China Plugs Mosquito Subgenus Some Species Of Molecular Systematics (diptera: Culicidae)

Subgenus Cellia is a kind of important medicinal insect in the Anopheles(Dipeter:Culicidae), some species are important malaria vector.There are 30 species record in China.At the present time, studies about the phylogeny and evolution of Cellia have focus on some complexs,groups and series, which cannot analysis phylogentic relationship of subgenus Cellia.we reconstruct the phylogentic relationship of subgenus Cellia mosquitoes from China inferred by mitochondrial DNA and ribosomal DNA.The stady can offer academic information for mosquito control,and improve the level of Chinese mosquito biology and vector biology process study.The samples of Cellia mosquitoes were collected from distribution sites from China.Genomic DNA was extracted from single specimens,the mtDNA-COⅡ,rDNA-28S-D3 and rDNA-ITS2 were amplified and sequenced.At least two individuals each species were chosen to analyze.All the sequences were aligned by Clustal X software.Sequence statistics were performed by MEGA V3.1 software.Several statistical approaches were used to evaluate the level on phylogenetic signal in the DNA sequence data sets.Phylogenetic analyses under the Neighbor Joining(NJ),maximum parsimony(MP) and maximum likelihood(ML) optimal criteria were performed by PAUP*4.0b10 software.Bayesian inference also was employed to reconstruct the phylogenetic tree by MrBayes V3.0 software.Twenty-three of COⅡ,23 of D3 and 15 of ITS2 sequences from Cellia mosquitoes were obtained in this study.The phylogenetic signal was strong in all datasets,including COⅡ,D3,ITS2, D3+ITS2,COⅡ+D3,COⅡ+ITS2 and COⅡ+D3+ITS2.By the criterion of Incongruence Length Difference(ILD) test,only COⅡ+ITS2 datasets congruence cannot be rejected.We reconstructed 28 phylogenetic trees inferred by 7 datasets.The trees were compares and evaluated based on the outgroup and the clusters.The results showed that the 50%consense tree between MP,NJ and ML by COⅡand COⅡ+D3+ITS2,and ML tree by D3 were more reasonable.The evolution relationship among Cellia mosquitoes was as follows:1.The relationship of Myzomyia Series could be described as:(((An.minimus/An.minimus A, An.harrisoni/An.minimus C) An.aconitus) An.culicifacies).The analysis by D3 with An. pampanai,showed that was closer to An.aconitus.The position of An.jeyporiensis could not be confirmed.2.In the Neomyzomyia Series,An.dirus/An.dirus A was all close to An.baimaii/An.dirus D, but the places of An.kochi and An.tessellates was not stable. 3.The relationship among Neocellia Series was complicated.The monophyly of An.maculatus complex could not be confirmed completely,because the position of An.pseudowillmori was unfixed.In An.annularis group,it was showed((An.nivipes,An.philippinensis) An.annularis) and cluster with An.splendidus by ITS2 and COⅡ+ITS2,but the position of An.splendidus was different by other markers.The Neocellia Series was not monophyly caused by the position of An.splendidus,An.ramsayi and An.philippinensis in this study.4.For An.indefiniyus and An.vagus of Pyretophorus Series,only involved in COⅡ,could not be explained completely.5.All the trees supported the paraphyly of Neoccllia Series and Neomyzomyia Series,cluster with Myzomyia Series afterwards.Only by D3,the Pyretophorus Series cluster before Myzomyia Series.