Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions

Abstract Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression leve...

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Autores principales:	Min-dong Chen, Bin Wang, Yong-ping Li, Mei-juan Zeng, Jian-ting Liu, Xin-ru Ye, Hai-sheng Zhu, Qing-fang Wen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b629d974140348aca7c5db4d8ef12ab3
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spelling	oai:doaj.org-article:b629d974140348aca7c5db4d8ef12ab32021-12-02T10:44:14ZReference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions10.1038/s41598-021-81524-w2045-2322https://doaj.org/article/b629d974140348aca7c5db4d8ef12ab32021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81524-whttps://doaj.org/toc/2045-2322Abstract Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression levels in luffa plants exposed to various simulated abiotic stresses [i.e., cold, drought, heat, salt, H2O2, and abscisic acid (ABA) treatments] were analyzed by qRT-PCR. The stability of the reference gene expression levels was validated using the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. The results indicated that EF-1α was the most stably expressed and suitable reference gene overall and for the heat, cold, and ABA treatments. Additionally, UBQ expression was stable following the salt treatment, whereas TUB was identified as a suitable reference gene for H2O2 and drought treatments. The reliability of the selected reference genes was verified by analyzing the expression of copper/zinc superoxide dismutase (Cu/Zn-SOD) gene in luffa. When the most unstable reference genes were used for data normalizations, the resulting expression patterns had obvious biases when compared with the expression patterns for the most ideal reference genes used alone or combined. These results will be conducive to more accurate quantification of gene expression levels in luffa.Min-dong ChenBin WangYong-ping LiMei-juan ZengJian-ting LiuXin-ru YeHai-sheng ZhuQing-fang WenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Min-dong Chen Bin Wang Yong-ping Li Mei-juan Zeng Jian-ting Liu Xin-ru Ye Hai-sheng Zhu Qing-fang Wen Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
description	Abstract Selecting suitable internal reference genes is an important prerequisite for the application of quantitative real-time PCR (qRT-PCR). However, no systematic studies have been conducted on reference genes in luffa. In this study, seven reference genes were selected, and their expression levels in luffa plants exposed to various simulated abiotic stresses [i.e., cold, drought, heat, salt, H2O2, and abscisic acid (ABA) treatments] were analyzed by qRT-PCR. The stability of the reference gene expression levels was validated using the geNorm, NormFinder, BestKeeper, and RefFinder algorithms. The results indicated that EF-1α was the most stably expressed and suitable reference gene overall and for the heat, cold, and ABA treatments. Additionally, UBQ expression was stable following the salt treatment, whereas TUB was identified as a suitable reference gene for H2O2 and drought treatments. The reliability of the selected reference genes was verified by analyzing the expression of copper/zinc superoxide dismutase (Cu/Zn-SOD) gene in luffa. When the most unstable reference genes were used for data normalizations, the resulting expression patterns had obvious biases when compared with the expression patterns for the most ideal reference genes used alone or combined. These results will be conducive to more accurate quantification of gene expression levels in luffa.
format	article
author	Min-dong Chen Bin Wang Yong-ping Li Mei-juan Zeng Jian-ting Liu Xin-ru Ye Hai-sheng Zhu Qing-fang Wen
author_facet	Min-dong Chen Bin Wang Yong-ping Li Mei-juan Zeng Jian-ting Liu Xin-ru Ye Hai-sheng Zhu Qing-fang Wen
author_sort	Min-dong Chen
title	Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
title_short	Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
title_full	Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
title_fullStr	Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
title_full_unstemmed	Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions
title_sort	reference gene selection for qrt-pcr analyses of luffa (luffa cylindrica) plants under abiotic stress conditions
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b629d974140348aca7c5db4d8ef12ab3
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Reference gene selection for qRT-PCR analyses of luffa (Luffa cylindrica) plants under abiotic stress conditions

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