摘要
In this preliminary experiment two kinds of crypprotectants were used: (1).10% DMSO+ 0.5 mol/L sorbitol; (2).7.5% DMSO +5% glycerin +5% sucrose.Calli samples weretaken at day-10 and day-15 after subculture respectively.The specimens were cooled at a rateof l℃/min from 0℃,kept -40℃ for 2h,put into liquid nitrogen( - 196℃) and then thawedin 40℃ water bath.Calli which have been subcultured for 10 days showed normal growthby using cryoprotectant 2 and slow-freezing method mentioned above which indicated thatthey were effective methods for cryopreservation of Arnebia euchoma.Normal growth of thecallus after cryopreservation could be restored,but the initial growth rate was somehow retarded.Using polycrylamide gel electrophoresis analysis no difference in esterase and peroxidaseisozymes was found in both types of the treated calli.The present experiment indicates it ispossible to use this slow-freezing method for germplasm cryopreservation of Arnebia euchroma.
In this preliminary experiment two kinds of crypprotectants were used: (1).10% DMSO + 0.5 mol/L sorbitol; (2).7.5% DMSO +5% glycerin +5% sucrose.Calli samples were taken at day-10 and day-15 after subculture respectively.The specimens were cooled at a rate of l℃/min from 0℃,kept -40℃ for 2h,put into liquid nitrogen( - 196℃) and then thawed in 40℃ water bath.Calli which have been subcultured for 10 days showed normal growth by using cryoprotectant 2 and slow-freezing method mentioned above which indicated that they were effective methods for cryopreservation of Arnebia euchoma.Normal growth of the callus after cryopreservation could be restored,but the initial growth rate was somehow retarded. Using polycrylamide gel electrophoresis analysis no difference in esterase and peroxidase isozymes was found in both types of the treated calli.The present experiment indicates it is possible to use this slow-freezing method for germplasm cryopreservation of Arnebia euchroma.
基金
国家“七五”攻关项目
关键词
新疆紫草
愈伤组织
超低温保存
Arnebia euchroma
Calli
Cryoprotectant
Cryopreservation
