利用DNA条形码技术对口岸白纹伊蚊的分子溯源研究

Application of DNA barcoding technology in molecular tracing of Aedes albopictus at ports

目的 建立国境口岸白纹伊蚊的分子溯源方法,区分不同地理种群的白纹伊蚊。方法 选择国境口岸采集点,采集白纹伊蚊,经形态学鉴定后,单只蚊提取基因组DNA,进行PCR扩增COI基因序列,产物经电泳鉴定后,进行双向测序。序列经分析后,去除引物序列和不稳定序列,拼接后提交至NCBI和BOLD数据库进行比对,并使用MAGE 7.0建立系统进化树进行分析。结果 获得青岛、厦门、磨憨3个国境口岸共86只白纹伊蚊蚊的COI基因序列,经分析3个口岸白纹伊蚊COI基因序列基本可以区分开。结论 本研究初步建立了国境口岸白纹伊蚊的分子溯源方法。

Objective To establish molecular traceability method for Aedes albopictus at frontier ports to distinguish different geographical populations.Methods Ae.albopictus were collected at frontier ports.After morphological identification, the genomic DNA was extracted from a single mosquito, PCR products aimed at cytochrome oxidase I(CO1) sequences were identified by electrophoresis and bidirectional sequencing was performed.After sequencing, primers and unstable sequences of the CO1 sequences were removed, spliced and submitted to NCBI and BOLD database for comparison, and the phylogenetic tree was built using MAGE 7.0.Results The COI gene sequences of 86 Ae.albopictus accepted from three ports of Qingdao, Xiamen and Mohan ports were distinguished clearly.Conclusion This study preliminarily established the molecular tracing method of Ae.albopictus at the frontier ports.