“矮丰”扁桃休眠芽玻璃化超低温保存条件优化

Cryopreservation of Dormant Buds by Vitrification in Almond Cultivars 'Aifeng'

以扁桃休眠芽为试材,采用玻璃化超低温保存的方法,研究了休眠芽大小、蔗糖预处理浓度/时间、装载处理时间、PVS2脱水处理时间对扁桃休眠芽超低温保存后的影响,以期为扁桃种质资源的保存提供参考依据。结果表明:从健康的扁桃植株上剥取0.6~1.5mm(含有1~2个叶原基)休眠芽茎尖,放入含MS的0.5mol·L^(-1)蔗糖预处理液中预培养1d后,室温下用装载液(含2mol·L^(-1)甘油和0.4mol·L^(-1)蔗糖)装载20min,0℃下用PVS2处理70min,加入少量新鲜的PVS2后迅速投入液氮罐,保存15d后取出,在40℃水浴锅中化冻2~3min,用1.2mol·L^(-1)蔗糖+MS的洗涤液卸载30min,无菌纸吸干洗涤液后转至恢复培养基中,在24℃条件下暗培养7d后进行正常光照培养,2周后成活率可达61.01%。

In order to provide a theoretical basis for the preservation of germplasm resources,the effects of sucrose pretreatment time,pretreatment time,loading time and PVS2 dehydration treatment time on the preservation of almond dormant buds were studied by using the method of vitrification and cryopreservation.The results showed that the dormant buds were removed from healthy plants,and the dormant buds were stripped to 0.6-1.5 mm（containing 1-2 leaf primordia）and cultured in MS containing 0.5 mol·L^-1 sucrose pretreatment solution 1 day,at room temperature with the liquid loadingof dormant buds loaded 20 minutes,dehydrated 70 minutes under the condition of 0℃.Then added a small amount of fresh PVS2,quickly put into liquid nitrogen tank,saved the 15 days out,the dormant buds were thawed 2-3 minutes in a 40℃ celsius water bath,uninstalled 30 minutes using 1.2 mol·L^-1 sucrose washing solution containing MS medium,after the aseptic paper sucked the washing liquid,it went to the recovery medium.Under the condition of 24℃celsius,the dormant buds were cultured in dark 7 days,and then cultured in normal light.After 2 weeks,the survival rate was 61.01%.