基于复制酶基因序列的黄瓜绿斑驳花叶病毒系统进化及生物信息学分析  

Phylogenetic and Bioinformatics Analysis of Replicase Gene Sequence of Cucumber Green Mottle Mosaic Virus

作　　者：梁超琼[1] 孟嫣[1] 罗来鑫[1] 刘鹏飞[1] 李健强[1] 

机构地区：[1]中国农业大学植物病理学系种子病害检验与防控北京市重点实验室,北京100193

出　　处：《病毒学报》2015年第6期620-628,共9页Chinese Journal of Virology

基　　金：公益性行业(农业)科研专项(201303028);国家自然科学基金项目(31371910)

摘　　要：本研究对江苏、浙江、湖南和北京等四个地区的葫芦、西瓜、黄瓜、西葫芦和甜瓜上的黄瓜绿斑驳花叶病毒（Cucumber green mottle mosaic virus,CGMMV）129kD和57kD蛋白复制酶基因分别进行扩增、测序和序列分析。结果显示,供试CGMMV分离物CGMMV-No.1、CGMMV-No.2、CGMMV-No.3、CGMMV-No.4和CGMMVNo.5的129kD和57kD蛋白复制酶基因序列相似性分别为99.64%和99.74%,其中CGMMV-No.1、CGMMVNo.3和CGMMV-No.4的相似性较高,三者的129kD和57kD蛋白复制酶基因序列相似性分别为99.95%和99.94%,而CGMMV-No.2的129kD和57kD蛋白复制酶基因序列与其余四个参比序列的相似性相对较低,分别为99.16%~99.27%和99.04%~99.18%;供试样品在基于129kD和57kD蛋白复制酶基因序列构建的NJ系统发育树中均被分为6个类群,CGMMV-No.1、CGMMV-No.3、CGMMV-No.4与GenBank上已报道的中国山东分离物（Accession No.KJ754195）聚为一支,CGMMV-No.5与中国辽宁分离物（Accession No.EF611826）聚为一支,而浙江的CGMMV-No.2西瓜分离物在129kD蛋白复制酶基因系统发育树中与韩国西瓜分离物（Accession No.AF417242）聚为一支,其在57kD蛋白复制酶基因系统发育树中独立成群。本研究中供试CGMMV分离物的129kD和57kD蛋白均无显著疏水性,也无高度卷曲螺旋部位,ProtParam预测显示仅CGMMV-No.4的129kD蛋白为稳定蛋白,其余均为不稳定蛋白,CGMMV-No.1、CGMMV-No.2、CGMMV-No.3和CGMMV-No.5的129kD蛋白分别有6个、6个、2个和4个可能的跨膜结构区域,CGMMV-No.2、CGMMV-No.4和CGMMV-No.5的57kD蛋白分别有13个、13个和5个可能的跨膜结构域,供试样品129kD蛋白的糖基化位点分别有2个、4个、4个、4个和4个,57kD蛋白的糖基化位点分别有2个、5个、2个、5个和2个,其129kD和57kD蛋白的紊乱区、球蛋白区、磷酸化位点以及B细胞抗原表位位点均存在差异。综合分析认为,供试CGMMV分离物复制酶基因序列相似性较高,种内稳定且保守,种间差异�The replicase genes of five isolates of Cucumber green mottle mosaic virus from Jiangsu,Zhejiang,Hunan and Beijing were amplificated,sequenced and analyzed.The similarities of nucleotide acid sequences indicated that 129 kD and 57 kD replicase genes of CGMMV-No.1,CGMMV-No.2,CGMMVNo.3,CGMMV-No.4and CGMMV-No.5 were 99.64% and 99.74%,respectively.The similarities of129 kD and 57 kD replicase genes of CGMMV-No.1,CGMMV-No.3and CGMMV-No.4 were 99.95%and 99.94%,while they were lower between CGMMV-No.2and the rest of four reference sequences,just from 99.16%to 99.27% and from 99.04%to 99.18%.All reference sequences could be divided into six groups in neighbor-joining（NJ）phylogenetic trees based on the replicase gene sequences of 129 kD,57kD protein respectively.CGMMV-No.1,CGMMV-No.3and CGMMV-No.4 were clustered together with Shandong isolate（Accession No.KJ754195）in two NJ trees;CGMMV-No.5was clustered together with Liaoning isolate（Accession No.EF611826）in two NJ trees;CGMMV-No.2was clustered together with Korea watermelon isolate（Accession No.AF417242）in phylogenetic tree of 129 kD replicase gene of CGMMV;Interestingly,CGMMV-No.2was classified as a independent group in phylogenetic tree of 57 kD replicase gene of CGMMV.There were no significant hydrophobic and highly coiled coil regions on 129 kD and 57 kD proteins of tested CGMMV isolates.Except 129 kD protein of CGMMV-No.4,the rest were unstable protein.The number of transmembrane helical segments（TMHs）of 129 kD protein of CGMMVNo.1,CGMMV-No.2,CGMMV-No.3and CGMMV-No.5were 6,6,2and 4,respectively,which were13,13 and 5on the 57 kD protein of CGMMV-No.2,CGMMV-No.4and CGMMV-No.5.The glycosylation site of 129 kD protein of tested CGMMV isolates were 2,4,4,4and 4,and that of 57 kD protein were 2,5,2,5and 2.There were difference between the disorders,globulins,phosphorylation sites and B cell antigen epitopes of 129 kD and 57 kD proteins of tested CGMMV isolates.Thecurrent results that there was no significant difference between the replicase gene sequen

关 键 词：黄瓜绿斑驳花叶病毒 复制酶基因 系统进化树 生物信息学 

分 类 号：S432.4[农业科学—植物病理学]