Biochemical characterization of diterpene synthases of Taiwania cryptomerioides expands the known functional space of specialized diterpene metabolism in gymnosperms

• Published in: The Plant journal : for cell and molecular biology Vol. 100; no. 6; pp. 1254 - 1272
• Main Authors: Ma, Li‐Ting, Lee, Yi‐Ru, Tsao, Nai‐Wen, Wang, Sheng‐Yang, Zerbe, Philipp, Chu, Fang‐Hua
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01.12.2019
• Subjects: Abietatriene; Angiospermae; Angiosperms; biochemical enzyme characterization; Biological evolution; Biosynthesis; Chemical defense; chemical defenses; cupressaceae; decay resistance; diterpene metabolism; diterpene synthase; Diterpenes; diterpenoids; Divergence; divergent evolution; durability; Enzymes; Fungicides; genes; Gymnosperms; Metabolism; Metabolites; Methyl jasmonate; Molecular evolution; Organic chemistry; plant specialized metabolism; Taiwania cryptomerioides; Terpenes; trees; wood; diterpene synthase; biochemical enzyme characterization; gymnosperms; plant specialized metabolism; Taiwania cryptomerioides; diterpene metabolism; methyl jasmonate; cupressaceae
• ISSN: 0960-7412; 1365-313X; 1365-313X
• DOI: 10.1111/tpj.14513

Summary Taiwania cryptomerioides is a monotypic gymnosperm species, valued for the high decay resistance of its wood. This durability has been attributed to the abundance of terpenoids, especially the major diterpenoid metabolite ferruginol, with antifungal and antitermite activity. Specialized diterpenoid metabolism in gymnosperms primarily recruits bifunctional class‐I/II diterpene synthases (diTPSs), whereas monofunctional class‐II and class‐I enzymes operate in angiosperms. In this study, we identified a previously unrecognized group of monofunctional diTPSs in T. cryptomerioides, which suggests a distinct evolutionary divergence of the diTPS family in this species. Specifically, five monofunctional diTPS functions not previously observed in gymnosperms were characterized, including monofunctional class‐II enzymes forming labda‐13‐en‐8‐ol diphosphate (LPP, TcCPS2) and (+)‐copalyl diphosphate (CPP, TcCPS4), and three class‐I diTPSs producing biformene (TcKSL1), levopimaradiene (TcKSL3) and phyllocladanol (TcKSL5), respectively. Methyl jasmonate (MeJA) elicited the accumulation of levopimaradiene and the corresponding biosynthetic diTPS genes, TcCPS4 and TcKSL3, is consistent with a possible role in plant defense. Furthermore, TcCPS4 and TcKSL3 are likely to contribute to abietatriene biosynthesis via levopimaradiene as an intermediate in ferruginol biosynthesis in Taiwania. In conclusion, this study provides deeper insight into the functional landscape and molecular evolution of specialized diterpenoid metabolism in gymnosperms as a basis to better understand the role of these metabolites in tree chemical defense. Significance Statement Taiwania cryptomerioides TcCPS2 and TcCPS4 were characterized as monofunctional class‐II diTPSs that have not previously been reported in gymnosperms. Phylogenetic analysis indicates that these TcdiTPSs evolved from bifunctional (TPS‐d group) and monofunctional (TPS‐e/f group) ancestors. The (+)‐CPS (TcCPS4) and levopimaradiene synthase (TcKSL3) are likely to be involved in abietatriene biosynthesis in Taiwania, which is a possible precursor of ferruginol and taiwaniaquinonoids.