Optimization of reference genes for qRT-PCR of Pleurotus ostreatus un-der different lighting treatments
HU Mengdan
ZHU Mengke
ZHANG Peng
GAO Yuqian
QIU Liyou
LI Yanan
Abstract：To identify suitable reference genes for analyzing the specific gene expression of Pleurotus ostreatus under dif-ferent lighting treatments,the mycelium of P.ostreatus was used as the material,and 12 genes were screened from the transcriptome data of mycelium under different lighting treatments as candidate reference genes for qRT-PCR of P.ostrea-tus,and their expression abundance and expression stability were evaluated.The results indicated that the primers of 12 candidate reference genes had good specificity in qPCR,with single peaks in the melting curve,and the CT values of the genes were between 18 and 26,demonstrating good gene expression abundance.The expression stability analysis of ge-Norm,NormFinder and BestKeeper showed that the expression stability of eukaryotic translation initiation factorl encod-ing gene(eIF1),ubiquitin-conjugating enzyme encoding gene(UBC),40S ribosomal protein encoding gene(40SRP)and actinl encoding gene(Actinl)were at the forefront in the three analyses.The expression abundance of these four genes un-der different lighting treatments from high to low was UBC,eIF1,Actin1 and 40SRP.eIF1 and UBC can be used as the best reference gene combination at the high abundance expression level of target genes.40SRP and Actin1 can be used as the best reference gene combination at medium and low abundance expression levels of target genes.
Keywords：Pleurotus ostreatusReference geneLighting treatmentqRT-PCR
Publication Date：2025-06-05
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：9( 35-43 )