Honey is the natural sweet substance produced by Apis mellifera honeybees in Europe. Depending on the country/region, the A. mellifera subspecies native to Europe belong to three different lineages: A (A. m. iberiensis), M (A. m. iberiensis and A. m. mellifera) and C (A. m. ligustica and A. m. carnica). Portugese researchers have developed two assays based on mitochondrial markers to identify the different sub-species. A cytb specific PCR assay was proposed to identify A-lineage honeybees, while a second method based on real-time PCR coupled to high resolution melting analysis targeting the COI gene, was developed to differentiate C- and M-lineages honeybees. The assays were validated on authentic honeys from known origin, and then applied to 20 commercial samples from different Euroean countries. The results highlight the predominance of honeys from C-lineage honeybees in Europe, except in Iberian Peninsula countries where honey from A-lineage honeybees predominates.
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