Genetic variation in Turkish honeybees Apis mellifera anatoliaca, A. m. caucasica, A. m. meda (Hymenoptera: Apidae) inferred from RFLP analysis of three mtDNA regions (16S rDNA-COI-ND5)

Abstract

Abstract. In this study, the genetic structure of Turkish honey bee (Apis mellifera L.) populations, mainly obtained from the Central Anatolian region, were investigated at three different mitochondrial regions. A total of 165 worker bees were collected from 15 different populations in ten different locations. Portions of the mitochondrial 16S ribosomal RNA (16S rDNA), cytochrome C oxidase I (COI) and NADH dehydrogenase 5 (ND5) genes were amplified by PCR and then subjected to RFLP pattern analysis using 18 restriction enzymes (these having at least one recognition site in each region were used). Nucleotide polymorphisms were revealed using restriction enzymes Bsp143I, DraI and SspI in 16S rDNA and TaqI in the COI gene segment. The polymorphisms were subsequently confirmed by direct DNA sequencing with sequences thereafter deposited in Genbank. In this study, six novel composite genotypes (haplotypes) were found in Turkish honey bee populations. The most common haplotype, type 1, was found in 12 of the sampled populations and overall accounted for 85.5% of the samples. TCS spanning network of haplotypes revealed that type 1 was the basal haplotype. Genetic distance (D) values were found to be low (0.0-0.0112) within Turkish honey bee populations. The average haplotype diversity (h) within populations was 0.082. Molecular phylogenetic analysis revealed that Konya/Sızma, Antalya/Elmalı and Konya/Selçuklu populations were the most distant from all the other Turkish honey bee populations surveyed.

Genetic Variation in Iranian Honey bees, Apis mellifera meda Skorikow, 1829, (Hymenoptera: Apidae) Inferred from PCR-RFLP Analysis of two mtDNA Gene Segments (COI and 16S rDNA)

“…In comparison our results with Bouga et al (2005), Kekeçoglu et al (2009) and Ozdil et al (2012), we find similar restriction profiles with generally small base differences in the fragments, except for Sau3A I, Ssp I and Taq I and Bsp143I, Ssp I and Dra I digestions in COI and 16S rDNA gene regions, respectively (Table 2). In these regions, different restriction profiles were detected in our samples or populations.…”

“…Sau3A I, Ssp I and Taq I restriction in COI revealed six or five, five or four and four or three restriction sites in this study, respectively, whereas in others subspecies such as A. m. anatoliaca, A. m. caucasica and A. m. meda (in Turkey), restriction analysis were previously reported to have revealed four or six, three or two and two restriction sites for these enzymes, respectively (Bouga et al, 2005, Kekeçoglu et al, 2009, Ozdil et al, 2012. Correspondingly, the restriction of the 16S rDNA gene region with Sau3A I, Ssp I and Dra I revealed two or one, three or two and seven or six sites (Table 2), compared with the three or two, two and two or seven as reported in Turkish honey bees (Bouga et al, 2005, Kekeçoglu et al, 2009, Ozdil et al, 2012.…”

“…Correspondingly, the restriction of the 16S rDNA gene region with Sau3A I, Ssp I and Dra I revealed two or one, three or two and seven or six sites (Table 2), compared with the three or two, two and two or seven as reported in Turkish honey bees (Bouga et al, 2005, Kekeçoglu et al, 2009, Ozdil et al, 2012. Comparing our findings with these of Ozdil et al (2012), honey bees in Iran and Turkey (specially A. m. meda) were found to be well discriminated; it is noted that for the first time are reported diagnostic patterns that distinguish the honey bee populations of these neighboring countries. Honey bees from Iran are also discriminating from other subspecies of honey bees in Turkey, Iraq and Syria.…”

“…m. anatoliaca, A. m. caucasica and A. m. meda (in Turkey), restriction analysis were previously reported to have revealed four or six, three or two and two restriction sites for these enzymes, respectively (Bouga et al, 2005, Kekeçoglu et al, 2009, Ozdil et al, 2012. Correspondingly, the restriction of the 16S rDNA gene region with Sau3A I, Ssp I and Dra I revealed two or one, three or two and seven or six sites (Table 2), compared with the three or two, two and two or seven as reported in Turkish honey bees (Bouga et al, 2005, Kekeçoglu et al, 2009, Ozdil et al, 2012. Comparing our findings with these of Ozdil et al (2012), honey bees in Iran and Turkey (specially A. m. meda) were found to be well discriminated; it is noted that for the first time are reported diagnostic patterns that distinguish the honey bee populations of these neighboring countries.…”

Phylogenetic relationship of Turkish Apis mellifera subspecies based on sequencing of mitochondrial cytochrome C oxidase I region

“…Although barcoding, a DNA-based identification system (Hebert et al, 2003), has mainly focused on sequence diversity in the COI region, we have little knowledge about the mitochondrial COI region beyond the tRNA leu -COII region in A. mellifera subspecies. Only the RFLP profile of the COI region (Bouga et al, 2005;Kekeçoğlu et al, 2009;Özdil et al, 2012) and the phylogenetic relationships of A. mellifera subspecies based on sequencing of this region have been reported (Tanaka et al, 2001;Marino et al, 2002a,b;Sheffield et al, 2009;Martimianakis et al, 2011).…”