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从杭州田间表现典型花叶症的甘蔗病叶中得到病毒分离物SC-ZJ,按照SCMV亚组中甘蔗花叶病毒(Sugarcanemosaic virus,SCMV)、玉米矮花叶病毒(Maize dwarf mosaic Virus,MDMV)、高粱花叶病毒(Sorghum mosaic virus,SrMV)、约翰逊草花叶病毒(Johnsongrass mosaic virus,JGMV)等4种病毒基因组序列设计特异性引物,对SC-ZJ进行IC-RT-PCR扩增,结果用SrMV特异性引物扩增到1条长约1.1 kb的片段,而用其它3种病毒特异性引物扩增不出任何片段。扩增片段经克隆后测定序列,序列包含1135核苷酸(nt),编码378个氨基酸(aa),取其中的近全长外壳蛋白(CP)氨基酸序列(312 aa),与SrMV、MDMV、SCMV、JGMV等4种病毒对应的序列进行同源性比较,结果核苷酸序列同源性依次为95.3％、75.2％、61.2％和53.6％,氨基酸序列同源性依次为97.1％、78.1％、72.9％和56.2％。根据IC-RT-PCR扩增鉴定及序列同源性比较结果,将SC-ZJ鉴定为SrMV。并根据CP氨基酸序列的同源性进一步分析了4种病毒的亲缘关系。
The virus isolate SC-ZJ was obtained from the sugarcane leaves of typical mosaic disease in Hangzhou field. According to SCMV, SCMV, Maize dwarf mosaic virus (MDMV) , Sorghum mosaic virus (SrMV), Johnsongra mosaic virus (JGMV) and other four kinds of virus genome sequence specific primers designed for SC-ZJ IC-RT-PCR amplification, the results with SrMV-specific primers amplified to a length of about 1.1 kb fragments, with the other three virus-specific primers did not amplify any fragment. The amplified fragment was cloned and sequenced. The sequence contained 1135 nucleotides (nt) and encoded a polypeptide of 378 amino acids. The full-length CP protein (312 aa) SCMV and JGMV. The homology of nucleotide sequences was 95.3%, 75.2%, 61.2% and 53.6%, and the amino acid sequence homologies were 97.1%, 78.1% , 72.9% and 56.2% respectively. According to the results of IC-RT-PCR amplification and sequence homology comparison, SC-ZJ was identified as SrMV. According to the homology of CP amino acid sequence, the genetic relationship of four viruses was further analyzed.