金银花细胞悬浮培养生物积累咖啡酰奎宁酸的诱导调控及其分子机制研究

咖啡酰奎宁酸具有较强的抑菌、抗氧化、预防肿瘤等功能，在食品、保健品、医药中广泛应用。前个国家基金研究表明咖啡酰奎宁酸是金银花叶抑菌的主要物质，本项目针对其产量匮乏现状，拟利用细胞悬浮培养和次生代谢调控技术，研究金银花细胞悬浮培养生物积累咖啡酰奎宁酸的诱导调控及分子机制。采用响应面法优化培养条件建立悬浮细胞培养体系，通过比较生物、物理、化学单因素及交互因素作用效果系统筛选高产咖啡酰奎宁酸的诱导子，综合分析基因表达、酶活性与产物积累的相关性探索诱导信号产生、传导分子机制，建立基于HPLC-MS法咖啡酰奎宁酸指纹图谱，并通过连续测量计算产物积累动力学方程。通过本项目研究筛选出金银花细胞悬浮培养体系诱导子、阐明诱导作用机制和产物积累规律，对诱导调控其他植物细胞悬浮培养体系高效积累次生代谢产物提供理论依据。项目的开展和完成为突破咖啡酰奎宁酸深度开发的资源瓶颈拓宽思路。

Caffeoylquinic acid is commonly used in the food, healthy products and medicine because of possessing bacteriostatic effects, antioxidant, cancer prevention and so on. It was also suggested caffeoylquinic acid is the most responsible for the bacteriostatic activity of Lonicera japonica leaves by the completed National Science Foundation of China of the applicant. This project aim at elicitation and induction mechanism of caffeoylquinic acid biosynthesis in L. japonica cell suspension cultures using suspension culture plant cell and secondary metabolic elicitation technology to overcome production lack. The suspension cell culture system will be established by culture conditions optimization applying response surface methodology. The elicitor for enhancement accumulation of caffeoylquinic acid will be screened by comparing treatment effect of single and interaction factors among biological, physical and chemical factors. Mechanisms of inducing signal generation and conduction will be explored by comprehensive analysis the relevance between gene expression, enzyme activity and product accumulation. The chromatogram fingerprint will be established based on HPLC-MS method and the kinetic equations of product accumulation will be got through continuous measuring and calculation. Elicitor, induction mechanism and accumulation kinetic equations of caffeoylquinic acid biosynthesis in L. japonica cell suspension cultures will be clarified in this project, which can provide value theoretical basis on elicitation other plant cell suspension cultures for efficient biosynthesis functional secondary metabolites. The launch and completion of this project will widen ideas to break resources bottleneck for further development and utilization caffeoylquinic acid.