安徽农业大学学报2025，Vol.52Issue(5)：779-787,9.DOI:10.13610/j.cnki.1672-352x.20250519.016
神农香菊萜类合酶基因CaTPS30的克隆及功能验证
Cloning and functional verification of terpene synthase gene CaTPS30 from Chrysanthemum aromaticum
摘要
Abstract
[Objective]Chrysanthemum aromaticum is an important wild ornamental plant renowned for its unique fragrance.Terpene synthases(TPSs)are key enzymes catalyzing the formation of terpenoids in C.aro-maticum.This study aimed to functionally characterize the critical TPS gene responsible for β-ocimene biosynthe-sis in C.aromaticum.[Method]The functional characterization of a terpene synthase gene,CaTPS30,was inves-tigated by integrating volatile metabolomics analysis,gene expression analysis,bioinformatics analysis,protein prokaryotic expression,and in vitro enzyme activity assay.[Result]Nine volatile monoterpenes were detected across different tissues of C.aromaticum,with β-ocimene(1.299%)and γ-terpinene(1.572%)exhibiting the high-est relative abundances in the leaves.Quantitative PCR revealed that CaTPS30 expression was highest in the leaves and lowest in the flowers,correlating strongly with β-ocimene accumulation patterns across or-gans.CaTPS30 was identified as a protein of 584 amino acids(67.55 kDa),classified as an unstable hydrophilic protein.Phylogenetic analysis placed CaTPS30 within the TPS-b subfamily,featuring conserved DDXXD and NSE/DTE motifs.In vitro enzymatic assays demonstrated that recombinant CaTPS30 catalyzed geranyl diphosphate(GPP)to produce β-ocimene as the primary product,with trace amounts of allo-ocimene.[Conclusion]This study elucidates the molecular mechanisms underlying terpenoid biosynthesis in C.aromaticum and provides a theoretical foundation for improving aromatic traits in chrysanthemum cultivars.关键词
神农香菊/萜类合酶/原核表达/功能验证/β-罗勒烯Key words
Chrysanthemum aromaticum/terpene synthase/prokaryotic expression/functional verification/β-ocimene分类
农业科技引用本文复制引用
陈昱媛,钟剑,张鹏,岳肖雨,徐玉娴,孙明..神农香菊萜类合酶基因CaTPS30的克隆及功能验证[J].安徽农业大学学报,2025,52(5):779-787,9.基金项目
国家自然科学基金项目(31971708,32271947) （31971708,32271947）
雄安新区科技创新专项(2022XAGG0100) （2022XAGG0100）
中央高校优秀青年团队项目(QNTD202306) （QNTD202306）
