High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain

The accurate varietal identification is an essential requirement for every process involved in the exploitation of grapevine resources and derived products. The advancements achieved during the last years allowed the simultaneous analysis of multiple molecular markers capable of identifying grapevin...

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Detalhes bibliográficos
Autores: Barrias, Sara, Ibáñez Marcos, Javier, Martins-Lopes, Paula
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2024
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositório:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/359014
Acesso em linha:http://hdl.handle.net/10261/359014
https://api.elsevier.com/content/abstract/scopus_id/85184575695
Access Level:Acceso aberto
Palavra-chave:Authenticity
Melting analysis
Molecular markers
Must
Simple sequence repeats
Wine
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spelling High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chainBarrias, SaraIbáñez Marcos, JavierMartins-Lopes, PaulaAuthenticityMelting analysisMolecular markersMustSimple sequence repeatsWineThe accurate varietal identification is an essential requirement for every process involved in the exploitation of grapevine resources and derived products. The advancements achieved during the last years allowed the simultaneous analysis of multiple molecular markers capable of identifying grapevine varieties. Despite the establishment of a recommended set of nine microsatellite (SSR) markers for this purpose, their effective application with DNA extracted from must and wine samples remains a challenging task. This work aimed to develop High Resolution Melting (HRM) assays based on SSR markers applicable for grapevine varietal identification using leaf, must and wine samples. The grapevine varieties used were Cabernet Sauvignon, Touriga Franca, Touriga Nacional and Rufete. A total of 12 SSR markers were used to screen the varieties: nine markers recommended by the OIV (VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VVS2, VrZAG62 and VrZAG79) and three markers selected based on their repeat motif and length (VvIv35, VChr5c and VChr9a). The results from multiplex PCR amplification of DNA from wine samples revealed that these three markers performed better than the nine established SSR markers. HRM assays were developed targeting markers VvIv35, VChr5c and VChr9a, successfully discriminating the varietal composition in must DNA samples. Promising results were obtained using wine DNA, where assay HRM-VChr9a proved to have the highest discriminant power. The HRM-SSR assays need to be applied in a larger number of varieties, to explore its suitability for grapevine fingerprinting applications throughout the wine chain. Overall, the proposed small SSR makers can be more suitable for wine DNA analysis. The HRM-SSR approach presented here provides fast results, allowing the complete discrimination of varietal composition in must DNA. It also shows to be a promising tool to discriminate the varieties using wine DNA, a task usually hampered by the inherent complexity of wine samples.This research was supported by the Portuguese Foundation for Science and Technology (FCT) through the project Biosensor Development for Wine Traceability in the Douro Region – WineBioCode PTDC/AGR-ALI/117341/2010-FCOMP-01-0124-FEDER-019439, PhD grant of Sara Barrias (SFRH/BD/146346/2019) and BioISI center grants (UIDB/04046/2020 (DOI: 10.54499/UIDB/04046/2020) and UIDP/04046/2020 (DOI: 10.54499/UIDP/04046/2020)).Centre grants).Peer reviewedElsevierFundação para a Ciência e a Tecnologia (Portugal)European CommissionBarrias, Sara [0000-0003-2536-2107]Ibáñez Marcos, Javier [0000-0002-6286-5638]Martins-Lopes, Paula [0000-0001-8229-0117]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/359014https://api.elsevier.com/content/abstract/scopus_id/85184575695reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttps://doi.org/10.1016/j.foodcont.2024.110368Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3590142026-05-22T06:33:51Z
dc.title.none.fl_str_mv High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
title High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
spellingShingle High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
Barrias, Sara
Authenticity
Melting analysis
Molecular markers
Must
Simple sequence repeats
Wine
title_short High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
title_full High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
title_fullStr High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
title_full_unstemmed High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
title_sort High resolution melting analysis of microsatellite markers applied to grapevine varietal fingerprinting throughout the wine production chain
dc.creator.none.fl_str_mv Barrias, Sara
Ibáñez Marcos, Javier
Martins-Lopes, Paula
author Barrias, Sara
author_facet Barrias, Sara
Ibáñez Marcos, Javier
Martins-Lopes, Paula
author_role author
author2 Ibáñez Marcos, Javier
Martins-Lopes, Paula
author2_role author
author
dc.contributor.none.fl_str_mv Fundação para a Ciência e a Tecnologia (Portugal)
European Commission
Barrias, Sara [0000-0003-2536-2107]
Ibáñez Marcos, Javier [0000-0002-6286-5638]
Martins-Lopes, Paula [0000-0001-8229-0117]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Authenticity
Melting analysis
Molecular markers
Must
Simple sequence repeats
Wine
topic Authenticity
Melting analysis
Molecular markers
Must
Simple sequence repeats
Wine
description The accurate varietal identification is an essential requirement for every process involved in the exploitation of grapevine resources and derived products. The advancements achieved during the last years allowed the simultaneous analysis of multiple molecular markers capable of identifying grapevine varieties. Despite the establishment of a recommended set of nine microsatellite (SSR) markers for this purpose, their effective application with DNA extracted from must and wine samples remains a challenging task. This work aimed to develop High Resolution Melting (HRM) assays based on SSR markers applicable for grapevine varietal identification using leaf, must and wine samples. The grapevine varieties used were Cabernet Sauvignon, Touriga Franca, Touriga Nacional and Rufete. A total of 12 SSR markers were used to screen the varieties: nine markers recommended by the OIV (VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VVS2, VrZAG62 and VrZAG79) and three markers selected based on their repeat motif and length (VvIv35, VChr5c and VChr9a). The results from multiplex PCR amplification of DNA from wine samples revealed that these three markers performed better than the nine established SSR markers. HRM assays were developed targeting markers VvIv35, VChr5c and VChr9a, successfully discriminating the varietal composition in must DNA samples. Promising results were obtained using wine DNA, where assay HRM-VChr9a proved to have the highest discriminant power. The HRM-SSR assays need to be applied in a larger number of varieties, to explore its suitability for grapevine fingerprinting applications throughout the wine chain. Overall, the proposed small SSR makers can be more suitable for wine DNA analysis. The HRM-SSR approach presented here provides fast results, allowing the complete discrimination of varietal composition in must DNA. It also shows to be a promising tool to discriminate the varieties using wine DNA, a task usually hampered by the inherent complexity of wine samples.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/359014
https://api.elsevier.com/content/abstract/scopus_id/85184575695
url http://hdl.handle.net/10261/359014
https://api.elsevier.com/content/abstract/scopus_id/85184575695
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv https://doi.org/10.1016/j.foodcont.2024.110368
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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