Please use this identifier to cite or link to this item: http://www.arca.fiocruz.br/handle/icict/18597
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dc.contributor.authorMafra, Valéria-
dc.contributor.authorKubo, Karen S.-
dc.contributor.authorFerreira, Marcio Alves-
dc.contributor.authorAlves, Marcelo Ribeiro-
dc.contributor.authorStuart, Rodrigo M.-
dc.contributor.authorBoava, Leonardo P.-
dc.contributor.authorRodrigues, Carolina M.-
dc.contributor.authorMachado, Marcos A.-
dc.date.accessioned2017-04-20T15:33:37Z-
dc.date.available2017-04-20T15:33:37Z-
dc.date.issued2012-
dc.identifier.citationMAFRA, Valéria; et al. Reference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions. Plos One, v.7, n.2, e31263, 11p, Feb. 2012.pt_BR
dc.identifier.issn1932-6203pt_BR
dc.identifier.urihttp://www.arca.fiocruz.br/handle/icict/18597-
dc.language.isoengpt_BR
dc.publisherPublic Library of Sciencept_BR
dc.rightsopen accesspt_BR
dc.subject.otherGenótipos de citrospt_BR
dc.subject.otherDiferentes Condições Experimentaispt_BR
dc.subject.otherGenes de referênciapt_BR
dc.subject.otherNormalização Exata de Transcriçãopt_BR
dc.titleReference genes for accurate transcript normalization in citrus genotypes under different experimental conditionspt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1371/journal.pone.0031263-
dc.description.abstractenReal-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.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.pt_BR
dc.creator.affilliationUniversidade Federal do Rio de Janeiro. Instituto de Biologia. Departamento de Genética. Laboratório de Genética Molecular Vegetal. Rio de Janeiro, RJ, Brasil.pt_BR
dc.creator.affilliationFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ. Brasil.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.pt_BR
dc.creator.affilliationCentro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.pt_BR
dc.subject.enReference Genespt_BR
dc.subject.enCitrus Genotypespt_BR
dc.subject.enDifferent Experimental Conditionspt_BR
dc.subject.enAccurate Transcript Normalizationpt_BR
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