Selection of Novel Reference Genes by RNA-Seq and Their Evaluation for Normalising Real-Time qPCR Expression Data of Anthocyanin-Related Genes in Lettuce and Wild Relatives

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dc.bibliographicCitation.titleInternational Journal Of Molecular Sciencesen
dc.bibliographicCitation.volume24(3)es_ES
dc.contributor.authorMedina Lozano, Inéses_ES
dc.contributor.authorArnedo Andrés, María Soledades_ES
dc.contributor.authorGrimplet, Jérômees_ES
dc.contributor.authorDíaz Bermúdez, Auroraes_ES
dc.coverage.spatialCiencia vegetales_ES
dc.date.accessioned2023-02-08T11:45:18Z
dc.date.available2023-02-08T11:45:18Z
dc.date.issued2023es_ES
dc.description.abstractLettuce is a popular vegetable source of bioactive compounds, like anthocyanins, powerful antioxidants present in red and semi-red varieties. Selection of reliable reference genes (RGs) for the normalization of real-time quantitative PCR (qPCR) data is crucial to obtain accurate gene expression results. Among the genes with totally unrelated biological functions, six candidate RGs (ADF2, CYB5, iPGAM, SCL13, TRXL3-3, and VHA-H) with low variation in expression according to RNA-seq analyses, were selected for future expression studies of anthocyanin-related genes in three different experiments: leaf colour comparison (green vs. red) in commercial varieties; tissue comparison (leaf vs. stem) in a wild relative; and drought stress experiment in commercial and traditional varieties, and a wild relative. Expression profiles of the candidate RGs were obtained by qPCR and their stability was assessed by four different analytical tools, geNorm, NormFinder, BestKeeper, and Delta Ct method, all integrated in RefFinder. All results considered, we recommend CYB5 to be used as RG for the leaf colour experiment and TRXL3-3 for the tissue and drought stress ones, as they were the most stable genes in each case. RNA-seq is useful to preselect novel RGs although validation by qPCR is still advisable. These results provide helpful information for gene expression studies in Lactuca spp. under the described conditions.en
dc.description.statusPublishedes_ES
dc.identifier.citationInternational Journal of Molecular Sciences, vol. 24, num. 3, (2023)
dc.identifier.urihttp://hdl.handle.net/10532/6307
dc.language.isoenes_ES
dc.relation.doi10.3390/ijms24033052es_ES
dc.relation.urihttps://www.mdpi.com/1422-0067/24/3/3052es_ES
dc.rightsAtribución-NoComercial-SinDerivadas 3.0 España*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/3.0/es/*
dc.subject.agrovocLactuca sativaes
dc.subject.agrovocEstrés de sequiaes
dc.subject.agrovocExpresión génicaes
dc.subject.agrovocNormalizaciónes
dc.subject.agrovocPCRes
dc.titleSelection of Novel Reference Genes by RNA-Seq and Their Evaluation for Normalising Real-Time qPCR Expression Data of Anthocyanin-Related Genes in Lettuce and Wild Relativesen
dc.typeJournal Contribution*
dc.type.refereedRefereedes_ES
dc.type.specifiedArticlees_ES

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