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Reference gene validation for quantitative RT-PCR during biotic and abiotic stresses in Vitis vinifera

2014Journal article Open access
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dc.contributor.authorBorges, Alexandre Filipe
dc.contributor.authorFonseca, Catarina
dc.contributor.authorFerreira, Ricardo Boavida
dc.contributor.authorLourenço, Ana Maria
dc.contributor.authorMonteiro, Sara
dc.date.accessioned2019-05-09T13:53:14Z
dc.date.available2019-05-09T13:53:14Z
dc.date.issued2014
dc.description.abstractGrapevine is one of the most cultivated fruit crop worldwide with Vitis vinifera being the species with the highest economical importance. Being highly susceptible to fungal pathogens and increasingly affected by environmental factors, it has become an important agricultural research area, where gene expression analysis plays a fundamental role. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is currently amongst the most powerful techniques to perform gene expression studies. Nevertheless, accurate gene expression quantification strongly relies on appropriate reference gene selection for sample normalization. Concerning V. vinifera, limited information still exists as for which genes are the most suitable to be used as reference under particular experimental conditions. In this work, seven candidate genes were investigated for their stability in grapevine samples referring to four distinct stresses (Erysiphe necator, wounding and UV-C irradiation in leaves and Phaeomoniella chlamydospora colonization in wood). The expression stability was evaluated using geNorm, NormFinder and BestKeeper. In all cases, full agreement was not observed for the three methods. To provide comprehensive rankings integrating the three different programs, for each treatment, a consensus ranking was created using a non-weighted unsupervised rank aggregation method. According to the last, the three most suitable reference genes to be used in grapevine leaves, regardless of the stress, are UBC, VAG and PEP. For the P. chlamydospora treatment, EF1, CYP and UBC were the best scoring genes. Acquaintance of the most suitable reference genes to be used in grapevine samples can contribute for accurate gene expression quantification in forthcoming studiespt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationBorges AF, Fonseca C, Ferreira RB, Lourenc¸o AM, Monteiro S (2014) Reference Gene Validation for Quantitative RT-PCR during Biotic and Abiotic Stresses in Vitis vinifera. PLoS ONE 9(10): e111399pt_PT
dc.identifier.doi10.1371/journal.pone.0111399pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.5/17786
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherPlos ONEpt_PT
dc.relationPTDC/AGR PRO/112340/2009pt_PT
dc.relationDIRIGENT PROTEINS - MODULATION OF PLANT DEFENSE BY ENANTIOSELECTIVITY REGULATION
dc.subjectVitis viniferapt_PT
dc.subjectbiotic stresspt_PT
dc.subjectabiotic stresspt_PT
dc.subjectRT-PCRpt_PT
dc.titleReference gene validation for quantitative RT-PCR during biotic and abiotic stresses in Vitis viniferapt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleDIRIGENT PROTEINS - MODULATION OF PLANT DEFENSE BY ENANTIOSELECTIVITY REGULATION
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBD%2F61903%2F2009/PT
oaire.citation.titlePLoS ONEpt_PT
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isProjectOfPublicationed3e9783-0c87-425b-b09d-d28c77566816
relation.isProjectOfPublication.latestForDiscoveryed3e9783-0c87-425b-b09d-d28c77566816

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