Mitochondrial DNA analysis of the neotropical malaria vector Anopheles nuneztovari

Restriction endonucleases were used to test an hypothesis, based on cytological data, proposing the existence of two races in Anopheles nuneztovari, one Venezuelan-Colombian and the other Amazonian, and to examine variation in the mitochondrial DNA (mtDNA) of 263 individual A. nuneztovari mosquitoes from 12 sites in Bolivia, Brazil, Colombia, Suriname, and Venezuela. Cladistic analysis of the mtDNA data using three outgroups suggests that A. nuneztovari is monophyletic. However, this analysis is poorly resolved by parsimony, probably caused by extensive homoplasy. Analysis of the relationships among haplotypes, based on genetic distances, results in five haplotype lineages, three of which are in the Amazon Basin. Relationships among the 12 populations reveal distinctive lineages, one in Venezuela-Colombia and two within the Amazon Basin. Molecular variance components and F statistics (phi) among neotropical biomes, among populations/biomes, among cytotypes, among populations/cytotypes, and within populations all demonstrate significant subdivision and strongly support multiple lineages, which could be interpreted as recently evolved species, within A. nuneztovari. The Mantel analysis showed no significant correlation between genetic and geographic distances, suggesting that it is biomes and cytotypes that contribute most strongly to subdivision.