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皱木复合病(rugose wood complex disease)是一种发生普遍、危害严重的葡萄病毒病,该病与葡萄病毒属(Vitivirus)病毒的侵染密切相关,葡萄病毒E(Grapevine virus E,GVE)是葡萄病毒属的新成员。为明确我国GVE侵染状况以及不同分离物的基因变异情况,本研究针对GVE的MP和CP基因设计了两对引物,RT-PCR结果表明,两对引物均能从带毒样品中扩增到目的基因片段。采用上述引物检测了211株葡萄样品,GVE检出率为5.7%。对7个GVE分离物的外壳蛋白基因进行测序和系统进化树分析,结果显示,上述分离物克隆分别处于2个明显分化的组群(Group I和Group II)。研究测定了GVE中国分离物GFM G-1的全基因组序列,该分离物与日本分离物Tv AQ7核苷酸序列相似度高达98%,但与日本分离物Tv P15、南非分离物SA94、美国分离物WAHH2及波兰分离物59PE核苷酸序列相似性仅为70%~75%。本研究为进一步明确我国GVE发生、分布及分子检测提供了依据。
The rugose wood complex disease is a common and serious grapevirus disease. The disease is closely related to the infection of the genus Vitivirus. Grapevine virus E (GVE) is a New member of the genus Vaccinia. In order to clarify the status of GVE infection in China and the genetic variation of different isolates, we designed two pairs of primers for the MP and CP genes of GVE. RT-PCR results showed that both pairs of primers could amplify from the infected samples to Target gene fragment. A total of 211 grape samples were tested using the above primers. The detection rate of GVE was 5.7%. The coat protein genes of seven GVE isolates were sequenced and phylogenetically analyzed. The results showed that the isolates cloned in two distinctly differentiated cohorts (Group I and Group II), respectively. The complete genome sequence of GVE Chinese isolate GFM G-1 was determined. The similarity of this isolate to the Japanese isolate Tv AQ7 was as high as 98%, but it was separated from the Japanese isolate Tv P15, the South African isolate SA94, and the United States The WAHH2 and Polish isolate 59PE nucleotide sequence similarity was only 70% -75%. This study provides a basis for further clarifying the occurrence, distribution and molecular detection of GVE in China.