Reference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions

dc.creatorMafra, V
dc.creatorKubo, KS
dc.creatorAlves-Ferreira, M
dc.creatorRibeiro-Alves, M
dc.creatorStuart, RM
dc.creatorBoava, LP
dc.creatorRodrigues, CM
dc.creatorMachado, MA
dc.date2012
dc.dateFEB 9
dc.date2014-08-01T18:32:09Z
dc.date2015-11-26T17:01:40Z
dc.date2014-08-01T18:32:09Z
dc.date2015-11-26T17:01:40Z
dc.date.accessioned2018-03-28T23:49:32Z
dc.date.available2018-03-28T23:49:32Z
dc.identifierPlos One. Public Library Science, v. 7, n. 2, 2012.
dc.identifier1932-6203
dc.identifierWOS:000302730200016
dc.identifier10.1371/journal.pone.0031263
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/80361
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/80361
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1278767
dc.descriptionReal-time reverse transcription PCR (RT-qPCR) has emerged as an accurate and widely used technique for expression profiling of selected genes. However, obtaining reliable measurements depends on the selection of appropriate reference genes for gene expression normalization. The aim of this work was to assess the expression stability of 15 candidate genes to determine which set of reference genes is best suited for transcript normalization in citrus in different tissues and organs and leaves challenged with five pathogens (Alternaria alternata, Phytophthora parasitica, Xylella fastidiosa and Candidatus Liberibacter asiaticus). We tested traditional genes used for transcript normalization in citrus and orthologs of Arabidopsis thaliana genes described as superior reference genes based on transcriptome data. geNorm and NormFinder algorithms were used to find the best reference genes to normalize all samples and conditions tested. Additionally, each biotic stress was individually analyzed by geNorm. In general, FBOX (encoding a member of the F-box family) and GAPC2 (GAPDH) was the most stable candidate gene set assessed under the different conditions and subsets tested, while CYP (cyclophilin), TUB (tubulin) and CtP (cathepsin) were the least stably expressed genes found. Validation of the best suitable reference genes for normalizing the expression level of the WRKY70 transcription factor in leaves infected with Candidatus Liberibacter asiaticus showed that arbitrary use of reference genes without previous testing could lead to misinterpretation of data. Our results revealed FBOX, SAND (a SAND family protein), GAPC2 and UPL7 (ubiquitin protein ligase 7) to be superior reference genes, and we recommend their use in studies of gene expression in citrus species and relatives. This work constitutes the first systematic analysis for the selection of superior reference genes for transcript normalization in different citrus organs and under biotic stress.
dc.description7
dc.description2
dc.descriptionInstituto Nacional de Ciencia e Tecnologia de Genomica para Melhoramento de Citros
dc.languageen
dc.publisherPublic Library Science
dc.publisherSan Francisco
dc.publisherEUA
dc.relationPlos One
dc.relationPLoS One
dc.rightsaberto
dc.sourceWeb of Science
dc.subjectTime Rt-pcr
dc.subjectSinensis L. Osbeck
dc.subjectCandidatus Liberibacter-asiaticus
dc.subjectPolymerase-chain-reaction
dc.subjectInternal Control Genes
dc.subjectFunctional-characterization
dc.subjectReal
dc.subjectExpression
dc.subjectSelection
dc.subjectValidation
dc.titleReference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions
dc.typeArtículos de revistas


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