Abstract
A large-scale survey of the Iberian honey bee (Apis mellifera iberiensis) diversity patterns, using sequence data of the tRNAleu-cox2 mitochondrial DNA (mtDNA) region, demonstrates that earlier studies based on the DraI test missed significant components of genetic variation. Based on results from this survey, existing haplotype names were revised and updated following a nomenclature system established earlier and extended herein for the intergenic region. A more complete picture of the complex diversity patterns of IHBs is revealed that includes 164 novel haplotypes, 113 belonging to lineage A and 51 to lineage M and within lineage A and 69 novel haplotypes that belong to sub-lineage AI, 13 to AII, and 31 to AIII. Within lineage M, two novel haplotypes show a striking architecture with features of lineages A and M, which based on sequence comparisons and relationships among haplotypes are seemingly ancestral. These data expand our knowledge of the complex architecture of the tRNAleu-cox2 intergenic region in Apis mellifera and re-emphasizes the importance of Iberia as a source of honey bee mtDNA diversity.
Highlights
The natural distribution of the honey bee, Apis mellifera L., encompasses Europe, Africa, and the Middle East (Ruttner 1988)
The Iberian honey bee (IHB, A. m. iberiensis ), which was allocated by Ruttner (1988) to lineage M, has been one of the most surveyed for genetic diversity due to its complex patterns of diversity and evolutionary history
GENEALEX 6.5 (Peakall and Smouse 2012) was used to estimate mean number of haplotypes per locus (Na), effective number of haplotypes (Ne), number of private haplotypes (Np), and Sequence data of the tRNAleu-cox2 intergenic region generated from 812 honey bee individuals collected in Iberia (N = 711), Western Europe and north of the Pyrenees (N = 34), Eastern Europe (N = 36), and North Africa (N = 31) revealed 212 haplotypes, of which 182 are reported here for the first time (Table S1)
Summary
The natural distribution of the honey bee, Apis mellifera L., encompasses Europe, Africa, and the Middle East (Ruttner 1988). MtDNA analyses of IHBs digested the entire molecule with a battery of restriction endonucleases (reviewed by Meixner et al 2013) This approach was soon replaced by the Dra I test, which consists of a single digestion, with the restriction enzyme Dra I, of the PCRamplified tRNAleu-cox intergenic fragment (Garnery et al 1993). The clinal pattern has been confirmed by a recent study using genomewide nuclear SNPs (Chávez-Galarza et al 2015), strengthening the hypothesis of an ancestral secondary contact (Smith et al 1991)
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