生姜玻璃化法超低温保存技术

Ginger Vitrification Cryopreservation Technology

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%。

Taking buds obtained from the shoot of ginger by vitrification virus-free cuvette plantlet cultured 25days in medium as material,the optimal vitrification procedure of cryogenic preservation of ginger was explored.The results showed that the shoot were subcultured by 45days and detached the size of 2～3mm,shoot was precultured on MS medium with 0.5mol/L sucrose for 2days,then it was dehyrated with 60% PVS2solution for 5minutes at room temperature,then with 100%PVS2solution for 30minutes at 0℃.The materials were plunged into liquid nitrogen.After48h,the material was taken off from liquid nitrogen and thawed in 37～40℃2minute,then it was cultivated on MS＋6-BA 0.5mg/L＋NAA 0.1mg/L＋GA30.3mg/L＋sucrose 30g/L＋agar 7g/L,the highest survival rate was 57.7%.