Genotyping north American black flies by means of mitochondrial ribosomal RNA sequences

Subregions in the mitochondrial small (12S) and large (16S) subunits of rRNA genes were amplified by polymerase chain reaction (PCR) from 14 species of North American black flies and assayed using directed heteroduplex analysis (DHDA). All 14 species could be distinguished from one another on the basis of the diagnostic mobilities of their heteroduplex products in either 12S or 16S DHDAs, utilizing PCR products derived from Simulium bivittatum as probes. Only minimal intraspecific variations were noted for two sibling species of Simulium vittatum s.l. collected from 10 locations, while S. luggeri appeared to have multiple mitochondrial haplotypes. DHDA banding patterns based on mitochondrially encoded sequences are shown to be an informative and easily interpreted taxonomic character, which should prove useful for future studies of black fly taxonomy and ecology.