Abstract
Blumea balsamifera, a wooden plant belonging to the family Asteraceae, is a medicinal herb with anticancer, antiviral, and multiple pharmacological effects, which are believed to be caused by its essential oil. The essential oil from B. balsamifera is comprised of mono- and sesqui-terpenes as the majority. Unfortunately, this plant has been facing the challenge of resource shortage, which could be effectively alleviated by biological engineering. Therefore, the identification of key elements involved in the biosynthesis of active ingredients becomes an indispensable prerequisite. In this study, candidate genes encoding monoterpene synthase were screened by transcriptome sequencing combined with metabolomics profiling in the roots, stems, and leaves of B. balsamifera. Then, these candidates were successfully cloned and verified by heterologous expression and in vitro enzyme activity assays. As a result, six candidate BbTPS genes were isolated from B. balsamifera, of which three encoded single-product monoterpene synthases and one encoded a multi-product monoterpene synthase. Among them, BbTPS1, BbTPS3, and BbTPS4 could catalyze the formation of D-limonene, α-phellandrene, and L-borneol, respectively. Meanwhile, BbTPS5 functioned in catalyzing GPP into terpinol, β-phellandrene, β-myrcene, D-limonene, and 2-carene in vitro. In general, our results provided important elements for the synthetic biology of volatile terpenes in B. balsamifera, which laid a foundation for subsequent heterologous production of these terpenoids through metabolic engineering and increasing their yield, as well as promoting sustainable development and utilization of B. balsamifera.
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The sequencing raw data of RNA-seq has been deposited on the Sequence Read Archive (SRA) of NCBI, with an accession number PRJNA950886. The peptide sequences of those enzymes authenticated in this study have been deposited onto the NCBI database as well, with accession numbers listed below: BbTPS1 (OQ599523), BbTPS3 (OQ599524), BbTPS4 (OQ599525), and BbTPS5 (OQ599526). All the data are presented in the main manuscript and additional supplementary materials.
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This research was supported by the National Natural Science Foundation of China (Grant number: 81903741).
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TZ carried out the experiments, analyzed the data, and wrote the manuscript. FL, JL, and LL participated in the experiments and revised the manuscript. ZY and CX guided the experiments and participated in bioinformatics analysis. HW reviewed the draft. All authors read and approved the manuscript. XZ conceived and designed the experiments, and revised the manuscript.
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Zhan, T., Li, F., Lan, J. et al. Functional characterization of four mono-terpene synthases (TPSs) provided insight into the biosynthesis of volatile monoterpenes in the medicinal herb Blumea balsamifera. Physiol Mol Biol Plants 29, 459–469 (2023). https://doi.org/10.1007/s12298-023-01306-8
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DOI: https://doi.org/10.1007/s12298-023-01306-8