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对红豆杉愈伤组织超低温保存中几个主要因素进行了比较 ,试验证明 :预培养时间、预培养基中蔗糖浓度、保护剂的组合以及冰冻降温方法与超低温保存后的相对细胞活力密切相关。试验结果表明 ,在含 8%蔗糖的 62号液体培养基中振荡预培养 6d,红豆杉愈伤组织在超低温保存后细胞活力可保持最高。有效的冷冻保护剂为 1 0 %山梨醇 + 1 0 % DMSO,冷冻方法以分步冷冻和慢冻较为适宜 ,而经快冻的愈伤组织复苏后活力低下
The main factors in cryopreservation of callus were compared. The results showed that the pre-culture time, the concentration of sucrose in pre-culture medium, the combination of protective agent and the method of freezing and cooling were closely related to the relative cell viability after cryopreservation. The results showed that in the liquid medium containing 8% sucrose 62 pre-cultured shaking 6d, callus callus survival after cryopreservation maintained the highest cell viability. The effective cryoprotectant is 10% sorbitol + 10% DMSO. The freezing method is more suitable for fractional freezing and slow freezing, while the viable callus is less viable after resuscitation