Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress
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...
| Main Authors: | , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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MDPI
2022
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| Online Access: | http://hdl.handle.net/20.500.12123/12078 https://www.mdpi.com/2223-7747/10/8/1555 https://doi.org/10.3390/plants10081555 |
| _version_ | 1855484905207627776 |
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| author | Ulrich, Maria Noelia Muñiz-Padilla, Esteban Corach, Alejandra Hopp, Horacio Esteban Tosto, Daniela Sandra |
| author_browse | Corach, Alejandra Hopp, Horacio Esteban Muñiz-Padilla, Esteban Tosto, Daniela Sandra Ulrich, Maria Noelia |
| author_facet | Ulrich, Maria Noelia Muñiz-Padilla, Esteban Corach, Alejandra Hopp, Horacio Esteban Tosto, Daniela Sandra |
| author_sort | Ulrich, Maria Noelia |
| collection | INTA Digital |
| 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. |
| format | Artículo |
| id | INTA12078 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | INTA120782022-06-13T10:51:22Z Validation of reference genes for quantitative PCR in Johnsongrass (Sorghum halepense L.) under glyphosate stress Ulrich, Maria Noelia Muñiz-Padilla, Esteban Corach, Alejandra Hopp, Horacio Esteban Tosto, Daniela Sandra Weeds Herbicide Resistance PCR Gene Expression Glyphosate Stress Malezas Resistencia a los Herbicidas Expresión Génica Sorghum halepense Glifosato Estres 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 2022-06-13T10:46:56Z 2022-06-13T10:46:56Z 2021-08 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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. 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) application/pdf MDPI Plants 10 (8) : 1555 (Agosto 2021) |
| spellingShingle | Weeds Herbicide Resistance PCR Gene Expression Glyphosate Stress Malezas Resistencia a los Herbicidas Expresión Génica Sorghum halepense Glifosato Estres Ulrich, Maria Noelia Muñiz-Padilla, Esteban Corach, Alejandra Hopp, Horacio Esteban Tosto, Daniela Sandra 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 |
| 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_short | 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 |
| topic | Weeds Herbicide Resistance PCR Gene Expression Glyphosate Stress Malezas Resistencia a los Herbicidas Expresión Génica Sorghum halepense Glifosato Estres |
| url | http://hdl.handle.net/20.500.12123/12078 https://www.mdpi.com/2223-7747/10/8/1555 https://doi.org/10.3390/plants10081555 |
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