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Harnessing mitochondrial divergence in Apis mellifera to determine the origin of mediterranean honey

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datacite.subject.fosCiências Naturais
datacite.subject.fosCiências Agrárias::Biotecnologia Agrária e Alimentar
datacite.subject.sdg04:Educação de Qualidade
datacite.subject.sdg13:Ação Climática
datacite.subject.sdg15:Proteger a Vida Terrestre
dc.contributor.authorHonrado, Mónica
dc.contributor.authorHenriques, Dora
dc.contributor.authorSantos, Joana
dc.contributor.authorYadró Garcia, Carlos A.
dc.contributor.authorPinto, M. Alice
dc.contributor.authorAmaral, Joana S.
dc.date.accessioned2026-02-13T16:01:44Z
dc.date.available2026-02-13T16:01:44Z
dc.date.issued2025
dc.description.abstractThe Mediterranean region holds one of the highest levels of Apis mellifera subspecific diversity, exhibiting a rich evolutionary history shaped by long-term geographic isolation, climatic variation, and, more recently, by human-driven processes. As part of the international project MEDIBEES, we developed a DNA-metabarcoding approach to characterise the entomological origin of honey, focusing on mitochondrial lineages A (African), M (Western European), C (Eastern European), and O (Middle Eastern). To build a robust reference database, DNA was extracted from 1280 honeybees representing 16 A. mellifera subspecies (A.m. sahariensis, A.m. intermisa, A.m. siciliana, A.m. ruttneri, A.m. iberiensis, A.m. ligustica, A.m. macedonica, A.m. adami, A.m. cecropia, A.m. cypria, A.m. caucasica, A.m. meda, A.m. anatoliaca, A.m. syriaca, A.m. jemenitica, A.m. lamarcki) and whole genomes were sequenced. Mitochondrial genomes were assembled using MitoZ, resulting in 769 assemblies, which were individually aligned against the reference mitochondrial genome using MEGA. Only mitogenomes corresponding to native subspecific ancestry were retained, producing a final curated database of 355 sequences. This database was then used to calculate the fixation index (FST) pairwise values, and a 400 bp sliding window was used to identify single-nucleotide polymorphisms (SNPs) that effectively differentiate (FST>0.98) the four lineages. Three promising regions emerged for primer design: one in the COI gene, one in the ND1 gene, and one in the CYTB gene. The resulting primers were first validated using 36 honeybee samples of known lineage representing all 16 subspecies and subsequently applied to 83 honey samples from Lebanon, Jordan, Türkiye, Italy, Algeria, Malta, UAE, and Oman. Among the three markers, ND1 showed the highest discriminatory power, correctly assigning all reference honeybee samples to their respective lineage. The COI and CYTB primers also showed strong potential, and the observed error rates (5.8% in COI and 1.7% for CYTB) provide useful thresholds for interpreting the DNA-metabarcoding results in honey samples. Overall, this study demonstrates how patterns of mitochondrial divergence shaped by long-term evolutionary processes can be harnessed for applied DNA-metabarcoding on food authenticity.eng
dc.description.sponsorshipM. Honrado and C. Yadró acknowledge the PhD scholarship from FCT (2021.08119.BD, 2021.06948.BD). This research was funded by the project MEDIBEES - Monitoring the Mediterranean Honey Bee Subspecies and their Resilience to Climate Change for the Improvement of Sustainable Agro-Ecosystems. MEDIBEES is part of the PRIMA programme supported by the European Union. This research was also supported by national funds through FCT/MCTES (PIDDAC): CIMO UID/00690/2025 (10.54499/UID/00690/2025) and UID/PRR/00690/2025 (10.54499/UID/PRR/00690/2025); SusTEC, LA/P/0007/2020 (DOI: 10.54499/LA/P/0007/2020).
dc.identifier.citationHonrado, Mónica; Henriques, Dora; Santos, Joana; Yadró Garcia, Carlos A.; Pinto, M. Alice; Amaral, Joana S. (2025). Harnessing mitochondrial divergence in Apis mellifera to determine the origin of mediterranean honey. XXI International Meeting of the Portuguese Association for Evolutionary Biology. p. 97
dc.identifier.isbn978-972-745-368-9
dc.identifier.urihttp://hdl.handle.net/10198/35749
dc.language.isoeng
dc.peerreviewedyes
dc.relationAssociate Laboratory for Sustainability and Tecnology in Mountain Regions
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectApis mellifera
dc.subjectHoney
dc.subjectMitochondrial DNA
dc.subjectEntomological origin
dc.titleHarnessing mitochondrial divergence in Apis mellifera to determine the origin of mediterranean honeyeng
dc.typeconference object
dspace.entity.typePublication
oaire.awardNumberLA/P/0007/2020
oaire.awardTitleAssociate Laboratory for Sustainability and Tecnology in Mountain Regions
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/LA%2FP%2F0007%2F2020/PT
oaire.citation.conferenceDate2025
oaire.citation.conferencePlaceBragança, Portugal
oaire.citation.startPage97
oaire.citation.titleXXI International Meeting of the Portuguese Association for Evolutionary Biology
oaire.fundingStream6817 - DCRRNI ID
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85
person.familyNameHenriques
person.familyNameYadró Garcia
person.familyNamePinto
person.familyNameAmaral
person.givenNameDora
person.givenNameCarlos A.
person.givenNameM. Alice
person.givenNameJoana S.
person.identifier.ciencia-id291F-986F-07DA
person.identifier.ciencia-idF71F-B08E-EC39
person.identifier.ciencia-idF814-A1D0-8318
person.identifier.ciencia-id5319-7DE8-BEDA
person.identifier.orcid0000-0001-7530-682X
person.identifier.orcid0000-0002-6916-3647
person.identifier.orcid0000-0001-9663-8399
person.identifier.orcid0000-0002-3648-7303
person.identifier.scopus-author-id55761737300
person.identifier.scopus-author-id8085507800
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
relation.isAuthorOfPublicationd2abd09f-a90c-4cfb-9a60-7fc32f56184d
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relation.isAuthorOfPublication42be2cf4-adc4-4e7f-ac60-7aab515b38cd
relation.isAuthorOfPublication.latestForDiscovery59212a6c-fc6a-45fd-b37a-51e90926c9e3
relation.isProjectOfPublication6255046e-bc79-4b82-8884-8b52074b4384
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