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

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.