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

Detalhes bibliográficos
Autor(a) principal: Mafra, Valéria
Data de Publicação: 2012
Outros Autores: Kubo, Karen S., Ferreira, Marcio Alves, Alves, Marcelo Ribeiro, Stuart, Rodrigo M., Boava, Leonardo P., Rodrigues, Carolina M., Machado, Marcos A.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/18597
Resumo: Marcelo 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.
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spelling Mafra, ValériaKubo, Karen S.Ferreira, Marcio AlvesAlves, Marcelo RibeiroStuart, Rodrigo M.Boava, Leonardo P.Rodrigues, Carolina M.Machado, Marcos A.2017-04-20T15:33:37Z2017-04-20T15:33:37Z2012MAFRA, 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. 2012.1932-6203https://www.arca.fiocruz.br/handle/icict/1859710.1371/journal.pone.0031263Marcelo 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.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Biologia. Departamento de Genética. Laboratório de Genética Molecular Vegetal. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil / Universidade Estadual de Campinas. Instituto de Biologia. Campinas, SP, Brasil.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.Centro de Citricultura Sylvio Moreira. Laboratório de Biotecnologia. Cordeirópolis, SP, Brasil.Real-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.engPublic Library of ScienceGenótipos de citrosDiferentes Condições ExperimentaisGenes de referênciaNormalização Exata de TranscriçãoReference GenesCitrus GenotypesDifferent Experimental ConditionsAccurate Transcript NormalizationReference genes for accurate transcript normalization in citrus genotypes under different experimental conditionsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; 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dc.title.pt_BR.fl_str_mv Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
title Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
spellingShingle Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
Mafra, Valéria
Genótipos de citros
Diferentes Condições Experimentais
Genes de referência
Normalização Exata de Transcrição
Reference Genes
Citrus Genotypes
Different Experimental Conditions
Accurate Transcript Normalization
title_short Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
title_full Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
title_fullStr Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
title_full_unstemmed Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
title_sort Reference genes for accurate transcript normalization in citrus genotypes under different experimental conditions
author Mafra, Valéria
author_facet Mafra, Valéria
Kubo, Karen S.
Ferreira, Marcio Alves
Alves, Marcelo Ribeiro
Stuart, Rodrigo M.
Boava, Leonardo P.
Rodrigues, Carolina M.
Machado, Marcos A.
author_role author
author2 Kubo, Karen S.
Ferreira, Marcio Alves
Alves, Marcelo Ribeiro
Stuart, Rodrigo M.
Boava, Leonardo P.
Rodrigues, Carolina M.
Machado, Marcos A.
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Mafra, Valéria
Kubo, Karen S.
Ferreira, Marcio Alves
Alves, Marcelo Ribeiro
Stuart, Rodrigo M.
Boava, Leonardo P.
Rodrigues, Carolina M.
Machado, Marcos A.
dc.subject.other.pt_BR.fl_str_mv Genótipos de citros
Diferentes Condições Experimentais
Genes de referência
Normalização Exata de Transcrição
topic Genótipos de citros
Diferentes Condições Experimentais
Genes de referência
Normalização Exata de Transcrição
Reference Genes
Citrus Genotypes
Different Experimental Conditions
Accurate Transcript Normalization
dc.subject.en.pt_BR.fl_str_mv Reference Genes
Citrus Genotypes
Different Experimental Conditions
Accurate Transcript Normalization
description Marcelo 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.
publishDate 2012
dc.date.issued.fl_str_mv 2012
dc.date.accessioned.fl_str_mv 2017-04-20T15:33:37Z
dc.date.available.fl_str_mv 2017-04-20T15:33:37Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv MAFRA, 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.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/18597
dc.identifier.issn.pt_BR.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0031263
identifier_str_mv MAFRA, 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.
1932-6203
10.1371/journal.pone.0031263
url https://www.arca.fiocruz.br/handle/icict/18597
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/18597/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/18597/2/marcelo_alves_etal_IOC_2012.pdf
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