Evaluation of Reference Genes Suitable for Gene Expression during Root Enlargement in Cherry Radish Based on Transcriptomic Data

Abstract

Reliable reference genes (RGs) are of great significance for the normalization of quantitative data. RGs are often used as a reference to ensure the accuracy of experimental results to detect gene expression levels by reverse transcription–quantitative real-time PCR (RT-qPCR). To evaluate the normalized RGs that are suitable for studying the expression of genes during the process of radish stele enlargement, based on the functional annotations and fragment per kilobase of transcript per million mapped reads (FPKM) values in the transcriptome data, three traditional RGs (GAPDH, 18SrRNA, and ACTIN7) and seven commonly used RGs (UBQ11, TUA6, TUB6, EF-1b1, EF-1a2, PP2A11, and SAND) were obtained. In the study, the results of geNorm, NormFinder, and BestKeeper from RefFinder comprehensively analyzed the stability ranking of candidate RGs. The results showed that compared with the traditional RGs, the common RGs show higher and more stable expression. Among the seven commonly used RGs, PP2A11 is recommended as the optimal RG for studying cherry radish stele enlargement. This research provides a useful and reliable RG resource for the accurate study of gene expression during root enlargement in cherry radishes and facilitates the functional genomics research on root enlargement.