中国媒介生物学及控制杂志 ›› 2017, Vol. 28 ›› Issue (6): 530-534.DOI: 10.11853/j.issn.1003.8280.2017.06.004

• 论著 • 上一篇    下一篇

9种蚊虫细胞色素C氧化酶亚基Ⅰ基因的分子进化分析

郭秀霞, 程鹏, 刘丽娟, 史琦琪, 王海防, 张崇星, 王怀位, 公茂庆   

  1. 山东省医学科学院, 山东省寄生虫病防治研究所, 山东省医药卫生媒介生物学重点实验室, 山东 济宁 272033
  • 收稿日期:2017-06-20 出版日期:2017-12-20 发布日期:2017-12-20
  • 通讯作者: 公茂庆,Email:maoqingg@yahoo.com
  • 作者简介:郭秀霞,女,硕士,助理研究员,从事病原生物学研究,Email:guoguoxiua@163.com
  • 基金资助:
    国家自然科学基金(81471985);山东省医学科学院医药卫生科技创新工程

Molecular evolution analysis of mitochondrial DNA cytochrome C oxidase subunitⅠgene among nine mosquito species

GUO Xiu-xia, CHENG Peng, LIU Li-juan, SHI Qi-qi, WANG Hai-fang, ZHANG Chong-xing, WANG Huai-wei, GONG Mao-qing   

  1. Shandong Institute of Parasitic Diseases, Shandong Academy of Medical Sciences, Shandong Province Key Laboratory of Health and Vector Biology, Jining 272033, Shandong Province, China
  • Received:2017-06-20 Online:2017-12-20 Published:2017-12-20
  • Supported by:
    Supported by the National Natural Science Foundation of China(No. 81471985)and the Innovation Project of Shandong Academy of Medical Science

摘要: 目的 分析常见的9种蚊虫线粒体DNA细胞色素C氧化酶亚基Ⅰ(mtDNA-COⅠ)基因序列特征,了解其分子系统进化关系。方法 2016年采用人饵帐诱捕法在济宁市北湖地区采集蚊虫,经实验室种类鉴定后,提取淡色库蚊、三带喙库蚊、二带喙库蚊、白纹伊蚊、刺扰伊蚊、中华按蚊、骚扰阿蚊、常型曼蚊和黄色轲蚊的DNA,PCR扩增mtDNA-COⅠ基因并测序。在GenBank中进行同源性比对,采用Bioedit 7.0软件及DNAman软件对测序结果进行比对分析,通过DNAStar、ClustalX 1.81和Mega 6.0软件分析序列特征、颠换率、分化年代并构建系统进化树,探讨系统发生关系。结果 9种蚊虫的mtDNA-COⅠ基因成功扩增并测序,获得长度均为528 bp的基因片段,(A+T)含量为65.72%~69.32%,有375个保守位点,153个变异位点,104个简约信息位点,49个单态位点。不同蚊种间的颠换率为3.409%~9.470%,分化年代为1.482×106a~4.117×106a。不同蚊种间序列同源性为84.28%~92.94%。系统进化关系显示,所有蚊虫的COⅠ基因分子鉴定与形态学鉴定一致,同属间聚集,不同蚊种为独立分支。结论 mtDNA-COⅠ基因可用于蚊虫属和种的区分,为蚊虫的分子鉴定提供科学依据。

关键词: 蚊虫, 线粒体DNA, 细胞色素C氧化酶亚基Ⅰ, 分子进化

Abstract: Objective To explore gene characteristics of mitochondrial DNA cytochrome C oxidase subunitⅠ (mtDNA-COⅠ) in nine common mosquito species and analyze the corresponding phylogenetic relationship of these mosquitoes. Methods Nine species of mosquitoes (Culex pipiens pallens, Cx. tritaeniorhynchus, Cx. bitaeniorhynchus, Aedes albopictus, Ae. vexans, Anopheles sinensis, Armigeres subalbatus, Mansonia uniformis, Coquillettidia ochracea) were collected with man-bait net trap from Beihu area of Jining district in 2016 and identified in the laboratory. Single mosquito genomic DNA was extracted from each species and mtDNA-COⅠ was specifically amplified by PCR and sequenced. Gene sequences were blasted in GenBank, compared and analyzed by Bioedit 7.0 and DNAman. The gene traits, transversion rates, and divergence times were analyzed by DNAStar、ClustalX 1.81 and Mega 6.0. The phylogenetic tree was constructed. Results The amplified mtDNA-COⅠfragments of nine species of mosquitoes were 528 bp with (A+T)contents 65.72%-69.32%. There were 375 conserved sites, 153 variable sites, 104 parsimony informative sites and 49 singleton sites in the sequences. The transversion rates were from 3.409% to 9.470%, and the divergence times were from 1.482×106a to 4.117×106a. The nucleotide sequence homology among the different mosquito species were 84.28%-92.94%. Molecular phylogenetic study showed that different species were clustered at their own branch at genus level, which was consistent with traditional morphological identification. Conclusion mtDNA-COⅠ can serve as a useful classification and identification molecular marker for different mosquito species and genus, providing a molecular method for the identification and phylogenetic analysis of the mosquitoes.

Key words: Mosquito, Mitochondrial DNA, Cytochrome C oxidase subunit Ⅰ, Molecular evolution

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