3种苹果潜隐病毒多重RT-PCR检测体系的建立  

Multiplex RT-PCR Assay for Simultaneous Detection of Three Latent Apple Viruses

作　　者：范旭东[1] 董雅凤[1] 张尊平[1] 李丽丽[2] 裴光前[1] 

机构地区：[1]中国农业科学院果树研究所,辽宁兴城125100 [2]沈阳农业大学园艺学院,沈阳110161

出　　处：《园艺学报》2009年第12期1821-1826,共6页Acta Horticulturae Sinica

基　　金：2008年农业科技跨越计划项目;中央级公益性科研院所基本科研业务费专项项目(082060302-07)

摘　　要：研究建立了能同时检测苹果茎沟病毒(Applestem grooving virus,ASGV)、苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus,ACLSV)和苹果茎痘病毒(Apple stem pitting virus,ASPV)的多重RT-PCR方法。以复合感染3种病毒的苹果‘望山红’组培苗为试材,对影响多重PCR的反应条件进行了一系列的调整和优化。多重RT-PCR体系灵敏性测验显示,最低能从RNA总量187.5ng的样品中检测3种病毒的存在。多重RT-PCR产物的序列与报道的病毒序列有较高的同源性,12个田间苹果样品的多重RT-PCR检测结果与单一PCR的结果一致,初步证明了多重RT-PCR检测的准确性。The multiplex RT-PCR assay was established to simuhaneously detect Apple stem grooving virus （ASGV）, Apple chlorotic leaf spot virus （ACLSV） and Apple stem pitting virus （ASPV）. Using tissue culture apple seedling （Wangshanhong） infected by three latent apple viruses as sample, a series of factors affecting the multiplex RT-PCR were optimized. Sensibility test showed that it could simultaneously detect out ASGV, ACLSV and ASPV in 187.5 ng total RNA. The muhiplex RT-PCR products were cloned and sent for sequencing, and the sequences show high homologies to the published virus sequences. Consistent results for virus detection of 12 apple samples in field were obtained by the multiplex RT-PCR and single RT-PCR sys- tem, suggesting the accuracy of the system in ASGV, ACLSV and ASPV detection.

关 键 词：苹果 多重RT-PCR 检测 苹果茎沟病毒 苹果褪绿叶斑病毒 苹果茎痘病毒 

分 类 号：S661.1[农业科学—果树学]