Selection of suitable reference genes for qRT-PCR expression analysis of Codonopsis pilosula under different experimental conditions

文献类型： 外文期刊

第一作者： Yang, Jing

作者： Yang, Jing;Li, Bin;Lu, Xiayang;Cao, Xiaoyan;Kang, Jiefang;Yang, Jing;Yang, Xiaozeng;Kuang, Zheng;Lu, Xiayang;Kuang, Zheng;Kuang, Zheng

作者机构：

关键词： Codonopsis pilosula; Developmental stages; Tissues; Stress treatments; qRT-PCR; Reference genes

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

ISSN： 0301-4851

年卷期： 2020 年 47 卷 6 期

页码：

收录情况： SCI

摘要： Codonopsis pilosula is a well-known medicinal plant. Although its transcriptome sequence has been published, suitable reference genes have not been systematically identified for conducting expression analyses via quantitative real-time polymerase chain reaction (qRT-PCR). To screen appropriate genes for use with this species, we applied four different methods-GeNorm, NormFinder, BestKeeper, and RefFinder-to evaluate the stability of 13 candidates: CpiEF1Bb, CpiCACS, CpiF-Box, Cpi beta-Tubulin, CpiGAPDH, CpiActin2, CpiAPT1, CpiActin7, CpiActin8, CpiRPL6, CpiHAF1, CpiTubulin6, and CpiUBQ12. Expression was examined by qRT-PCR for various tissue types, chemical treatments, and developmental stages. For all tested samples, CpiGAPDH proved to be the most stable. Comprehensive analysis indicated that the most stable internal reference genes were CpiF-Box and CpiCACS in different tissues and at different developmental stages, respectively. Under NaCl stress, CpiAPT1 was the best internal reference gene. For methyl jasmonate and abscisic acid treatments, CpiGAPDH and CpiF-Box, respectively, presented the highest degree of expression stability. Based on these findings, we chose CpiSPL9 as the target gene for validating the suitability of these selected reference genes. All of these results provide a foundation for accurate quantification of expression levels by genes of interest in C. pilosula.

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