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目的对蝇类mtDNA 523bp COI和347bp ND5基因片段进行序列分析,评价其在以嗜尸性蝇类种属鉴定中应用的可行性。方法从广州(广东省)、湛江(广东省)、韶关(广东省)、沈丘(河南省)及蜂蛹寨(四川省)采集7种嗜尸性蝇类标本,进行形态学种属鉴定,取其腹部肌肉提取DNA,利用基因特异性引物对线粒体COI、ND5基因进行PCR扩增,产物经纯化后进行测序,MEGA 3.0软件对DNA序列进行碱基组成、进化分歧率和系统发育分析。结果进化分歧率ND5基因种内小于1.83%,种间大于2.62%;种间与种内进化分歧率范围间没有交叉;COI基因种间在0.48%～14.8%之间,种内在0.24%～8.3%之间,种内进化分歧范围与种间进化分歧范围存在交叉。结论 ND5基因片段可在种水平有效鉴别常见嗜尸性蝇类,也可鉴别近缘种。而单独运用COI基因不能有效进行种属鉴定。
Objective To analyze the sequence of flies mtDNA 523 bp COI and 347 bp ND5 gene and evaluate their feasibility in the identification of sarcophagous fly species. Methods Seven species of sarcophagous flies were collected from Guangzhou (Guangdong), Zhanjiang (Guangdong), Shaoguan (Guangdong), Shenqiu (Henan) and bee pupai (Sichuan Province) for morphological identification The DNA was extracted from the abdominal muscles, and the mitochondrial COI and ND5 genes were amplified by PCR using gene-specific primers. The products were purified and sequenced. The MEGA 3.0 software was used to analyze the base composition, evolutionary divergence rate and phylogenetic analysis of DNA sequences. Results The divergence rates of ND5 were less than 1.83% and the inter-species was more than 2.62%. There was no crossover between the divergence rates of interspecific and in-species divergences. The COI genes were between 0.48% and 14.8% within 0.24% and 8.3% %, There is a cross between the scope of divergence within species and the scope of divergence between species. Conclusion The ND5 gene fragment can identify common sarcoid flies at species level and identify related species. However, the COI gene alone can not effectively identify species.