A simple test using restricted PCR-amplified mitochondrial DNA to study the genetic structure ofApis mellifera L

The COI-COII intergenic region ofApis mellifera mitochondrial DNA contains an important length polymorphism based on a variable number of copies of a 192–196 bp sequence (Q) and the completer or partial deletion of 67 pb sequence (Po). This length variability has been combined with a restriction site polymorphism to produce a rapid and simple test for the characterization of mtDNA haplotypes. This test included the amplification by the polymerase chain reaction of the COI-COII region followed by aDraI restriction of the amplified fragment. In a survey of 302 colonies belonging to 12 subspecies, 21 different haplotypes have been found which have been unambiguously allocated to one of the 3 mtDNA lineages of the species. Although all colonies of lineage C exhibit the same pattern (C1), each one of lineages A and M presents up to 10 different haplotypes, opening the way to studies on the genetic structure and the evolution of a large fraction of the species. This test also differentiates southern Spanish and South African colonies, which can be of great interest for the Africanized bee problem.