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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%。
Dormancy shoots of Venus Seedless grape varieties with typical leaf curl symptoms during the growing season in Xingcheng, Liaoning Province were collected, and four Grapevine leafroll-associated viruses (GLRaVs) were detected by RT-PCR. The complete sequence of the major coat protein (CP) gene of two viruses, GLRaV-2 and GLRaV-3, was amplified and sequenced (GenBank Accession numbers FJ786017 and FJ786016, respectively). This indicates that the grapevine plant was co-infected with GLRaV-2 and GLRaV-3 Liaoning isolates (GLRaV-2-LN and GLRaV-3-LN). According to the results, the minor capsid protein, p19 (19-kDa protein) and p24 (24-kDa protein) genes of GLRaV-2-LN genome were cloned (GenBank accession numbers are FJ786018, FJ786019 and FJ786018, respectively) . Sequence analysis showed that the full-length CP gene of GLRaV-3-LN was 942 nt in length, which was 89.8% -91.8% homologous to the full-length CP gene sequence reported from other isolates at home and abroad. The amino acid sequence homology was 94.9% ~ 97.4%. The full length of CP, CPm, p19 and p24 genes of GLRaV-2-LN were 597 nt, 672 nt, 486 nt and 618 nt, respectively. The nucleotide sequence homology was 88.3% -100.0%, 78.7% -99.9%, 75.1% -99.4% and 87.5% -99.5%, respectively, compared with that of the corresponding foreign gene sequences of several isolates reported abroad. The deduced amino acid sequence homologies were 92.9% -100.0%, 89.2% -100.0%, 73.9% -99.4% and 89.3% -99.0%, respectively.