16S mtDNA variation in Apis mellifera detected by PCR-RFLP

Phylogeographic and morphometric evidence can be used to cluster Apis mellifera subspecies into evolutionary lineages or branches. Mitochondrial DNA sequence and restriction site analyses have shown similar clustering of subspecies groups. Thus, mtDNA variation can be used to infer honey bee evolutionary relationships. In this paper, we describe three 16S mtDNA PCR-RFLP patterns, each one completely associated with a previously determined A, M, or C Dra I restriction pattern of the COI-COII region. These results indicate that the COI-COII and the 16S genes have had a very closely linked evolutionary history. Although distinct patterns were obtained with Eco RI, Alu I, Hinc II and Taq I, the best differentiation among the three patterns was observed with Dra I and Vsp I enzymes. Nucleotide sequence analysis of the 16S gene fragment displayed 10 sites of base substitution (1.35%) among the three patterns and two insertions in the A. m. scutellata pattern.