Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress

Autores
Ulrich, Maria Noelia; Muñiz-Padilla, Esteban; Corach, Alejandra; Hopp, Horacio Esteban; Tosto, Daniela Sandra
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Weeds are one of the main causes of the decrease in crop yields, with Johnsongrass (Sorghum halepense L.) being one of the most significant. Weeds can be controlled by herbicides, but some have developed resistance. Quantitative PCR is the technique of choice for studying gene expression related to herbicide resistance because of its high sensitivity and specificity, although its quantitative accuracy is highly dependent on the stability of the reference genes. Thus, in this study we evaluated the stability of different reference genes of glyphosate-resistant S. halepense. Nine genes frequently used as reference genes were selected: MDH, ADP, PP2A, EIF4α, ACT, ARI8, DnaJ, Hsp70, and ALS1, and their expression analyzed in susceptible and resistant biotypes at 0, 24 and 72 h post-application of glyphosate. The stability was analyzed with the geNorm, NormFinder, and BestKeeper software programs and using the ΔCt method. RefFinder was used to generate a comprehensive stability ranking. The results showed that PP2A and ARI8 were the most stable genes under the test conditions. EPSPS expression was also verified against the best two and the worst two reference genes. This study provides useful information for gene expression analysis under glyphosate stress and will facilitate resistance mechanism studies in this weed species.
Instituto de Biotecnología
Fil: Ulrich, Maria Noelia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Ulrich, Maria Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Muñiz-Padilla, Esteban. Universidad Nacional de Entre Ríos. Facultad de Ciencias Agropecuarias; Argentina
Fil: Corach, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Corach, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Hopp, Horacio Esteban. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Tosto, Daniela Sandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Tosto, Daniela Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Plants 10 (8) : 1555 (Agosto 2021)
Materia
Weeds
Herbicide Resistance
PCR
Gene Expression
Glyphosate
Stress
Malezas
Resistencia a los Herbicidas
Expresión Génica
Sorghum halepense
Glifosato
Estres
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/12078
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oai_identifier_str oai:localhost:20.500.12123/12078
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network_name_str INTA Digital (INTA)
spelling Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stressUlrich, Maria NoeliaMuñiz-Padilla, EstebanCorach, AlejandraHopp, Horacio EstebanTosto, Daniela SandraWeedsHerbicide ResistancePCRGene ExpressionGlyphosateStressMalezasResistencia a los HerbicidasExpresión GénicaSorghum halepenseGlifosatoEstresWeeds are one of the main causes of the decrease in crop yields, with Johnsongrass (Sorghum halepense L.) being one of the most significant. Weeds can be controlled by herbicides, but some have developed resistance. Quantitative PCR is the technique of choice for studying gene expression related to herbicide resistance because of its high sensitivity and specificity, although its quantitative accuracy is highly dependent on the stability of the reference genes. Thus, in this study we evaluated the stability of different reference genes of glyphosate-resistant S. halepense. Nine genes frequently used as reference genes were selected: MDH, ADP, PP2A, EIF4α, ACT, ARI8, DnaJ, Hsp70, and ALS1, and their expression analyzed in susceptible and resistant biotypes at 0, 24 and 72 h post-application of glyphosate. The stability was analyzed with the geNorm, NormFinder, and BestKeeper software programs and using the ΔCt method. RefFinder was used to generate a comprehensive stability ranking. The results showed that PP2A and ARI8 were the most stable genes under the test conditions. EPSPS expression was also verified against the best two and the worst two reference genes. This study provides useful information for gene expression analysis under glyphosate stress and will facilitate resistance mechanism studies in this weed species.Instituto de BiotecnologíaFil: Ulrich, Maria Noelia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Ulrich, Maria Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Muñiz-Padilla, Esteban. Universidad Nacional de Entre Ríos. Facultad de Ciencias Agropecuarias; ArgentinaFil: Corach, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Corach, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hopp, Horacio Esteban. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Tosto, Daniela Sandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; ArgentinaFil: Tosto, Daniela Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaMDPI2022-06-13T10:46:56Z2022-06-13T10:46:56Z2021-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/12078https://www.mdpi.com/2223-7747/10/8/15552223-7747https://doi.org/10.3390/plants10081555Plants 10 (8) : 1555 (Agosto 2021)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2019-PE-E6-I114-001/2019-PE-E6-I114-001/AR./Caracterización de la diversidad genética de plantas, animales y microorganismos mediante herramientas de genómica aplicada.info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:45:35Zoai:localhost:20.500.12123/12078instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:45:35.903INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
title Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
spellingShingle Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
Ulrich, Maria Noelia
Weeds
Herbicide Resistance
PCR
Gene Expression
Glyphosate
Stress
Malezas
Resistencia a los Herbicidas
Expresión Génica
Sorghum halepense
Glifosato
Estres
title_short Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
title_full Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
title_fullStr Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
title_full_unstemmed Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
title_sort Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
dc.creator.none.fl_str_mv Ulrich, Maria Noelia
Muñiz-Padilla, Esteban
Corach, Alejandra
Hopp, Horacio Esteban
Tosto, Daniela Sandra
author Ulrich, Maria Noelia
author_facet Ulrich, Maria Noelia
Muñiz-Padilla, Esteban
Corach, Alejandra
Hopp, Horacio Esteban
Tosto, Daniela Sandra
author_role author
author2 Muñiz-Padilla, Esteban
Corach, Alejandra
Hopp, Horacio Esteban
Tosto, Daniela Sandra
author2_role author
author
author
author
dc.subject.none.fl_str_mv Weeds
Herbicide Resistance
PCR
Gene Expression
Glyphosate
Stress
Malezas
Resistencia a los Herbicidas
Expresión Génica
Sorghum halepense
Glifosato
Estres
topic Weeds
Herbicide Resistance
PCR
Gene Expression
Glyphosate
Stress
Malezas
Resistencia a los Herbicidas
Expresión Génica
Sorghum halepense
Glifosato
Estres
dc.description.none.fl_txt_mv Weeds are one of the main causes of the decrease in crop yields, with Johnsongrass (Sorghum halepense L.) being one of the most significant. Weeds can be controlled by herbicides, but some have developed resistance. Quantitative PCR is the technique of choice for studying gene expression related to herbicide resistance because of its high sensitivity and specificity, although its quantitative accuracy is highly dependent on the stability of the reference genes. Thus, in this study we evaluated the stability of different reference genes of glyphosate-resistant S. halepense. Nine genes frequently used as reference genes were selected: MDH, ADP, PP2A, EIF4α, ACT, ARI8, DnaJ, Hsp70, and ALS1, and their expression analyzed in susceptible and resistant biotypes at 0, 24 and 72 h post-application of glyphosate. The stability was analyzed with the geNorm, NormFinder, and BestKeeper software programs and using the ΔCt method. RefFinder was used to generate a comprehensive stability ranking. The results showed that PP2A and ARI8 were the most stable genes under the test conditions. EPSPS expression was also verified against the best two and the worst two reference genes. This study provides useful information for gene expression analysis under glyphosate stress and will facilitate resistance mechanism studies in this weed species.
Instituto de Biotecnología
Fil: Ulrich, Maria Noelia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Ulrich, Maria Noelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Muñiz-Padilla, Esteban. Universidad Nacional de Entre Ríos. Facultad de Ciencias Agropecuarias; Argentina
Fil: Corach, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Corach, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Hopp, Horacio Esteban. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina
Fil: Tosto, Daniela Sandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina
Fil: Tosto, Daniela Sandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Weeds are one of the main causes of the decrease in crop yields, with Johnsongrass (Sorghum halepense L.) being one of the most significant. Weeds can be controlled by herbicides, but some have developed resistance. Quantitative PCR is the technique of choice for studying gene expression related to herbicide resistance because of its high sensitivity and specificity, although its quantitative accuracy is highly dependent on the stability of the reference genes. Thus, in this study we evaluated the stability of different reference genes of glyphosate-resistant S. halepense. Nine genes frequently used as reference genes were selected: MDH, ADP, PP2A, EIF4α, ACT, ARI8, DnaJ, Hsp70, and ALS1, and their expression analyzed in susceptible and resistant biotypes at 0, 24 and 72 h post-application of glyphosate. The stability was analyzed with the geNorm, NormFinder, and BestKeeper software programs and using the ΔCt method. RefFinder was used to generate a comprehensive stability ranking. The results showed that PP2A and ARI8 were the most stable genes under the test conditions. EPSPS expression was also verified against the best two and the worst two reference genes. This study provides useful information for gene expression analysis under glyphosate stress and will facilitate resistance mechanism studies in this weed species.
publishDate 2021
dc.date.none.fl_str_mv 2021-08
2022-06-13T10:46:56Z
2022-06-13T10:46:56Z
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/20.500.12123/12078
https://www.mdpi.com/2223-7747/10/8/1555
2223-7747
https://doi.org/10.3390/plants10081555
url http://hdl.handle.net/20.500.12123/12078
https://www.mdpi.com/2223-7747/10/8/1555
https://doi.org/10.3390/plants10081555
identifier_str_mv 2223-7747
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2019-PE-E6-I114-001/2019-PE-E6-I114-001/AR./Caracterización de la diversidad genética de plantas, animales y microorganismos mediante herramientas de genómica aplicada.
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv Plants 10 (8) : 1555 (Agosto 2021)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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