葡萄卷叶伴随病毒2号和3号辽宁分离物部分基因组的序列分析

 将采自辽宁兴城地区在生长期间具典型卷叶病症状的金星无核(Venus Seedless)葡萄品种休眠枝条,用RT-PCR检测4种葡萄卷叶伴随病毒(Grapevine leafroll-associated viruses,GLRaVs),扩增得到了葡萄卷叶伴随病毒2号(GLRaV-2)和葡萄卷叶伴随病毒3号(GLRaV-3)两种病毒的主要外壳蛋白(major coat protein,CP)基因的完整序列(GenBank登录号分别为FJ786017和FJ786016)。这表明该葡萄植株受到了GLRaV-2和GLRaV-3辽宁分离物(GLRaV-2-LN和GLRaV-3-LN)的复合侵染。根据检测结果,克隆了GLRaV-2-LN基因组3'端CPm (minor capsid protein)、p19(19-kDa protein)和p24(24-kDa protein)基因(GenBank登录号分别为FJ786018、FJ786019和FJ786018)。序列分析表明,GLRaV-3-LN的CP基因全长942 nt,与已报道的国内外其它分离物CP基因全序列相比,核苷酸序列同源性为89.8%~91.8%,由此推导的氨基酸序列同源性为94.9%~97.4%。GLRaV-2-LN的CP、CPm、p19和p24基因全长分别为597 nt、672 nt、486 nt和618 nt。与国外报道的几个分离物的相应蛋白基因全序列相比,核苷酸序列同源性分别为88.3%~100.0%、78.7%~99.9%、75.1%~99.4%和87.5%~99.5%;由此推导的氨基酸序列同源性分别为92.9%~100.0%、89.2%~100.0%、73.9%~99.4%和89.3%~99.0%。

Sequence analysis of the genes of two isolates of Grapevine leafroll-associated viruses from Liaoning

The dormant cutting of grapevine ( Vitis vinifera cv. Venus Seedless) with typical leafroll symptom originated from Xingcheng region in Liaoning Province was detected for Grapevine leafroll-associated vi-ruses (GLRaVs) by RT-PCR. The results showed that two complete sequences of major coat protein (CP) gene of GLRaV-2 (GenBank accession number FJ786017) and GLRaV-3 (FJ786016) were obtained, which indicated that the grapevine plant detected was co-infected with GLRaV-2 and GLRaV-3. The genes encoding CPm, pl9 and p24 located in 3'genomic RNA of GLRaV-2 Liaoning isolate (LN) were then cloned and se-quenced. The nucleotide sequence of GLRaV-3 CP was 942 nt, and sequence analysis showed that the identi-ties of nucleotide sequence and deduced amino acid sequence among GLRaV-3-LN and other previously repor-ted isolates including ten overseas and two domestic isolates were ranged from 89.8% to 91.8% and 94.9% to 97. 4%, respectively. The nucleotide sequences of CP, CPm (FJ786018), pl9 (FJ786019) and p24 (FJ786018) of GLRaV-2-LN were 597 nt, 672 nt, 486 nt and 618 nt in length, respectively. The relative se-quence analysis showed that the nucleotide identities of CP, CPm, pl9 and p24 among GLRaV-2-LN and oth-er isolates reported were ranged from 88.3% to 100.0% , 78.9% to 100.0% , 75.1% to 99.4% and 87.5% to 99.5% , and the identities of deduced amino acids were 92.9 to 100.0% , 89.2% to 100.0% , 73.9% to 100.0% and 89.3% to 99.0%, respectively.