园艺学报 ›› 2020, Vol. 47 ›› Issue (2): 310-320.doi: 10.16420/j.issn.0513-353x.2019-0721

• 研究论文 • 上一篇    下一篇

桂花‘莲籽丹桂’芳樟醇合酶基因 OfTPS5 的克隆及功能鉴定

刘 偲 1,4,*,席 婉 1,2,*,袁金梅 1,2,朱琳琳 1,2,陈洪国 ,3 邹晶晶 3,郑日如 1,2,**,王彩云   

  1. 1 华中农业大学园艺植物生物学教育部重点实验室,武汉 430070;2 农业农村部华中都市农业重点实验室,武汉430070;3湖北科技学院,湖北咸宁 437100;4 四川省林业和草原科学技术推广总站,成都 610000
  • 出版日期:2020-02-25 发布日期:2020-02-25
  • 基金资助:

Molecular Cloning and Functional Characterization of Linalool Synthase Gene OfTPS5 in Osmanthus fragrans‘Lianzi Dangui’Flowers

LIU Cai1,4,*,XI Wan1,2,*,YUAN Jinmei1,2,ZHU Linlin1,2,CHEN Hongguo3,ZOU Jingjing3,ZHENG Riru1,2,**,and WANG Caiyun1,2   

  1. 1Key Laboratory for Horticultural Plant Biology,Ministry of Education,Huazhong Agricultural University,Wuhan 430070,China;2Key Laboratory of Urban Agriculture in Central China,Ministry of Agriculture and Rural Affairs,P. R. China,Wuhan 430070,China;3Hubei University of Science and Technology,Xianning,Hubei437100,China;4Sichuan Science and Technology Extensive Station of Forestry and Grassland,Chengdu 610000,China
  • Online:2020-02-25 Published:2020-02-25

摘要: 以桂花(Osmanthus fragrans Lour.)‘莲籽丹桂’为材料,从基因组数据库中克隆到 1 个萜烯合酶基因 OfTPS5,鉴定其序列信息及功能,分析其表达规律及其与香气形成的关系。结果表明,OfTPS5(GenBank 登录号为 MN366471)的开放阅读框为 1 770 bp,编码 589 个氨基酸,其编码的蛋白质具有萜烯合酶基因家族的保守结构域,属于 TPS-g 亚家族。序列比对发现它与其他芳樟醇合酶基因的同源性在50%左右。OfTPS5 在盛花期高表达,且呈现明显的昼升夜降的节律性变化,其转录表达水平与单萜类物质芳樟醇及其衍生物的合成呈正相关。烟草瞬时表达试验证明,OfTPS5 可催化形成芳樟醇。由此推测,OfTPS5 在桂花萜类香气形成中发挥重要作用。

关键词: 桂花, 香气, 芳樟醇及其衍生物, 芳樟醇合酶基因, 表达分析, 功能鉴定

Abstract: This study was conducted to isolate a linalool synthase gene from Osmanthus fragrans Lour.‘Lianzi Dangui’based on genome sequences. Functional identification,expression analysis and the relation with aroma compound formation were investigated. The gene was designated as OfTPS5(GenBank accession No. MN366471). Sequence analysis demonstrated that the open reading frame(ORF)was 1 770 bp,and putatively encoded 589 amino acids. The OfTPS5 contained conserved domains and belonged to TPS-g subfamily. Multiple sequence alignment analysis demonstrated that it has approximate 50% homology with other plant TPSs. The transcript expression of OfTPS5 reached high level at the full bloom stage with a significant circadian rhythm,which also showed positive correlation with the aroma compound formation. Further transient expression in tobacco leaves proved that OfTPS5 was capable of producing linalool. Thus,OfTPS5 might play an important role in terpenoid biosynthesis in O. fragrans flowers.

Key words: Osmanthus fragrans, aroma, linalool and its derivatives, linalool synthase gene, expression analysis, functional characterization