摘要
药用植物是最古老的药物形式之一,次生代谢物的含量是判定药用植物品质的重要指标,但由于仅在特定组织中积累且含量较低,其应用受到限制.成簇的规律间隔的短回文重复序列及其相关蛋白(clusters of regularly spaced short palindromic repeats/CRISPR-associated proteins,CRISPR/Cas)基因编辑技术可以对目标基因实现定点替换、插入和缺失,从而影响基因的表达和功能.目前,CRISPR/Cas9技术已成为药用植物中探索基因功能和解析代谢途径的重要手段,在提高次生代谢物产量和改良药用植物品质方面具有至关重要的作用.而相对于农作物和园艺植物等,CRISPR/Cas9在药用植物中的应用仍然面临编辑效率低和应用范围小等问题.本文综述了CRISPR/Cas9技术在次生代谢物合成途径及转录调控方面的应用,结合CRISPR/Cas9面临的问题,对技术的发展方向进行了展望,以为其在药用植物领域的推广使用提供参考.
China is one of the countries with the richest resources of medicinal plants, and the discovery, use and cultivation ofmedicinal plants have a long history. Medicinal plants are one of the oldest forms of medicine, and since ancient times,people have used medicinal plants as a source of food, healing and industry. Secondary metabolites are a class of nonessential small-molecule organic compounds produced by secondary metabolism for the normal functioning of cell lifeactivities or plant growth and development, and their production and distribution are usually species, organ, tissue, andgrowth and development period specific. The content of secondary metabolites is an important index to determine thequality of medicinal plants, and secondary metabolites from medicinal plants have become potential clues to explore newdrugs to improve human mortality and morbidity. Due to the long growth cycle of medicinal plants and their local nature,the accumulation of secondary metabolites is spatiotemporally specific, which makes the content of secondary metabolitesin medicinal plants low, thus limiting their application in medicine. Classical molecular biology techniques such asoverexpression and gene silencing have been successfully applied to regulate plant secondary metabolism pathways toenhance the production of valuable secondary metabolites. So far, protein-based editing tools, namely zinc finger nucleases(ZFNs) and transcription activator-like endonuclease enzymes (TALENs) have been promoted for transcriptional genomemanipulation. Cluster of regularly spaced short palindromic repeats/CRISPR-associated proteins (CRISPR/Cas) is the thirdgeneration gene editing tool after the ZFNs and TALENs, and is a cheap, simple and efficient gene editing technique. Thistechnology mainly uses sequence-specific nucleases to identify and cut the target sites on the genome, causing DNA doublestrand breaks (DSBs), and further induces two repair mechanisms, Nonhomologous end-joining (NHEJ) and homologydirected Repair (HDR), to repair the broken DNA double strand, achieving site-specific substitutions, insertions anddeletions, thus affecting gene expression and function. It is widely used in metabolic engineering, synthetic biology andmedical research. At present, CRISPR/Cas9 technology has become an important tool for exploring gene function andanalyzing metabolic pathways in medicinal plants, and plays a crucial role in improving the production of secondarymetabolites and improving the quality of medicinal plants. Compared with crops and horticultural plants, the application ofCRISPR/Cas9 in medicinal plants still faces problems such as low editing efficiency and small application range. Thisstudy briefly describes the principle of CRISPR/Cas9 technology, focuses on the application of CRISPR/Cas9 technologyin the three main secondary metabolites: terpenoids, alkaloids and phenylpropanoids, and enumerates the studies onCRISPR/Cas9 technology for exploring gene function and analyzing metabolic pathways in medicinal plants. This papersummarizes the application of CRISPR/Cas9 technology in the analysis of secondary metabolic pathways and secondarymetabolite production, and points out that the application of CRISPR/Cas9 technology in medicinal plants faces problemssuch as wide application, PAM dependence, delivery efficiency and off-target effect. Finally, the research directions ofCRISPR/Cas9 technology for secondary metabolite production, variety improvement and germplasm innovation ofmedicinal plants were prospected. This review is expected to provide references for the widespread application of CRISPR/Cas9 technology in the field of medicinal plants.
作者
朱瑞艳
许莹
时敏
开国银
Ruiyan Zhu;Ying Xu;Min Shi;Guoyin Kai(College of Horticulture,Shenyang Agricultural University,Shenyang 110866,China;College of Pharmacy,Zhejiang Chinese Medicine University,Hangzhou 310053,China)
出处
《科学通报》
北大核心
2025年第16期2526-2541,共16页
Chinese Science Bulletin
基金
国家自然科学基金(82073963,82204554)
浙江省自然科学基金(LQ23H280010)资助。
作者简介
联系人:时敏,E-mail:kaiguoyin@163.com;联系人:开国银,E-mail:min_shi@zcmu.edu.cn。
