罗汉果茎尖玻璃化法超低温保存初探

对罗汉果试管苗茎尖玻璃化法超低温保存进行了初步研究。结果表明,切取继代15 d生长健壮的罗汉果试管苗茎尖,长2~3 mm,在25℃下用60%PVS2(玻璃化保护液)预处理40 min,再用100%PVS2在0℃处理50 min,更换新鲜PVS2后投入液氮保存24 h,于40℃水浴中快速解冻2 min,用MS+410.8 g/L蔗糖的液体培养基洗涤40 min,滤纸吸干后接种到MS+1.0 mg/L6-BA+0.05 mg/L NAA+0.1 mg/L GA3+0.7%琼脂粉+45 g/L蔗糖的固体培养基上,25℃暗培养7 d,转入正常光下培养。1周时的存活率最高可达94.56%。

Preliminary Study on Stem Apex's Cryopreservation of Fructus Momordicae using Vitrification Method

The cryopreservation of the stem apex's of Fructus Momordicae in vitro by vitrification was preliminarily researched.The results showed that the stem apexes(2-3mm) from healthy in vitro subcultured 15d were pretreated with 60% PVS2 for 40 min at 25℃,then treated with 100% PVS2 for 50 min at 0℃,and then protected with fresh 100% PVS2 and put them into the liquid nitrogen.After 24h,these stem apexes were thawed rapidly in water for 2min at 40℃,then rinsed in MS+410.8g/L sucrose for 40min,after absorbing the liquid using filter paper,inoculated it on the medium MS+1.0mg/L 6-BA+0.05mg/L NAA+0.1mg/L GA3+0.7% agar+45g/L sucrose,cultured in dark at 25℃ for 7d,then transferred the stem apexes into the condition of normal lighting.So,the highest survival rate was up to 94.56% after 1 week.