Evaluation of Reference Genes for Expression Normalization in Psoralea Corylifolia L. under Abiotic Stress

Abstract

Background: Psoralea corylifolia L., a traditional Chinese herbal medicine, has a wide range of pharmacological activities for the treatment of various diseases. To date, there have been no published reports of the systematic evaluation and selection of the best reference genes for biological analysis of P. corylifolia. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a method for quantifying gene expression. Selecting the appropriate reference genes is essential to ensure accurate normalization of RT-qPCR results.Results: In this study, 10 candidate reference genes, namely actin (ACT), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), protein phosphatase 2A subunit A3 (PP2A), nuclear cap-binding protein subunit 2 (NCBP2), cyclophilin 2 (CYP2), elongation factor 1-alpha (EF-1α), protein phosphatase 2A-2 (PP2A2), thioredoxin-like protein YLS8 (YLS8), polypyrimidine tract-binding protein (PTBP), and TIP41-like family protein (TIP41), were screened from the transcriptome database of P. corylifolia. The plants were treated under different abiotic stress conditions (H2O2, NaCl, CuSO4, high-temperature, low-temperature, PEG, and UV). Next, their expression stability was evaluated using four algorithm tools: BestKeeper, NormFinder, geNorm, and RefFinder. The results revealed that TIP41, CYP2, and YLS8 were the top three genes with the most stable expression levels under various abiotic stress conditions.Conclusion: We successfully selected appropriate reference genes for P. corylifolia, providing a reliable basis for further studies on the quantitative gene expression profile of the main active components in P. corylifolia.