天津地区番茄褪绿病毒的分子检测和鉴定

摘要

In the summer of 2014 ,tomato plants showed suspected symptoms caused by Tomato chlorosis virus ( ToCV) infection collected in greenhouse in Tianjin .In order to clarify the Pathogen of tomato plants ,specific prim-er pairs of ToCV coat protein (CP) and heat shock protein 70 (HSP70) were used for reversing transcription PCR ( RT-PCR ) detection, specific fragments were obtained and sequenced .HSP70 nucleotide sequences obtained 99.5% similarity with ToCV isolate Tochigi ( AB513442 ) , CP amino acid and nucleotide sequences obtained shared 99.2%and 99.3%similarity with ToCV isolate Tochigi (AB513443),indicating the tomato was infected by ToCV in Tianjin, the sequences of ToCV isolate between Tianjin and Japan with a highest similarity .As Tomato yellow leaf curl virus ( TYLCV) were widespread in survey areas ,the ToCV positive samples were also tested for TYLCV by specific primers .The result was positive and gene sequence analysis showed that the similarity between amplification products and TYLCV-IL isolates was more than 99.0%,indicating mixed infections in the field .It is the first report that ToCV infect tomato ,ToCV and TYLCV mixed infections tomato in Tianjin .%为明确2014年夏天在天津番茄种植区采集到疑似感染番茄褪绿病毒的番茄样品的侵染病原，利用ToCV外壳蛋白和热激蛋白的特异引物进行反转录PCR （ RT-PCR）检测，分别扩增得到特异核苷酸片段。序列分析表明， HSP70核苷酸序列与已登录的番茄褪绿病毒日本分离物Tochigi（AB513442）相似性为99．5％，CP氨基酸和核苷酸序列与已登录的番茄褪绿病毒日本分离物Tochigi（AB513443）相似性分别为99．2％和99．3％，表明天津地区的番茄受到ToCV侵染，且天津与日本ToCV分离物序列相似性最高。由于调查地区番茄黄化曲叶病毒发生普遍，针对ToCV阳性样品利用TYLCV特异性引物进行了分子检测，扩增产物序列与TYLCV以色列株系分离物的相似性均在99．0％以上，表明2种病毒在田间发生复合侵染，该研究首次明确天津地区的番茄受到ToCV侵染和ToCV与TYLCV的复合侵染。