Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover

Abstract The forage species Caucasian clover (Trifolium ambiguum M. Bieb.), a groundcover plant, is resistant to both cold and drought. However, reference genes for qRT-PCR-based analysis of Caucasian clover are lacking. In this study, 12 reference genes were selected on the basis of transcriptomic...

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Autores principales:	Xiujie Yin, Taotao He, Kun Yi, Yihang Zhao, Yao Hu, Jiaxue Liu, Xiaomeng Zhang, Lingdong Meng, Lina Wang, Haoyue Liu, Yonggang Li, Guowen Cui
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9251ac3fdd034d6eb3c5618f93373c0c
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spelling	oai:doaj.org-article:9251ac3fdd034d6eb3c5618f93373c0c2021-12-02T13:30:28ZComprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover10.1038/s41598-021-82633-22045-2322https://doaj.org/article/9251ac3fdd034d6eb3c5618f93373c0c2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82633-2https://doaj.org/toc/2045-2322Abstract The forage species Caucasian clover (Trifolium ambiguum M. Bieb.), a groundcover plant, is resistant to both cold and drought. However, reference genes for qRT-PCR-based analysis of Caucasian clover are lacking. In this study, 12 reference genes were selected on the basis of transcriptomic data. These genes were used to determine the most stably expressed genes in various organs of Caucasian clover under cold, salt and drought stress for qRT-PCR-based analysis. Reference gene stability was analyzed by geNorm, NormFinder, BestKeeper, the ∆Ct method and RefFinder. Under salt stress, RCD1 and PPIL3 were the most stable reference genes in the leaves, and NLI1 and RCD1 were the most stable references genes in the roots. Under low-temperature stress, APA and EFTu-GTP were the most stable reference genes in the leaves, and the RCD1 and NLI2 genes were highly stable in the roots. Under 10% PEG-6000 stress, NLI1 and NLI2 were highly stable in the leaves, and RCD1 and PPIL3 were the most stable in the roots. Overall, RCD1 and NLI2 were the most stable reference genes in organs under normal conditions and across all samples. The most and least stable reference genes were validated by assessing their appropriateness for normalization via WRKY genes.Xiujie YinTaotao HeKun YiYihang ZhaoYao HuJiaxue LiuXiaomeng ZhangLingdong MengLina WangHaoyue LiuYonggang LiGuowen CuiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Xiujie Yin Taotao He Kun Yi Yihang Zhao Yao Hu Jiaxue Liu Xiaomeng Zhang Lingdong Meng Lina Wang Haoyue Liu Yonggang Li Guowen Cui Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
description	Abstract The forage species Caucasian clover (Trifolium ambiguum M. Bieb.), a groundcover plant, is resistant to both cold and drought. However, reference genes for qRT-PCR-based analysis of Caucasian clover are lacking. In this study, 12 reference genes were selected on the basis of transcriptomic data. These genes were used to determine the most stably expressed genes in various organs of Caucasian clover under cold, salt and drought stress for qRT-PCR-based analysis. Reference gene stability was analyzed by geNorm, NormFinder, BestKeeper, the ∆Ct method and RefFinder. Under salt stress, RCD1 and PPIL3 were the most stable reference genes in the leaves, and NLI1 and RCD1 were the most stable references genes in the roots. Under low-temperature stress, APA and EFTu-GTP were the most stable reference genes in the leaves, and the RCD1 and NLI2 genes were highly stable in the roots. Under 10% PEG-6000 stress, NLI1 and NLI2 were highly stable in the leaves, and RCD1 and PPIL3 were the most stable in the roots. Overall, RCD1 and NLI2 were the most stable reference genes in organs under normal conditions and across all samples. The most and least stable reference genes were validated by assessing their appropriateness for normalization via WRKY genes.
format	article
author	Xiujie Yin Taotao He Kun Yi Yihang Zhao Yao Hu Jiaxue Liu Xiaomeng Zhang Lingdong Meng Lina Wang Haoyue Liu Yonggang Li Guowen Cui
author_facet	Xiujie Yin Taotao He Kun Yi Yihang Zhao Yao Hu Jiaxue Liu Xiaomeng Zhang Lingdong Meng Lina Wang Haoyue Liu Yonggang Li Guowen Cui
author_sort	Xiujie Yin
title	Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
title_short	Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
title_full	Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
title_fullStr	Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
title_full_unstemmed	Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover
title_sort	comprehensive evaluation of candidate reference genes for quantitative real-time pcr-based analysis in caucasian clover
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/9251ac3fdd034d6eb3c5618f93373c0c
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Comprehensive evaluation of candidate reference genes for quantitative real-time PCR-based analysis in Caucasian clover

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