超低温保存箱

以生姜的脱病毒试管苗在繁殖培养基上培养25d获得的丛生芽为试材,探讨生姜超低温玻璃化法保存的最优程序。结果表明:采用继代培养45d的丛生芽,切取2~3mm,于0.5mol/L蔗糖浓度的MS培养基内预培养2d,60%PVS2室温处理5min,100%PVS2 0℃下处理30min,迅速投入液氮,48h后,37~40℃水浴快速化冻2min,1.2mol/L蔗糖的MS培养液洗涤20min,在MS+0.5mg/L BA+0.1mg/L NAA+0.3mg/L GA3+30g/L蔗糖+7g/L琼脂的培养基上恢复培养的成活率最高,为57.7%。

A procedure on cryopreservation of shoot-tips of Eriobotrya japonica Lindl.had been studied in vitro by two steps vitrification.The effects of cold-hardening,preculture,cryoprotecten treatment,thawing and reculture on survival rate of the shoot-tips were tested.Optimal media for shoot-tip induction and propagation were 1/2 MS+1.0 mg·L~(-1) 6-BA+0.5 mg·L~(-1)NAA and MS+0.5 mg·L~(-1)6-BA+0.1 mg·L~(-1) NAA.Preculture for 30 min with 60% PVS_(2)(30% glycerol+15% ethylene glycol+15% DMSO+0.4 mol·L~(-1) sucrose) and treatment for 50 min by PVS_(2) were suitable to shoot-tip cryopreservation.After cryopreservationsome shoot-tips couldregenerate plantlets,the regenerate rate was 46.8%.There was no difference in regenerating plantlets and tube plantlets not stored by cryopreservation.