番茄黄化曲叶病毒侵染危害海南番茄及其分子特征  

Tomato yellow leaf curl virus infecting tomato in Hainan and its molecular characteristics

作　　者：汤亚飞[1,2] 周洋 张丽 李正刚[1] 佘小漫 于琳[1] 蓝国兵 邓铭光 何自福[1,2] TANG Ya-fei;ZHOU Yang;ZHANG Li;LI Zheng-gang;SHE Xiao-man;YU Lin;LAN Guo-bing;DENG Ming-guang;HE Zi-fu(Plant Protection Research Institute,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;Guangdong Provincial Key Laboratory of High Technology for Plant Protection,Guangzhou 510640,China;Agricultural Service Center of Dongfang,Dongfang,Hainan 572600,China)

机构地区：[1]广东省农业科学院植物保护研究所,广州510640 [2]广东省植物保护新技术重点实验室,广州510640 [3]东方市农业服务中心,海南东方572600

出　　处：《南方农业学报》2020年第8期1970-1976,共7页Journal of Southern Agriculture

基　　金：国家重点研发计划项目(2018YFD0201209);广东省基础与应用基础研究基金项目(2019A1515012150);广州市科技计划项目(201904010173);广东省现代农业产业共性关键技术研发创新团队建设项目(2019KJ134)。

摘　　要：【目的】明确引起海南三亚、东方和陵水番茄黄化曲叶病的病毒分子特征,为防控该病害提供科学依据。【方法】以2019年2月从海南三亚、东方和陵水采集的15份疑似番茄黄化曲叶病样本为材料,采用菜豆金色花叶病毒属病毒的通用简并引物AV494/CoPR对15份样本进行PCR检测,并对PCR检测为阳性的样本进行滚环扩增(RCA)及基因克隆,获得侵染海南三亚、陵水和东方番茄的病毒基因组全序列,进而在GenBank数据库BLASTh程序进行相似性比对,利用SDT 1.2的MUSCLE alignment进行序列相似性比较分析,采用MEGA 6.0的邻接法(Neighbor-joining,NJ)构建系统发育进化树,明确所获得病毒分离物与已报道分离物的亲缘关系。【结果】获得侵染海南三亚、陵水和东方番茄的3个番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)分离物基因组全序列,其大小均为2781 nt,编码6个开放阅读框(ORF),其中病毒链上AV1和AV2基因,互补链上AC1、AC2、AC3和AC4基因。相似性比对分析结果表明,TYLCV海南分离物与墨西哥和美国分离物的相似性在99.0%以上,与来自我国不同省(市)的14个TYLCV分离物的相似性在98.0%以上。系统发育进化分析显示,海南三亚、陵水和东方3个分离物与墨西哥、美国及我国北京和江苏分离物聚集在一个小分支,进一步与来自我国其他12个省(市)、韩国、哥斯达黎加和澳大利亚的分离物形成一个分支。【结论】侵染海南番茄的TYLCV可能来自我国大陆的番茄等种苗和烟粉虱带毒传入,也有可能来自墨西哥和美国等其他国家。【Objective】This study was to identify the molecular characterization of Tomato yellow leaf curl virus(TYLCV)that caused tomato yellow leaf curl disease in Sanya,Dongfang and Lingshui of Hainan,in order to provide scientific basis for prevention and control of the disease in the area.【Method】In February,2019,15 suspected diseased tomato samples were collected from Sanya,Dongfang and Lingshui of Hainan.The 15 samples were detected by PCR using the universal degenerate primers AV494/CoPR for Begomoviruses.The PCR positive samples were amplified by rolling circle amplification(RCA)and cloned to obtain whole genome sequences of virus isolated from tomato in Sanya,Lingshui and Dongfang.These obtained sequences identity was compared and analyzed using BLAST on GenBank database.Pairwise nucleotide sequence identities were determined using SDT 1.2 with MUSCLE alignment.Phylogenetic analysis was conducted using MEGA 6.0 with neighbor joining method to clarify the genetic relationship between isolates of obtained virus and the reported isolates.【Result】Three whole genome sequences of TYLCV isolated from tomato in Sanya,Lingshui and Dongfang of Hainan were 2781 nucleotides(nt)long and encoded six open reading frames(ORFs).The AV1 and AV2 were located on the virion sense DNA strand,whereas,the AC1,AC2,AC3,AC4 genes were located on the complementary sense strand.The results of identity comparison showed that TYLCV isolated from Hainan shared over 99.0%nt identities with Mexico isolate and USA isolate,and over 98.0%nt identities with the 14 isolates from different provinces or municipalities of China.The result of phylogenetic analysis showed that Sanya,Lingshui and Dongfang isolates clustered with isolates of TYLCV from Mexico,USA,Beijing and Jiangsu of China to form a unique clade,further clustered with all isolates from other 12 provinces or municipalities of China,South Korea,Costa Rica,Australia to form a clade.【Conclusion】TYLCV infecting tomato in Hainan should come from other areas of China,or Mexico,the

关 键 词：番茄黄化曲叶病毒 分子鉴定 序列分析 海南 

分 类 号：S432.41[农业科学—植物病理学]