Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions

dc.creatorMafra, Valéria
dc.creatorKubo, Karen S.
dc.creatorFerreira, Marcio Alves
dc.creatorAlves, Marcelo Ribeiro
dc.creatorStuart, Rodrigo M.
dc.creatorBoava, Leonardo P.
dc.creatorRodrigues, Carolina M.
dc.creatorMachado, Marcos A.
dc.date2017-04-20T15:33:37Z
dc.date2017-04-20T15:33:37Z
dc.date2012
dc.date.accessioned2023-09-26T23:45:03Z
dc.date.available2023-09-26T23:45:03Z
dc.identifierMAFRA, Valéria et al. Reference Genes for Accurate Transcript Normalization in Citrus Genotypes under Different Experimental Conditions. Plos One, v. 7, n. 2, p. 1-11, Feb. 2012.
dc.identifier1932-6203
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/18597
dc.identifier10.1371/journal.pone.0031263
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8893821
dc.descriptionMarcelo Ribeiro Alves. Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Documento produzido em parceria ou por autor vinculado à Fiocruz, mas não consta à informação no documento.
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.formatapplication/pdf
dc.languageeng
dc.publisherPublic Library of Science
dc.rightsopen access
dc.subjectGenótipos de citros
dc.subjectDiferentes Condições Experimentais
dc.subjectGenes de referência
dc.subjectNormalização Exata de Transcrição
dc.subjectReference Genes
dc.subjectCitrus Genotypes
dc.subjectDifferent Experimental Conditions
dc.subjectAccurate Transcript Normalization
dc.titleReference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
dc.typeArticle


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