包埋玻璃化法超低温保存沙生柽柳休眠茎尖的研究

Cryopreservation of Shoot-tips of Tamarix taklamakanensis Domant Buds by Encapsulation-vitrification

[目的]采用包埋玻璃化法,对沙生柽柳休眠茎尖成功地进行冷冻保存。[方法]将沙生柽柳休眠茎尖包埋在含3%褐藻酸钙的胶球中,研究了预培养中蔗糖浓度、培养时间和不同玻璃化保护液处理的时间对沙生柽柳休眠茎尖存活率的影响。[结果]含沙生柽柳茎尖的胶球在0.8mol/L蔗糖溶液中培养24h可获得较高的存活率;胶球经玻璃化保护液[40%甘油+45%(0.4mol/L)蔗糖+10%聚乙二醇(分子量4000)+5%二甲基亚砜]在25℃处理20min可以明显提高存活率,在优化条件下存活率可高达48.3%。[结论]蔗糖浓度和预培养时间是决定沙生柽柳茎尖存活率的关键因素。

[Objective]Shoot-tips of Tamarix taklamakanensis Domant buds were successfully cryopreserved by using encapsulation-vitrification.[Method]Domant shoot-tips were encapsulated 3% calcium alginate beads.The main factors influencing the viability such as sucrose concentration,preculture time, different vitrification solution and the time beads in vitrification solution were studied.[Result] High viability could be obtained when alginate beads were precultured for 24 h in 0.8 mol/L sucrose.The beads were dehydrated with vitrification solution [40%Gly ＋ 45% （0.4 mol/L） Suc ＋ 10%PEG（4 000）＋5%DMSO]for 20 min at 25 ℃.Under optimum conditions, the viability of domant shoot-tips could reach 48.3% at the most.[Conclusion] Sucrose concentration and preculture time were important factors influencing the viability of Tamarix taklamakanensis Domant buds.