郑州地区2种菊花病毒CP基因的克隆与序列分析

Cloning and Sequence Analysis of Coat Protein Genes of Two Chrysanthemum Viruses from Zhengzhou

为了探明郑州地区感病菊花上病毒的种类及来源,采集有明显病症的菊花样品,根据已知的番茄不孕病毒(TAV)和菊花B病毒(CVB)的基因序列设计特异性引物,采用RT-PCR扩增其外壳蛋白(CP)基因的方法对这2种病毒进行了检测,然后进行序列同源性及系统进化分析。结果显示,所得TAV的CP基因序列长度为481 bp,NCBI登录号为KU873003;所得3条CVB的CP基因序列长度均为547 bp,NCBI登录号分别为KU873004、KU873005、KU873006。将郑州分离物的CP基因与NCBI上已登录的其他分离物进行比对,TAV核苷酸序列同源性为91%~99%,氨基酸序列同源性为86%~100%;CVB核苷酸序列同源性为76%~99%,氨基酸序列同源性为77%~99%。系统进化分析表明,菊花TAV郑州分离物与北京分离物KP019201、黑龙江分离物KP019202、河南分离物KP019200和山西分离物KP019204的亲缘关系最近;CVB郑州分离物与印度分离物AJ812569、AJ871367的亲缘关系最近。

To investigate the species and origin of viruses that infect chrysanthemum in Zhengzhou area,some leaves of different infective chrysanthemum plant were sampled.According to the sequences submitted on NCBI web,the specific primers of coat protein(CP)genes of tomato aspermy virus(TAV)and chrysanthemum virus B(CVB)were designed to detect the two viruses by RT-PCR method,and then the homologous relationship and phylogenetic analysis of obtained isolates were carried out.The cloned CP gene fragment of TAV was 481 bp(GenBank accession:KU873003)and that of CVB was 547 bp(GenBank accession:KU873004,KU873005,KU873006).The sequence alignments of CP genes from Zhengzhou and the other isolates on NCBI web revealed that the TAV homology was 91%-99%at nucleotide level and 86%-100%at amino acid level,and was 76%-99%at nucleotide level and 77%-99%at amino acid level for CVB isolates.The phylogenetic analysis showed that the Zhengzhou isolate was closely related to Beijing isolate KP019201,Heilongjiang isolate KP019202,Henan isolate KP019200 and Shanxi isolate KP019204 of TAV,and India isolate AJ812569 and AJ871367 of CVB.