Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time
Plant stress studies are increasingly being based on gene expression. The analysis of gene expression requires sensitive, precise, and replicable measurements for specific mRNA sequences. Real-time RT-PCR is nowadays the most sensitive method for the detection of low abundance mRNA. A stable referen...
| Autores principales: | , , , , , |
|---|---|
| Formato: | info:eu-repo/semantics/article |
| Lenguaje: | Inglés |
| Publicado: |
2017
|
| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/941 https://journals-crea.4science.it/index.php/maydica/article/view/1571/1081 |
| _version_ | 1855034733163970560 |
|---|---|
| author | Decima Oneto, Cecilia Andrea Bossio, Adrian Ezequiel Faccio, Paula Daniela Beznec, Ailin Blumwald, Eduardo Lewi, Dalia Marcela |
| author_browse | Beznec, Ailin Blumwald, Eduardo Bossio, Adrian Ezequiel Decima Oneto, Cecilia Andrea Faccio, Paula Daniela Lewi, Dalia Marcela |
| author_facet | Decima Oneto, Cecilia Andrea Bossio, Adrian Ezequiel Faccio, Paula Daniela Beznec, Ailin Blumwald, Eduardo Lewi, Dalia Marcela |
| author_sort | Decima Oneto, Cecilia Andrea |
| collection | INTA Digital |
| description | Plant stress studies are increasingly being based on gene expression. The analysis of gene expression requires sensitive, precise, and replicable measurements for specific mRNA sequences. Real-time RT-PCR is nowadays the most sensitive method for the detection of low abundance mRNA. A stable reference gene is mandatory to obtain reliable quantitative real-time PCR (qPCR) analysis results. Real-time RT-PCR is referred with one or several internal control genes, which should not fluctuate during treatments. In this study, we have chosen eight genes
as candidates of possible reference genes for maize (Zea mays L) during water deficit stress at flowering time: a-tubulin, 3`phosphate glyceraldehyde dehydrogenase (GAPDH), 18S ribosomal subunit, protein 13S ribosomal, actin, zein, invertase, and starch synthase IIB. The eight reference genes candidates were tested on maize plants around flowering time, under three different conditions: before water deficit (BWD), under water deficit (WD) and after water deficit (AWD). Results from the three experimental conditions indicated that protein 13S ribosomal gene was the most stable among all the reference genes tested. This result suggests that protein 13S ribosomal gene can be used as internal control (housekeeping) for qPCR analysis in maize plants under water deficit stress during flowering time. |
| format | info:eu-repo/semantics/article |
| id | INTA941 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA9412018-02-26T13:07:48Z Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time Decima Oneto, Cecilia Andrea Bossio, Adrian Ezequiel Faccio, Paula Daniela Beznec, Ailin Blumwald, Eduardo Lewi, Dalia Marcela Genética Genes Estrés de Sequia Floración Genetics Frought Stress Flowering Genes de Limpieza Estres Hídrico Plant stress studies are increasingly being based on gene expression. The analysis of gene expression requires sensitive, precise, and replicable measurements for specific mRNA sequences. Real-time RT-PCR is nowadays the most sensitive method for the detection of low abundance mRNA. A stable reference gene is mandatory to obtain reliable quantitative real-time PCR (qPCR) analysis results. Real-time RT-PCR is referred with one or several internal control genes, which should not fluctuate during treatments. In this study, we have chosen eight genes as candidates of possible reference genes for maize (Zea mays L) during water deficit stress at flowering time: a-tubulin, 3`phosphate glyceraldehyde dehydrogenase (GAPDH), 18S ribosomal subunit, protein 13S ribosomal, actin, zein, invertase, and starch synthase IIB. The eight reference genes candidates were tested on maize plants around flowering time, under three different conditions: before water deficit (BWD), under water deficit (WD) and after water deficit (AWD). Results from the three experimental conditions indicated that protein 13S ribosomal gene was the most stable among all the reference genes tested. This result suggests that protein 13S ribosomal gene can be used as internal control (housekeeping) for qPCR analysis in maize plants under water deficit stress during flowering time. Inst. de Genética "Ewald A. Favret"- IGEAF Fil: Decima Oneto, Cecilia Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina Fil: Bossio, Adrian Ezequiel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina Fil: Faccio, Paula Daniela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina Fil: Beznec, Ailin. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Blumwald, Eduardo. University of California Davies. Department Plant Sciences; Estados Unidos Fil: Lewi, Dalia Marcela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Genética; Argentina 2017-08-10T12:00:47Z 2017-08-10T12:00:47Z 2017 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/941 https://journals-crea.4science.it/index.php/maydica/article/view/1571/1081 2279-8013 eng 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 Maydica 62 : 6 p. |
| spellingShingle | Genética Genes Estrés de Sequia Floración Genetics Frought Stress Flowering Genes de Limpieza Estres Hídrico Decima Oneto, Cecilia Andrea Bossio, Adrian Ezequiel Faccio, Paula Daniela Beznec, Ailin Blumwald, Eduardo Lewi, Dalia Marcela Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title | Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title_full | Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title_fullStr | Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title_full_unstemmed | Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title_short | Validation of housekeeping genes for qPCR in maize during water deficit stress conditions at flowering time |
| title_sort | validation of housekeeping genes for qpcr in maize during water deficit stress conditions at flowering time |
| topic | Genética Genes Estrés de Sequia Floración Genetics Frought Stress Flowering Genes de Limpieza Estres Hídrico |
| url | http://hdl.handle.net/20.500.12123/941 https://journals-crea.4science.it/index.php/maydica/article/view/1571/1081 |
| work_keys_str_mv | AT decimaonetoceciliaandrea validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime AT bossioadrianezequiel validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime AT facciopauladaniela validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime AT beznecailin validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime AT blumwaldeduardo validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime AT lewidaliamarcela validationofhousekeepinggenesforqpcrinmaizeduringwaterdeficitstressconditionsatfloweringtime |