侵染构树的菜豆金色黄花叶病毒属病毒的鉴定及基因组序列分析  

Molecular identification and full genome sequence analyses of begomoviruse infec-ting Broussonetia papyifera

作　　者：汤亚飞[1] 李正刚 佘小漫[1] 于琳 蓝国兵[1] 丁善文 何自福[1] TANG Yafei;LI Zhenggang;SHE Xiaoman;YU Lin;LAN Guobing;DING Shanwen;HE Zifu(Institute of Plant Protection,Guangdong Academy of Agricultural Sciences,Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China Ministry of Agriculture and Rural Affairs,Guangdong Provincial Key Laboratory of High Technology for Plant Protection,Guangzhou 510640,China)

机构地区：[1]广东省农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广东省植物保护新技术重点实验室,广州510640

出　　处：《植物病理学报》2024年第1期59-68,共10页Acta Phytopathologica Sinica

基　　金：国家自然科学基金(32072392);广东省自然科学基金(2019A1515012150);广东省现代农业产业技术体系创新团队项目(2021KJ134);科技创新战略专项资金“金颖之星”(R2019PY-JX005);广东省农业科学院“十四五”学科团队建设项目(202105TD)。

摘　　要：从广东省河源市连平县采集3份疑似双生病毒侵染引起的叶片表现黄花叶症状构树样品,提取总DNA,利用Bego-movirus通用引物AV494/CoPR进行PCR检测表明,疑似病样中均检测到菜豆金色黄花叶病毒属病毒。进一步选取PCR检测为阳性的样品进行RCA扩增、酶切、克隆及测序,获得侵染广东构树的病毒分离物基因组全长序列。广东构树分离物(GS-2021)为一个双组分病毒,包含DNA-A和DNA-B两组分。DNA-A组分(GS-2021-A)全长为2777 nt,编码7个ORFs;DNA-B组分(GS-2021-B)为2742 nt,编码2个ORFs。GS-2021与已报道的中国大青金色花叶病毒(clerodendrum golden mosaic China virus,ClGMCNV)各分离物的DNA-A、DNA-B均有较高的一致性,全长序列的一致性分别为93.0%~93.9%和86.3%~89.6%,其中与福建Fz7分离物DNA-A(GenBank登录号:FJ011668)和DNA-B(GenBank登录号:FJ011669)的相一致性最高,为93.9%和89.6%。GS-2021与ClGMCNV福建、浙江、江苏和美国的5个分离物亲缘关系近,同属一个分支,其中与福建Fz7分离物聚集在一个小分支,亲缘关系最近。基因重组分析显示,GS-2021无明显的基因重组事件存在。根据ICTV对菜豆金色黄花叶病毒属病毒最新分类标准,GS-2021属ClGMCNV的一个新株系。本研究首次在构树上检测到菜豆金色花叶病毒属病毒,获得其病毒基因组全序列,明确其为ClGMCNV的新株系。因此,构树是菜豆金色黄花叶病毒属病毒的新自然寄主。Three Broussonetia papyifera samples suspected to be infected by Begomovirus,with yellow mosaic leaves,were collected from Lianping county,Heyuan city,Guangdong province.Total DNA was extracted from suspected samples individually,and was used as template for PCR detection with degenerate Begomovirus pri-mers AV494/CoPR.The PCR detection result showed that three suspected samples were infected by Begomovirus.The full genome sequence of virus isolated from Broussonetia papyifera in Guangdong(GS-2021)was obtained by RCA amplification,followed by enzyme digestion,cloning and sequencing.GS-2021 was a bipartite virus,including DNA-A and DNA-B components.The full sequence of DNA-A(GS-2021-A)was 2777 nt in size,and encoded seven ORFs.The full sequence of DNA-B(GS-2021-B)was 2742 nt in size,and encoded two ORFs.GS-2021 shared the higher similarity with all isolates of clerodendrum golden mosaic China virus(ClGM-CNV).GS-2021-A shared a 93.0%-93.9%identity with DNA-A of all isolates of ClGMCNV,and the highest i-dentity(93.9%)is with the Fujian Fz7 isolate(GenBank accession number:FJ011668).GS-2021-B shared an 86.3%-89.6%identity with DNA-B of all isolates of ClGMCNV,and the highest identify(89.6%)is with the Fujian Fz7 isolate(GenBank accession number:FJ011669).GS-2021 was closely related to five isolates of ClG-MCNV from Fujian,Zhejiang,Jiangsu and the United States,which belonged to the same clade.In addition,GS-2021 clustered with Fz7 isolate from Fujian in a small clade,and had the closest relationship with it.Recom-bination analysis showed that there was no obvious gene recombination event in GS-2021.Based on the latest demarcation threshold for Begomovirus,GS-2021 was a new strain of ClGMCNV.In this study,Begomovirus was detected on Broussonetia papyifera for the first time.The full viral genome sequence of this virus was obtained and identified as a new strain of ClGMCNV.This result shows that Broussonetia papyifera is a newly discovered natural host for Begomovirus.

关 键 词：构树 菜豆金色黄花叶病毒属 中国大青金色花叶病毒 病毒基因组 新寄主 

分 类 号：S432.1[农业科学—植物病理学]