Molecular identification and phylogeny of the Maculatus group of Anopheles mosquitoes (Diptera: Culicidae) based on nuclear and mitochondrial DNA sequences

The Maculatus group of Anopheles mosquitoes (Diptera: Culicidae) comprises eight known species, including important malaria vectors in Southeast Asia. The sequences of the second internal transcribed spacer (ITS2) and third domain (D3) of ribosomal DNA, and cytochrome oxidase subunit II (COII) of mitochondrial DNA were obtained for five species of the group from China, as An. maculatus, An. willmori, An. pseudowillmori, An. sawadwongporni and An. dravidicus. The variation within taxon is much smaller than that between taxa. A diagnostic PCR assay for distinguishing the five members was developed based on the interspecific ITS2 variation. The phylogenetic relationships for the group were estimated on the ITS2 and D3 data. The Maculatus group appears monophyletic with An. pseudowillmori at a basal position, the Sawadwongporni subgroup and the Maculatus subgroup form sister clades. Our data concludes that An. dispar and An. greeni belong to the Maculatus subgroup, and An. willmori is not closer to either of the subgroups.