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Genetic characterization of Thrace honey bee populations of Turkey: restriction and sequencing of inter cytochrome C oxidase I-II (CoxI-CoxII) genes

In this study, the genetic characterization of Thrace honey bee populations of Turkey were identified using the mitochondrial gene region, 3′ end of cytochrome C oxidase I gene to 5′ end of cytochrome C oxidase II gene (inter CoxI-CoxII genes) by using XbaI PCR-RFLP and DNA sequencing. A total of 322 worker honey bees were collected from five different localities; 4 provinces (Tekirdağ, Kırklareli, Edirne and Çanakkale) and Gökçeada Island near Thrace. PCR RFLP and sequence data combined revealed three haplotypes. The most frequently observed haplotype (Type 1), revealed only one XbaI restriction site in inter CoxI-CoxII genes. Type 2 haplotype revealed additional XbaI restriction site and this digestion revealed three bands. The rarely seen third haplotype, which is first reported in this study, was only found in two of the samples that were collected from Gökçeada Island. According to Maximum Likelihood phylogenetic analysis which was constructed on the basis of the distance matrix, Type 1 haplotype was clustered in the same clade with A. m. caucasica and A. m. anatoliaca whereas Type 2 was clustered in the same clade with A. m. macedonica and A. m. carnica. These results may indicate that Carniolan or Macedonian honey bee subspecies or their ecotypes can be found in the Thrace region of Turkey, mainly in samples that were collected from Kırklareli and Edirne provinces near the borders with Greece and Bulgaria.