山东省沙地葡萄茎痘相关病毒的检测  

Detection of grapevine rupestris stem pitting associated virus occurred in Shandong province

作　　者：尚佑芬[1] 王升吉[1] 赵玖华[1] 张家魁 吕志华[3] 路兴波[1] 孙红炜[1] 杨崇良[1] 

机构地区：[1]山东省农业科学院植物保护研究所 [2]山东省葡萄研究所,济南250100 [3]山东省济宁市林业局经济林站,山东济宁272000

出　　处：《果树学报》2009年第2期158-162,共5页Journal of Fruit Science

基　　金：山东省自然科学基金(Y2006D32)

摘　　要：为明确沙地葡萄茎痘相关病毒(GRSPaV)在山东省发生的情况,2006-2007年在葡萄主栽区和葡萄品种资源圃进行了田间调查、样品采集和病原检测。应用GRSPaV外壳蛋白基因表达获得的多克隆抗体,经间接酶联免疫吸附试验(PTA-ELISA)和斑点酶联免疫吸附试验(Dot-ELISA)检测,在2a采集的298个样品中,125个样品为阳性反应,阳性率41.95%。125个阳性样品涉及43个品种,品种被侵染率74.14%。研究对检测的样品用RT-PCR法验证,有3对引物均扩增出了预期片段,分别为解旋酶基因340bp片段、外壳蛋白(CP)基因842bp片段和RNA聚合酶(RdRp)基因499bp片段。In order to investigate the infection situation of grapevine rupestris stem pitting associated virus （GRSPaV）in Shandong province, 298 grapevine samples of 58 cuhivars were collected from major grape-growing areas and germplasm collection plots of grapevines and tested for GRSPaV by plate trapped antigen indirect enzyme-linked immunosorbent assay （PTAELISA） and Dot-ELISA with a polyclonal antibody against prokaryotic expressed coat protein of GRSPaV. The results showed that 125 grapevine plants were positive to GRSPaV and the percentage was 41.95%. The positive samples involved 43 cultivars and the percentage was 74.14%. With 3 pairs of designed primers, anticipated fragments, 340 bp for the helicase gene, 842 bp fragment for CP gene and 499 bp fragment for RdRp gene, were amplified successfuly by RT-PCR from the samples showing positive reaction in ELISA tests. These results provide valuable information on the infection situation of the GRSPaV in Shandong province.

关 键 词：葡萄茎痘相关病毒 病原检测 ELISA RT-PCR 

分 类 号：S663.1[农业科学—果树学]