Objective To explore gene characteristics of D3 domain of 28S ribosomal DNA in nine common mosquito species of Shandong Province,so as to provide a basis for the molecular identification of the mosquitoes.Methods Mosquitoes were collected from Jining City and identified in the laboratory.Single mosquito genomic DNA was extracted from different species and 28S rDNA D3 was specifically amplified by PCR and sequenced.Gene sequences were blasted in GenBank,and the gene traits were analyzed by ClustalX 1.81,DNAMAN,DNASTAR and Mega6.The phylogenetic tree was constructed.Results The 28S rDNA D3 gene fragments of Culex pipiens pallens,Culex tritaeniorhynchus,Culex bitaeniorhynchus,Aedes albopictus,Aedes vexans,Anopheles sinensis,Armigeres obturbans,Mansonia uniformis,Coquillettidia ochracea were amplified,from 361 bp to 422 bp,GC contents were 52.9％to 57.4％.There were 297 conserved sites,96 variable sites,15 transitionsal pairs,17 transversional pairs,transition:transversion bias was 0.90,and the mean genetic distance was 0.073.The molecular identification of the species based on 28S rDNA D3 was consistent with the traditional morphological identification.The phylogenetic tree showed that the mosquitoes of different orders were in different branches.The genus Culex and genus Aedes belong to different branches.Mansonia uniformis and Coquillettidia ochraces were grouped together,and Anopheles sinensis was an independent branch.Conclusion 28S rDNA D3 can serve as an useful classification and identification molecular marker for different mosquito species and genus.
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