沙生蜡菊qRT-PCR内参基因筛选与验证OA

Five reference genes(ACT1,ACT7,CYP,HIS,TUB)were selected as candidate reference genes from the transcriptome data of Helichrysum arenarium,and qRT-PCR was used to detect the expression levels of candidate reference genes in different tissues and under freezing stress of Helichrysum arenarium.geNorm,NormFinder,BestKeeper and RefFinder were used to evaluate their stability.The stability of the reference gene was verified by using HaWRKY gene as the target gene.The expression of HaWRKY gene in Helichrysum arenarium was analyzed to verify the effect of reference genes on the experimental results.It was found that CYP and HIS were stably expressed in Helichrysum arenarium,but HIS was stably expressed after freezing stress.In summary,HIS can be used as an internal reference gene of Helichrysum arenarium.In this study,the suitable reference genes of Helichrysum arenarium were obtained,which provided reliable technical support for gene expression analysis of Helichrysum arenarium,and also laid a foundation for further study on the molecular mechanism of freezing tolerance and regulation of secondary metabolism.