贵阳地区常见尸食性蝇类mtDNA分子标记的检测

Using the mtDNA sequence identify the common species of necrophagous flies in Guiyang

目的应用mt DNA中COI（348bp）、COII（637bp）及16Sr DNA（513bp）基因序列鉴定贵阳地区常见尸食性蝇类的种属。方法收集贵阳地区常见尸食性蝇类标本,经形态学分类鉴定后提取胸肌DNA,扩增COI（348bp）、COII（637bp）及16Sr DNA（513bp）基因片段,测序后用DNAMAN 4.0序列分析软件进行序列拼接,NCBI中的BLAST、MEGA 5.2软件包对所得序列分析建立种内及种间进化分歧率,并构建系统发育树。结果 COI种间平均进化分歧率为12.55%,种间范围在3.0%-18.6%,种内范围在0%-0.7%;COII种间平均进化分歧率为13.73%,种间范围在5.3%-21.6%,种内范围在0%-0.6%;16Sr DNA种间平均进化分歧率为4.54%,种间范围在0.4%-7.6%,种内范围在0%-0.4%。结论 COI、COII及16Sr DNA可用于尸食性蝇类的种属鉴定,该方法可作为传统分类方法的补充手段。

Objective This research was aimed to identify the common species of necrophagous flies in Guiyang using mtDNA sequence analysis of COI gene （ 348 bp）, COII gene （637 bp ） and 16 SrDNA （ 513 bp）. Methods Necrophagous flies were collected and classified by entomologists with traditional morphological characteristics. DNA of necrophagousflies was extracted and the COI gene, COII gene and 16SrDNA of mtDNA were analyzed. DNAMAN 4.0 sequence analysis software, NCBI BLAST and MEGA 5.2 software package were used for sequence analysis, genetic distance computation and phylogenetic tree construction. Results The rate of cladogenesis between species in COI ranged from 3.0% to 18.6% with an average of 12.55% while the rate within species varied from 0% to 0.7%. The rate of cladogenesis between species varied from 0% to 0.6%. The rate of cladogenesis between species in 16SrDNA ranged from 0.4% to 7.6% with an average of 4.54% while the rate within species varied from 0% to 0.4%. Conclusion As a supplementary method to the traditional identification, sequence analysis of COI gene, COII gene and 16SrDNA could be used to identify necrophagousflies in Guivang.