银杏愈伤组织超低温保存的研究

THE STUDIES OF CRYOPRESERVATION OF CALLUS FOR MAIDENHAIR TREE(GIBGKO BILOBA L.)

本文对银杏子叶愈伤组织进行了超低温保存研究。结果表明 ,冰冻保护剂、冷冻速度对解冻后材料相对存活率影响较大 ,冻前预培养和预冻至某一温度停留一段时间也较为重要。在含高浓度山梨醇或甘露醇的培养基上预培养 2d的愈伤组织 ,用 10 %二甲基亚砜 +0 .5mol·L- 1 山梨醇作为冰冻保护剂 ,以 1℃·min- 1 速度降温 ,在 - 15℃、- 35℃分别停留 10min和 30min后 ,投入液氮中保存 1d ,细胞相对存活率可达 6 0 %以上。再培养时 ,细胞能恢复生长。

Studies of cryopreservarion of callus of maidenhair tree (Gingko biloba L.) Cotyledon were conducted.The relative viability of cryopreserved materials was effected by both of cryoprotectant and freezing rate.It was also important to undergo pre-growth and that material for a while at freezing to some low temperature. By pre-culturing on MS adding high concentration of sorbitol or mannitol for two days-with the combinative cryoprotectant 10% Dimethyl sulfoxide(DMSO)+ 0.5*!mol·L -1 sorbitol-slow freezing at 1*!℃·min -1-maintaining for 10 min at -15*!℃ and for 30*!min at -35*!℃-directly plunging into LN-stored for 1 day-rapidly thawing at 40*!℃ water. The relative viability (TTC value) was beyond 60*!%.The recovery growth of materials stored at ultra-temperature was observed.