摘要: |
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0.8cm,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。 |
关键词: 丹参 发根农杆菌 毛状根 响应面法 实验研究 |
DOI: |
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Optimization of induction conditions of Salvia miltiorrhiza hairy root based on the response surface method |
CAI Yuan,ZHONG Can,XIE Fang-yi |
(Institute of Material Medica,Hunan Academy of Chinese Medicine,Changsha 410013,Hunan,China) |
Abstract: |
Objective:To optimize the induction conditions of Salvia miltiorrhiza hairy root using the response surface method,and to improve induction rate.Methods:The aseptic seedlings of Salvia miltiorrhiza were infected with Agrobacterium rhizogene ATCC15834 to 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 miltiorrhiza hairy root(leaf size,Agrobacterium rhizogene activity,co-culture time,and antibiotic concentration) and screen out the optimal conditions.Results:Salvia miltiorrhiza hairy root had an autonomous 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,an Agrobacterium rhizogene OD600 of 1.0,a co-culture time of 2 days,an Agrobacterium rhizogene infection time of 5 minutes,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 miltiorrhiza hairy root and provides a basis for further exploitation and utilization of Salvia miltiorrhiza. |
Key words: Salvia miltiorrhiza Agrobacterium rhizogene hairy root response surface experimental study |