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Updated: Jun 14, 2026

A Customizable Approach for the Enzymatic Production and Purification of Diterpenoid Natural Products
Published on: October 4, 2019
LaWRKY23 and LaWRKY65 negatively regulate key terpene synthases (LaTPS39) to repress sesquiterpenoid biosynthesis in
Yujie Shi1, Shumeng Zhang2, Lijuan Wei3
1Zhejiang International Science and Technology Cooperation Base for Biomass Resources Development and Utilization, School of Life Sciences, Taizhou University, Taizhou, 318000, China; Zhejiang Key Laboratory for Restoration of Damaged Coastal Ecosystems, School of Life Sciences, Taizhou University, Taizhou, 318000, China.
Abstract:
Lindera aggregata is a traditional Chinese medicine with sesquiterpenoids as key bioactive components. Although its genome has been published, the pivotal metabolic genes involved in sesquiterpenoid biosynthesis and their transcriptional regulation remain to be elucidated. This study explored sesquiterpenoid synthesis regulation via multi-omics. Headspace Solid Phase Micro-extraction Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS) identified 69 sesquiterpenoids, and the roots showed the highest accumulation. Genome-wide analysis revealed 64 terpene synthase (TPS) genes. LaTPS25, LaTPS35, and LaTPS39 were functionally validated to catalyze farnesyl pyrophosphate into sesquiterpenes, such as β-selinene and germacrene D, through in vitro assays and molecular docking. Weighted Gene Co-expression Network Analysis (WGCNA) identified LaTPS39 as a hub gene regulated by three WRKY transcription factors. Yeast One-Hybrid (Y1H), Dual-Luciferase Reporter Assay, and Electrophoretic Mobility Shift Assay (EMSA) confirmed that LaWRKY23 and LaWRKY65 directly bind to the W-box within the LaTPS39 promoter, potentially negatively regulating its expression and inhibiting sesquiterpenoid synthesis. These findings clarify the regulatory mechanism underlying sesquiterpenoid biosynthesis in L. aggregata.
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