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目的测序并比较多斑按蚊复合体5成员种(伪威氏按蚊、多斑按蚊、威氏按蚊、达罗毗按蚊及塞沃按蚊)mtD-NA-COⅠ基因序列差异,并探讨其系统进化关系。方法多斑按蚊复合体经形态学和ITS2分子鉴定种型后,PCR扩增多斑按蚊复合体mtDNA-COⅠ基因序列,采用Chromas软件核对,必要时手动调整,采用Bioedit软件进行比对分析,采用Mega3.0软件构建系统进化树。结果 PCR扩增mtDNA-COⅠ基因序列全长约645～660bp,平均G+C含量为29.1%,A+T含量为70.9%。5种不同算法计算的5成员种成对种间距离有较小偏差,但种间遗传距离范围基本趋于一致,以塞沃按蚊、达罗毗按蚊及多斑按蚊亲缘关系最近,其后依次为威氏按蚊和伪威氏按蚊。结论根据mtDNA-COⅠ基因序列,采用不同方法构建的多斑按蚊复合体系统进化树结果基本一致,该基因序列适用于多斑按蚊复合体分析系统学分析。
Objective To sequence and compare the sequence differences of mtD-NA-COⅠ gene among 5 species of Anopheles maculatus complex (Anopheles pandanus, Anopheles stephensi, Anopheles stephensi, Anopheles stephensi and Anopheles stephensi) And explores its systematic evolutionary relationship. Methods The mtDNA-COⅠ gene sequences of Anopheles maculatus complex were amplified by morphological and ITS2 molecular typing. The sequence of mtDNA-COⅠ was identified by Chromas software, adjusted manually if necessary, and analyzed by Bioedit software , Using Mega3.0 software to build phylogenetic tree. Results The sequence of mtDNA-COⅠ gene was about 645-660 bp in length with average G + C content of 29.1% and A + T content of 70.9%. The five species calculated by five different algorithms showed a small deviation from the distance between pairs of species, but the range of genetic distances among species tended to be the same. The closest genetic relationship was found between Anopheles stephensi, Anopheles stephensi and Anopheles stephensi, Subsequent to the order Anopheles anisopliae and Anopheles pipiens pallets. Conclusion According to the mtDNA-COⅠ gene sequence, phylogenetic tree based on the phylogenetic tree of P. multocida complex constructed by different methods is basically the same. The sequence of the phylogenetic tree is suitable for the phylogenetic analysis of Anopheles maculatus complex.