【摘要】 目的研究碳源、氮源和磷源对丹参Salvia miltiorrhiza不定根生长及其有效成分丹参酮ⅡA和原儿茶醛量的影响。方法利用组织培养技术结合HPLC分析手段,通过改变培养基碳源种类和浓度、氮源和磷源浓度,研究其对丹参不定根生长和丹参酮ⅡA、原儿茶醛量的影响。结果碳源、氮源、磷源对于不定根培养是必需的。以蔗糖为碳源,添加30g/L蔗糖,培养20d,不定根增殖倍数最高,60g/L蔗糖最适合丹参酮ⅡA合成,低质量浓度蔗糖更利于原儿茶醛合成。间歇添加蔗糖至培养25d,不定根增殖率是对照的2.3倍,丹参酮ⅡA量是对照的2.4倍。培养基NH4+和NO3-总浓度保持60mmol/L,NH4+/NO3-为1∶4、1∶4、1∶1时分别得到最大的不定根增殖倍数、丹参酮ⅡA和原儿茶醛的量。改变培养基KH2PO4浓度时,丹参不定根生长均比对照组旺盛,但高浓度KH2PO4抑制了丹参酮ⅡA合成。结论本实验确定了丹参不定根悬浮培养的最佳碳源种类和浓度、氮源比例和磷源浓度,表明不同碳源、氮源和磷源对丹参不定根生长和次生代谢物合成有显著影响。更多还原

【Abstract】 Objective To study the effects of carbon, nitrogen, and phosphate sources on the growth of Salvia miltiorrhiza adventitious roots and the contents of tanshinone ⅡA and protocatechuic aldehyde. Methods The adventitious roots were obtained through tissue culture by manipulation of carbon, nitrogen, and phosphate sources and the contents of tanshinone ⅡA and protocatechuic aldehyde were determined by HPLC. Results Carbon, nitrogen, and phosphate sources were necessary for the culture of S. miltiorrhiza adventitious roots. The highest times of root multiplication were achieved at sucrose level of 30 g/L after 20 d culture, 60 g/L sucrose and low level sucrose were favorable for biosyntheses of tanshinone ⅡA and protocatechuic aldehyde, respectively. The highest root yield and tanshinone ⅡA content on day 25 were obtained by intermittent sugar adding during cultivation, and the production of adventitious roots and tanshinone ⅡA were 2.3-and 2.4-fold compared with those of control, respectively. The maximum root growth rate, contents of tanshinone ⅡA and protocatechuic aldehyde were achieved while NH4+-NO3-was 1∶4, 1∶4, and 1∶1, respectively when concentration of total nitrogen source was kept at 60 mmol/L. To compare with the control group, changing of KH2PO4 concentration could favor for the adventilious root growth, but high KH2PO4 concentration inhibited tanshinone ⅡA biosynthesis. ConclusionThe results show that various carbon, nitrogen, and phosphate sources have the significant effects on adventitious root culture of S. miltiorrhiza. The best carbon source and its concentration, nitrogen and phosphate sources for the growth of S. miltiorrhiza adventitious root and the synthesis of secondary metabolite are confirmed.更多还原