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Development of a new approach based on real - time PCR coupled with high resolution melting (HRM) analysis towards the entomological authentication of honey

2021Conference object Open access
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dc.contributor.authorHonrado, Mónica
dc.contributor.authorPinto, M. Alice
dc.contributor.authorLopes, Ana
dc.contributor.authorHenriques, Dora
dc.contributor.authorAmaral, Joana S.
dc.date.accessioned2021-07-28T09:16:36Z
dc.date.available2021-07-28T09:16:36Z
dc.date.issued2021
dc.descriptionThis work has received funding from the Programa Nacional Apícola 2020-2022 under the project “AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português”. The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020). D. Henriques is supported by the project BeeHappy (POCI-01-0145-FEDER-029871) funded by FEDER (Fundo Europeu de Desenvolvimento Regional) through the program COMPETE 2020—POCI (Programa Operacional para a Competividade e Internacionalização), and by Portuguese funds through FCT and A.R. Lopes by the PhD scholarship funded by the FCT (SFRH/BD/143627/2019).pt_PT
dc.description.abstractHoney is a natural product widely consumed around the globe, not only for its taste and nutritional value, but also for its health benefits. Being a product of high dietary relevance and increasing demand, it has also become a target of economically motivated adulteration. According to the 2014 European Parliament report on the food crisis, fraud in the food chain and the control thereof, honey is among the 10 food products most prone of being adulterated [1]. Up until now, honey authenticity was mainly focused on the issues of sugars addition and botanical and geographical origin. However, recently an increased attention has been paid to the entomological origin of honey. To this aim, different approaches have been proposed to differentiate honey produced by different Apis mellifera subspecies, including those from distinct mitochondrial (mt) DNA lineages [2]. This work aimed to develop a novel real-time PCR method coupled with HRM analysis that allows for the simultaneous differentiation of honeybee from maternal lineages A, M and C, for further application in honey authentication. In this sense, data previously obtained from the mitogenomes of a total of 112 honeybees of different lineages were considered for the development of new DNA markers. Considering the aim of further application in honey, new primer sets were designed to amplify short fragments that included different single nucleotide polymorphisms (SNPs) allowing for HRM application. Three primer sets were proposed, amsCOI-F/amsCOI-R targeting the Cytochrome oxidase I (COI) gene, amsND1-F-amsND1-R targeting the NADH-ubiquinone oxidoreductase chain 1 (ND1) gene and amsCox3-F/amsCox3-R targeting the Cytochrome oxidase subunit III (Cox3) gene. Each primer set was first tested using qualitative PCR using DNA extracted from honeybees of A, M and C mtDNA lineages. After optimizing the real-time PCR conditions, each primer set was tested using a series of mtDNA extracted from honeybees. While amsCOI-F/amsCOI-R allowed only for the separation of the honeybees in two clusters, with lineage C and M clustering together, both the amsCox3-F/amsCox3-R and amsND1-F/amsND1-R set of primers allowed to differentiate the three lineages in separate clusters, with high level of confidence. As future work, the methodology will be assayed in commercial honey samples.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationHonrado, Mónica; Pinto, M. Alice; Lopes, Ana Rita; Henriques, Dora; Amaral, Joana S. (2021). Development of a new approach based on real - time PCR coupled with high resolution melting (HRM) analysis towards the entomological authentication of honey. In 1st Natural products application: health, cosmetic and food: book of abstracts. Bragança: Instituto Politécnico. ISBN 978-972-745-286-6pt_PT
dc.identifier.isbn978-972-745-286-6
dc.identifier.urihttp://hdl.handle.net/10198/23784
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectHoneypt_PT
dc.subjectNatural productpt_PT
dc.titleDevelopment of a new approach based on real - time PCR coupled with high resolution melting (HRM) analysis towards the entomological authentication of honeypt_PT
dc.typeconference object
dspace.entity.typePublication
oaire.citation.conferencePlaceon-linept_PT
oaire.citation.titleNatural products application: Health, Cosmetic and Food 1st International Online Conference: Book of Abstractspt_PT
person.familyNameHonrado
person.familyNamePinto
person.familyNameLopes
person.familyNameHenriques
person.familyNameAmaral
person.givenNameMónica
person.givenNameM. Alice
person.givenNameAna
person.givenNameDora
person.givenNameJoana S.
person.identifier.ciencia-id4712-B40B-4B0E
person.identifier.ciencia-idF814-A1D0-8318
person.identifier.ciencia-idF014-6835-054D
person.identifier.ciencia-id291F-986F-07DA
person.identifier.ciencia-id5319-7DE8-BEDA
person.identifier.orcid0000-0002-5126-4693
person.identifier.orcid0000-0001-9663-8399
person.identifier.orcid0000-0002-1888-2262
person.identifier.orcid0000-0001-7530-682X
person.identifier.orcid0000-0002-3648-7303
person.identifier.scopus-author-id8085507800
person.identifier.scopus-author-id55761737300
rcaap.rightsopenAccesspt_PT
rcaap.typeconferenceObjectpt_PT
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