碳源、氮源和磷源对西洋参悬浮细胞生物量和活性成分积累的影响

 目的研究碳源、氮源和磷源对西洋参悬浮细胞生长,人参皂苷Re,Rb1以及西洋参多糖合成的影响。方法利用组织培养技术结合高效液相色谱法和紫外分光光度法,通过改变培养基碳源种类、氮源和磷源浓度,研究其对西洋参悬浮细胞生长,人参皂苷Re,Rb1以及西洋参多糖含量的影响。结果添加40g·L^-1蔗糖的细胞干重生长率最大,加入30g·L^-1蔗糖得到的人参皂苷Re、Rb1和多糖的含量最高,分别为0.531%,0.145%和3.62%。本实验保持氮源浓度60mmol·L^-1,当培养基中全部为NO₃^-时,细胞干重生长率最低,而人参皂苷Re、Rb1和多糖含量最高,分别为对照组的2.40,1.96和4.41倍。添加5.00mmol·L^-1的KH₂PO₄,细胞干重生长率最大。当培养基中KH₂PO₄浓度为0.63mmol·L^-1时,人参皂苷Re、Rb1和多糖的含量最高。结论本实验确定了西洋参细胞悬浮培养的最佳碳源种类、氮源比例和磷源浓度,表明不同碳源、氮源和磷源对西洋参细胞生长和活性成分有显著影响。

Effects of Carbon,Nitrogen and Phosphate Sources on Biomass and Active Components of Suspension Cells in Panax quinquefolium L.

OBJECTIVE To study the effects of carbon,nitrogen and phosphate sources on the growth of Panax quinquefolium suspension cells and the synthesis of ginsenosides Re,Rb1 and polysaccharide.METHODS The suspension cells were obtained through tissue culture by manipulation of carbon,nitrogen and phosphate sources.The contents of ginsenosides Re and Rb1 were determined by HPLC while the contents of polysaccharide were determined by ultraviolet spectrophotometry.RESULTS The maximum growth rate of suspension cells of P.quinquefolium was achieved with the addition of sucrose at level of 40 g·L^-1.The maximum contents of ginsenosides Re,Rb1 and polysaccharide were achieved with the addition of sucrose at level of 30 g·L^-1,and the contents were 0.531%,0.145% and 3.62%,respectively.In the case of NO3-as the sole N source,the cell growth rate was lowest when the concentration of total nitrogen source was kept at 60 mmol·L^-1.The maximum contents of ginsenosides Re,Rb1 and polysaccharide were increased by 2.40,1.96 and 4.41-fold compared with those of control,respectively.The maximum growth rate was achieved at 5.00 mmol·L^-1 KH₂PO₄ concentration.The maximum contents of ginsenosides Re,Rb1 and polysaccharide were achieved at 0.63 mmol·L^-1 KH₂PO₄.CONCLUSION The results show that various carbon,nitrogen,and phosphate sources have the significant effects on suspension cells growth of P.quinquefolium and active components.The sources of carbon,nitrogen and phosphate for the suspension cells culture of P.quinquefolium were optimized.