Total RNA of all samples was extracted using the Plant RNAprep Pure Kit (Tiangen, Beijing, China). cDNA synthesis from total RNA and qRT-PCR analyses of all gene transcripts were performed as previously described24. The housekeeping gene OpActin in O. pumila was used as an internal control gene in qRT-PCR for normalization of all samples. All gene-specific primer sequences of all camptothecin biosynthesis pathway genes and OpWRKY genes used for qRT-PCR analyses are listed in Table S1. The relative gene expression values in all samples were calculated using the 2−ΔΔCt method. All qRT-PCR analyses of each sample were performed for three biological replicates.

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