Evaluation and validation of reference genes in Cymbidium faberi for real-time quantitative PCR

For all organs at all Cymbidium faberi stages, ACT, UBQ3 and GAPDH can be selected as reference genes. For organs of the vegetative stage, UBQ2 and UBQ3 can be chosen for analysis of normalized gene expression. For the bud stage, ACT and UBQ3 can be used for analysis of gene expression. For the full blossom stage, ACT, UBQ3 and UBQ2 can be introduced into relative gene expression analysis. For vegetative organs, UBQ2 and ACT can be used as reference genes. For reproductive organs, ACT, UBQ3 and UBQ2 can be used as a reference for data processing. CfAG1 gene expression is more consistent when UBQ3, GAPDH and ACT are used as reference genes. Method summary To select reference genes in Cymbidium faberi for more accurate quantification of target genes, expression levels of candidate reference genes were detected by real-time quantitative PCR. Expression stability of eight candidate reference genes was evaluated using geNorm and NormFinder software. To further verify stability of the eight candidate reference genes, the expression of CfAG1 was detected.