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为明确我国葡萄中沙地葡萄茎痘相关病毒(GRSPaV)的感染情况及病毒外壳蛋白(coat protein,CP)基因的变异特点,从而为其致病性、病害的防治以及抗病毒基因工程等研究提供依据,本研究对采自我国16个省市自治区的65个葡萄品种305株葡萄样品中的GRSPaV进行RT-PCR检测,根据地区与品种差异选取了24个阳性样品进行cp基因克隆与测序分析,并对不同RNA提取方法进行了比较。结果显示,114株样品被GRSPaV侵染,平均带毒株率为37.4%;分离物间及同一分离物不同克隆间的序列差异较大,从24个分离物克隆获得的37条cp基因序列与来源于不同国家的12个GRSPaV分离物的核苷酸序列同源性为80.5%~99.7%,氨基酸序列同源性为88.8%~100%;各个分离物的遗传距离无明显地域差异;SiO2吸附法比SDS法和CTAB法更适宜葡萄样品RNA的提取。
In order to clarify the infection status of Grapevine pox-related virus (GRSPaV) and the variation of coat protein (CP) gene in grapevine in China, the research of its pathogenicity, disease prevention and anti-virus gene engineering According to the results, GRSPaV in 305 grape samples collected from 65 grapevines in 16 provinces of our country was detected by RT-PCR. 24 positive samples were selected for the cloning and sequencing of cp gene according to the differences of regions and varieties , And compared different RNA extraction methods. The results showed that 114 samples were infected with GRSPaV, with an average rate of 37.4%. The sequences of isolates and isolates were different among different clones. The sequences of 37 cp genes cloned from 24 isolates The nucleotide sequence homologies of 12 GRSPaV isolates from different countries were 80.5% -99.7%, and the amino acid sequence homologies were 88.8% -100%. There was no significant difference in the genetic distances among the isolates. The SiO2 adsorption Farby SDS method and CTAB method are more suitable for grape sample RNA extraction.