Study On Transformation And Biosynthesis Of Saikosaponin In Adventitious Root Culture Of Bupleurum Chinense DC.

Bupleurum chinense DC. is a well-known traditional chinese medicine, and was recently named as the original plant of authentic herbs by "People's Republic of China Pharmacopoeia". Its root contains saikosaponins as the main active components, saikosaponin-a and saikosaponin-d are the key pharmacologically active substances, which have anti-pyretic, sedative, analgesic, anti-bacterial, anti-liver injury and anti-virus activities.In this study, three types of adventitious roots of Bupleurum chinense DC. from the province of Shanxi are the main materials, discussing its saikosaponin content by means of adding precursors, elicitors as well as semi-continuous culture.The results showed that the contents of saikosaponin were affected by precursors and elicitors. The 1/2 MS combined 0.37 g/L magnesium sulfate,3.0 mmol/L sodium pyruvate,9.5 g/L potassium nitrate,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Zuoquan. The 1/2 MS combined 1.1 g/L magnesium sulfate,1.9 g/L potassium nitrate,3.0 mmol/L sodium pyruvate,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Taigu. The 1/2 MS combined 2.0 mmol/L sodium pyruvate, 1.85 g/L magnesium sulfate, 1.9 g/L potassium nitrate,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Fangshan.The contents of saikosaponins reached to 0.291% by continuing to add 1.00 mmol/L glycine,0.10 mmol/L citric acid and 0.01 mmol/L benzoic acid to the most suitable medium above for Bupleurum chinense from Zuoquan, compared to increased 0.142%.The 1/2 MS combined 1.25 mg/L CoCl2,0.01 mmol/L AgNO3, 0.22 g/L CaCl2,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Zuoquan. The 1/2 MS combined 12.5 mg/L CoCl2,0.22 g/L CaCl2, 0.10 mmol/LAgNO3,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Taigu. The 1/2 MS combined 0.01 mmol/L AgNO3,12.5 mg/L CoCl2,0.44 g/L CaCl2,0.5 mg/L double acid calcium diphenylurea and 50 g/L sucrose was the most suitable medium for Bupleurum chinense from Fangshan.The Bupleurum chinense from Zuoquan and Taigu had the similar requirements for the concentration of salicylic acid, either proliferation or the amount of saikosaponins have reached the maximum when the concentration is 0.001 mmol/L.The proliferation and the content of total saikosaponin of adventitious roots reached a maximum when the sugar concentration of 50 g/L by means of multi-step controlling sucrose method.When fresh medium was added at day 30, many new young roots developed and grew, and an increase in both root weight and the total amount of the saikosaponins was observed at day 60 in the adventitious roots of Bupleurum chinense from Taigu and Fangshan, the root weight and the total amount of the saikosaponins of Bupleurum chinense from Zuoquan was to the maximum at day 80.The results of this experimental system provide an experimental basis for the production of main secondary metabolites of adventitious roots of Bupleurum chinense DC, it also provide a technical support to the industrial production and has a significant effect on modernization of chinese medicine.