越橘离体茎尖包埋玻璃化超低温保存  

Cryopreservation of Shoot Tips of Blueberry (Vaccinium) in vitro byEncapsulation-Vitrification

作　　者：毛宇金 李娴 张志东 苏原慧 梁杉 孙海悦 李亚东 MAO Yujin;LI Xian;ZHANG Zhidong;SU Yuanhui;LIANG Shan;SUN Haiyue;LI Yadong(College of Horticulture,Jilin Agricultural University,Jilin Provincial Engineering Center of Genetic Breeding and Innovative Utilization of Small Fruits,Changchun 130118,China)

机构地区：[1]吉林农业大学园艺学院,吉林省小浆果遗传育种与创新利用工程中心,长春130118

出　　处：《吉林农业大学学报》2023年第2期155-162,共8页Journal of Jilin Agricultural University

基　　金：吉林省科技发展计划项目(20170414023GH,20180201076NY)。

摘　　要：为越橘种质资源保存提供新途径,试验研究了越橘组培苗茎尖包埋玻璃化法超低温保存方法。结果显示适宜的条件包括:剥取顶芽茎尖(长1.0~1.5 mm),在(25±2)℃黑暗条件下预培养1 d;使用移液枪吸取茎尖与藻酸盐溶液,滴入氯化钙溶液中形成包埋球(直径4 mm);包埋球在加载液中加载1.5 h(室温);包埋球转入PVS2溶液处理4 h(0℃);将装有包埋球的冷冻管投入液氮保存1 h;取出液氮中的冷冻管迅速转入37℃水浴锅中解冻1~2 min,在室温下取出包埋球,在卸载液中卸载20 min;卸载的茎尖在再生培养基上(25±2)℃黑暗培养5 d;之后在正常光照条件下培养30 d。所测试的8个越橘基因型平均再生率为57.9%,最高再生率为78.5%,最低为41.7%。In this study,shoots of blueberry in viro were used as materials to study the cryopreservation of its shoot tips by encapsulation-vitrification,which provided a new way for the preservation of Vaccinium germplasm resources.The results showed that the suitable conditions are as follows:The shoot tips(1.0-1.5 mm length)excised from shoots were pre-cultured for 1d at(25±2)℃ in the dark.The shoot tips and alginate solution were absorbed with a pipette.The alginate pellets(4 mm dia-meter)were formed by dripping into the calcium chloride solution.The pellets were treated in the loading solution for 1.5h at room temperature.Then the pellets were treated with PVS2 solution at 0℃ for 4 h.The cryotubes containing alginate pellets were put into liquid nitrogen and stored for 1h.The cryotube was removed from liquid nitrogen and transferred quickly to the 37℃ water bath to defrost for 1 to 2 min.Alginate pellets were removed from the freezing tube at room temperature and then placed in the unloading solution for 20 min.The unloaded shoot tips were cultured in dark on regeneration medium(25±2)℃ for 5 d.They were then cultured under normal light for 30 d.The average regeneration rate of 8 genotypes tested was 57.9%,and the highest regeneration rate was 78.5%while the lowest was 41.7%.

关 键 词：越橘 超低温保存 小滴玻璃化 组培苗茎尖 

分 类 号：S663.9[农业科学—果树学]