Abstract:
To screen stable reference genes for studies on gene expression of aroma release and the circadian clock changes in
Cestrum nocturnum,six traditional reference genes,
Actb7,EF-1A,GAPDH,TUA,TUB2,and
UBQ,were isolated from flowers and leaves in the blooming stage by homology cloning and RACE technology.The expression patterns of
18s rRNA and the six reference genes were analysed by real-time quantitative PCR (qRT-PCR) and the expression stabilities were analysed by Bestkeeper,geNorm,and NormFinder.Results showed that the expression of
Actb7 was stable in flowers;
EF-1A and
UBQ were stable in leaves;and
EF-1A was stable in both tissues.According to the analysis of the three groups by geNorm,the optimal number of reference genes was two and the best combination was
Actb7 and
EF-1A.The stable reference genes screened in this study provide a basis for accurate detection of the expression of genes related to petal rhythm movement,aroma release,and circadian clock changes.