The Ginkgo biloba microRNA160–ERF4 module participates in terpene trilactone biosynthesis.

This article investigates the regulatory role of the gb-miR160–GbERF4 module in the biosynthesis of terpene trilactones (TTLs), important secondary metabolites unique to Ginkgo biloba with medicinal properties. The study demonstrates that gb-miR160 negatively regulates TTL synthesis by targeting and cleaving the transcription factor GbERF4, which itself positively regulates TTL biosynthesis by binding to and activating promoters of key genes in both the mevalonate (MVA) and methylerythritol phosphate (MEP) pathways, including HMGS1, AACT1, DXS1, GGPPS2, and LPS2. Transgenic overexpression of GbERF4 in tobacco and transient expression in G. biloba increased terpenoid content and upregulated structural genes, while overexpression of gb-miR160 reduced terpenoid accumulation. These findings elucidate a molecular mechanism by which the gb-miR160–GbERF4 regulatory module modulates TTL biosynthesis, providing potential gene targets for genetic improvement of G. biloba to enhance TTL content.