The investigation of the biological diversity of honey bee (Apis mellifera L.) in duzce province by genetic and morpometric methods

In this study, the honey bee biodiversity of Düzce province, which stands out in beekeeping with Rhododendron and chestnut flora and Yığılca ecotype in the Western Black Sea Region was studied by using morphometric and genetic methods. For this purpose, a totally 1440 worker bee samples were collected from 24 apiaries, 72 colonies representing all districts of Düzce city. The lengths of the proboscis, wing and leg segments were measured as well as 31 morphometric characters of the right fore wings of the worker honey bee samples. In addition to these morphometric characters, 5 gene regions (CoxI- CoxII, 16srDNA, ND5, Cytb and CoxI) in mtDNA were evaluated with 10 different restriction endonuclease enzymes based on the PCR-RFLP method and by performing DNA sequence analysis. According to Discriminant Function Analysis (DFA) of morphometric data, Merkez, Akçakoca and Cumayeri districts were clearly separated from each other and other districts on the two dimensional plane. It was observed that the group centers of Gümüşova-Çilimli and Kaynaşlı-Yığılca districts overlapped in pairs and clustered closely with Gölyaka District. In the UPGMA dendrogram created according to the mahalanobis distances between the populations based on the colony averages; Akçakoca, Yığılca, Merkez, Çilimli and Gümüşova grouped together, while Cumayeri, Gölyaka and Kaynaşlı formed a group together. A worker bee sample from each colony was analysed regarding to the CoxI-CoxII intergenic region with DraI, XbaI and HınfI; CoxI gene with SspI, XhoI, DraI; 16srDNA gene with SspI, SwaI, DraI, EcoRI and HincII; the ND5 gene with HincII, DraI and SspI; Cytb gene with BglII, ClaI, HınfI and DraI restriction endonuclease enzymes. As a result of the cleavage of the restriction endonuclease enzymes, two different mitotypes were formed in the CoxI-CoxII /XbaI, HınfI, while three different mitotypes were observed in CoxI-CoxII / DraI. Two types were formed in the 16srDNA/SspI segment, type 2 was only seen in one sample from each of Cumayeri, Gümüşova, Kaynaşlı and Merkez districts, while type 2 was seen only in two samples of Yığılca in the 16srDNA/HincII segment. Two different mitotypes were formed in the Cytb/DraI segment. Type 2 profile was seen in 6 samples of Gümüşova and 2 samples of Merkez. No difference was observed as a result of cleavage of CoxI and ND5 genes with restriction endonuclease enzymes. DNA sequence analyzes were applied on the samples which were different according to the PCR/RFLP results. The results showed that mitochondrial DNA type common in Düzce province was compatible with the Anatolian honey bee (A. m. anatoliaca) mitotype; however, honey bee biodiversity of Düzce province was affected by the queen bee trade and migratory beekeeping activities.