The PatERF061 transcription factor activates patchoulol synthase gene expression and positively regulates jasmonate-induced patchoulol biosynthesis in Pogostemon cablin

The sesquiterpene patchoulol, the main active constituent of the Lamiaceae herbal plant Pogostemon cablin, is broadly applied in the pharmaceutical and cosmetic industries for its excellent pharmacological and biological activities. However, research on the biosynthesis and regulation of patchoulol remains significantly limited. In this study, PatERF061, a member of AP2/ERF transcription factor family, was identified as a PatPTS promoter ( PatPTSpro )-binding protein. Subcellular localization assay revealed that PatERF061 was mainly located in the nucleus and cytoplasm. The expression of PatERF061 was predominantly observed in patchouli leaves and was induced by methyl jasmonate (MeJA). Combination with the results of yeast one-hybrid assay, dual-luciferase reporter assays, and transient overexpression assay, we demonstrated that PatERF061 could directly bind to the PatPTSpro and positively regulate patchoulol biosynthesis via modulating the expression of patchoulol biosynthetic genes. Additionally, the patchouli jasmonate ZIM-domain 4 (PatJAZ4) was found to interact with PatERF061 and repress patchoulol biosynthesis. Moreover, the interaction between PatERF061 and PatSWC4 was also identified; the resulting complex of PatERF061-PatSWC4 was capable of binding to and activating the PatPTSpro , ultimately enhancing patchoulol production. However, PatJAZ4 could repress the transcriptional activation capability of the PatERF061-PatSWC4 complex, and this interference could be reversed by JA. Overall, this study revealed the positive regulated roles of PatERF061 and the PatERF061-PatSWC4 complex in synthesizing patchoulol. These findings contribute significantly to our understanding of the regulatory network governing patchoulol biosynthesis.