成都机场口岸常见媒介蝇类的分子鉴定

Molecular Identification of the Common Fly Species in the Air frontier port in ChengDu

目的:建立以线粒体COI基因作为分子标记的蝇类分子鉴定方法。方法:设计COI基因引物对成都双流机场口岸常见4科10属14种卫生蝇类进行PCR扩增和序列测定,并与不同地区蝇类的COI基因片段比较,分析其碱基差异和系统发育关系。结果:部分不同地理来源的种类种内碱基差异变化较大,中国四川与马来西亚棕尾别麻蝇碱基差异为3.4%,中国四川与中国广东巴浦绿蝇碱基差异为3.3%;其他种类种内碱基差异较小(<1.0%)。系统发育分析显示所有种类的COI基因分子鉴定结果和形态学结果相一致。麻蝇科种类聚为一个分支,而丽蝇科、花蝇科和蝇科种类没有聚集形成单系。结论:可以将COI基因片段作为分子标记,建立一种适用于口岸的蝇类种类之间分子鉴定方法。

Objective： To establish the molecular method of cytochrome coxidase subunit （COl） for the identification of the fly with traditional morphology appraisal mutually. Methods： The specific PCR primers were designed to amplify sequences about 14 species in 4 families, 10 genus. The base difference and phylogenetic relationship among the species based on sequence data of the CO1 gene were analyzed. Results： The intraspecific variation from the different regions was large. The sequence base difference was calculated at 3.4% about Boettcherisca peregrine between SiChuan, China and Malaysia, 3.3% about Lucilia papuensis between SiChuan and GuangDong, China, less 1.0% about others. The result of phylogeny showed that the CO1 molecular identification was consistent with the morphological characteristics among all the species. Though the Sarcophagidae family formed a clade, three families, Calliphoridae, Anthomyiidae and Muscidae diverged into no momophyletic. Conclusions： The COI gene as a molecular marker could be used to identify the fly species in frontier ports. However, through the phylogenetic analysis COI gene was suitable for the interspecific discrimination, not applicable to the higher order.