黄瓜花叶病毒贵州辣椒分离物外壳蛋白基因序列分析

Sequence analysis of the coat protein gene of Cucumber mosaic virus isolated from Guizhou pepper

【目的】分析贵州辣椒主产区黄瓜花叶病毒(Cucumber mosaic virus,CMV)分离物的分子变异情况,为抗病毒辣椒品种选育及辣椒病毒病的防控提供科学依据。【方法】以来自贵州省8个县(榕江、大方、石阡、安龙、绥阳、德江、关岭和惠水)的16个辣椒CMV分离物为材料,采用逆转录-聚合酶链反应(RT-PCR)扩增辣椒CMV分离物的CP基因序列,利用DNAMAN和BioEdit对CMV分离物的CP基因序列进行比对分析,用MAGE 7.0的邻接法(Neighborjoining,NJ)构建系统发育进化树,分析贵州辣椒CMV分离物的遗传多样性。【结果】将16个贵州辣椒CMV分离物的CP基因核苷酸序列及编码蛋白氨基酸序列与CMV亚组代表株系进行一致性比对,结果表明,贵州辣椒CMV分离物与CMV亚组分离物的CP基因核苷酸序列一致性为89.7%~100.0%、CP蛋白氨基酸序列一致性为96.8%~100.0%。系统进化分析结果表明,CMV-DF、CMV-RJ、CMV-SQ、CMV-SY、CMV-DJ与CMV I代表株系聚为一个亚支,但更趋近于CMV亚组IB株系,CMV-HS、CMV-GL、CMV-AL与CMV亚组Ⅱ株系聚为另一个亚支,说明侵染贵州辣椒的CMV株系有两大组群,其中大方、榕江、绥阳、石阡和德江CMV分离物归属于CMV亚组IB株系,惠水、关岭和安龙CMV分离物归属于CMV亚组Ⅱ株系。【结论】不同地域的贵州辣椒CMV株系发生了变异。

【Objective】The molecular variation of Cucumber mosaic virus(CMV)pepper isolates in the main producing areas of pepper in Guizhou was analyzed to provide theoretical basis for the breeding of antiviral pepper varieties and preventive measures of CMV infection.【Method】Taking 16 pepper CMV isolates from 8 counties(Rongjiang,Dafang,Shiqian,Anlong,Suiyang,Dejiang,Guanling and Huishui)of Guizhou Province as materials,reverse transcription-polymerase chain reaction(RT-PCR)amplification,cloning,sequencing and other methods were used to obtain the full length CP gene of CMV isolates from pepper.The DNAMAN and BioEdit were used to analyze and compare the CP gene sequence of CMV isolates.The adjacency algorithm of MAGE 7.0(Neighbor-joining,NJ)constructed a phylogenetic tree and analyzed the genetic diversity of Guizhou pepper CMV isolates.【Result】The CP nucleotide sequences and the amino acid sequences encoding the protein of 16 CMV isolates from Guizhou pepper were compared with the representative strains of CMV subgroup.The results showed that the nucleotides of CMV isolate and CMV subgroup isolate sequence identity was 89.7%-100.0%,amino acid identity was 96.8%-100.0%.Phylogenetic analysis showed that the CMVDF,CMV-RJ,CMV-SQ,CMV-SY,CMV-DJ and CMV I representative lines were clustered into one subbranch,but closer to the CMV subgroup IB line;CMV-HS,CMV-GL,CMV-AL and CMV subgroupⅡstrains were clustered into another subbranch.These results showed that there were two major groups of CMV strains infected with pepper in Guizhou,Dafang,Rongjiang,Suiyang,Shiqian and Dejiang CMV isolates belonged to CMV subgroup IB strain,Huishui,Guanling and Anlong CMV isolates belonged to CMV subgroupⅡstrain.【conclusion】Variation happens on CMV Guizhou pepper isolates in different geographical origins.