Validation of Terpenoid Pathway Genes in Rosa chinensis

Stage 5 petals and different tissues from OB and OBL were collected and processed for total RNA extraction and first-strand cDNA synthesis as previously described [51 (link)]. The key DEGs involved in the terpenoid metabolic pathway and RcWRKY70 were subjected to validation by RT–qPCR using RcACTIN as the reference transcript. Primer sequences are provided in Table S11. qPCR was performed with a Green^® Premix Ex Taq™ II (RR420Q, Takara, Tokyo, Japan) and the CFX Connect Real-Time PCR Detection System (Bio–Rad, Hercules, CA, USA). Each sample was tested as three technical replicates. Relative gene expression levels were obtained by the comparative 2^−ΔΔCt method [52 ]. All results were plotted in Origin9 (OriginLab, Northampton, MA, USA).