红豆杉愈伤组织超低温保存有关因素的研究

Studies on Factors in Cryopreservation of Taxus Callus

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 。

The experiment compares the main factors of cryopreservation of Taxus callus. The results show that with the differences of preculture time, saccharose content of culture medium, cryoprotectants and freezing method, the cell vitality varies remarkably. After Taxus callus was precultured in No.62 medium with 8% saccharose for 6 days, the TTC value of Taxus calluses after freezing and storage in LN(liquid nitrogen) was high. The most effective cryoprotectant is the mixture of 10% DMSO and 10% sorbital. The effectiveness of the step freezing and slow freezing methods was much better than that of the rapid freezing method.