Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum

doi:10.3390/ijms23020738

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	Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum
	Dong, Xiu-Mei; Zhang, Wei; Zhang, Shi-Bao
	2022
发表期刊	INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
卷号	23 期号:2 页码:738
摘要	The development and tissue-dependent color formation of the horticultural plant results in various color pattern flowers. Anthocyanins and carotenoids contribute to the red and yellow colors, respectively. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) is used to analyze the expression profiles of anthocyanin and carotenoids biosynthesis genes in Cymbidium lowianum (Rchb.f.) Rchb.f. Appropriate reference gene selection and validation are required before normalization of gene expression in qRT-PCR analysis. Thus, we firstly selected 12 candidate reference genes from transcriptome data, and used geNorm and Normfinder to evaluate their expression stability in lip (divided into abaxial and adaxial), petal, and sepal of the bud and flower of C. lowianum. Our results show that the two most stable reference genes in different tissues of C. lowianum bud and flower are EF1 delta and 60S, the most unstable reference gene is 26S. The expression profiles of the CHS and BCH genes were similar to FPKM value profiles after normalization to the two most stable reference genes, EF1 delta and 60S, with the upregulated CHS and BCH expression in flower stage, indicating that the ABP and CBP were activated across the stages of flower development. However, when the most unstable reference gene, 26S, was used to normalize the qRT-PCR data, the expression profiles of CHS and BCH differed from FPKM value profiles, indicating the necessity of selecting stable reference genes. Moreover, CHS and BCH expression was highest in the abaxial lip and adaxial lip, respectively, indicating that the ABP and CBP were activated in abaxial and adaxial lip, respectively, resulting in a presence of red or yellow segments in abaxial and adaxial lip. This study is the first to provide reference genes in C. lowianum, and also provide useful information for studies that aim to understand the molecular mechanisms of flower color formation in C. lowianum.
关键词	Cymbidium lowianum flower organ development reference gene selection and validation qRT-PCR transcriptome data geNorm and Normfinder software POLYMERASE-CHAIN-REACTION EXPRESSION NORMALIZATION QUANTIFICATION
DOI	10.3390/ijms23020738
收录类别	SCI
WOS记录号	WOS:000746022100001
引用统计
文献类型	期刊论文
条目标识符	http://ir.kib.ac.cn/handle/151853/74711
专题	中国科学院昆明植物研究所
推荐引用方式 GB/T 7714	Dong, Xiu-Mei,Zhang, Wei,Zhang, Shi-Bao. Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum[J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,2022,23(2):738.
APA	Dong, Xiu-Mei,Zhang, Wei,&Zhang, Shi-Bao.(2022).Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum.INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,23(2),738.
MLA	Dong, Xiu-Mei,et al."Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum".INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 23.2(2022):738.

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