biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 64:798-806, 2020 | DOI: 10.32615/bp.2020.154

Transcriptome-based screening and the optimal reference genes for real-time quantitative PCR in Rehmannia chingii and R. henryi

X. ZUO, F.-Q. WANG*, X.-R. LI, M.-M. LI
College of Agronomy, Henan Agricultural University, Zhengzhou 450002, P.R. China

Real time quantitative PCR (qPCR) is a powerful tool for studying the expression of specific genes. The accuracy and reliability of qPCR analysis data require the selection of reference genes with stable expression. However, the reference genes that can be used for qPCR of Rehmannia chingii and R. henryi have not yet been identified. In this study, based on the transcriptome data of R. chingii and R. henryi, we initially selected genes with relatively stable expression in different samples. We screened six candidate reference genes in R. chingii and R. henryi and calculated their expression abundance by real time qPCR. Their expression stability was evaluated by three algorithms geNorm, NormFinder, and BestKeeper. Although the results obtained by different algorithms were not completely consistent, R. chingii type 2A phosphatase activator TIP41 and R. chingii 18S ribosomal RNA had the highest expression stability in six different samples of R. chingii, and R. henryi 18S ribosomal RNA and R. henryi actin showed the most stable expression in different samples of R. henryi. In addition, based on transcriptome data, four genes were screened in R. chingii and R. henryi, and the expression stability of the selected reference genes was further verified. This study laid the foundation for further analysis and verification of the functions of important genes in R. chingii and R. henryi.

Keywords: BestKeeper, gene expression stability, geNorm, NormFinder.

Received: July 13, 2020; Revised: October 18, 2020; Accepted: October 19, 2020; Published online: December 11, 2020  Show citation

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ZUO, X., WANG, F.-Q., LI, X.-R., & LI, M.-M. (2020). Transcriptome-based screening and the optimal reference genes for real-time quantitative PCR in Rehmannia chingii and R. henryi . Biologia plantarum, 64, Article 798-806. https://doi.org/10.32615/bp.2020.154
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