玻璃化法超低温高效保存‘Gala’苹果组培苗  

Cryopreservation of'Gala'Apple in Vitro by Vitrification

作　　者：徐娜[1] 陈桂玲[1] 杨磊 朱子涵 陈丽[1] 韩亚丁 刘飞 XU Na;CHEN Gui-ing;YANG Lei;ZHU Zi-han;CHEN Li;HAN Ya-ding;LIU Fei(College of Biological Sciences/Jining Medical University,Rizhao 276800,China)

机构地区：[1]济宁医学院,山东日照276800

出　　处：《山东农业大学学报（自然科学版）》2023年第5期718-723,共6页Journal of Shandong Agricultural University：Natural Science Edition

基　　金：国家自然科学基金(32000194);山东省自然科学基金(ZR2017BC081);作物生物学国家重点实验室开放项目(2017KF08);山东省高等学校青年创新团队(2022KJ102);山东省教育科学规划创新素养专项课题(2022CYB210);济宁医学院教师国内访学项目。

摘　　要：本试验以苹果‘Gala’苹果为材料,从低温锻炼时间、预培养基蔗糖浓度、预培养时间和玻璃化溶液及处理时间4个方面优化玻璃化超低温保存体系,获得‘Gala’苹果茎尖超低温保存的最佳体系。结果表明,将生长30 d的组培苗在4℃的条件下低温锻炼5周,剥取茎尖,将其置于含0.4 mol/L蔗糖的预培养基中培养5 d,渗透装载液渗透30 min,0℃的条件下用PVS3玻璃化溶液处理60 min,放入液氮中冷冻24 h,取出后置于40℃水浴中1 min快速化冻,卸载液清洗2次,每次10 min,然后在恢复培养基上培养,茎尖的再生率最高达到96.6%且生长稳定性好。本试验成功建立了‘Gala’苹果的玻璃化法超低温保存体系,为苹果种质资源的长期保存提供了一条有效途径。We optimized the vitrified cryopreservation system from low-temperature trained time,sucrose concentration of pre-culture medium,pre-culture time,vitrification solution and treatment time for cryopreservation of'Gala'apple stem tip.Tissue culture seedlings grown for 30 days were subjected to cold acclimation at 4℃for 5 weeks;the shoot tips were stripped,cultured in the preculture solution containing 0.4 mol/L sucrose for 5 days;followed by permeating in loading solution for 30 min;vitrified by PVS3 for 60 min at 0℃;frozen in liquid nitrogen for 24 h;thawed in water bath at 40℃for 1 min;washed twice by dilution solution,10 min each time;finally recovered in the medium,the shoot tips grew stably and the regeneration rate was up to 96.6%.The study has successfully established effective vitrification cryopreservation protocol of'Gala'shoot tips,and provided an innovative way for long-term conservation of apple germplasm.

关 键 词：‘Gala’苹果 超低温处理 组培 

分 类 号：S602.4[农业科学—园艺学]