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Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress
Quantitative real-time reverse-transcriptase PCR (qRT-PCR) has been frequently used for detecting gene expression. To obtain reliable results, selection of suitable reference genes is a fundamental and necessary step. Garlic (Allium sativum), a member from Alliaceae family, has been used both as a f...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
PeerJ Inc.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6640627/ https://www.ncbi.nlm.nih.gov/pubmed/31341748 http://dx.doi.org/10.7717/peerj.7319 |
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author | Wang, Guanglong Tian, Chang Wang, Yunpeng Wan, Faxiang Hu, Laibao Xiong, Aisheng Tian, Jie |
author_facet | Wang, Guanglong Tian, Chang Wang, Yunpeng Wan, Faxiang Hu, Laibao Xiong, Aisheng Tian, Jie |
author_sort | Wang, Guanglong |
collection | PubMed |
description | Quantitative real-time reverse-transcriptase PCR (qRT-PCR) has been frequently used for detecting gene expression. To obtain reliable results, selection of suitable reference genes is a fundamental and necessary step. Garlic (Allium sativum), a member from Alliaceae family, has been used both as a food flavoring and as a traditional medicine. In the present study, garlic plants were exposed to salt stress (200 mM NaCl) for 0, 1, 4 and 12 h, and garlic roots, bulbs, and leaves were harvested for subsequent analysis. The expression stability of eight candidate reference genes, eukaryotic translation initiation factor 4α (eIF-4α), actin (ACTIN), tubulin β-7 (TUB7), TAP42-interacting protein of 41 kDa (TIP41), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), SAND family protein (SAND), elongation factor 1 alpha (EF-1α), and protein phosphatase 2A (PP2A) were evaluated by geNorm, NormFinder, and BestKeeper. All genes tested displayed variable expression profiles under salt stress. In the leaf and root group, ACTIN was the best reference gene for normalizing gene expression. In garlic clove, ACTIN and SAND were the least variable, and were suitable for gene expression studies under salt stress; these two genes also performed well in all samples tested. Based on our results, we recommend that it is essential to use specific reference genes in different situations to obtain accurate results. Using a combination of multiple stable reference genes, such as ACTIN and SAND, to normalize gene expression is encouraged. The results from the study will be beneficial for accurate determination of gene expression in garlic and other plants. |
format | Online Article Text |
id | pubmed-6640627 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | PeerJ Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-66406272019-07-24 Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress Wang, Guanglong Tian, Chang Wang, Yunpeng Wan, Faxiang Hu, Laibao Xiong, Aisheng Tian, Jie PeerJ Agricultural Science Quantitative real-time reverse-transcriptase PCR (qRT-PCR) has been frequently used for detecting gene expression. To obtain reliable results, selection of suitable reference genes is a fundamental and necessary step. Garlic (Allium sativum), a member from Alliaceae family, has been used both as a food flavoring and as a traditional medicine. In the present study, garlic plants were exposed to salt stress (200 mM NaCl) for 0, 1, 4 and 12 h, and garlic roots, bulbs, and leaves were harvested for subsequent analysis. The expression stability of eight candidate reference genes, eukaryotic translation initiation factor 4α (eIF-4α), actin (ACTIN), tubulin β-7 (TUB7), TAP42-interacting protein of 41 kDa (TIP41), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), SAND family protein (SAND), elongation factor 1 alpha (EF-1α), and protein phosphatase 2A (PP2A) were evaluated by geNorm, NormFinder, and BestKeeper. All genes tested displayed variable expression profiles under salt stress. In the leaf and root group, ACTIN was the best reference gene for normalizing gene expression. In garlic clove, ACTIN and SAND were the least variable, and were suitable for gene expression studies under salt stress; these two genes also performed well in all samples tested. Based on our results, we recommend that it is essential to use specific reference genes in different situations to obtain accurate results. Using a combination of multiple stable reference genes, such as ACTIN and SAND, to normalize gene expression is encouraged. The results from the study will be beneficial for accurate determination of gene expression in garlic and other plants. PeerJ Inc. 2019-07-16 /pmc/articles/PMC6640627/ /pubmed/31341748 http://dx.doi.org/10.7717/peerj.7319 Text en ©2019 Wang et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited. |
spellingShingle | Agricultural Science Wang, Guanglong Tian, Chang Wang, Yunpeng Wan, Faxiang Hu, Laibao Xiong, Aisheng Tian, Jie Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title | Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title_full | Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title_fullStr | Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title_full_unstemmed | Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title_short | Selection of reliable reference genes for quantitative RT-PCR in garlic under salt stress |
title_sort | selection of reliable reference genes for quantitative rt-pcr in garlic under salt stress |
topic | Agricultural Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6640627/ https://www.ncbi.nlm.nih.gov/pubmed/31341748 http://dx.doi.org/10.7717/peerj.7319 |
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