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植物研究 ›› 2023, Vol. 43 ›› Issue (5): 779-786.doi: 10.7525/j.issn.1673-5102.2023.05.014

• 分子生物学 • 上一篇    下一篇

龙牙楤木Aeβ-AS基因的表达对烟草中皂苷含量的影响

霍清清, 夏雨新, 李佳乐, 韩薇, 张书雅, 张哲, 夏美玲, 郭雯华, 由香玲()   

  1. 东北林业大学生命科学学院东北盐碱植被恢复与重建教育部重点实验室,哈尔滨 150040
  • 收稿日期:2023-03-19 出版日期:2023-09-20 发布日期:2023-09-05
  • 通讯作者: 由香玲 E-mail:youxiangling@nefu.edu.cn
  • 作者简介:霍清清(2002—),女,本科,主要从事药用植物细胞工程研究。
  • 基金资助:
    东北林业大学本科生创新项目(202210225097)

Effect of Expression of Aeβ-AS gene from Aralia elata on the Content of Saponins in Nicotiana tabacum L.

Qingqing HUO, Yuxin XIA, Jiale LI, Wei HAN, Shuya ZHANG, Zhe ZHANG, Meiling XIA, Wenhua GUO, Xiangling YOU()   

  1. Laboratory of Saline-alkali Vegetation Ecology Restoration,Ministry of Education,College of Life Sciences,Northeast Forestry University,Harbin 150040
  • Received:2023-03-19 Online:2023-09-20 Published:2023-09-05
  • Contact: Xiangling YOU E-mail:youxiangling@nefu.edu.cn
  • About author:HUO Qingqing(2002—),female,undergraduate student,research direction is medicinal plant cell engineering.
  • Supported by:
    Undergraduate Innovation Project of Northeast Forestry University(202210225097)

摘要:

为探讨龙牙楤木(Aralia elata)三萜皂苷合成的关键酶基因β-香树素合成酶基因(Aeβ-AS)对三萜皂苷合成的影响。利用基因克隆技术对Aeβ-AS基因进行克隆,并对烟草进行遗传转化,分析转基因烟草中Aeβ-AS基因在各部位的表达差异,检测Aeβ-AS基因及上下游关键酶基因表达量和总三萜含量。结果表明:成功构建了植物过表达载体pROKⅡ-Aeβ-AS,并转入了野生型烟草,获得7个转基因株系,并在mRNA水平表达,且在叶子中的表达量要高于根和茎;在转基因烟草中,Aeβ-AS基因及其上下游的关键酶基因表达量均高于野生型,其中株系L21的NtFPS、NtSS基因的相对表达量最高,株系L30的NtSE、Aeβ-AS基因的相对表达量最高;与野生型烟草相比,转基因烟草的总三萜含量显著升高(1.1~1.6倍)。试验结果证明合成Aeβ-AS基因并在烟草中进行异源转化,可以使转基因烟草内的总三萜含量显著升高。

关键词: 龙牙楤木, Aeβ-AS基因, 烟草, 三萜含量

Abstract:

To investigate the effect of β-amyrin synthase gene(Aeβ-AS) on triterpenoid saponin synthesis of Aralia elata, the Aeβ-AS gene was cloned and genetically transformed into tobacco. The expression difference of Aeβ-AS gene in different parts of transgenic tobacco was analyzed, and the expression levels of Aeβ-AS gene, upstream and downstream key enzyme genes and total triterpene content were detected respectively. The results showed that the plant overexpression vector PROKⅡ-Aeβ-AS was successfully constructed and transferred into wild type tobacco, and seven transgenic lines were obtained and expressed at the mRNA level, and the expression level in leaves was higher than that in roots and stems. In transgenic tobacco, the expression levels of Aeβ-AS gene and its upstream and downstream key enzyme genes were higher than those of wild type, and the relative expression levels of NtFPS and NtSS genes of strain L21 were the highest, and the relative expression levels of NtSE and Aeβ-AS genes of strain L30 were the highest. Compared with wild-type tobacco, the total triterpene content of GM tobacco was significantly increased(1.1-1.6 times). The results showed that synthesis of Aeβ-AS gene and heterologous transformation in tobacco could significantly increase the total triterpene content in transgenic tobacco.

Key words: Aralia elata, Aeβ-AS gene, Nicotiana tabacum, total triterpenoids content

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