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: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biotecnológicas; Argentina.
Fil: Acevedo, Raúl Maximiliano. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.
Fil: Avico, Edgardo Hernán. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.
Fil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Investigaciones Biotecnológicas; Argentina.
Fil: Ruiz, Oscar Adolfo. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín"; Argentina.
Fil: Sansberro, Pedro Alfonso. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.
Fuente
Biologia Plantarum, 2018, vol. 62, no. 1, p. 89-96.
Materia
Abiotic stress
Gene expression
House-keeping genes
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
Institución
Universidad Nacional del Nordeste
OAI Identificador
oai:repositorio.unne.edu.ar:123456789/9149
Acceder
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oai_identifier_str oai:repositorio.unne.edu.ar:123456789/9149
network_acronym_str RIUNNE
repository_id_str 4871
network_name_str Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
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 stressGene expressionHouse-keeping genesReverse 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 speciesFil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biotecnológicas; Argentina.Fil: Acevedo, Raúl Maximiliano. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.Fil: Avico, Edgardo Hernán. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.Fil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.Fil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Investigaciones Biotecnológicas; Argentina.Fil: Ruiz, Oscar Adolfo. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín"; Argentina.Fil: Sansberro, Pedro Alfonso. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.Springer2018info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfp. 89-96application/pdfAcevedo, Raúl Maximiliano, et al, 2018. Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought. Biologia Plantarum. República Checa: Czech Academy of Sciences. Institute of Experimental Botany, vol. 62, no. 1, p. 89-96. ISSN 1573-8264.0006-3134http://repositorio.unne.edu.ar/handle/123456789/9149Biologia Plantarum, 2018, vol. 62, no. 1, p. 89-96.reponame:Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)instname:Universidad Nacional del Nordesteenghttps://bp.ueb.cas.cz/pdfs/bpl/2018/01/09.pdfinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/2.5/ar/Atribución-NoComercial-SinDerivadas 2.5 Argentina2025-09-29T14:29:13Zoai:repositorio.unne.edu.ar:123456789/9149instacron:UNNEInstitucionalhttp://repositorio.unne.edu.ar/Universidad públicaNo correspondehttp://repositorio.unne.edu.ar/oaiososa@bib.unne.edu.ar;sergio.alegria@unne.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:48712025-09-29 14:29:14.168Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE) - Universidad Nacional del Nordestefalse
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 stress
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 stress
Gene expression
House-keeping genes
topic Abiotic stress
Gene expression
House-keeping genes
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: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Biotecnológicas; Argentina.
Fil: Acevedo, Raúl Maximiliano. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Acevedo, Raúl Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.
Fil: Avico, Edgardo Hernán. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Avico, Edgardo Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica del Nordeste; Argentina.
Fil: Ruiz, Oscar Adolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Instituto de Investigaciones Biotecnológicas; Argentina.
Fil: Ruiz, Oscar Adolfo. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. Instituto de Investigaciones Biotecnológicas "Dr. Raúl Alfonsín"; Argentina.
Fil: Sansberro, Pedro Alfonso. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias; Argentina.
Fil: Sansberro, Pedro Alfonso. Consejo Nacional de Investigaciones Científicas y Técnicas. 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
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 Acevedo, Raúl Maximiliano, et al, 2018. Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought. Biologia Plantarum. República Checa: Czech Academy of Sciences. Institute of Experimental Botany, vol. 62, no. 1, p. 89-96. ISSN 1573-8264.
0006-3134
http://repositorio.unne.edu.ar/handle/123456789/9149
identifier_str_mv Acevedo, Raúl Maximiliano, et al, 2018. Assessment of reference genes for real-time quantitative PCR normalization in Ilex paraguariensis leaves during drought. Biologia Plantarum. República Checa: Czech Academy of Sciences. Institute of Experimental Botany, vol. 62, no. 1, p. 89-96. ISSN 1573-8264.
0006-3134
url http://repositorio.unne.edu.ar/handle/123456789/9149
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://bp.ueb.cas.cz/pdfs/bpl/2018/01/09.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Atribución-NoComercial-SinDerivadas 2.5 Argentina
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Atribución-NoComercial-SinDerivadas 2.5 Argentina
dc.format.none.fl_str_mv application/pdf
p. 89-96
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Biologia Plantarum, 2018, vol. 62, no. 1, p. 89-96.
reponame:Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
instname:Universidad Nacional del Nordeste
reponame_str Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
collection Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE)
instname_str Universidad Nacional del Nordeste
repository.name.fl_str_mv Repositorio Institucional de la Universidad Nacional del Nordeste (UNNE) - Universidad Nacional del Nordeste
repository.mail.fl_str_mv ososa@bib.unne.edu.ar;sergio.alegria@unne.edu.ar
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score 12.559606

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