摘要
红豆草(Onobrychis viciaefolia Scop.)组织培养物在5%DMSO+10%甘油+8%蔗糖的冰冻保护剂及以1℃/分钟的速度降温到-35—-40℃,停留2小时后,投入液氮,40℃水浴快速化冻等条件下,存活率达60—70%,并保持了高的分化能力。电子显微镜的观察结果表明,快速冰冻和1℃/分钟慢速冰冻至-35℃—40℃不停留,对细胞结构造成严重的致死性破坏;-35℃停留30分钟对细胞结构的损伤是可逆性的;停留2小时的其超微结构基本上与对照材料无明显差别。
Sainfoin(Onobrychis viciaefolia Scop)tissue cultures were frozen using the cryoprotectant of 10% DMSO+10% glycerin+8% sucrose at a cooling rate of 1℃/min from 0℃ to -35℃— -40℃,kept for 2 hrs followed by storage in liquid nitrogen,and then rapidly thawed in a 40℃ water bath.The survival rate of the stored specimens could be reached to 60— 70%,and these cultures still maintained their high differentiation ability. The ultrastructural changes occurred markedly with the different freezing methods:The rapid-freezing and slowfreezing at the cooling rate of 1℃/min to -35℃—-40℃,but with out keeping samples for certain time,led cell ultrastructure to be lethally ruptured.The cell injury was reversible when the samples were kept for 30 min at terminal temperature -35℃. The ultrastructure of the samples kept for 2 hrs at -35℃—-40℃ was basically similar to that of the control material.The results further demonstrate that the technological system established previously by us is reasonable and could be used effectively the long-term pre- servation of sainfoin germplasm.
基金
国家<七五>计划攻关课题。
关键词
红豆草
种质
超低温保存
组织培养
Sainfoin
Germplasm cryopreservation
Tissue culture
Ultrastructure
