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Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium
Pleurotus tuber-regium is a sclerotium-producing mushroom with pharmacological and nutritional value. Many researches applied molecular biological strategies that allow analyzing numerous genes expressed in response to different growth stresses or tissues. In order to capture accurate and reliable r...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The Mycological Society of Japan
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9721512/ https://www.ncbi.nlm.nih.gov/pubmed/37089465 http://dx.doi.org/10.47371/mycosci.2021.05.001 |
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author | Sun, Xueyan Wu, Junyue Mo, Cuiyuan Ma, Aimin |
author_facet | Sun, Xueyan Wu, Junyue Mo, Cuiyuan Ma, Aimin |
author_sort | Sun, Xueyan |
collection | PubMed |
description | Pleurotus tuber-regium is a sclerotium-producing mushroom with pharmacological and nutritional value. Many researches applied molecular biological strategies that allow analyzing numerous genes expressed in response to different growth stresses or tissues. In order to capture accurate and reliable results in gene expression studies, it is necessary to select appropriate internal reference genes. They must have relatively uniform expression in the majority of tested samples, thereby reducing the impact of the sample itself on the RT-qPCR results. The selection and validation of 14 candidate reference genes, including α-tub, β-tub, γ-tub, GAPDH, Tif-5a, Tef-1α, ATPase, acyl-CoA, U2ribo, S/G, Wlp, VtpAsE, E3upl, and Sdh, were carried out for gene expression analysis in P. tuber-regium mycelia in response to different temperatures, drought levels, and salinity shifts, fruitbody, and sclerotium. Four statistical algorithms, NormFinder, geNorm, BestKeeper, and RefFinder, were recruited to evaluate the transcription stability of candidate reference genes. The RT-qPCR expression stability analysis indicated that the E3upl and Tif-5a were the most stable expressed genes among all experimental samples, so the combination of these two reference genes is suitable for the analysis of gene expression patterns in P. tuber-regium. |
format | Online Article Text |
id | pubmed-9721512 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | The Mycological Society of Japan |
record_format | MEDLINE/PubMed |
spelling | pubmed-97215122023-04-20 Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium Sun, Xueyan Wu, Junyue Mo, Cuiyuan Ma, Aimin Mycoscience Full Paper Pleurotus tuber-regium is a sclerotium-producing mushroom with pharmacological and nutritional value. Many researches applied molecular biological strategies that allow analyzing numerous genes expressed in response to different growth stresses or tissues. In order to capture accurate and reliable results in gene expression studies, it is necessary to select appropriate internal reference genes. They must have relatively uniform expression in the majority of tested samples, thereby reducing the impact of the sample itself on the RT-qPCR results. The selection and validation of 14 candidate reference genes, including α-tub, β-tub, γ-tub, GAPDH, Tif-5a, Tef-1α, ATPase, acyl-CoA, U2ribo, S/G, Wlp, VtpAsE, E3upl, and Sdh, were carried out for gene expression analysis in P. tuber-regium mycelia in response to different temperatures, drought levels, and salinity shifts, fruitbody, and sclerotium. Four statistical algorithms, NormFinder, geNorm, BestKeeper, and RefFinder, were recruited to evaluate the transcription stability of candidate reference genes. The RT-qPCR expression stability analysis indicated that the E3upl and Tif-5a were the most stable expressed genes among all experimental samples, so the combination of these two reference genes is suitable for the analysis of gene expression patterns in P. tuber-regium. The Mycological Society of Japan 2021-09-20 /pmc/articles/PMC9721512/ /pubmed/37089465 http://dx.doi.org/10.47371/mycosci.2021.05.001 Text en 2021, by The Mycological Society of Japan https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open-access paper distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivative 4.0 international license (CC BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Full Paper Sun, Xueyan Wu, Junyue Mo, Cuiyuan Ma, Aimin Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title | Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title_full | Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title_fullStr | Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title_full_unstemmed | Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title_short | Selection and validation of endogenous reference genes for RT-qPCR normalization in different stresses and tissues of the tiger milk mushroom, Pleurotus tuber-regium |
title_sort | selection and validation of endogenous reference genes for rt-qpcr normalization in different stresses and tissues of the tiger milk mushroom, pleurotus tuber-regium |
topic | Full Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9721512/ https://www.ncbi.nlm.nih.gov/pubmed/37089465 http://dx.doi.org/10.47371/mycosci.2021.05.001 |
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