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目的检测嗜尸性蝇类线粒体DNA(mtDNA)细胞色素氧化酶辅酶I(COI)中278bp序列,鉴定蝇科3属5种嗜尸性蝇,解决其形态学种类鉴定的难题,为死亡时间推断提供技术支持。方法从15省、市(地区)室外草地家兔尸体上采集蝇科3属5种共计18个蝇类成蝇样本,提取mtDNA进行PCR扩增,序列测定且上传GENBANK,并使用MEGA软件进行序列比对及进化距离分析,构建不同地区各种内和种间系统发育树,尝试区分常见嗜尸性蝇类。结果基因序列分析结果显示,上述蝇类mtDNA上COI基因序列的种内进化分歧均数在0%～0.8%,种间进化分歧均数在7.6%～13.4%之间;进化距离分析显示,在种间水平,家蝇和孕幼家蝇、斑趾黑蝇和家蝇、厩螫蝇和厚环黑蝇Bootstrap检验的可信度分别为56%、31%、53%;种内水平除了家蝇和厚环黑蝇外,其他Bootstrap检验可信度均为100%。结论嗜尸性蝇类mtDNA中COI序列分析能够区分常见嗜尸性蝇类,作为形态学方法的补充手段,实现精确的蝇种鉴定。
Objective To detect the 278bp sequence of mitochondrial DNA (mtDNA) cytochrome oxidase coenzyme I (COI) in moths and to identify 5 species of sarcophagous flies in 3 genera of Muscidae, to solve the problem of their morphological identification and to estimate the time of death provide technical support. Methods A total of 18 adult flies from 3 genera and 3 genera of Muscidae were collected from the dead bodies of rabbits in 15 provinces and municipalities. The mtDNA was extracted for PCR amplification, sequenced and GENBANK was sequenced and sequenced using MEGA software Comparison and evolutionary distance analysis, to build a variety of different types of internal and interspecific phylogenetic trees, trying to distinguish common sarcoides. Results The results of gene sequence analysis showed that the average divergences of COI gene sequences in flies were between 0% and 0.8%, and the average divergence between species was between 7.6% and 13.4%. The evolution distance analysis showed that The confidence level of the Bootstrap test was 56%, 31%, 53% for interspecific levels, housefly and pregnant-child housefly, black-flies and housefly, black flies and black flies, respectively. Flies and black-throated blackflies, all other Bootstrap tests have a 100% confidence level. Conclusion The COI sequence analysis of mtDNA in sarcophagous flies can distinguish common sarcophagous flies, as a complement to morphological methods, to achieve accurate identification of fly species.