Chinese Anopheles Genus Anopheles Subgenus Molecular Systematic Studies (diptera: Culicidae)

Anopheles subgenus (Cellia) belongs to Diptera, Nematocera Culicidae mosquitoes subfamily (Anophelinae) Anopheles, and 36 species of eight kinds of groups of it have been recorded in China. In this study, we collect the mosquito species Anopheles subgenus of China and genomic DNA is extracted from single specimens. The mtDNA-COII, mtDNA-COI,rDNA-28S-D3 and rDNA-ITS2 are amplified and sequenced. All the sequences are aligned by Bioedit V5.0.9 and Clustal X1.8.3 software. Sequence statistics are performed using MEGA V4.0 software. The level on phylogenetic signal in the DNA sequence data sets is evaluated based on several statistical approaches. Phylogenetic analyses under the Neighbor Joining (NJ), maximum parsimony (MP) and maximum likelihood (ML) optimal criteria are performed by PAUP*4.0b10 software. Bayesian inference is also employed to reconstruct the phylogenetic tree by MrBayes V3.1 software. Moreover, the identification of Anopheles hyrcanus group from Liaoning Province of China is implemented based on molecular characteristics. The main achievements of our research are as follows:1. We have obtained 15 kinds of mtDNA-COâ…¡sequences of 41 individuals,11 kinds of mtDNA-COâ… sequences of 33 individuals,16 kinds of rDNA-28S-D3 sequences of 69 individuals and 16 kinds of rDNA-ITS2 sequences of 105 individuals, respectively.2. Based on analysis of the mtDNA-COâ…¡gene sequence, it is shown that the Anopheles species are with good resolution and the reconstruction of phylogenetic relationships is divided into four:Firstly, (((An. sinensis+ An. belenrae)+An. kleini+An. hyrcanus)+((An. lesteri+An. kweiyangensis)+(An. kunmingensis+An. liangshanensis)))+An.peditaeniatus+(An.pullus+ An. junlianensis) [An. hycranus group]; Secondly:An. messeae; Thirdly:(An. freyi+An. lindesayi); Lastly:An. barbirostris.3. Based on rDNA-ITS2 gene sequence reconstruction of the Anopheles species of the phylogenetic relationships between the analysis is also divided into four:Firstly:(((((An. belenrae+ An. sinensis)+An. kleini)+An. lesteri)+((An. kunmingensis+An. liangshanensis)+ An. kweiyangensis))+(An. crawfordi+An.peditaeniatus))+((An.junlianensis+An.pullus)+An. hyrcanus)); Secondly:An. messeae; Thirdly:(An.freyi+An.koreicus); Lastly:An. barbirostris. Anopheles hycranus group's An. hyrcanus,An. lesteri and An. kweiyangensis different location from COâ…¡-based, other mosquito species have the same location.4. The analysis of COâ… and D3 sequences reveales that the two markers are not suitable for reconstruction of subgenus Anopheles species phylogeny. 5. In this study molecular identification is implemented on one hundred sixty eight specimens belonging to Anopheles hyrcanus group from Liaonong Province of China including An. sinensis (n=14), An. lesteri (n=4), An. yatsushiroensis (n=113), An. belenrae (n=2) and An. kleini (n=35). Among these species, An. kleini is new record in China.