Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L

Abstract Iris germanica L. is a perennial herbaceous plant that has been widely cultivated worldwide and is popular for its elegant and vibrantly colorful flowers. Selection of appropriate reference genes is the prerequisite for accurate normalization of target gene expression by quantitative real-t...

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Autores principales:	Yinjie Wang, Yongxia Zhang, Qingquan Liu, Haiying Tong, Ting Zhang, Chunsun Gu, Liangqin Liu, Suzhen Huang, Haiyan Yuan
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/aaeb94a8180748249aa6eca117309f2a
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spelling	oai:doaj.org-article:aaeb94a8180748249aa6eca117309f2a2021-12-02T14:35:39ZSelection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L10.1038/s41598-021-89100-y2045-2322https://doaj.org/article/aaeb94a8180748249aa6eca117309f2a2021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89100-yhttps://doaj.org/toc/2045-2322Abstract Iris germanica L. is a perennial herbaceous plant that has been widely cultivated worldwide and is popular for its elegant and vibrantly colorful flowers. Selection of appropriate reference genes is the prerequisite for accurate normalization of target gene expression by quantitative real-time PCR. However, to date, the most suitable reference genes for flowering stages have not been elucidated in I. germanica. In this study, eight candidate reference genes were examined for the normalization of RT-qPCR in three I. germanica cultivars, and their stability were evaluated by four different algorithms (GeNorm, NormFinder, BestKeeper, and Ref-finder). The results revealed that IgUBC and IgGAPDH were the most stable reference genes in ‘00246’ and ‘Elizabeth’, and IgTUB and IgUBC showed stable expression in ‘2010200’. IgUBC and IgGAPDH were the most stable in all samples, while IgUBQ showed the least stability. Finally, to validate the reliability of the selected reference genes, the expression patterns of IgFT (Flowering Locus T gene) was analyzed and emphasized the importance of appropriate reference gene selection. This work presented the first systematic study of reference genes selection during flower bud development and provided guidance to research of the molecular mechanisms of flowering stages in I. germanica.Yinjie WangYongxia ZhangQingquan LiuHaiying TongTing ZhangChunsun GuLiangqin LiuSuzhen HuangHaiyan YuanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yinjie Wang Yongxia Zhang Qingquan Liu Haiying Tong Ting Zhang Chunsun Gu Liangqin Liu Suzhen Huang Haiyan Yuan Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
description	Abstract Iris germanica L. is a perennial herbaceous plant that has been widely cultivated worldwide and is popular for its elegant and vibrantly colorful flowers. Selection of appropriate reference genes is the prerequisite for accurate normalization of target gene expression by quantitative real-time PCR. However, to date, the most suitable reference genes for flowering stages have not been elucidated in I. germanica. In this study, eight candidate reference genes were examined for the normalization of RT-qPCR in three I. germanica cultivars, and their stability were evaluated by four different algorithms (GeNorm, NormFinder, BestKeeper, and Ref-finder). The results revealed that IgUBC and IgGAPDH were the most stable reference genes in ‘00246’ and ‘Elizabeth’, and IgTUB and IgUBC showed stable expression in ‘2010200’. IgUBC and IgGAPDH were the most stable in all samples, while IgUBQ showed the least stability. Finally, to validate the reliability of the selected reference genes, the expression patterns of IgFT (Flowering Locus T gene) was analyzed and emphasized the importance of appropriate reference gene selection. This work presented the first systematic study of reference genes selection during flower bud development and provided guidance to research of the molecular mechanisms of flowering stages in I. germanica.
format	article
author	Yinjie Wang Yongxia Zhang Qingquan Liu Haiying Tong Ting Zhang Chunsun Gu Liangqin Liu Suzhen Huang Haiyan Yuan
author_facet	Yinjie Wang Yongxia Zhang Qingquan Liu Haiying Tong Ting Zhang Chunsun Gu Liangqin Liu Suzhen Huang Haiyan Yuan
author_sort	Yinjie Wang
title	Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
title_short	Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
title_full	Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
title_fullStr	Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
title_full_unstemmed	Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L
title_sort	selection and validation of appropriate reference genes for rt-qpcr analysis of flowering stages and different genotypes of iris germanica l
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/aaeb94a8180748249aa6eca117309f2a
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Selection and validation of appropriate reference genes for RT-qPCR analysis of flowering stages and different genotypes of Iris germanica L

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