目的 建立水飞蓟Silybum rnarianum悬浮培养体系,测定不同影响因子对水飞蓟宾量的影响。方法 采用正交试验确定水飞蓟悬浮培养体系,采用HPLC法测定水飞蓟宾的量。结果 激素配比6-BA 1.5 mg/L+NAA 1.0 mg/L+ZT 1.5 mg/L的MS培养基、光照12 h/d、pH值为7、转数110 r/min,为水飞蓟悬浮培养的最佳生长条件。当水杨酸、壳聚糖、硝普钠质量浓度分别为0.05、3、1 μg/L时水飞蓟增长量达到最大值,水飞蓟宾量也达到最大值。结论 建立的悬浮培养体系可用于水飞蓟的快速繁殖,适宜质量浓度的壳聚糖和硝普钠利于悬浮细胞的生长及水飞蓟宾的积累。

Objective To establish the suspension culture system of Silybum marianum cell and study the effects of different factors on silybin content. Methods Orthogonal test was adopted to determine S. marianum cell suspension culture system and HPLC to silybin content.. Results The best growth cycle of cell suspension culture was defined when cultured in liquid MS medium with 6-BA 1.5 mg/L + NAA 1.0 mg/L + ZT 1.5 mg/L, light 12 h/d, pH value = 7, revolution = 110 r/min. The effects of salicylic acid, chitosan, and sodium nitroprusside on cell suspension culture were investigated. When the concentration of three impact factors were 0.05, 3, and 1 μg/L, respectively, the growth amount of S. marianum cell reached the maximum, i.e. At the same time, the silybin content also reached the maximum. Conclusion The established cell suspension culture system is suitable to use for the rapid propagation of S. marianum. Chitosan and sodium nitroprusside at proper concentration are benifit to the growth of suspensious cells and accumulation of silybin.

中国博士后科学基金项目(20090461042)