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Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress
BACKGROUND: Real-time quantitative PCR (RT-qPCR) is the most commonly used method for accurately detecting gene expression patterns. As part of RT-qPCR analysis, normalization of the data requires internal control gene(s) that display uniform expression under different biological conditions. However...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5644265/ https://www.ncbi.nlm.nih.gov/pubmed/29075311 http://dx.doi.org/10.1186/s13007-017-0238-7 |
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author | Tang, Xun Zhang, Ning Si, Huaijun Calderón-Urrea, Alejandro |
author_facet | Tang, Xun Zhang, Ning Si, Huaijun Calderón-Urrea, Alejandro |
author_sort | Tang, Xun |
collection | PubMed |
description | BACKGROUND: Real-time quantitative PCR (RT-qPCR) is the most commonly used method for accurately detecting gene expression patterns. As part of RT-qPCR analysis, normalization of the data requires internal control gene(s) that display uniform expression under different biological conditions. However, no invariable internal control gene exists, and therefore more than one reference gene is needed to normalize RT-qPCR results. Identification of stable reference genes in potato will improve assay accuracy for selecting stress-tolerance genes and identifying molecular mechanisms conferring stress tolerance in this species. RESULTS: In the experiment, we assessed the expression of eight candidate internal control genes, namely elongation factor-1alpha (EF1α), actin, tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), adenine phosphoribosyl transferase (APRT), 60S ribosomal protein L8 (L8), Cullin 3A (CUL3A), and exocyst complex component sec3 (sec3), in a diverse set of potato samples representing drought stress and osmotic stress challenges, and using geNorm, NormFinder, BestKeeper and RefFinder softwares. CONCLUSIONS: The results indicated that EF1α and sec3 were the most stably expressed genes in the potato under drought and osmotic stress conditions. This work will facilitate future work on gene expression studies in potato and also benefit other species of the Solanaceae, such as tomato. |
format | Online Article Text |
id | pubmed-5644265 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-56442652017-10-26 Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress Tang, Xun Zhang, Ning Si, Huaijun Calderón-Urrea, Alejandro Plant Methods Research BACKGROUND: Real-time quantitative PCR (RT-qPCR) is the most commonly used method for accurately detecting gene expression patterns. As part of RT-qPCR analysis, normalization of the data requires internal control gene(s) that display uniform expression under different biological conditions. However, no invariable internal control gene exists, and therefore more than one reference gene is needed to normalize RT-qPCR results. Identification of stable reference genes in potato will improve assay accuracy for selecting stress-tolerance genes and identifying molecular mechanisms conferring stress tolerance in this species. RESULTS: In the experiment, we assessed the expression of eight candidate internal control genes, namely elongation factor-1alpha (EF1α), actin, tubulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), adenine phosphoribosyl transferase (APRT), 60S ribosomal protein L8 (L8), Cullin 3A (CUL3A), and exocyst complex component sec3 (sec3), in a diverse set of potato samples representing drought stress and osmotic stress challenges, and using geNorm, NormFinder, BestKeeper and RefFinder softwares. CONCLUSIONS: The results indicated that EF1α and sec3 were the most stably expressed genes in the potato under drought and osmotic stress conditions. This work will facilitate future work on gene expression studies in potato and also benefit other species of the Solanaceae, such as tomato. BioMed Central 2017-10-16 /pmc/articles/PMC5644265/ /pubmed/29075311 http://dx.doi.org/10.1186/s13007-017-0238-7 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Tang, Xun Zhang, Ning Si, Huaijun Calderón-Urrea, Alejandro Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title | Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title_full | Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title_fullStr | Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title_full_unstemmed | Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title_short | Selection and validation of reference genes for RT-qPCR analysis in potato under abiotic stress |
title_sort | selection and validation of reference genes for rt-qpcr analysis in potato under abiotic stress |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5644265/ https://www.ncbi.nlm.nih.gov/pubmed/29075311 http://dx.doi.org/10.1186/s13007-017-0238-7 |
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