欢迎访问《植物研究》杂志官方网站,今天是 分享到:

植物研究 ›› 2018, Vol. 38 ›› Issue (4): 611-618.doi: 10.7525/j.issn.1673-5102.2018.04.017

• 研究报告 • 上一篇    下一篇

珠子参中法尼基焦磷酸合酶(FPS)对皂苷生物合成的影响研究

刘美佳, 于怡琳, 姜森, 张翔, 刘迪秋, 葛锋   

  1. 昆明理工大学生命科学与技术学院, 昆明 650500
  • 收稿日期:2018-02-28 出版日期:2018-07-15 发布日期:2018-07-21
  • 通讯作者: 葛锋 E-mail:panaxcordyceps@163.com
  • 作者简介:刘美佳(1991-)女,硕士研究生,主要从事分子生物学研究。
  • 基金资助:
    国家自然科学基金项目(81560616)

Effect of Farnesyl-pyrophosphate Synthase(FPS) on the Biosynthesis of Saponins in Panax japonicus

LIU Mei-Jia, YU Yi-Lin, JIANG Sen, ZHANG Xiang, LIU Di-Qiu, GE Feng   

  1. Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500
  • Received:2018-02-28 Online:2018-07-15 Published:2018-07-21
  • Supported by:
    National Natural Science Foundation of China(81560616)

摘要: 珠子参作为名贵中草药已有上千年应用历史,珠子参皂苷是珠子参的主要活性成分,由达玛烷型和齐墩果烷型三萜皂苷组成。目前,对珠子参皂苷的生物合成了解甚少,有必要开展与皂苷合成相关的基础研究工作。已有报道表明,法尼基焦磷酸合酶(FPS)可能是植物中皂苷合成的关键酶。本研究克隆了珠子参FPS基因(PjFPS)的cDNA序列,并对其进行了生物信息学分析。基于PjFPS序列构建了珠子参过表达载体pCAMBIA2300s-PjFPS,将其转化到珠子参细胞中,成功获得阳性转基因细胞;测定了阳性细胞系中PjFPS基因的相对表达量、FPS酶活、皂苷以及植物甾醇含量的变化。结果表明,与野生型珠子参细胞相比,PjFPS转基因珠子参细胞系中,PjFPS基因的相对表达量、FPS酶活以及皂苷含量均有不同程度的提高。而且,由于皂苷合成代谢流的增加,也促进了相关重要关键酶基因的表达。在效果最好的阳性细胞中,PjFPS基因的相对表达量、FPS酶活、皂苷含量分别为对照的12、4和3倍;同时,转基因细胞系的植物甾醇含量也有所升高。本研究通过对皂苷合成途径关键酶基因的调控实现了对皂苷生物合成的调节,为获得高效、稳定的珠子参皂苷合成调控技术提供理论参考和依据。

关键词: 珠子参, 人参属, 人参皂苷, 三萜, 过表达

Abstract: Panax japonicus recognized as a valuable Chinese herb has been used for thousands of years. P.japonicus saponins are the main bioactive ingredients composed by dammarane-type and oleanane-type triterpenoid saponins. So far, because of the lack of study about the biosynthesis of P.japonicus saponins, it's necessary to carry out the basic research on saponins biosynthesis. Farnesyl pyrophosphate synthase(FPS) may be a key enzyme in the synthetic pathway of saponins in plants. The cDNA sequence of PjFPS was cloned and bioinformatics analysis was conducted. Overexpression vector pCAMBIA2300s-PjFPS was constructed, and then transferred into the P.japonicus cells to obtain transgenic cell lines successfully. The relative expression levels of PjFPS, the enzyme activities of PjFPS, the contents of PJS and phytosterols in transgenic cell lines were determined. All of them in PjFPS transgenic cells achieved enhancements compared with the control. Furthermore, the expressions of key enzyme genes were promoted because the metabolic flux of saponins was increased. Especially, the expression level of PjFPS, the enzyme activity and total P.japonicus saponins content of the best-performing positive cell line were 12, 4 and 3 times of those in control, respectively. Meanwhile, the content of phytosterol in transgenic cell lines was also enhanced. This study realized the biosynthetic regulation of saponins by controlling the expression of a key enzyme gene, and thus to provide theoretical reference and basis for acquiring an efficient and stable technique of regulating the P.japonicus saponins biosynthesis.

Key words: Panax japonicus, Panax, ginsenoside, triterpenoid, overexpression

中图分类号: