Reference Gene Selection for Quantitative Real-Time PCR Analysis of Hymenopellis radicata under Abiotic Stress

Abstract

Abstract Housekeeping genes are genes that are expressed stably in organisms. It is essential to select appropriate reference genes prior to conducting gene expression in order to obtain accurate results. Hymenopellis radicata (H. radicata) is an edible fungus rich in protein and mineral elements, with high edible and medical value. And reference genes suitable for normalization of qRT-PCR data from this species have not been investigated. In this study, therefore, we selected 11 housekeeping genes common in biology (CYP, GAPDH, PP2A, RPL6, UBC, UBQ10, ACT, EF, RPB2, TUA, TUB). The expression levels of these housekeeping genes were measured in three different tissues (pileus, stipe and root) and six different abiotic stress treatments in mycelium. They were evaluated for expression stability using online tools (GeNorm, NormFinder, BestKeeper and RefFinder). This study is the first to screen the most suitable genes as internal references in H. radicata and analyze the expression levels of H. radicata genes under abiotic stress and in different tissues. This project provides a reference for further research on the molecular mechanism of H. radicata under abiotic stress. Additionally, the results of this study can be used to select internal reference genes for expression analysis in other species.