Intrabreed Diversity and Relationships between Races of the Honey Bee Apis mellifera carpathica and Apis mellifera caucasica

Based on the analysis of 94 sequences of the mtDNA CO1 gene, the intrabreed diversity and evolutionary relationships of bees from Russia, Ukraine, Armenia, Kyrgyzstan, Tajikistan, and Poland were studied. We revealed two main haplotypes corresponding to the races A. m. carpathica and A. m. caucasica, which are common in the regions studied. However, in Armenia bees of the Caucasian race predominate, and in Poland, the Carpathian A. m. caucasica has a higher haplotype diversity: one of its haplotypes is formed by samples of bees from the Crimean mountain breed, possibly an independent race A. m. taurica. There are two clusters on the phylogenetic tree: one cluster includes A. m. mellifera and A. m. iberica (evolutionary line M), the other (line C), A. m. ligustica, A. m. carpathica, and A. m. caucasica. Haplotypes of A. m. caucasica have more substitutions per site, indicating their earlier origin compared to A. m. ligustica and A. m. carpathica. According to the time of divergence of A. mellifera and A. cerana at 6 Ma, the divergence of all races of line C is estimated from 1.3 to 0.6 Ma; races A. m. caucasica at 0.35-0.25 Ma and A. m. carpathica at 0.2-0.04 Ma. When using universal primers with optimization of amplification conditions for the mtDNA CO1 gene region, a nonsynonymous G/A SNP was detected in position 4 (680 bp) which can be used to distinguish A. m. carpathica from A. m. ligustica.