河北省设施番茄褪绿病毒分子检测和鉴定研究  

Molecular Detection and Identification of Tomato chlorosis virus Infecting Greenhouse-grown Tomato Plants in Hebei Province

作　　者：孙国珍[1] 高利利[1] 陆文利[2] 王勇[3] 张安胜[4] 竺晓平[1] 

机构地区：[1]山东农业大学植物保护学院,山东泰安271018 [2]山东农业大学园艺科学与工程学院,山东泰安271018 [3]天津市植物保护研究所,天津300381 [4]山东省农业科学院植物保护研究所,山东济南251000

出　　处：《北方园艺》2015年第9期95-98,共4页Northern Horticulture

基　　金：国家公益性(农业)行业科研专项资助项目(201003065);山东省科技发展计划资助项目(2014GNC111008);山东省自然科学基金资助项目(ZR2012CM032)

摘　　要：以2014年夏季在河北7个县市设施番茄种植区田间采集的疑似番茄褪绿病毒(Tomato chlorosis virus,ToCV)感病植株为试材,采用RT-PCR检测、BLAST比对和MEGA 5.0系统树分析等方法,研究河北设施番茄上ToCV的发生和分子鉴定以及ToCV与另一种严重发生的番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLC)复合侵染情况。结果表明:以ToCV外壳蛋白(Coat Protein,CP)和热激蛋白(Heat Shock Protein 70,HSP70)共3对特异引物对其中4个典型地区ToCV疑似样品进行检测,分别扩增到约1 032、943、911bp的特异条带,同时针对ToCV阳性样品利用TYLCV特异引物从部分样品中扩增得到约833bp的核苷酸序列,4个样品均确定为ToCV阳性,样品与ToCV北京分离物(KC887999)CP基因序列一致性最高,为99.7%,同时确定田间存在ToCV和TYLCV复合侵染。Taking tomato samples showing symptom similar to that caused by Tomato chlorosis virus（ToCV）found and collected from 7regions in Hebei Province in the summer of 2014 as material,using RT-PCR detection,BLAST alignment and phylogenetic analysis based on MEGA 5.0as research methods to detect and identify ToCV occurrence and mixed infection on ToCV and another economic important virus Tomato yellow leaf curl virus（TYLCV）.The results showed that three pairs of specific primers targeting the ToCV coat protein（CP）and heat shock protein（HSP70）genes were used for detection,specific 1 032 bp,943bp and 911 bp fragments were amplified from the symptomatic samples,respectively.The ToCV positive samples were also tested for presence of TYLCV,specific primers of TYLCV were used and a 833 bp specific fragments were amplified and sequenced.Plant samples from Hebei were infected by Tomato chlorosis virus,the sequence analyses showed the nucleotide sequence of Hebei samples had the highest identity of 99.7%with a Beijing isolate of ToCV（KC887999）.The mixed infection of ToCV and TYLCV occurred in the field was also confirmed.

关 键 词：番茄褪绿病毒 HSP70 CP RT-PCR检测 番茄黄化曲叶病毒 复合侵染 

分 类 号：S641.2[农业科学—蔬菜学]