【摘要】 本研究分析了水稻悬浮细胞的生理状态和各种冷冻前处理等因素对超低温保存后细胞存活率的影响。结果表明,继代后培养3—5天,处于对数生长期的细胞,采用二步冷冻法,超低温保存后存活率最高。电镜超微结构观察显示,0.5mol/L山梨醇预培养,10%DMSO+0.5mol/L山梨醇复合保护剂处理,液泡显著变小,数目明显减少,从而降低了细胞内自由水含量,增强细胞的抗冷冻能力。在上述合适的前处理和冷冻-化冻条件下,超低温保存水稻悬浮细胞的恢复生长率为58%,恢复生长的细胞转移到分化培养基上,可再生健壮绿苗, 移植到盆钵,在温室中长成正常结实的植株。更多还原

【Abstract】 Cryopreservation of plant cells and tissues combines conventional germplasm conservation with morden biotechnology. This technique, by which cells and tissues are frozen undercontrolled conditions and stored in liquid nitrogen, has proven to be a reliable method for long-term preservation of cell suspensions.The effects of various factors, such as physiological states and different pre-treatments, on the survival of cryopreser-ved cells were studied. The highest survival was obtained in the exponential phase cells with two-step freezing method. Ul-trastructure study showed that vacuolesbecame smaller and fewer in the cells pre-cultured in 0.5 mol/L sorbitol or treated with 10%DMSO + 0.5 mol/L sorbitol. The resultes of decreased free water led to resistant to freezing and thawing injury.In suitable conditions, the regrowth ratio of cryopreserved suspensions was 58%. The recovered cells, subcultured one time, were transferred to the regeneration media supplemented with 2 mg/L 6-BA, 2 mg/L KT and 0.5 mg/L NAA in light. After 30 days, strong and green plantlets were obtained. Plants transplated to soil fertiled in the greenhouse.更多还原