沈阳市番茄褪绿病毒与番茄黄化曲叶病毒复合侵染的分子鉴定  

Molecular Identification on Mixed Infection of Tomato chlorosis virus and Tomato yellow leaf curl virus in Shenyang

作　　者：赵秀香[1] 王春阳 厉彦芳 黄欣阳 吴俊清 吴元华[1] ZHAO Xiu-xiang;WANG Chun-yang;LI Yan-fang;HUANG Xin-yang;WU Jun-qing;WU Yuan-hua(College of Plant Protection,Shenyang Agricultural University,Shenyang 110161,China;Shenyang Modern Agricultural Research and Development Service Center,Shenyang 110034,China;Shenyang Daguang Agricultural Service Center,Shenyang 110200,China)

机构地区：[1]沈阳农业大学植物保护学院,沈阳110161 [2]沈阳市现代农业研发服务中心,沈阳110034 [3]沈阳市大光农业服务中心,沈阳110200

出　　处：《沈阳农业大学学报》2019年第4期470-475,共6页Journal of Shenyang Agricultural University

基　　金：“十三五”国家重点研发计划项目(2016YFD0201004);辽宁省自然基金面上项目(201602671)

摘　　要：在辽宁省沈阳市辽中设施蔬菜产区采集到疑似感染番茄褪绿病毒(Tomato chlorosis virus,ToCV)和番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的番茄样品,利用ToCV的特异性引物ToC5/ToC6进行RT-PCR检测,结果从4份病样中检测到ToCV预期大小的特异片段。利用ToCV外壳蛋白(coat protein, CP)和热激蛋白(heat shock protein 70, HSP70)基因的特异性引物进行CP基因和HSP70基因扩增及序列测定,结果得到ToCV辽中分离物CP基因和HSP70基因全长分别为774bp(GenBank登录号:MF278016)和1665bp(GenBank登录号:MF278017),序列分析结果表明ToCV辽中分离物的CP核苷酸序列与北京ToCV分离物(KC887999)和河北的ToCV分离物(KP217196)的CP 核苷酸序列的相似性最高,均为99.87%;HSP70核苷酸序列与山东ToCV分离物(KC709510)的HSP70核苷酸序列的相似性最高,为99.76%。同时该样品利用背靠背引物TY-F/TY-R对TYLCV的DNA-A分子进行全基因序列扩增及序列测定,结果得到TYLCV辽中分离物的DNA-A分子全基因长度为2784 bp,序列分析结果表明TYLCV辽中分离物与山东TYLCV分离物(GU199587)的DNA-A分子全基因核苷酸序列的相似性为99.64%。从而证实了沈阳市辽中区的番茄受到ToCV与TYLCV复合侵染。Tomato leaf samples that may infected by Tomato chlorosis virus(ToCV) or Tomato yellow leaf curl virus (TYLCV) were collected in Liaozhong district, Shenyang, Liaoning province. To identify whether these samples were infected by ToCV or TYLCV, specific primer pairs of ToC5/ToC6 specific to ToCV was used to detect ToCV and the expected DNA fragments were amplified from four infected samples. The coat protein gene (CP) and heat shock protein 70 (HSP70) gene of ToCV were cloned and analyzed, respectively. 774 bp (GenBank accession number:MF278016) and 1665 bp(GenBank accession number:MF278017) specific fragments were obtained from the infected sample. Further sequence blast analysis showed that CP gene of ToCV fragment from LNLZ isolate has high nucleotide identity at 99.87% with ToCV isolates from Beijing (KC887999) and Hebei (KP217196), while HSP70 of ToCV fragment from LNLZ isolate has high nucleotide identity at 99.76% with isolate from Shandong (KC709510). In addition, specific fragment of TYLCV was amplified by PCR using TY-F/TY-R primers. Sequence blast analysis showed that the DNA-A sequence of TYLCV from LNLZ isolate has high nucleotide identity at 99.64% with TYLCV isolate from Shandong(GU199587). All of the above results indicated that the candidate samples from Liaozhong district of Shenyang were mixed infected by ToCV and TYLCV.

关 键 词：番茄褪绿病毒 番茄黄化曲叶病毒 复合侵染 辽中分离物 序列分析 

分 类 号：S436.418[农业科学—农业昆虫与害虫防治]