LcERF134 increases the production of monoterpenes by activating the terpene biosynthesis pathway in Litsea cubeba

Abstract

Litsea cubeba, an aromatic species of the Lauraceae family, produces a diverse array of monoterpenes. The biosynthesis of monoterpenes is regulated by transcriptional factors (TFs), such as APETALA2/ethylene response factor (AP2/ERF). However, the regulatory mechanisms that control the AP2/ERF gene responsible for the biosynthesis of monoterpenes in L. cubeba have yet to be elucidated. Here, we identified an AP2/ERF gene, LcERF134, as an activator for the accumulation of citral and other monoterpenes. The expression level of LcERF134 was consistent with terpene synthase LcTPS42 in the pericarp. The transient overexpression of LcERF134 significantly increased monoterpene production in L. cubeba as well as the expression of rate-limiting genes involved in the monoterpene biosynthesis pathway. Furthermore, yeast one-hybrid, dual-luciferase and electrophoretic mobility shift assays demonstrated that LcERF134 activated the monoterpene biosynthesis pathway by directly binding to the GCC-box elements of the LcTPS42 and LcGPPS.SSU1 promoters. However, the overexpression of LcERF134 in tomatoes had no impact on the synthesis of monoterpenes, thus indicating that LcERF134 is a species-specific TF. Our research demonstrated that LcERF134 significantly increased the biosynthesis of monoterpenes by inducing the expression of LcTPS42 and LcGPPS.SSU1, thus offering insight into how to enhance the flavor of L. cubeba essential oil.