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初步研究了水母雪莲愈伤组织的超低温保存方法. 结果表明,预培养、保护剂、预处理、冰冻后处理对愈伤组织存活率都有一定影响. 水母雪莲愈伤组织预培养基为添加5%二甲基亚砜的MS培养基,优化的冰冻保护剂是15%二甲基亚砜+15%乙二醇+30%甘油的0.4 mol/L蔗糖液,冰冻保护剂的预处理温度是15oC,时间为10 min,解冻温度25~35oC;在25oC水浴中用含1.2 mol/L蔗糖的MS溶液反复洗3次,每次10 min. 用TTC法测定细胞存活率可达58.5%.
The cryopreservation of callus from Saussurea nigra was preliminary studied.The results showed that pre-culture, protective agent, pre-treatment and post-freezing treatment had some effects on the survival rate of callus.The pre-culture medium of Saussurea fruticosa callus was 5 % Dimethyl sulfoxide MS medium, the optimized cryoprotectant is 0.4 mol / L sucrose solution of 15% dimethyl sulfoxide + 15% ethylene glycol + 30% glycerol, the pre-treatment temperature of the cryoprotectant is 15oC for 10 min, thawing temperature 25 ~ 35oC, and repeatedly washed three times with MS solution containing 1.2 mol / L sucrose for 10 min at 25oC in water bath, and the cell survival rate was 58.5% by TTC method.