Dual-LUC assays were performed as reported previously14. The primer pairs used to clone the promoters of genes involved in phenolic acid biosynthesis are listed in Supplementary Table S1. The promoter regions of candidate genes (~2000 bp) were inserted into A pGreen 0800-LUC vector driving a firefly LUC reporter gene, and the Renilla luciferase (REN) gene was controlled by the 35S promoter (Fig. S4). The recombinant vectors were subsequently transferred into A. tumefaciens (GV3101) together with a helper plasmid (pSoup-P19) encoding a cosuppressor. N. benthamiana leaves were then transiently transformed with the constructs by infiltrating the Agrobacterium mixture into the abaxial sides of the leaves. The infected area was harvested, extracts were evaluated using a Dual-Luciferase Reporter Assay System (Promega, Madison, USA), and, after 48 h, the fluorescence values of LUC and REN were detected with a luminometer. The LUC/REN ratio indicated the ability of SmMYB1 to bind to and activate gene promoters.

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