Stable Internal Reference Genes for Normalizing Real-Time Quantitative PCR in Baphicacanthus cusia under Hormonal Stimuli and UV Irradiation, and in Different Plant Organs

Frontiers in Plant Science
Yuxiang HuangYong Diao

Abstract

Baphicacanthus cusia (Nees) Bremek, the plant source for many kinds of drugs in traditional Chinese medicine, is widely distributed in South China, especially in Fujian. Recent studies about B. cusia mainly focus on its chemical composition and pharmacological effects, but further analysis of the plant's gene functions and expression is required to better understand the synthesis of its effective compounds. Real-time quantitative polymerase chain reaction (RT-qPCR) is a powerful method for gene expression analysis. It is necessary to select a suitable reference gene for expression normalization to ensure the accuracy of RT-qPCR results. Ten candidate reference genes were selected from the transcriptome datasets of B. cusia in this study, and the expression stability was assessed across 60 samples representing different tissues and organs under various conditions, including ultraviolet (UV) irradiation, hormonal stimuli (jasmonic acid methyl ester and abscisic acid), and in different plant organs. By employing different algorithms, such as geNorm, NormFinder, and BestKeeper, which are complementary approaches based on different statistical procedures, 18S rRNA was found to be the most stable gene under UV irradiation and hormona...Continue Reading

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Citations

Jun 6, 2020·The Journal of Organic Chemistry·Ling-Juan ZhuHong-Wei Liu

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Datasets Mentioned

BETA
SRR4428209

Methods Mentioned

BETA
environmental stresses
RNA-Seq
electrophoresis
Hi-Seq
PCR

Software Mentioned

RefFinder
geNorm
Primer3
GAPDH
tblastx
BestKeeper
Excel
NormFinder
RNA

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