红松胚性愈伤组织的超低温保存技术

Cryopreservation of Korean Pine Somatic Calli

本文初步研究报道了红松胚性愈伤组织的超低温保存方法,结果表明,预处理、保护剂与解冻方式对胚性愈伤组织存活率均有一定影响。效果较好的方法为:利用0.4mol·L-1Sorb(山梨醇)预处理愈伤组织24h,5%DMSO(二甲基亚砜)+0.4mol·L-1Sorb为冷冻保护剂,冷冻方式为程序降温盒在-70℃停留3h后投入液氮罐中保存,解冻方式为20℃流水冲洗。胚性愈伤组织解冻后再培养,其存活率可达71.4%。

Preliminary study on cryopreservation of Korean pine somatic calli showed that the pretreatment, the cryoprotectant and thawing method had certain influences on the survival rate of calli. The better-performing method was established： calli were pretreated with 0.4 mol·L^-1 Sorb for 24 h, and then kept in programmed cooling box Nalgene Mr. Frosty at -70 ℃ for 3 h with 5 % DMSO and 0.4 mol·L^-1 Sorb as cryoprgtectant, at last put into liquid nitrogen container. While thawed in 20 ℃ flowing water, the survival rate of thawed calli was 71.4%.