Overexpression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase enhances the monoterpene content in Litsea cubeba

Abstract

Monoterpenes are important components of plant essential oils and have long been used as raw materials for spices and food flavorings. Litsea cubeba is an economically aromatic plant species, the fruits of which produce essential oil with monoterpenes as the dominant components. As a branch point of carbon flow in the methyl erythritol phosphate (MEP) biosynthesis pathway, 1-deoxy-D-xylo-5-phosphate reductoisomerase (DXR) is a key rate-limiting enzyme that catalyzes the MEP pathway’s second committed step. Therefore, DXR has become an effective regulation target to improve the biosynthesis of plant monoterpenes. In this study, we identified a DXR from L. cubeba, which was highly expressed in fruits, induced by MeJA and repressed by darkness. An enzyme assay showed that recombination LcDXR protein catalyzed DXP with NADPH as the cofactor. Transient overexpression of LcDXR significantly increased the content of monoterpenes in L. cubeba. Furthermore, LcDXR-overexpressing tobaccos were conducted and showed almost 5.9-fold increase in monoterpenes production, including limonene, α-pinene, eucalyptol, linalool, terpineol and camphor. Overexpression of LcDXR activated the metabolic flux of monoterpene biosynthesis through crosstalk and feedback mechanism. In addition, LcDXR-overexpressing tobaccos had no effect on phenotype of transgenic tobaccos. Our results demonstrate that LcDXR is a critical regulator of the monoterpene production in L. cubeba and other plants.

Keywords

• litsea cubeba
• monoterpene
• dxp reductoisomerase
• mep pathway