侵染云南苹果的苹果褪绿叶斑病毒的RT-PCR检测和序列分析

RT-PCR Detection and Sequence Analysis of Apple Chlorotic Leaf Spot Virus (ACLSV) Infecting Apple in Yunnan

以云南昭通、昆明、曲靖等地采集的苹果病毒病样品为试材,对其是否携带苹果褪绿叶斑病毒(ACLSV)进行RT-PCR检测,将本试验获得3个云南ACLSV分离物(ZT,ML,TJX)和GenBank上已登录的24个分离物进行序列比对分析。所有的ACLSV分离物的核苷酸序列相似性为81.3%～99.7%,它们可以被分成两大组群,组群A,包括了大部分不同寄主、地理来源的分离物,所有的ACLSV苹果分离物位于组A;组群B,包括了ACLSV全部的寄主为桃的分离物。本研究获得的3个苹果云南分离物属于组群A,组群A可分成两个亚组,其中,分离物ML和TJX位于同一个亚组,而分离物ZT位于另一个亚组。结果表明ACLSV苹果分离物与寄主种类存在一定的相关性,该研究为株系划分、分子流行和寄主抗性分析提供依据。

In this study,using the apple virus samples from Zhaotong,Kunming and Qujing in Yunnan as test materials,ACLSV were detected by RT-PCR and the gene sequences of three isolates（ZT,ML,TJX） obtained in this study along with 24 isolates logged on GenBank were comparatively analyzed.All the isolates of the nucleotide sequence showed 81.3 % ～ 99.7 % similarities and could be grouped into two clusters.Group A,including the most isolates from different hosts and geographic locations,all the apple isolates located in Group A.Group B,including all peach isolates.Three isolates from our study were grouped into Group A.Among the group A,the isolates ML and TJX showed the close genetic relationship and located in the same subgroups,whereas ZT isolate and TJX,ML isolates showed the further genetic relationship and grouped into another subgroup.The results proved that the ACLSV molecular variation had some correlation with host.The study provided the basis for study strains,molecular epidemiology of ACLSV disease and resistance of apple for ACLSV.