基于响应面法对丹参毛状根诱导条件的优化研究

Optimization of induction conditions of Salvia miltiorrhiza hairy root based on the response surface method

目的:应用响应面法优化丹参毛状根诱导条件,提高其诱导率。方法:采用发根农杆菌ATCC15834侵染丹参无菌苗获得毛状根,在单因素实验的基础上,利用Box-Benhnkcn设计和响应面分析方法对丹参毛状根诱导过程的培养条件(叶片大小、农杆菌活力、侵染时间、共培养时间、抗生素浓度)进行分析,优选最佳培养条件。结果:丹参毛状根在无外源激素的MS培养基上自主生长,表现出典型的发根特征。毛状根的最优培养条件为:叶片大小0. 8cm2,农杆菌OD600为1. 0,共培养时间2d,农杆菌侵染时间5min,头孢噻吩浓度400ng/L。在此条件下,丹参毛状根的诱导率为69. 8%。结论:该研究优化了丹参毛状根的培养条件,为丹参的进一步开发利用提供了依据。

Objective:To optimize the induction conditions of Salvia mihiorrhiza hairy root using the response surface method,and to improve induction rate.Methods :The aseptic seedlings of Salvia mihiorrhiza were infected with Agrobacterium rhizogene ATCC15834to obtain hairy root.On the basis of single-factor experiment,the Box Benhnkcn design and the response surface analysis were used to analyze the culture and induction conditions of Salvia raihiorrhiza hairy root (leaf size,Agrobacterium rhizogene activity,co cuhure time,and antibiotic concentration)and screen out the optimal conditions.Resuhs:Salvia miltiorrhiza hairy root had anautonomous growth on MS medium without exogenous hormone and showed the typical rooting features.The optimal culture conditions of hairy root were a leaf size of 0.8cm^2,an Agrobacterium rhizogene OD600of 1.0,a co culture time of 2days,an Agrobacterium rhizogene infection time of 5minutes,and a cefalotin concentration of 400ng/L.Under these conditions,the induction rate of Salvia miltiorrhiza hairy root was 69.8%.Conclusion :This study optimizes the culture conditions of Salvia mihiorrhiza hairy root and provides a basis for further exploitation and utilization of Salvia mihiorrhiza.