪婷 王亚琴[2] 王占旗 LI Jiapeng;LIU Mengyang;WU Xiaoyin;YE Zengkang;ZHONG Xueting;WANG Yaqin;WANG Zhanqi(Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province,College of Life Sciences,Huzhou University,Huzhou 313000,China;State Key Laboratory of Rice Biology,Institute of Biotechnology,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]湖州师范学院,浙江省媒介生物学与病原控制重点实验室,湖州313000 [2]浙江大学生物技术研究所,水稻生物学国家重点实验室,杭州310058

出　　处：《植物病理学报》2023年第5期831-840,共10页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(32102162);浙江省自然科学基金资助项目(LQ19C140001);浙江省公益研究计划(LGN19C140002);大学生创新创业训练计划(202201269)。

摘　　要：为明确菊花R病毒(chrysanthemum virus R, CVR)的分子特征及其进化和重组关系,利用RNA-Sequencing技术与3′/5′-RACE(rapid amplification of cDNA ends)技术对CVR浙江湖州分离物CVR-ZJHU1和CVR-ZJHU2进行克隆并获得其全基因组序列(GenBank登录号:ON137989和ON137990),并采用MEGA 11.0和RDP 4.1软件进行系统进化树构建和重组分析。结果表明:CVR-ZJHU基因组全长为8 873 nt(不包括polyA尾巴),包含6个开放阅读框(open reading frames, ORFs),其中ORF1编码一个RNA复制酶,ORFs 2-4编码3个运动相关蛋白(TGBp1、TGBp2和TGBp3),ORF5编码病毒衣壳蛋白(coat protein, CP),ORF6编码一个富半胱氨酸蛋白(cysteine-rich protein, CRP)。序列分析和系统进化分析结果显示,浙江湖州分离物(CVR-ZJHU1和CVR-ZJHU2)与浙江桐乡分离物(CVR-TX)同源性最高、亲缘关系最近,他们与CVR-TX分离物的核苷酸序列同源性分别为94.2%和93.7%。重组分析结果表明,CVR-ZJHU1为重组体,其主要父本为CVR-TX,次要父本为CVR-ZJHU2;而CVR-ZJHU2未与其他CVR分离物发生重组,且他们与中国北京分离物(CVR-BJ)以及俄罗斯分离物(CVR-X6、CVR-X13和CVR-X21)均无重组关系,说明CVR现在为小规模、局部范围内传播。To clarify the molecular characterization,evolution,and recombination of chrysanthemum virus R(CVR),genome-wide cloning of two CVR isolates from Zhejiang,China,CVR-ZJHU1 and CVR-ZJHU2(GenBank accession numbers:ON137989 and ON137990),was carried out using RNA-Sequencing and 3′/5′rapid amplification of cDNA ends(3′/5′-RACE)technology.Subsequently,MEGA 11.0 and RDP 4.1 software were employed to construct the phylogenetic tree and to analyze the molecular recombination.The results showed that the full-length genome of CVR-ZJHU isolates was 8873 nt(excluding the polyA tail),including six open reading frames(ORFs),in which ORF1 encoded a viral replicase,ORFs 2-4 encoded movement-related proteins(TGBp1,TGBp2,and TGBp3),ORF5 encoded a viral coat protein(CP),and ORF6 encoded a cysteine-rich protein(CRP).Sequence alignment and phylogenetic tree analyses showed that CVR-ZJHU1 and CVR-ZJHU2 had the highest homology and closest relationship with Zhejiang Tongxiang isolate of CVR(CVR-TX),and their nucleotide sequence homology with CVR-TX was 94.2%and 93.7%,respectively.The recombination analysis showed that CVR-ZJHU1 was a recombinant whose major parent was CVR-TX and minor parent was CVR-ZJHU2;whilst CVR-ZJHU2 did not recombine with other CVR isolates.Neither CVR-ZJHU1 nor CVR-ZJHU2 recombined with the Beijing isolate(CVR-BJ)and the Russian isolates(CVR-X6,CVR-X13,and CVR-X21),indicating that CVR is now spreading on a small and local scale.

关 键 词：菊花R病毒 基因组 系统进化分析 重组分析 

分 类 号：S432.41[农业科学—植物病理学]