玻璃化法超低温保存猕猴桃离体茎尖及其植株再生

 以中华猕猴桃矮型种质为离体培养材料, 建立试管无性系, 对其离体茎尖的玻璃化法超低温保存技术进行研究, 探讨猕猴桃种质长期保存的适宜途径。结果表明, 含1～2个叶原基(1.5～2.5 mm) 的茎尖, 在5%二甲基亚砜(DMSO) + 5%蔗糖+MS培养基上预培养4 d后, 于室温下用2 mol/L甘油+ 0.4 mol/L蔗糖预处理30 min, 再在0℃下用玻璃化液(PVS2 ) 脱水处理40 min并迅速投入液氮中, 保存24 h后接种至继代培养基上再培养, 成活率和再生率分别为56.7%和51.6%。再生植株生根后可移栽成活。

Cryopreservation of in Vitro Cultured Kiwifruit Shoot-tips by Vitrification and their Regeneration

Experiments were conducted to provide an appropriate means for long-term p reservation of kiwifruit germp lasm using cryopreservation by vitrification. Shoot-tips of a dwarf genotype of kiwifruit were cultured in vitro to obtain aseptic clones. The results showed that shoot-tips 1.5 - 2.5 mm in length with one or two leaf primordium were precultured in 5% DMSO + 5% sucrose + MS medium for 4 days. Pretreated shoottips with 2 mol/L glycerol + 0.4 mol/L sucrose for 30 min at room temperature, followed by dehydration with PVS2 (30% glycerol + 15% ethylene glycol + 15% DMSO + 0.4 mol/L sucrose) for 40 min at 0℃ and rapidly put into liquid nitrogen for 24 h. The defrosted shoot-tips were then inoculated onto subculture medium.The survival and regeneration rate of shoot-tips treated in thisway was 56.7% and 51.6% respectively. Theplantlets could normally root and survive after transplanting.