葡萄卷叶伴随病毒Ⅲ宁夏分离物外壳蛋白基因的克隆及序列分析

Clone and Sequence Analysis of Coat Protein Gene of Grapevine Leafroll Associated-virus Ⅲ from Ningxia

用酶联免疫吸附法对宁夏贺兰山东麓地区不同葡萄品种进行卷叶伴随病毒Ⅲ(GLRaVⅢ)的测定,检测率为37.1%,与田间自然发病率调查结果基本一致.设计GLRaVⅢ外壳蛋白(CP)基因序列引物对,进行RT-PCR扩增,获得1.1 kb的特异片段.其中,CP基因长942 bp,推导的编码蛋白含有313个氨基酸.通过对GLRaVⅢCP基因同源性比较,结果表明,GLRaVⅢ宁夏分离物的CP基因与四川分离物SL-10 CP基因的亲缘关系最近,核苷酸和所编码的氨基酸序列同源性分别为98.8%和98.1%;与巴西分离物PET-4和智利分离物C1-817 CP基因亲缘关系较远,核苷酸序列同源性低于95.0%,所编码的氨基酸序列同源性低于97.0%,认为GLRaVⅢ株系间的CP基因存在差异.

Through enzyme-linked immunosorbent assay(ELISA),of rapevine leafroll associated-virus Ⅲ(GLRaVⅢ) on different grapevine varieties from the Helan mountain east areas of Ningxia was detected,the detected rate 37.1%,which was in relative accordance with the incidence rate in the field.Using primers for coat protein(CP) gene of GLRaVⅢ,a 1.1kb DNA frgment was amplified by RT-PCR in RNA samples extracted from the infected grapevine of Ningxia.The results showed that CP gene of GLRaVⅢ from Ningxia is 942bp in leng...