Genetic divergence and phylogenetic relationships of honey bee Apis mellifera (Hymenoptera: Apidae) populations from Greece and Cyprus using PCR – RFLP analysis of three mtDNA segments

Abstract

-The genetic structure and phylogenetic relationships among six honey bee populations were studied using RFLP analysis on three PCR-amplified mtDNA gene segments (16s rDNA, CO I, and ND 5). The populations were sampled from various areas of Greece and Cyprus and correspond to Apis mellifera adami, A. m. macedonica, A. m. cecropia, and A. m. cypria races, based on origin (Ruttner, 1988). Seven, eight and seven restriction enzymes were found to have at least one recognition site at the 16s rDNA, CO I, and ND 5 segments, respectively. Seven different haplotypes were detected and diagnostic patterns enabled us to discriminate A. m. macedonica from the rest of the populations (races). The estimated net nucleotide sequence divergence among the populations examined was found to range from 0.00 to 1.18 with the highest value observed to be between A. m. macedonica and non-A. m. macedonica populations. The trees obtained (by UPGMA and Dollo parsimony methods) revealed that the most distant population was that of A. m. macedonica.Apis mellifera / honey bee / mtDNA / genetic variability / Greece

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)

“…Mitochondrial regions were amplified according to Bouga et al (2005). Three sets of primers were used for amplifying 16S rDNA, COI and ND5 gene regions, respectively.…”

“…PCR-RFLP with DraI restriction enzyme analysis of the cytochrome C oxidase COI and COII (COI-COII) intergenic region revealed more than 50 different haplotypes, mainly in lineages A and M and with less in lineage C (Garnery et al, 1992(Garnery et al, , 1993Franck et al, 2000Franck et al, , 2001Palmer et al, 2000;De la Rúa et al, 2006;Suppasat et al, 2007;Munoz et al, 2009;Nedic et al, 2009;Özdil et al, 2009a;Solorzano et al, 2009;Magnus & Szalanski, 2010). More recently, the other mitochondrial regions including 16S ribosomal DNA (rDNA) (Bouga et al, 2005;Collet et al, 2007;Kekeço lu et al, 2009), NADH dehydrogenase-2 (ND2) (Arias & Sheppard, 1996;Kandemir et al, 2006), and NADH dehydrogenase-5 (ND5) regions (Bouga et al, 2005;Martimianakis et al, 2011;Özdil et al, 2012), along with the COI region (Tanaka et al, 2001;Bouga et al, 2005;Kekeço lu et al, 2009;Sheffield et al, 2009;Martimianakis et al, 2011), were used to differentiate evolutionary lineages and groups of honey bee subspecies.…”

Genetic characterization of the cyprian honey bee (Apis mellifera cypria) based on microsatell ites and mitochondrial DNA polymorphisms

“…Bouga et al (2005b) performed isoenzymatic analysis of honey bees from Greece and the southern part of Cyprus, and their results showed that Cyprian honey bee populations are well discriminated from the Greek populations based on the ADH and LAP enzymatic systems. MtDNA analysis of the same samples (Bouga et al, 2005a;Martimianakis et al, 2011) demonstrated that honey bees from Cyprus belong to the East Mediterranean C lineage. Additionally, results based on geometric morphometry show that honey bee populations located near the northern part of Cyprus are different from those located in the southernmost part, suggesting a possible introgression within these populations from the north (Lykoudis et al, 2004).…”

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

“…The COI gene segments were amplified according to the method of Bouga et al (2005). The PCR products were purified using a gel purification kit (QIAGEN) and sequenced in both directions on an ABI Prism 3130 automated sequencer (Applied Biosystems) using standard protocols.…”

“…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).…”