应用茎尖滴冻法超低温保存香蕉资源研究(英文)Study on the cryopreservation of in vitro shoot tips of Musa germplasm by droplet vitrification
李建国;张守梅;陈厚彬;徐春香;王泽槐;
摘要(Abstract):
采用正交试验探讨了滴冻法中装载液处理时间、玻璃化溶液处理时间和过渡培养时间的最优组合;另外还研究了不同冰冻保护剂、茎尖大小、恢复培养基中大量元素浓度对滴冻法保存效果的影响。通过试验筛选出了最佳的滴冻法保存方案:剥取含有1~2个叶原基的茎尖,长约1.5~2.5 mm,先在室温下用装载液预处理20~120 min,再用玻璃化溶液PVS2于0℃下处理30~50 min,接着将PVS2溶液滴于铝箔上,每个液滴中放一个茎尖,迅速投入液氮。保存1 h后,将铝箔取出投入卸载液(Suc.1.2 mol.L-1+MS)中快速解冻。15 min后取出茎尖,用滤纸吸去水分,转入过渡培养基上(Suc 0.3 mol.L-1+MS)暗培养24 h。再转到恢复培养基上,暗培养1周后转移到光下常规培养。采用滴冻法成功保存了分属于5个基因组型的15份香蕉资源,每种基因组型的平均成活率为:AAA 78.0%,AAB 57.8%,ABB72.1%,AA 43.3%,野生蕉42.2%。
关键词(KeyWords): 香蕉;种质资源;茎尖;滴冻法;超低温保存
基金项目(Foundation): 国家科技支撑计划(2008BAD92B06-4);; 广东省农业科技攻关项目(2002A2080101102)
作者(Author): 李建国;张守梅;陈厚彬;徐春香;王泽槐;
Email:
DOI: 10.13925/j.cnki.gsxb.2010.05.017
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