新的柑橘叶斑驳病毒桑树分离株的基因组克隆及传播方式分析  

Analysis of a New Citrus Leaf Blotch Virus Isolate from Mulberry

作　　者：唐佳璇 方苗 张健 尹珍妮[1] 韩沛羽 韩涛涛 张朋 卢全有 Tang Jiaxuan;Fang Miao;Zhang Jian;Yin Zhenni;Han Peiyu;Han Taotao;Zhang Peng;Lu Quanyou(School of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 212100,China;Key Laboratory of Silkworm and Mulberry Genetic Improvement,Ministry of Agriculture and Rural Affairs,Sericultural Scientific Research Center,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212100,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212100 [2]农业农村部蚕桑遗传改良重点实验室,中国农业科学院桑蚕科学研究中心,江苏镇江212100

出　　处：《蚕业科学》2024年第1期1-8,共8页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(32271885)。

摘　　要：柑橘叶斑驳病毒(citrus leaf blotch virus,CLBV)可侵染多种植物。2018年,通过对叶片表现为皱缩、变窄症状的桑叶样品进行高通量测序后发现,桑树中存在CLBV的侵染。通过RT-PCR、5′-和3′-RACE等方法确定了CLBV桑树分离株(CLBV-MA)的全基因组序列。CLBV-MA基因组全长8617 nt(不包含3′polyA尾),共包含3个开放阅读框,分别编码复制酶蛋白、运动蛋白和外壳蛋白。CLBV-MA与其他CLBV分离株编码的复制酶蛋白以及外壳蛋白的氨基酸序列相似性分别为79.0%~89.1%、87.6%~94.7%,是一个新分离株。基于全基因组核苷酸序列的系统进化分析显示,CLBV-MA与CLBV-Actinidia、CLBV-HZ、CLBV-XL、CLBV-Actinidia-V20以及CLBV-ML聚类到一个分支,亲缘关系最近,并且CLBV-MA更为原始。根据桑树的繁殖方式、修剪管理方式、树龄以及CLBV-MA在桑园中的分布情况分析,CLBV-MA主要通过嫁接传播,也可以通过被污染的刀具传播,但传播效率低下。CLBV-MA全基因组序列及其传播方式的确定,为建立CLBV-MA分子检测方法及制定有效的防控措施奠定了基础。Citrus leaf blotch virus(CLBV)has the potential to infect a wide range of species.In 2018,CLBV was detected in wrinkled and narrowed mulberry leaves through next-generation sequencing.The complete genome sequence of mulberry isolate of CLBV(CLBV-MA)was determined by the combination of RT-PCR,5′-and 3′-RACE.It is 8617 nucleotides long(excluding the polyA tail),which contains three open reading frames encoding replicase proteins(RP),movement protein(MP)and coat protein(CP),respectively.The amino acid sequence identities of RP and CP between CLBV-MA and other isolates in the genus Citrivirus are 79.0%to 89.1%and 87.6%to 94.7%,respectively,suggesting that CLBV-MA is a new isolate.Phylogenetic analysis based on whole genomic nucleotide squences revealed that CLBV-MA,CLBV-Actinidia,CLBV-HZ,CLBV-XL,CLBV-Actinidia-V20 and CLBV-ML cluster into a branch,indicating that they are genetically related to each other.Additionally,CLBV-MA dates back further than others in this branch.Depending on the propagation mode,pruning management,age,and distribution of CLBV-MA in mulberry field,CLBV-MA is transmitted mainly through grafting and at a low rate by contaminated knives.The determination of the complete genome sequence of CLBV-MA and its transmission manner lays the foundation to develop molecular detection methods and formulate effective prevention and control strategies.

关 键 词：桑树 柑橘叶斑驳病毒 全基因组序列 传播方式 

分 类 号：S888.712[农业科学—特种经济动物饲养]