Taxonomic And Phylogenetic Study On The Subgenus Cellia From Southern China(Diptera:Culicidae:Anopeles)

Mosquitoes of the subgenus Cellia in the Anopheles (Diptera:Culicidea) are mainly distributed in Oriental, Afrotropic, Australasian and Palearctic Region. Up to now,28species have been reported from China, which are classified into4Series. Many species in the subgenus are important medicinal insects, especially vectors of malaria. Many species are actually distinguishable in morphology (cryptic species, named as Complex), which have seriously hindered the study of taxonomy and phylogeny. The fauna and species of Cellia have had a large change with the changes of natural environment and climate in the past decades, and the phylogenetics in the subgenus is still unresolved. Therefore, further investigations will be needed using molecular marker to elucidate the phylogeny of subgenus Cellia.The study reviewed the morphology, phylogeny, and molecular taxonomy of Cellia in China based on literatures, and revised the list and cited references. In the meantime,14Cellia species (238mosquitoes) was identified by morphological examination from8379mosquitoes, which were collected from48geographical sites of13provinces of Southern China from2010-2013. Then, mitochondrial DNAs were extracted from102Anopheles mosquitoes and used for COⅡ amplification and sequencing.22COⅡ sequences of Cellia were obtained by Blast analysis. Nucleotide composition and phylogenetic relationship were further analyzed. Nine of twenty two sequences combined with28Cellia COⅡ sequences from GenBank were retrived and used for phylogenic analysis. A maximum likelihood (ML) tree was constructed by MEGA5.0using Culex pipiens COⅡ as outgroup. Then, genetic relationship and taxonomic status of subgenus Cellia in China were inferred based on the phylogenetic tree.The conclusions are as follows:1. The new species list of Cellia in China (up to2012) consists of4series,6groups,6subgroups,6complexs and28species. An.harrison and An. baimaii was added to the list. The morphological description with corresponding figures of An. baimaii were provided. An. ramsayi was renominated as An. pseudojamesi. 2. It was found that Genus Culex and subgenus Anopheles are dominant species in the investigated localizations. Subgenus Cellia, including An.minimus, An.tessellatus and An.indefintus, are mainly distributed in Hainan and Yunnan provinces. It was inconsist with the previous report that Subgenus Cellia widely distribute in Sichuan, Guangxi, Guizhou and other areas. Taken together, our results showed that the distribution of mosquitoes fauna varied with environmental change.3. Morphological taxonomy results were further confirmed by mtDNA COⅡ. Sequence and phylogenic analysis revealed some differences in phylogenetic relationships stemed from morphological and molecular analysese. Therefore, further studies is needed to elucidate it using other molecular makers.4. The phylogenetic tree based on25species mtDNA COⅡ of Cellia showed that Pyretophorus series was grouped with Neocellia series, and Myzomyia series was clustered with Neomyzomyia series. It supported the view held by Foley and Morgan that Neocellia Series should be grouped with Pyretophorus series. The result also revealed that An.pseudowillmori of Maculatus Group was clustered with An.annularis of Annularis Group, which confimed Walton and Mayajun’s opinon that An.pseudowillmori should be reclassified. The results showed that the phylogenetic tree based on COⅡ could clearly distinguish the members of Cellia.