利用PCR技术检测草莓镶脉病毒

Detection of Strawberry Vein Banding Virus in Strawberries by PCR

采用特异引物的PCR扩增方法 ,对 30个草莓品种进行了草莓镶脉病毒的检测 ,同时用指示植物小叶嫁接法对被检植株进行了病毒检测 ,两种方法结果一致。回收、克隆PCR扩增出的特异DNA片段 ,获得了携有草莓镶脉病毒CP基因片段的载体。测序结果表明 ,得到的特异DNA片段同美国草莓镶脉病毒的株系ATCC 4 5 0 5 8CP基因片段相比 ,同源性为 90 .94 %。

Total DNA, isolated from 30 strawberry cultivars, was used as template to amplify strawberry vein banding virus (SVBV) CP gene fragment by polymerase chain reaction (PCR). Meanwhile, the tested plants were detected by indicator leaf grafting technique. The plants with typical band in PCR showed vein banding symptoms in indicator UC5 ( Fragaria vesca ), EMC ( F. vesca ), and those without typical band didn't showed symptoms. Results were identical in two detection methods. The amplified fragment by PCR was recycled, cloned in pMD18 T vector, and then sequenced. It is suggested that the expected SVBV CP gene fragment was obtained and is 90.94 % identical to that of American isolate ATCC 45058.