林业科学2009，Vol.45Issue(10)：49-54,6.
二乔刺槐愈伤组织超低温保存及适宜降温方法
Methods of Lowering Temperature for Cryopreservation of Calli of Robinia bella-rosea
摘要
Abstract
Robinia bella-rosea calli were initiated from the leaves on MS medium supplemented with 5 mg·L~(-1) 6-BA. The calli were cooled down at 4℃ for 2 h in a cryoprotectant (MS +10% DMSO + 0.5 mol·L~(-1) Sucrose), and further refrigerated to-7 ℃ at a rate of 1 ℃· min~(-1) by using a programmed temperature-changing freezer. After one hour at - 7 ℃ for 1 h, the calli in the cryoprotectant were further refrigerated to - 20℃ at a rate of 0.1 X.min-1.Being incubated for another hour at -20 ℃ ,the calli were cooled down to - 40 t at a rate of 0.3℃ min-1' and then the calli were transferred and preserved in liquid nitrogen for a day. The low temperature treated calli were thawed at 38 t water bath, and the thawed calli were washed with liquid MS medium with 6-BA 5 mg·L~(-1) and 30 g·L~(-1) sucrose, then transferred to solid medium and cultured in darkness. Two weeks later, the calli were cultured in light, and showed able to produce fresh and green new callus after 3 d, and the survival rate of the low temperature treated calli reached to 52 % . New-produced calli were able to regenerate plantlets.关键词
二乔刺槐/愈伤组织/超低温保存/降温方法Key words
Robinia bella-rosea/callus/cryopreservation/freezing method分类
农业科技引用本文复制引用
翟晓巧,程斐,朱延林..二乔刺槐愈伤组织超低温保存及适宜降温方法[J].林业科学,2009,45(10):49-54,6.基金项目
林业公益性行业科研专项(200804028) （200804028）
林业科技支撑计划(2006BAD01A1601). （2006BAD01A1601）
