宁夏地区葡萄卷叶伴随病毒遗传变异的PCR-SSCP分析

宁夏贺兰山东麓酿酒葡萄卷叶病病株率较高,严重影响了酿酒葡萄产业的健康发展,但有关其病原葡萄卷叶伴随病毒(Grapevine leafroll-associated virus, GLRaVs)的遗传变异情况则鲜有报道.利用RT-PCR技术对宁夏酿酒葡萄不同种植区的7个主栽品种的GLRaV-1和GLRaV-3进行分子检测,利用SSCP技术对扩增片段进行遗传变异分析.试验结果共获得8个GLRaV-1和13个GLRaV-3的阳性样本. 8个GLRaV-1分离物基于CP基因分析差异明显,可分为3类; 13个GLRaV-3分离物基于RdRp基因分析差异明显,可分为3类,而基于CP和HSP70基因分析可分为2类.说明宁夏酿酒葡萄不同种植区、不同品种的GLRaV-1和GLRaV-3种群分子特性存在差异且遗传变异明显.这有助于宁夏地区酿酒葡萄GLRaVs流行学调查和分子诊断分析,为该病害的防控提供理论依据.

PCR-SSCP Analysis of Genetic Variation Among GLRaVs in Ningxia

The rate of diseased plants of grapevine leaf roll disease is relatively high in the eastern Helan mountain region of Ningxia, which seriously hinders the healthy development of the wine grape industry, but the genetic variation of grapevine leafroll-associated viruses (GLRaVs), i. e. the pathogen of this disease, is poorly understood. In an experiment reported in this paper, PT-PCR technique was used to detect GLRaV-1 and GLRaV-3 of 7 widely cultivated wine grape cultivars in different growing regions of Ningxia, and the SSCP technology was employed to analyze the genetic variation of the amplified fragments. A total of 8 positive samples of GLRaV-1 and 13 positive samples of GLRaV-3 were obtained in the test. Based on the CP gene analysis, the 8 GLRaV-1 isolates showed significant differences, which were divided into 3 categories. The 13 GLRaV-3 isolates also showed significant differences, and they were divided into 3 categories based on RdRp gene analysis and into 2 categories based on CP and HSP70 gene analyses. The above results showed that there were significant differences and genetic variations in the population molecular characteristics of GLRaV-1 and GLRaV-3 which originated from different wine grape cultivars and from different grape-growing areas in Ningxia.