萜类化合物是结构多样的天然产物,已广泛用于制药,食物,和化妆品行业。研究表明,真菌产生多种萜类化合物,然而,真菌萜合酶仍未彻底探索。在这项研究中,tps1基因,萜烯合成途径的关键组成部分,通过基因组挖掘从木霉HB20111中分离。通过构建tps1基因缺失和过表达工程菌株,并在转录水平上评估tps1基因的表达差异,研究了该基因在萜烯合成途径中的功能。HS-SPME-GC-MS分析显示野生型中萜烯代谢物的显着差异,tps1-删除(Δtps1),和tps1过表达(Otps1)菌株;例如,大多数倍半萜挥发性有机化合物(VOCs)在Δtps1菌株中显著减少或不存在,而Nerolidol,β-acorenol,和愈创烯特别由Otps1菌株产生。然而,与野生型相比,Δtps1和Otps1菌株都产生了新的萜烯代谢产物,这表明tps1基因在萜烯合成中起重要作用,但不是T.atrovirideHB20111涉及的唯一基因。通过生物学信息和进化树分析,tps1基因编码的TPS1蛋白可以作为倍半萜环化酶发挥作用。此外,真菌抑制试验和小麦生长促进试验结果表明,tps1基因的缺失或过表达对真菌抑制活性的影响最小,促进植物生长,和发展,以及应激反应。这意味着T.atrovirideHB20111的这些活性可能来自多种代谢物的组合,而不是仅仅依赖于一种特定的代谢物。本研究为今后研究萜类化合物的合成机制提供了理论指导,为真菌中萜类化合物代谢途径的相关研究奠定了基础。
Terpenoids are structurally diverse natural products that have been widely used in the pharmaceutical, food, and cosmetic industries. Research has shown that fungi produce a variety of terpenoids, yet fungal terpene synthases remain not thoroughly explored. In this study, the tps1 gene, a crucial component of the terpene synthetic pathway, was isolated from Trichoderma atroviride HB20111 through genome mining. The function of this gene in the terpene synthetic pathway was investigated by constructing tps1-gene-deletion- and overexpression-engineered strains and evaluating the expression differences in the tps1 gene at the transcript level. HS-SPME-GC-MS analysis revealed significant variations in terpene metabolites among wild-type, tps1-deleted (Δtps1), and tps1-overexpressed (Otps1) strains; for instance, most sesquiterpene volatile organic compounds (VOCs) were notably reduced or absent in the Δtps1 strain, while nerolidol, β-acorenol, and guaiene were particularly produced by the Otps1 strain. However, both the Δtps1 and Otps1 strains produced new terpene metabolites compared to the wild-type, which indicated that the tps1 gene played an important role in terpene synthesis but was not the only gene involved in T. atroviride HB20111. The TPS1 protein encoded by the tps1 gene could function as a sesquiterpene cyclase through biological information and evolutionary tree analysis. Additionally, fungal inhibition assay and wheat growth promotion assay results suggested that the deletion or overexpression of the tps1 gene had a minimal impact on fungal inhibitory activity, plant growth promotion, and development, as well as stress response. This implies that these activities of T. atroviride HB20111 might result from a combination of multiple metabolites rather than being solely dependent on one specific metabolite. This study offers theoretical guidance for future investigations into the mechanism of terpenoid synthesis and serves as a foundation for related studies on terpenoid metabolic pathways in fungi.