韭葱黄条病毒黑龙江分离物外壳蛋白基因克隆与序列分析

Cloning and sequences analysis of CP gene of Leek yellow stripe virus Heilongjiang isolate

韭葱黄条病毒（Leek yellow stripe virus,LYSV）是大蒜上的重要病毒病害,几乎分布于全国所有大蒜种植区域,严重影响大蒜的产量和品质。对其建立RT-PCR分子检测体系以及对其外壳蛋白（CP）基因进行序列分析尤为重要。根据已报道的LYSVCP基因的核苷酸序列设计合成1对引物,以带毒植株总RNA为模板,应用RT-PCR方法,克隆了LYSV黑龙江分离物（LYSV-HLJ）CP基因全长cDNA序列。结果表明,LYSV-HLJCP基因全长885bp,编码295个氨基酸残基;与GenBank中其他不同分离物的CP核苷酸序列同源性为79.7%～88.6%,氨基酸序列同源性为83.7%～93.1%。依据LYSVCP氨基酸序列建立系统进化树,将LYSV不同分离物划分为4个类群,LYSV-HLJ与韩国和巴西大蒜上的LYSV有较近的亲缘关系,同属于ⅳ类,表现出一定的寄主相关性。

Leek yellow stripe virus （LYSV） is the important virus disease in garlic, which distributes nearly in all the garlic planted area, and can effect the yield and quality of garlic, so erecting RT-PCR molecule detecting system and CP sequence analysising are important. A pair of primers was designed based on the reported CP gene of LYSV the hole CP gene sequence of LYSV Heilongjiang isolate was cloned by reverse transcription polymerase chain reaction （RT-PCR）. The results showed that CP gene of LYSV-HLJ was made up of 885 nucleic acids, coding 295 amino acids. The CP nucleic acid sequence homology of LYSV-HLJ and other isolates of LYSV in GenBank was between 79.7%-88.6%, the CP amino acid sequence homology was between 83.7%-93.1%. The phylogenic tree which established based on LYSV CP amino acid was clustered into four groups, the isolate of LYSV-HLJ has nearer sibship with the isolates of South Korea and Japan, both in IV group, represented host correlation.