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DNA mini-barcodes coupled to high resolution melting (hrm) analysis for the botanical authentication of rosemary honey

dc.contributor.authorSoares, Sónia
dc.contributor.authorCosta, Joana
dc.contributor.authorAmaral, Joana S.
dc.contributor.authorOliveira, Beatriz
dc.contributor.authorMafra, Isabel
dc.date.accessioned2018-03-27T13:16:53Z
dc.date.available2018-03-27T13:16:53Z
dc.date.issued2016
dc.description.abstractHoney is a natural product highly consumed for its taste, nutritional value and health benefits. Monofloral honeys are the most appreciated by consumers and frequently attain high market values, thus being prone to fraudulent practices. Therefore, the development of methodologies to assess and authenticate the botanical origin of honey is of utmost importance. For this purpose, traditional methods based on pollen identification by microscopic analysis are still being used, but they are time‐consuming and greatly dependent on the experience/skill of trained analysts. As an alternative, the use of DNA markers represents promising approach for the identification of botanical species in honey. Currently, DNA barcoding has been regarded with increasing interest for the taxonomic identification of plants, with two plastidial genes (matK and rbcL) being proposed for their differentiation (Bruni et al., 2012). Thus, the objective of this work was to identify the botanical species in rosemary honey using mini‐barcode regions coupled to high resolution melting (HRM) analysis. For this purpose, different plant species (Lavandula spp.) and ten mono‐ and multifloral honeys were used. Three DNA barcoding loci, namely the plastidial coding genes rbcL and matK and the noncoding intergenic trnH‐psbA region, were used to design primers targeting Lavandula spp. (GenBank Z37408.1, KJ196360.1 and HQ902822.1). DNA from plants and honeys was extracted with NucleoSpin Plant II kit (method A), according to Soares et al. (2015). The specificity and sensitivity of the designed primers were assayed by qualitative polymerase chain reaction (PCR) and real‐time PCR. Prior to the specific amplifications, DNA extracts were positively tested targeting a universal eukaryotic sequence (18S rRNA gene). Results from specific PCR assays were further confirmed by real‐time PCR amplification using EvaGreen fluore scence dye. The application of HRM analysis allowed discriminating Lavandula spp. into distinct clusters with high level of confidence. When applying the developed methodology to rosemary honey, samples were classified on the same cluster of Lavandula stoechas (endemic species in Portugal), therefore confirming its botanical origin. To our knowledge, this is the first study using HRM analysis for the rapid discrimination of plant species in honey.pt_PT
dc.description.sponsorshipThis work was supported by FCT grant no. LAQV UID/QUI/50006/2013. Joana Costa and Sónia Soares are grateful to FCT grants (SFRH/BPD/102404/2014 and SFRH/BD/75091/2010, respectively) financed by POPH‐QREN (subsidized by FSE and MCTES). The authors are grateful for the kind supply of samples from Bank of Plant Germplasm of Tucson, USA and from Jardim de Serralves, Porto.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationSoares, Sónia; Costa, Joana; Amaral, J.S.; Oliveira, M.B.P.P.; Mafra, Isabel (2016). DNA mini-barcodes coupled to high resolution melting (hrm) analysis for the botanical authentication of rosemary honey. In FoodIntegrity 2016 - Assuring the integrity of the food chain: fighting food fraud. Praguept_PT
dc.identifier.urihttp://hdl.handle.net/10198/16572
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.relationStructural features and immunoreactivity of plant food allergens: impact of technological food processing and in vitro digestibility
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.titleDNA mini-barcodes coupled to high resolution melting (hrm) analysis for the botanical authentication of rosemary honeypt_PT
dc.typeconference object
dspace.entity.typePublication
oaire.awardTitleStructural features and immunoreactivity of plant food allergens: impact of technological food processing and in vitro digestibility
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FQUI%2F50006%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F102404%2F2014/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F75091%2F2010/PT
oaire.citation.conferencePlacePraguept_PT
oaire.citation.titleFoodIntegrity 2016 - Assuring the integrity of the food chain: FIGHTING FOOD FRAUDpt_PT
oaire.fundingStream5876
oaire.fundingStreamSFRH
person.familyNameAmaral
person.givenNameJoana S.
person.identifier.ciencia-id5319-7DE8-BEDA
person.identifier.orcid0000-0002-3648-7303
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typeconferenceObjectpt_PT
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relation.isAuthorOfPublication.latestForDiscovery42be2cf4-adc4-4e7f-ac60-7aab515b38cd
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