茉莉酸甲脂诱导雷公藤愈伤组织中雷公藤红素合成的影响研究

发布时间：2021-04-27 05:24

　　雷公藤（Tripteygium wilfordii）卫矛科雷公藤属植物,是中国传统医学中广为人知的一种,原产于中国南方,其医疗用途在《滇南奔草》中有所记录。卫矛科植物中含有的380多种代谢物,根据其结构特征评估了它们的生物活性,其中95%是萜类化合物。雷公藤是一种产生萜烯的植物,其生物活性化合物主要是二萜类化合物（例如雷公藤甲素）和三萜类化合物（雷公藤红素）。由于其具有抗炎,免疫抑制,抗囊肿作用和抗癌等活性,雷公藤植物的次生代谢物引起了人们广泛的兴趣。然而,这些化合物在雷公藤植物体内含量很低,且它们的生物合成途径尚未得到完全解析,如何大规模合成这些化合物一直是令人困扰的关键科学问题。植物愈伤是生产次生代谢物的良好体系,本研究摸索了雷公藤植物不同组织器官的愈伤诱导体系,并基于诱导的愈伤组织,研究了信号分子茉莉酸甲酯对雷公藤红素生物合成的影响,取得了以下结果:1、发现了雷公藤植物叶器官最为适合诱导愈伤组织,根与茎器官愈伤诱导效率低下,并且容易遭受真菌或细菌污染;同时,在黑暗条件下培养更容易获得质地疏松的愈伤组织.2、在所诱导的雷公藤植物愈伤组织中检测到了雷公藤红素,未能检测到雷公藤甲素;经... 

【文章来源】：中国科学院大学(中国科学院武汉植物园)湖北省

【文章页数】：79 页

【学位级别】：硕士

【文章目录】：
摘要
Abstract
Chapter 1 Introduction
    1.1 Research background and literature review
        1.1.1 Tripterygium
        1.1.2 Terpenoids
        1.1.3 Classification of terpenoids
        1.1.4 Functions of Terpenoids
        1.1.5 Signature compounds of Terpenoids from Tripterygium
        1.1.6 Terpenoid biosynthesis
        1.1.7 Feasible methods for terpenoid production
    1.2 The basis and significance of this study
    1.3 The main research results of predecessors in the present study topic
    1.4 The content of this study
    1.5 Purpose of this study
Chapter 2 Materials and methods
    2.1 Plant material and growth conditions
    2.2 Composition of the Murashige and Skoog （MS） basal medium
    2.3 Callus culture in Tripterygium wilfordii
    2.4 Methyl Jasmonate （MJ） induction
    2.5 Preparations of the calli for RNA and metabolite analysis
    2.6 Preparation of standard stock solutions
    2.7 HPLC conditions
    2.8 LC-MS analysis
    2.9 RNA isolation and synthesis of cDNA
    2.10 Real-Time quantitative PCR （qRT-PCR）
    2.11 Statistical analysis
    2.12 Sequences used in the present study
Chapter 3 Results
    3.1 Callus induction
        3.1.1 Callus induction from root explants
        3.1.2 Callus induction from leaf explants
    3.2 Effect of different combination of hormones on callus initiation
    3.3 Effect of light and dark conditions on callus initiation
        3.3.1 Calli cultured in 15h light 9 h dark condition
        3.3.2 Calli cultured in 24 h dark condition
    3.4 Metabolite identification and quantification by HPLC
    3.5 Metabolite determination by LC-MS
    3.6 Expression of genes potentially involved in celastrol biosynthesis
Chapter 4 Discussion
    4.1 Callus culture
    4.2 Effect of the source of explant on callus establishment in
    4.2 Effect of different combination of hormones on callus establishment
    4.4 MJ treatment and metabolite analysis
    4.5 Gene expression of genes relevant to celastrol biosynthesis
Chapter 5 Conclusions and perspectives
References
Appendices
Acknowledgement
Author profile and publications


【参考文献】：
期刊论文
[1]雷公藤MCT基因RNAi对雷公藤萜类活性成分生物合成的影响[J]. 宋雅迪,赵瑜君,陈上,胡添源,张睿,王家典,卢鋆,王秀娟,高伟,黄璐琦.  药学学报. 2018(08)
[2]TwHMGR过表达对雷公藤甲素和雷公藤红素生物合成的影响[J]. 王家典,赵瑜君,张逸风,胡添源,卢鋆,周家伟,马宝伟,张睿,高伟,黄璐琦.  药学学报. 2018(08)
[3]Celastrol targets IRAKs to block Toll-like receptor 4-mediated nuclear factor-κB activation[J]. Yu-fan Shen,Xue Zhang,Ying Wang,Fan-fan Cao,Georges Uzan,Bin Peng,Deng-hai Zhang.  Journal of Integrative Medicine. 2016(03)



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