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›› 2006, Vol. 6 ›› Issue (5): 794-798.

• 生化工程专栏 • 上一篇    下一篇

肉苁蓉愈伤组织的超低温保藏方法

葛锋,王晓东,赵兵,王玉春   

  1. 昆明理工大学生物与化学工程学院
  • 出版日期:2006-10-20 发布日期:2006-10-20

Cryopreservation of Cistanche deserticola Callus

GE Feng,WANG Xiao-dong,ZHAO Bing,WANG Yu-chun   

  1. Faculty of Biological and Chemical Engineering, Kunming University of Science and Technology
  • Online:2006-10-20 Published:2006-10-20

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被引次数

27

摘要： 提出了一个优化的简单的超低温保藏方法，成功地运用于肉苁蓉愈伤组织的低温保藏. 为了获得最佳的实验结果，肉苁蓉愈伤组织首先在添加了6%二甲亚砜的B5培养基中进行预培养，然后用玻璃化保护剂在25℃处理20 min，最后投入液氮中进行冷冻. 玻璃化保护剂的成分为30%(j)甘油+15%(j)乙二醇+10%(j)二甲亚砜+0.5 mol/L蔗糖. 冷冻后的愈伤组织在30℃的水浴中迅速解冻，接着用25℃的1.0 mol/L蔗糖溶液洗净愈伤组织上附着的玻璃化保护剂，最后在B5培养基上对愈伤组织进行恢复性培养. 经过上述冻存处理的肉苁蓉愈伤组织存活率可达86％. 恢复培养5个月后，愈伤组织中苯乙醇糖甙类化合物的含量和产量分别达到冷冻前的97%和95%.

关键词: 肉苁蓉, 愈伤组织, 超低温保存, 玻璃化

Abstract: A simple and optimized cryopreservation process was applied to Cistanche deserticola callus. To obtain optimal results, the callus was pre-cultured on B5 medium supplemented 6% (j) dimethyl sulfoxide (DMSO) and then treated with a vitrification solution at 25℃ for 20 min before immersion in liquid nitrogen. The vitrification solution contained 30% (j) glycerol, 15% (j) ethylene glycol, 10% (j) DMSO and 0.5 mol/L sucrose. After cryopreservation and rapid thawing in a water bath at 30℃, the callus was washed with 1.0 mol/L sucrose solution at 25℃. The survival rate of callus was 86% under the above cryopreservation process. After the cryopreserved callus was cultured on B5 medium for five months, the content and production of PeG were 97% and 95% of those in the non-frozen callus.

Key words: Cistanche deserticola, callus, cryopreservation, vitrification