Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought

Autores
Acevedo, Raúl Maximiliano; Avico, Edgardo Hernán; Ruiz, Oscar Adolfo; Sansberro, Pedro Alfonso
Año de publicación
2018
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Reverse transcription of RNA followed by real-time quantitative PCR (qPCR) is to date the most reliable method for gene expression studies. However, to control the errors introduced along the numerous experimental procedures, it requires a normalization using internal reference genes with stable expression. To address this issue, nine candidate reference genes were investigated in Ilex paraguariensis leaves subjected to water stress. To facilitate the selection, we analysed the real-time qPCR data with three different software programs. The obtained results support the conclusion that RNA polymerase associated protein rtf1 homolog (RTF) combined with any of the following pairs is the most suitable triad of genes to compute a normalization factor: elongation factor 1-alpha + tubulin alpha chain (EF1a + α-Tub), actin + cyclophilin 38 (ACT + CYP38), or cyclophilin 38 + vacuolar protein sorting-associated protein 18 homologs (CYP38 + VPS). Our analysis constitutes the first in-depth study to identify the appropriate reference genes for the quantification of transcription in Ilex paraguariensis leaves during drought and provides essential information for further gene expression studies in this tree species.
Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
Fil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
Fil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
Materia
ABIOTIC STRESSES
GENE EXPRESSION
HOUSE-KEEPING GENES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/89612
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/89612
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during droughtAcevedo, Raúl MaximilianoAvico, Edgardo HernánRuiz, Oscar AdolfoSansberro, Pedro AlfonsoABIOTIC STRESSESGENE EXPRESSIONHOUSE-KEEPING GENEShttps://purl.org/becyt/ford/4.4https://purl.org/becyt/ford/4Reverse transcription of RNA followed by real-time quantitative PCR (qPCR) is to date the most reliable method for gene expression studies. However, to control the errors introduced along the numerous experimental procedures, it requires a normalization using internal reference genes with stable expression. To address this issue, nine candidate reference genes were investigated in Ilex paraguariensis leaves subjected to water stress. To facilitate the selection, we analysed the real-time qPCR data with three different software programs. The obtained results support the conclusion that RNA polymerase associated protein rtf1 homolog (RTF) combined with any of the following pairs is the most suitable triad of genes to compute a normalization factor: elongation factor 1-alpha + tubulin alpha chain (EF1a + α-Tub), actin + cyclophilin 38 (ACT + CYP38), or cyclophilin 38 + vacuolar protein sorting-associated protein 18 homologs (CYP38 + VPS). Our analysis constitutes the first in-depth study to identify the appropriate reference genes for the quantification of transcription in Ilex paraguariensis leaves during drought and provides essential information for further gene expression studies in this tree species.Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); ArgentinaFil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaCzech Academy of Sciences. Institute of Experimental Botany2018-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/89612Acevedo, Raúl Maximiliano; Avico, Edgardo Hernán; Ruiz, Oscar Adolfo; Sansberro, Pedro Alfonso; Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought; Czech Academy of Sciences. Institute of Experimental Botany; Biologia Plantarum; 62; 1; 1-2018; 89-960006-31341573-8264CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1007/s10535-017-0754-3info:eu-repo/semantics/altIdentifier/url/https://bp.ueb.cas.cz/artkey/bpl-201801-0009_assessment-of-reference-genes-for-real-time-quantitative-pcr-normalization-in-ilex-paraguariensis-l.phpinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:42:17Zoai:ri.conicet.gov.ar:11336/89612instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:42:17.253CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
title Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
spellingShingle Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
Acevedo, Raúl Maximiliano
ABIOTIC STRESSES
GENE EXPRESSION
HOUSE-KEEPING GENES
title_short Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
title_full Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
title_fullStr Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
title_full_unstemmed Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
title_sort Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought
dc.creator.none.fl_str_mv Acevedo, Raúl Maximiliano
Avico, Edgardo Hernán
Ruiz, Oscar Adolfo
Sansberro, Pedro Alfonso
author Acevedo, Raúl Maximiliano
author_facet Acevedo, Raúl Maximiliano
Avico, Edgardo Hernán
Ruiz, Oscar Adolfo
Sansberro, Pedro Alfonso
author_role author
author2 Avico, Edgardo Hernán
Ruiz, Oscar Adolfo
Sansberro, Pedro Alfonso
author2_role author
author
author
dc.subject.none.fl_str_mv ABIOTIC STRESSES
GENE EXPRESSION
HOUSE-KEEPING GENES
topic ABIOTIC STRESSES
GENE EXPRESSION
HOUSE-KEEPING GENES
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.4
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Reverse transcription of RNA followed by real-time quantitative PCR (qPCR) is to date the most reliable method for gene expression studies. However, to control the errors introduced along the numerous experimental procedures, it requires a normalization using internal reference genes with stable expression. To address this issue, nine candidate reference genes were investigated in Ilex paraguariensis leaves subjected to water stress. To facilitate the selection, we analysed the real-time qPCR data with three different software programs. The obtained results support the conclusion that RNA polymerase associated protein rtf1 homolog (RTF) combined with any of the following pairs is the most suitable triad of genes to compute a normalization factor: elongation factor 1-alpha + tubulin alpha chain (EF1a + α-Tub), actin + cyclophilin 38 (ACT + CYP38), or cyclophilin 38 + vacuolar protein sorting-associated protein 18 homologs (CYP38 + VPS). Our analysis constitutes the first in-depth study to identify the appropriate reference genes for the quantification of transcription in Ilex paraguariensis leaves during drought and provides essential information for further gene expression studies in this tree species.
Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
Fil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
Fil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús). Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín" (sede Chascomús); Argentina
Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; Argentina
description Reverse transcription of RNA followed by real-time quantitative PCR (qPCR) is to date the most reliable method for gene expression studies. However, to control the errors introduced along the numerous experimental procedures, it requires a normalization using internal reference genes with stable expression. To address this issue, nine candidate reference genes were investigated in Ilex paraguariensis leaves subjected to water stress. To facilitate the selection, we analysed the real-time qPCR data with three different software programs. The obtained results support the conclusion that RNA polymerase associated protein rtf1 homolog (RTF) combined with any of the following pairs is the most suitable triad of genes to compute a normalization factor: elongation factor 1-alpha + tubulin alpha chain (EF1a + α-Tub), actin + cyclophilin 38 (ACT + CYP38), or cyclophilin 38 + vacuolar protein sorting-associated protein 18 homologs (CYP38 + VPS). Our analysis constitutes the first in-depth study to identify the appropriate reference genes for the quantification of transcription in Ilex paraguariensis leaves during drought and provides essential information for further gene expression studies in this tree species.
publishDate 2018
dc.date.none.fl_str_mv 2018-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/89612
Acevedo, Raúl Maximiliano; Avico, Edgardo Hernán; Ruiz, Oscar Adolfo; Sansberro, Pedro Alfonso; Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought; Czech Academy of Sciences. Institute of Experimental Botany; Biologia Plantarum; 62; 1; 1-2018; 89-96
0006-3134
1573-8264
CONICET Digital
CONICET
url http://hdl.handle.net/11336/89612
identifier_str_mv Acevedo, Raúl Maximiliano; Avico, Edgardo Hernán; Ruiz, Oscar Adolfo; Sansberro, Pedro Alfonso; Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought; Czech Academy of Sciences. Institute of Experimental Botany; Biologia Plantarum; 62; 1; 1-2018; 89-96
0006-3134
1573-8264
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1007/s10535-017-0754-3
info:eu-repo/semantics/altIdentifier/url/https://bp.ueb.cas.cz/artkey/bpl-201801-0009_assessment-of-reference-genes-for-real-time-quantitative-pcr-normalization-in-ilex-paraguariensis-l.php
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Czech Academy of Sciences. Institute of Experimental Botany
publisher.none.fl_str_mv Czech Academy of Sciences. Institute of Experimental Botany
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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