曼陀罗毛状根的诱导及其悬浮培养合成天麻素初探

利用A4和R1601两种发根农杆菌菌株分别感染曼陀罗的嫩叶及嫩茎诱导产生毛状根。探讨了不同菌株、不同外植体、蔗糖预处理、外植体预培养与染菌时间以及外植体二次染菌等条件对转化频率的影响,并建立毛状根悬浮培养体系及转化外源底物对羟基苯甲醇合成天麻素。结果表明:A4转化频率高于R1601;嫩叶转化频率最高;嫩叶最佳预培养时间为4 d;最佳染菌时间为5 min。HPLC法分析表明,悬浮培养毛状根能转化外源底物对羟基苯甲醇合成天麻素。

Study on the Hairy Root Induced from Datura tatula L. and Production of Gastrodin through Suspension Culture of Hairy Root

The effects of different strains of Agrobacterium rhizogenes,different explants,pretreatment of sucrose,pre-culture and infection time of explants and secondary infection of explants on the genetic transformation rate of hair root were studied and the hairy root suspension culture was established in this paper.The hairy roots were induced from the leaves and stems of Datura tatula infected by A4 and R1601 strains of A.rhizogenes.The production of gastrodin through biotransformation of p-hydroxybenzyl alcohol using hairy root suspension culture of D.tatula was also conducted.The results showed that the optimal transformation condition was A4 strain,using leaf as the explant,pre-culture for 4d and infection for 5 min.The analysis of HPLC indicated that the hairy root suspension culture could convert the exogenous substrate p-hydroxybenzyl alcohol into gastrodin.