Global identification and functional validation of transcription factors regulating sesquiterpenoid metabolism in Pogostemon cablin

Patchouli is a medicinal plant belonging to the Pogostemon genus within the Lamiaceae family. It is composed mainly of sesquiterpenoid metabolites, and has various pharmacological effects. The mechanism underlying the transcriptional regulation of the patchouli alcohol has been reported, but the mechanism underlying the regulation of transcription factors (TFs) involved in the sesquiterpenoid metabolic pathway is still unknown. In this study, a regulatory network was constructed for the sesquiterpenoid metabolic pathway in patchouli by using a gene expression database and WGCNA. It was found that 286 TFs have potential regulatory relationships with 28 genes that encode enzymes in the sesquiterpenoid metabolic pathway. The regulatory mechanism of this network was successfully verified by yeast one-hybrid (Y1H), dual luciferase reporter assay (dual-LUC), transient overexpression, and electrophoretic mobility shift assay (EMSA). In conclusion, a regulation network was constructed for sesquiterpenoid metabolic pathways in Pogostemon cablin, and all the TFs that may have regulatory functions in sesquiterpenoid metabolic pathways were identified, and the regulatiory functions of transcription factors were successfully verified through bench experiments.