Molecular variation and phylogeny of members of the Minimus Group of Anopheles subgenus Cellia (Diptera: Culicidae)

Intra- and interspecific molecular variation were investigated in four members of the Minimus Group of Anopheles subgenus Cellia: An. aconitus, An. varuna, An. minimus A and An. minimus C. DNA sequence divergence between these species at a mitochondrial locus (cytochrome oxidase II) and at three nuclear loci (ITS2 and D3 regions of rDNA and guanylate cyclase) is reported. The data confirm the presence of two cryptic species, A and C, within An. minimus and provide evidence for the existence of a third species. Anopheles minimus A and C are estimated to have diverged 0.57-1.5 million years ago. The discrepancy observed using the guanylate cyclase intron, which is the fastest evolving region known in the Gambiae Complex but is relatively slowly evolving in the Minimus Group, is discussed. The long-term effective population sizes of An. minimus A and C are estimated to be in their millions, with that of species A being approximately twice the size of species C. This implies that An. minimus C has a much wider distribution than currently recognized, with possible widespread implications for vector control. No evidence was found for population structuring in either species A or C: there was greater variation of mitochondrial haplotypes within than among localities. The phylogenetic relationships of Oriental members of the Myzomyia Series are reconstructed.