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Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys

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Resumo:As defined by the Codex Alimentarius, honey is the natural sweet substance produced by honeybees from the nectar of plants.1 This natural product is widely appreciated but is also considered one of the foods most prone to adulteration. The increasing demand for monofloral honey and those with protected designation of origin (PDO) has led to increased fraud by mislabeling botanical and geographical origin.2 Verifying the geographical origin of honey is a challenging endeavor. Recently, attention has been paid to the entomological origin, as it aligns with the geographical patterns of honeybee subspecies. The Mediterranean region is a hot spot of Apis mellifera subspecific diversity shaped by thousands of years of evolution. Although contemporary human-mediated movements of queens have impacted the native subspecific distribution, several PDO honeys specify the subspecies that produce those honeys, thus offering a unique avenue for authentication. As part of the European PRIMA project MEDIBEES, we aim to develop a DNA-metabarcoding approach to authenticate honey's entomological origin, focusing on mitochondrial lineages A, M, C, and O. To achieve this, the DNA of 1280 honeybees representing 16 subspecies and the four lineages (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) was extracted, and their whole genomes were sequenced. The MitoZ software was used to assemble the mitochondrial genomes, resulting in 769 mitochondrial genomes successfully assembled. Subsequently, each of these genomes was aligned individually with a reference genome using MEGA software, and mitogenomes not specific to Apis mellifera were discarded. Of these, only the mitogenomes corresponding to the native ancestry were retained, resulting in a final set of 355 mitogenomes in the database. A phylogenetic analysis was conducted with the final 355 mitochondrial sequences, revealing four distinct clusters corresponding to the four maternal lineages. This dataset was used for calculating the fixation index (FST) pairwise values, and a sliding window of 400 bp was used to identify single nucleotide polymorphisms (SNPs) that effectively differentiate (FST>0.98) the four lineages, enabling the identification of promising regions for primer design. This work resulted in the discovery of three promising regions for discriminating the four maternal lineages: one in the COI gene, one in the ND1 gene, and one in the CYTB gene (Fig. 1).
Autores principais:Honrado, Mónica
Outros Autores:Henriques, Dora; Santos, Joana; Yadró Garcia, Carlos A.; Medibees Consortium; Pinto, M. Alice; Amaral, Joana S.
Assunto:Mitogenomes Apis mellifera Authentication Mediterranean Honey Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
Ano:2024
País:Portugal
Tipo de documento:documento de conferência
Tipo de acesso:acesso aberto
Instituição associada:Instituto Politécnico de Bragança
Idioma:inglês
Origem:Biblioteca Digital do IPB
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author Honrado, Mónica
author2 Henriques, Dora
Santos, Joana
Yadró Garcia, Carlos A.
Medibees Consortium
Pinto, M. Alice
Amaral, Joana S.
author2_role author
author
author
author
author
author
author_facet Honrado, Mónica
Henriques, Dora
Santos, Joana
Yadró Garcia, Carlos A.
Medibees Consortium
Pinto, M. Alice
Amaral, Joana S.
author_role author
contributor_name_str_mv Biblioteca Digital do IPB
country_str PT
creators_json_txt [{\"Person.name\":\"Honrado, Mónica\",\"Person.identifier.orcid\":\"0000-0002-5126-4693\"},{\"Person.name\":\"Henriques, Dora\",\"Person.identifier.orcid\":\"0000-0001-7530-682X\"},{\"Person.name\":\"Santos, Joana\"},{\"Person.name\":\"Yadró Garcia, Carlos A.\",\"Person.identifier.orcid\":\"0000-0002-6916-3647\"},{\"Person.name\":\"Medibees Consortium\"},{\"Person.name\":\"Pinto, M. Alice\",\"Person.identifier.orcid\":\"0000-0001-9663-8399\"},{\"Person.name\":\"Amaral, Joana S.\",\"Person.identifier.orcid\":\"0000-0002-3648-7303\"}]
datacite.contributors.contributor.contributorName.fl_str_mv Biblioteca Digital do IPB
datacite.creators.creator.creatorName.fl_str_mv Honrado, Mónica
Henriques, Dora
Santos, Joana
Yadró Garcia, Carlos A.
Medibees Consortium
Pinto, M. Alice
Amaral, Joana S.
datacite.date.Accepted.fl_str_mv 2024-01-01T00:00:00Z
datacite.date.available.fl_str_mv 2024-10-29T15:14:57Z
datacite.date.embargoed.fl_str_mv 2024-10-29T15:14:57Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.subjects.subject.fl_str_mv Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
datacite.titles.title.fl_str_mv Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
dc.contributor.none.fl_str_mv Biblioteca Digital do IPB
dc.creator.none.fl_str_mv Honrado, Mónica
Henriques, Dora
Santos, Joana
Yadró Garcia, Carlos A.
Medibees Consortium
Pinto, M. Alice
Amaral, Joana S.
dc.date.Accepted.fl_str_mv 2024-01-01T00:00:00Z
dc.date.available.fl_str_mv 2024-10-29T15:14:57Z
dc.date.embargoed.fl_str_mv 2024-10-29T15:14:57Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10198/30492
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv Sociedade Portuguesa de Química
dc.rights.cclincense.fl_str_mv http://creativecommons.org/licenses/by/4.0/
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
dc.title.fl_str_mv Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_c94f
description As defined by the Codex Alimentarius, honey is the natural sweet substance produced by honeybees from the nectar of plants.1 This natural product is widely appreciated but is also considered one of the foods most prone to adulteration. The increasing demand for monofloral honey and those with protected designation of origin (PDO) has led to increased fraud by mislabeling botanical and geographical origin.2 Verifying the geographical origin of honey is a challenging endeavor. Recently, attention has been paid to the entomological origin, as it aligns with the geographical patterns of honeybee subspecies. The Mediterranean region is a hot spot of Apis mellifera subspecific diversity shaped by thousands of years of evolution. Although contemporary human-mediated movements of queens have impacted the native subspecific distribution, several PDO honeys specify the subspecies that produce those honeys, thus offering a unique avenue for authentication. As part of the European PRIMA project MEDIBEES, we aim to develop a DNA-metabarcoding approach to authenticate honey's entomological origin, focusing on mitochondrial lineages A, M, C, and O. To achieve this, the DNA of 1280 honeybees representing 16 subspecies and the four lineages (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) was extracted, and their whole genomes were sequenced. The MitoZ software was used to assemble the mitochondrial genomes, resulting in 769 mitochondrial genomes successfully assembled. Subsequently, each of these genomes was aligned individually with a reference genome using MEGA software, and mitogenomes not specific to Apis mellifera were discarded. Of these, only the mitogenomes corresponding to the native ancestry were retained, resulting in a final set of 355 mitogenomes in the database. A phylogenetic analysis was conducted with the final 355 mitochondrial sequences, revealing four distinct clusters corresponding to the four maternal lineages. This dataset was used for calculating the fixation index (FST) pairwise values, and a sliding window of 400 bp was used to identify single nucleotide polymorphisms (SNPs) that effectively differentiate (FST>0.98) the four lineages, enabling the identification of promising regions for primer design. This work resulted in the discovery of three promising regions for discriminating the four maternal lineages: one in the COI gene, one in the ND1 gene, and one in the CYTB gene (Fig. 1).
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funding.funder.identifier_str_mv http://doi.org/10.13039/501100001871
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identifier.url.fl_str_mv http://hdl.handle.net/10198/30492
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organization_str_mv urn:organizationAcronym:ipb
person_str_mv Honrado, Mónica
Honrado, Mónica
https://www.ciencia-id.pt/4712-B40B-4B0E
4712-B40B-4B0E
http://orcid.org/0000-0002-5126-4693
0000-0002-5126-4693
Henriques, Dora
Henriques, Dora
https://www.ciencia-id.pt/291F-986F-07DA
291F-986F-07DA
http://orcid.org/0000-0001-7530-682X
0000-0001-7530-682X
Santos, Joana
Yadró Garcia, Carlos A.
Yadró Garcia, Carlos A.
https://www.ciencia-id.pt/F71F-B08E-EC39
F71F-B08E-EC39
http://orcid.org/0000-0002-6916-3647
0000-0002-6916-3647
Medibees Consortium
Pinto, M. Alice
Pinto, M. Alice
https://www.ciencia-id.pt/F814-A1D0-8318
F814-A1D0-8318
http://orcid.org/0000-0001-9663-8399
0000-0001-9663-8399
Amaral, Joana S.
Amaral, Joana S.
https://www.ciencia-id.pt/5319-7DE8-BEDA
5319-7DE8-BEDA
http://orcid.org/0000-0002-3648-7303
0000-0002-3648-7303
publishDate 2024
publisher.none.fl_str_mv Sociedade Portuguesa de Química
reponame_str Biblioteca Digital do IPB
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spelling engSociedade Portuguesa de Químicapt_PTAs defined by the Codex Alimentarius, honey is the natural sweet substance produced by honeybees from the nectar of plants.1 This natural product is widely appreciated but is also considered one of the foods most prone to adulteration. The increasing demand for monofloral honey and those with protected designation of origin (PDO) has led to increased fraud by mislabeling botanical and geographical origin.2 Verifying the geographical origin of honey is a challenging endeavor. Recently, attention has been paid to the entomological origin, as it aligns with the geographical patterns of honeybee subspecies. The Mediterranean region is a hot spot of Apis mellifera subspecific diversity shaped by thousands of years of evolution. Although contemporary human-mediated movements of queens have impacted the native subspecific distribution, several PDO honeys specify the subspecies that produce those honeys, thus offering a unique avenue for authentication. As part of the European PRIMA project MEDIBEES, we aim to develop a DNA-metabarcoding approach to authenticate honey's entomological origin, focusing on mitochondrial lineages A, M, C, and O. To achieve this, the DNA of 1280 honeybees representing 16 subspecies and the four lineages (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) was extracted, and their whole genomes were sequenced. The MitoZ software was used to assemble the mitochondrial genomes, resulting in 769 mitochondrial genomes successfully assembled. Subsequently, each of these genomes was aligned individually with a reference genome using MEGA software, and mitogenomes not specific to Apis mellifera were discarded. Of these, only the mitogenomes corresponding to the native ancestry were retained, resulting in a final set of 355 mitogenomes in the database. A phylogenetic analysis was conducted with the final 355 mitochondrial sequences, revealing four distinct clusters corresponding to the four maternal lineages. This dataset was used for calculating the fixation index (FST) pairwise values, and a sliding window of 400 bp was used to identify single nucleotide polymorphisms (SNPs) that effectively differentiate (FST>0.98) the four lineages, enabling the identification of promising regions for primer design. This work resulted in the discovery of three promising regions for discriminating the four maternal lineages: one in the COI gene, one in the ND1 gene, and one in the CYTB gene (Fig. 1).application/pdfpt_PTExploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeysPersonalHonrado, MónicaDSpacehttp://dspace.org/items/87f8840d-04b1-427a-bca9-d37eadfc0e9bDSpacehttp://dspace.org/items/87f8840d-04b1-427a-bca9-d37eadfc0e9bHonradoMónicaCiência IDhttps://www.ciencia-id.pt4712-B40B-4B0EORCIDhttp://orcid.org0000-0002-5126-4693PersonalHenriques, DoraDSpacehttp://dspace.org/items/d2abd09f-a90c-4cfb-9a60-7fc32f56184dDSpacehttp://dspace.org/items/d2abd09f-a90c-4cfb-9a60-7fc32f56184dHenriquesDoraCiência IDhttps://www.ciencia-id.pt291F-986F-07DAORCIDhttp://orcid.org0000-0001-7530-682XScopus Author IDhttps://www.scopus.com55761737300Santos, JoanaPersonalYadró Garcia, Carlos A.DSpacehttp://dspace.org/items/59212a6c-fc6a-45fd-b37a-51e90926c9e3DSpacehttp://dspace.org/items/59212a6c-fc6a-45fd-b37a-51e90926c9e3GarcíaCarlos A. YadróCiência IDhttps://www.ciencia-id.ptF71F-B08E-EC39ORCIDhttp://orcid.org0000-0002-6916-3647Medibees ConsortiumPersonalPinto, M. AliceDSpacehttp://dspace.org/items/0667fe04-7078-483d-9198-56d167b19bc5DSpacehttp://dspace.org/items/0667fe04-7078-483d-9198-56d167b19bc5PintoM. AliceCiência IDhttps://www.ciencia-id.ptF814-A1D0-8318ORCIDhttp://orcid.org0000-0001-9663-8399Scopus Author IDhttps://www.scopus.com8085507800PersonalAmaral, Joana S.DSpacehttp://dspace.org/items/42be2cf4-adc4-4e7f-ac60-7aab515b38cdDSpacehttp://dspace.org/items/42be2cf4-adc4-4e7f-ac60-7aab515b38cdAmaralJoana S.Ciência IDhttps://www.ciencia-id.pt5319-7DE8-BEDAORCIDhttp://orcid.org0000-0002-3648-7303HostingInstitutionOrganizationalBiblioteca Digital do IPBe-mailmailto:dspace@ipb.ptdspace@ipb.pt2024-10-29T15:14:57Z20242024-01-01T00:00:00ZHandlehttp://hdl.handle.net/10198/30492http://purl.org/coar/access_right/c_abf2open accessMitogenomesApis melliferaAuthenticationMediterraneanHoneyResearch Subject Categories::TECHNOLOGY::Chemical engineering::Food technology6279752 bytesFundação para a Ciência e a TecnologiaMountain Research Center6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaAssociate Laboratory for Sustainability and Tecnology in Mountain Regions6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871other research producthttp://purl.org/coar/resource_type/c_c94fconference object2024http://creativecommons.org/licenses/by/4.0/http://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://bibliotecadigital.ipb.pt/bitstreams/c8cc29b7-d82c-499c-8c93-2261213114d0/downloadXVII Encontro Nacional Química dos Alimentos117117Vila Real
spellingShingle Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
Honrado, Mónica
Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
status SINGLETON
subject.fl_str_mv Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
title Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
title_full Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
title_fullStr Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
title_full_unstemmed Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
title_short Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
title_sort Exploiting the mitogenomes of Apis mellifera subspecies to authenticate the origin of Mediterranean honeys
topic Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
topic_facet Mitogenomes
Apis mellifera
Authentication
Mediterranean
Honey
Research Subject Categories::TECHNOLOGY::Chemical engineering::Food technology
url http://hdl.handle.net/10198/30492
visible 1