| 大苞鞘石斛原球茎玻璃化超低温保存技术的研究 |
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| 引用本文： | 吴元玲,申晓辉.大苞鞘石斛原球茎玻璃化超低温保存技术的研究[J].细胞生物学杂志,2011(3). |
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| 作者姓名： | 吴元玲 申晓辉 |
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| 作者单位： | 上海交通大学农业与生物学院农业部都市农业(南方)重点开放实验室; |
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| 基金项目： | 花卉种质创新和产业发展关键技术的研究-上海农委重点攻关(No.2006-4-9)资助项目~~ |
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| 摘 要： | 本文研究建立了大苞鞘石斛(Dendrobium wardianum Warner)原球茎玻璃化法超低温保存的技术体系。结果发现,预处理和玻璃化溶液(plant vitrification solution 2,PVS2)装载脱水是影响大苞鞘石斛原球茎相对存活率的两个关键步骤,高渗与低温-高渗两种预处理方法测定的相对存活率具有显著性差异;玻璃化溶液的种类以及脱水时间对冻后存活率具有重要的影响。基于此,建立了大苞鞘石斛原球茎的超低温保存体系,即:以继代培养60 d的大苞鞘石斛原球茎为材料,1/2MS+0.8 mol/L蔗糖的培养基上4℃低温预处理6 d后,转至1/2 MS+2 mol/L甘油+0.4 mol/L蔗糖的装载液中室温下装载40 min,在0℃下装载PVS2脱水40 min,然后转入装有新鲜PVS2冷冻管中并迅速投入液氮。在液氮保存1 h后放在40℃水浴中快速解冻1 min,利用含1.2 mol/L蔗糖的1/2MS培养液洗涤3次,每次间隔10 min;待恢复培养30 d后统计存活率,可使大苞鞘石斛原球茎超低温保存后存活率达到20.0%。
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| 关 键 词： | 大苞鞘石斛 原球茎 玻璃化 超低温保存 |
Cryopreservation of Dendrobium wardianum Warner.Protocorms by Vitrification |
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| Yuan-Ling Wu,Xiao-Hui Shen Ministry of Agriculture,Shanghai ,China.Cryopreservation of Dendrobium wardianum Warner.Protocorms by Vitrification[J].Chinese Journal of Cell Biology,2011(3). |
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| Authors: | Yuan-Ling Wu Xiao-Hui Shen Ministry of Agriculture Shanghai China |
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| Institution: | Yuan-Ling Wu,Xiao-Hui Shen~* (School of Agriculture and Biology,Shanghai Jiaotong University,Key Laboratory of Urban Agriculture(South) Ministry of Agriculture,Shanghai 200240,China) |
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| Abstract: | Cryopreservation system of Dendrobium wardianum Warner.protocorms were successfully established by using vitrification cryopreservation method.The results indicated that pretreatment and plant vitrification solutions(PVS2) dehydration played crucial roles for relative survival rate in the whole process.In addition, room temperature and low temperature hypertonic treatments presented significant survival differences.Furthermore, the different plant vitrification solutions(PVS) and dehydration times even infl... |
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| Keywords: | Dendrobium wardianum Warner protocorms vitrification cryopreservation |
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