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目的为罗汉果Siraitia grosvenorii种质资源的保存提供一条新途径。方法以继代15d生长健壮的罗汉果试管苗0.8～1cm长嫩梢进行预培养,剥取2～3mm茎尖,25℃下60%PVS2装载,再用100%PVS2在0℃脱水处理,更换新鲜100%PVS2后投入液氮保存24h,化冻,用MS+1.2mol/L蔗糖的液体培养基洗涤,滤纸吸干后接种于MS+1.0mg/L6-BA+0.05mg/L NAA+0.1mg/L GA3+0.8%琼脂粉+45g/L蔗糖的恢复培养基上,25℃暗培养7d,转入正常光下培养。再生苗生根培养基为1/2MS+0.2mg/L NAA+30g/L蔗糖。结果嫩梢于MS+0.7mol/L蔗糖的培养基上预培养3d,茎尖装载40min,脱水50min,液氮保存后于40℃水浴中快速化冻,洗涤40min,恢复培养1周时存活率最高可达100%,30d时再生率最高可达78.33%。再生苗转入生根培养基可形成完整植株。结论罗汉果种质资源的玻璃化法超低温保存操作简单、成活率高、再生植株正常。
Objective To provide a new way to preserve Siraitia grosvenorii germplasm resources. Methods The young shoots of Luohanguo, which grew robust for 15d, were pre-cultured in 0.8-1 cm long tender shoots, stripped of 2 ~ 3mm shoot tips, loaded with 60% PVS2 at 25 ℃, dehydrated at 100 ℃ with 100% PVS2, 100% PVS2 was stored in liquid nitrogen for 24h, thawed, washed with MS + 1.2mol / L sucrose liquid medium, the filter paper was dried and inoculated into MS + 1.0mg / L6-BA + 0.05mg / L NAA + 0.1mg / L GA3 + 0.8% agar powder + 45g / L sucrose, cultured in dark at 25 ℃ for 7 days and then transferred to normal light. Regeneration seedling rooting medium is 1 / 2MS + 0.2mg / L NAA + 30g / L sucrose. Results The shoots were preincubated for 3 days on MS medium supplemented with 0.7mol / L sucrose. The tips were loaded for 40min and dehydrated for 50min. After being stored in liquid nitrogen, the shoots were rapidly frozen in water bath at 40 ℃ for 40min. Up to 100%, 30d regeneration rate up to 78.33%. Regeneration seedlings into rooting medium can form a complete plant. Conclusion The vitrification method of Mangosteen germplasm resources is easy to be preserved by cryopreservation with high survival rate and normal regenerated plants.