种甜叶菊毛状根诱导和培养生产绿原酸和二咖啡酰奎宁酸的方法

本发明属于基因工程领域，具体涉及种采用发根农杆菌遗传转化甜叶菊建立毛状根培养体系生产绿原酸及二咖啡酰奎宁酸的方法。内容包括甜叶菊外植体制备、发根农杆菌活化培养、悬浮培养及绿原酸和二咖啡酰奎宁酸的生产。经PCR检测，证明甜叶菊毛状根为转化产生，HPLC检测表明甜叶菊毛状根能够生产绿原酸和二咖啡酰奎宁酸。由于所使用的毛状根具备在无激素培养基上快速生长的特性，因此该方法操作简单、成本较低、不受气候、土地等自然条件的限制等优点。为利用毛状根培养生产绿原酸和二咖啡酰奎宁酸提供了种稳定、可持续的新型药源和食品功能因子，为今后应用生物反应器进行绿原酸和二咖啡酰奎宁酸规模化生产提供可靠的来源和物质基础。 The invention belongs to the field of genetic engineering, specifically to a production method for chlorogenic acid and dicaffeoylquinic acid by establishing a hairy root culture system through genetic transformation of stevia rebaudiana with agrobacterium rhizogenes. The method comprises the following steps: preparation of an explant of stevia rebaudiana, activation and culture of agrobacterium rhizogenes, suspension culture and production of chlorogenic acid and dicaffeoylquinic acid. PCR detection results show that hairy roots of stevia rebaudiana are generated through transformation; HPLC detection results show that the hairy roots of stevia rebaudiana can produce chlorogenic acid and dicaffeoylquinic acid. Since