玻璃化法优化保存川贝母组织培养物研究

Research on Cryopreservation of Cultured Fritillaria Cirrhosa D.Don Tissues by Vitrification

为筛选出川贝母组织培养物中最适于用玻璃化法进行超低温保存的材料及其最适的玻璃化溶液脱水处理时间,将继代15 d的优质川贝母芽、愈伤组织和鳞茎接入MS+30 g/L蔗糖+5%二甲基亚砜的固体培养基中预培养6 d后,用60%玻璃化溶液II常温装载25 min,再于2℃下用100%玻璃化溶液II分别进行不同时间的脱水处理后投入液氮中保存.结果显示,在玻璃化超低温保存过程中,川贝母芽、愈伤组织和鳞茎的最佳脱水处理时间均为60 min,其中愈伤组织细胞保存后的相对存活率最高为80.08%.

To filter the most appropriate cultured Fritillaria cirrhosa D. Don tissue for cryopreservation by vitrification and its best dehydration treatment time bud, bulbs and callus of cultured Fritillaria cirrhosa D. Don which have been subcultured for 15 days and precultured in solid medium MS ＋ 30 g/L sucrose ＋ 5 % DMSO for 6 days were loaded with 60% PVS2 for 25 min under room temperature, and dehydrated with 100% PVS2 for different time at 2 ℃ respectively, then put into the liquid nitrogen for preservation. The results show that the best dehydration treatment time of bud, bulbs and callus of cultured Fritillaria cirrhosa D. Don all are 60 min, and the highest survival rate of callus reaches 80.08 %.