果树学报2013，Vol.30Issue(2)：197-201,5.
三种葡萄病毒的RT-PCR检测和系统进化分析
RT-PCR detections and phylogenetic studies on three viruses from grapevine
摘要
Abstract
[Objective]The objective of the study is to develop a sensitive detection technique on grapevine viruses and understand the phylogenetic relationships of Sichuan isolates with other geographic origins. [Method]Comparison with three different RNA extraction protocols, the Reverse Transcription-Poly-merase Chain Reaction (RT-PCR) were optimized to detect three different grapevine viruses. Partial genomes of three different viruses isolated from Sichuan were cloned and the phylogenetic studies of each isolate were conducted, respectively. [Result]The results indicated high quality RNA was obtained quickly from the mature phloem of shoots by the modified silica particles extraction only. RT-PCR was successfully used to amplify partial fragments of the GLRaV-2, GVB and GVA with 18S RNA of grapevine as internal control, respectively. In addition, phylogenetic analysis and nucleotide identities studies clearly clustered the 16 CP genes of GLRaV-2 isolates collected from different geographic origins into five phylogenetic groups. The nucleotide identities of GVB CP genes between 7 isolates of distinct geographic origins were calculated to 79.1%-98.7%. Five isolates of GVA from Sichuan were clustered into 4 clades based on the phylogenetic structure, so it was suggested that the virus populations of GVA from Sichuan consisted of genetic diversity of variants. [Conclusion]The studies suggested that GLRaV-2, GVA and GVB had genetic variability at level of nucleotide acids. Furthermore, GVA populations in Sichuan had significant genetic diversity and included 4 different strains.关键词
葡萄卷叶伴随2型病毒/葡萄B病毒/葡萄A病毒/病毒外壳蛋白基因/病毒沉默抑制子基因Key words
Grapevine leafroll associated virus-2/ Grapevine virus B/ Grapevine virus A/ Coat protein gene/ RNA silence suppressor分类
农业科技引用本文复制引用
王建辉,刘建军,陈克玲,李洪雯,何建,关斌..三种葡萄病毒的RT-PCR检测和系统进化分析[J].果树学报,2013,30(2):197-201,5.基金项目
"十二五"四川省财政基因工程:果树生物技术研究与创新(2011JYGC05-018) （2011JYGC05-018）
