福建省常见尸食性蝇类的COⅠ及16S rDNA序列鉴定

COⅠand 16S rDNA Sequence Identification of Common Necrophagous Flies in Fujian Province

目的利用线粒体细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunitⅠ,COⅠ)以及16S核糖体脱氧核糖核酸(ribosomal deoxyribonucleic acid,rDNA)基因片段序列对福建省常见尸食性蝇类种属进行分子鉴定,探讨两种遗传标记的鉴别效力。方法收集福建9个地区案件现场捕获的常见尸食性蝇类样本22只,经形态学鉴定后提取DNA,扩增COⅠ及16S rDNA基因片段,测序后上传GeneBank数据库,使用BLAST、MEGA 10.0软件进行序列比对、同源性分析以及种内和种间遗传距离分析,并采用非加权组平均法(unweighted pair-group method with arithmetic means,UPGMA)分别构建福建省常见尸食性蝇类COⅠ及16S rDNA基因片段序列的系统发育树。结果经形态学鉴定,共收集到3科5属6种常见尸食性蝇类。基因序列分析结果显示,16S rDNA基因片段序列的种间遗传距离均数为1.8%~8.9%,种内遗传距离均数为0.0%~2.4%;COⅠ基因片段序列的种间遗传距离均数为7.2%~13.6%,种内遗传距离均数为0.0%~6.3%。结论COⅠ和16S rDNA基因片段序列均能够对不同蝇种的种属进行鉴别,16S rDNA对于福建省常见丽蝇科尸食性蝇类的种属鉴别价值更高。

Objective To identify the species of common necrophagous flies in Fujian Province by gene fragment sequences of mitochondrial cytochrome c oxidase subunitⅠ(COⅠ)and 16S ribosomal deoxyribonucleic acid(16S rDNA),and to explore the identification efficacy of these two molecular markers.Methods In total 22 common necrophagous flies were collected from the death scenes in 9 different regions in Fujian Province and DNA was extracted from the flies after morphological identification.The gene fragments of COⅠand 16S rDNA were amplified and sequenced.All the sequences were uploaded to GeneBank and BLAST and MEGA 10.0 software were used to perform sequence alignment,homology analysis and intraspecific and interspecific genetic distance analysis.The phylogenetic trees of DNA fragment sequences of COⅠand 16S rDNA of common necrophagous flies in Fujian Province were established by unweighted pair-group method with arithmetic means(UPGMA),respectively.Re⁃sults The flies were classified into 6 species,5 genera and 3 families by morphological identification.The results of gene sequence analysis showed that the average number of interspecific and intraspecific genetic distance of 16S rDNA ranged from 1.8%to 8.9%and 0.0%to 2.4%,respectively.The average number of interspecific and intraspecific genetic distance of COⅠranged from 7.2%to 13.6%and 0.0%to 6.3%,respectively.Conclusion The gene sequences of COⅠand 16S rDNA can accurately identify the species of different necrophagous flies,and 16S rDNA showed higher value in species identification of common calliphoridae necrophagous flies in Fujian Province.