目的研究以秋水仙碱处理的长春花突变细胞在继代培养过程中的生长规律、吲哚生物碱积累的规律及营养成分的适宜浓度等方面的特征,期望得到适于工业化细胞培养的理想材料。方法将突变细胞培养于MS固体培养基上,定期称取其鲜质量,用有机溶剂萃取吲哚生物碱,应用RP-HPLC法检测药用成分阿玛碱和长春质碱。结果突变细胞培养至第30代时,生长速度和吲哚总碱积累得最快,培养至第45代时,两者均明显下降。而药用成分的量于第20代达到最高;在培养基中添加适量的色氨酸能提高药用成分的量;培养基中Ca²⁺和Zn²⁺的质量浓度分别为1760和12.6mg/L时,明显促进了药用成分的积累。结论长春花突变细胞可能是一种适于工业化细胞培养的理想材料。

Objective To investigate the characteristics of growth, accumulation of indole alkaloid, and the optical concentration of the nutrition components in the Catharanthus roseus mutant cells induced by the colchicine, looking forward to getting the ideal material producing alkaloids which was suited for industrialized cell cultivation. Methods Mutant cells cultivated on the MS medium were harvested according to experiment designs,and the fresh weight was got.The indole alkaloid as extracted by organic solvent. The ajmalicine and catharanthine extracted from the samples were analyzed by RP-HPLC. Results The growth rate was faster and the indole alkaloid accumulation was more when the mutant cells were cultivated generations 30 and the both were sharply decreased at generations 45, while the contents of medical compounds amounted to the peak at the generations 20. The content of medical components and alkaloid accumulation were evidently prompted by the tryptophan added in the media, especially at the concentration of Ca²⁺ 1 760 and Zn²⁺ 12.6 mg/L, respectively. Conclusion C.roseus mutant cells probably is an ideal material for industrialized cell cultivation.

山东省科技攻关项目(2005GG4402011)；南开大学科技创新基金