基于mtDNA-COI基因序列分析我国中华按蚊种群的遗传结构  

Analysis of population genetic structure of Anopheles sinensis based on mitochondrial DNA cytochrome oxidase subunit I gene fragment

作　　者：常雪莲[1] 钟代斌[2] 李小聪 黄亚铭[4] 朱国鼎[5] 魏星[1] 夏惠[1] 陈晓光 方强[1] 

机构地区：[1]蚌埠医学院病原生物学教研室,安徽蚌埠233030 [2]美国加州大学欧文分校健康科学院公共卫生部,加州尔湾92617 [3]广东普通高校新发传染病防治重点实验室,广东广州510515 [4]广西壮族自治区疾病预防控制中心,广西南宁530028 [5]江苏省血吸虫病防治研究所,江苏无锡214064

出　　处：《南方医科大学学报》2015年第2期234-238,247,共6页Journal of Southern Medical University

基　　金：美国国立卫生研究院NIH项目(D43TW009527-01);广东省普通高校新发传染病防治重点实验室开放基金项目(KLB09007);安徽省自然科学基金(1308085MH160);蚌埠医学院自然科学基金重点项目(BYKY1403ZD)

摘　　要：目的探讨我国中华按蚊种群遗传多样性、遗传分化及种群系统发育关系。方法本实验采用2010-2012年间在我国山东,安徽,江苏,贵州,广西和云南等6个不同地理环境采集的中华按蚊样品,进行线粒体COI基因扩增并测序。采用Bioedit 7.0软件对测序结果进行比对分析;运用Dna SP 5.0软件确定种群遗传结构特点;应用Arlequin 3.1软件计算种群的遗传距离及遗传分化系数;通过IBDWS在线软件来明确遗传距离与地理距离的相关性;使用PHYLIP 3.6软件构建种群系统发育进化树。结果六个中华按蚊种群共123个雌蚊个体的PCR产物成功扩增和测序。PCR扩增中华按蚊线粒体COI基因序列大小为814 bp,平均A+T含量(71.2%)>平均G+C含量(28.8%)。线粒体COI序列分析结果显示,种群具有丰富的遗传多样性;分子变异等级分析表明中华按蚊遗传分化主要来自于种群内部;种群间存在一定的地理隔离现象。中性检验,错配分析及聚类进化树结果显示:中华按蚊种群发展经历扩张状态,而云南种群相对其他种群较独立,在系统树中为孤立一支。结论线粒体COI基因可以作为研究中华按蚊种群遗传结构和系统进化的理想分子标志。云南种群遗传发育较其他地理种群具有一定特殊性。Objective To study the population genetic variation, genetic diversity and phylogenesis of Anopheles sinensis in China. Methods Anopheles sinensis samples collected from Shandong, Anhui, Jiangsu, Guizhou, and Yunnan Provinces and Guangxi Zhuang Autonomous Region with different geographical conditions between 2010 and 2012 were analyzed by mitochondrial DNA cytochrome oxidase subunit I （mtDNA-COI） gene amplification and sequencing. Bioedit 7.0 and DnaSP 5.0 software was used to compare the gene sequences and analyze the population genetic structure, respectively. Arlequin 3.1 was used to calculate the genetic distance and parameters of population differentiation. The relationship between the geographic and genetic distances was analyzed using IBD Web Service. PHYLIP 3.6 was used to construct the phylogenetic tree. Results PCR amplification and sequencing was performed successfully for 6 Anopheles sinensis populations containing 123 female mosquitoes. The length of mtDNA-COI gene fragment was 841 bp with an average A＋T content of 71.2% and G＋C content of 28.8%. High nucleotide diversity and genetic differentiation were observed among the Anopheles sinensis populations based on mtDNA-COI gene. Analysis of the molecular variance revealed a greater variation between populations than that within populations with isolation by distance between the populations. The Anopheles sinensis populations appeared to have undergone expansion, but the Yunnan population constituted an isolated branch in the phylogenetic tree. Conclusion mtDNA-COI can serve as the molecular marker to analyze population genetic variation and phylogenesis of Anopheles sinensis. The Yunnan population shows a phylogenetic difference from the other populations analyzed in this study.

关 键 词：中华按蚊 mtDNA-COI 种群遗传 

分 类 号：Q963[生物学—昆虫学]