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Reference genes selection and normalization of oxidative stress responsive genes upon different temperature stress conditions in Hypericum perforatum L

2014Journal article Open access
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dc.contributor.authorVelada, Isabel
dc.contributor.authorRagonezi, Carla
dc.contributor.authorArnholdt-Schmitt, Birgit
dc.contributor.authorCardoso, Hélia
dc.date.accessioned2022-02-08T14:27:03Z
dc.date.available2022-02-08T14:27:03Z
dc.date.issued2014
dc.description.abstractReverse transcription-quantitative real-time PCR (RT-qPCR) is a widely used technique for gene expression analysis. The reliability of this method depends largely on the suitable selection of stable reference genes for accurate data normalization. Hypericum perforatum L. (St. John’s wort) is a field growing plant that is frequently exposed to a variety of adverse environmental stresses that can negatively affect its productivity. This widely known medicinal plant with broad pharmacological properties (anti-depressant, anti-tumor, anti-inflammatory, antiviral, antioxidant, anti-cancer, and antibacterial) has been overlooked with respect to the identification of reference genes suitable for RT-qPCR data normalization. In this study, 11 candidate reference genes were analyzed in H. perforatum plants subjected to cold and heat stresses. The expression stability of these genes was assessed using GeNorm, NormFinder and BestKeeper algorithms. The results revealed that the ranking of stability among the three algorithms showed only minor differences within each treatment. The best-ranked reference genes differed between cold- and heat-treated samples; nevertheless, TUB was the most stable gene in both experimental conditions. GSA and GAPDH were found to be reliable reference genes in cold-treated samples, while GAPDH showed low expression stability in heat-treated samples. 26SrRNA and H2A had the highest stabilities in the heat assay, whereas H2A was less stable in the cold assay. Finally, AOX1, AOX2, CAT1 and CHS genes, associated with plant stress responses and oxidative stress, were used as target genes to validate the reliability of identified reference genes. These target genes showed differential expression profiles over time in treated samples. This study not only is the first systematic analysis for the selection of suitable reference genes for RT-qPCR studies in H. perforatum subjected to temperature stress conditions, but may also provide valuable information about the roles of genes associated with temperature stress responses.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationVelada, I., Ragonezi, C., Arnholdt-Schmitt, B., & Cardoso, H. (2014). Reference genes selection and normalization of oxidative stress responsive genes upon different temperature stress conditions in Hypericum perforatum L. PLoS One, 9(12), e115206.pt_PT
dc.identifier.doi10.1371/journal.pone.0115206pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.13/4061
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherPublic Library of Sciencept_PT
dc.relationStrategic Project - UI 115 - 2011-2012
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectHypericum perforatum L.pt_PT
dc.subjectOxidative stress responsive genespt_PT
dc.subjectTemperature stress conditionspt_PT
dc.subject.pt_PT
dc.subjectEscola Superior de Tecnologias e Gestãopt_PT
dc.titleReference genes selection and normalization of oxidative stress responsive genes upon different temperature stress conditions in Hypericum perforatum Lpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleStrategic Project - UI 115 - 2011-2012
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876-PPCDTI/PTDC%2FAGR-GPL%2F099263%2F2008/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6820 - DCRRNI ID/PEst-C%2FAGR%2FUI0115%2F2011/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/PEst-OE%2FAGR%2FUI0115%2F2014/PT
oaire.citation.issue12pt_PT
oaire.citation.startPagee115206pt_PT
oaire.citation.titlePLoS ONEpt_PT
oaire.citation.volume9pt_PT
oaire.fundingStream5876-PPCDTI
oaire.fundingStream6820 - DCRRNI ID
oaire.fundingStream5876
person.familyNameRagonezi
person.givenNameCarla
person.identifier.ciencia-idFC12-D3F0-EF3D
person.identifier.orcid0000-0002-1822-5473
person.identifier.scopus-author-id36460033800
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.typearticlept_PT
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relation.isAuthorOfPublication.latestForDiscoverya51cc988-63ce-40e5-9b71-a27b1eadc937
relation.isProjectOfPublication576b6af5-017d-4190-ad20-013bff9aca14
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