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不同贮藏条件下棉花叶片DNA提取效果分析
作者：郑子漂 曲延英 王丽丽等 来源：《山东农业科学》2015年第08期
摘要：针对棉叶贮藏方法不恰当导致DNA提取得率及纯度较差的问题，本研究以新鲜棉叶为对照，比较了5种贮藏条件下的棉叶DNA提取效果，以期筛选出一种棉叶贮藏的较优模式，从而提高DNA的提取得率及纯度。结果表明，-20℃冷冻3 d的棉叶DNA得率最高且纯度较好，甚至比新鲜叶片的DNA得率高51.4%； 3 d冷冻贮藏的DNA得率明显高于3 d冷鲜贮藏的；7 d液氮冷冻贮藏的DNA得率明显优于7 d硅胶干燥贮藏的和冷冻贮藏的；7 d液氮速冻贮藏的DNA纯度与7 d硅胶干燥贮藏的相近。综上所述，短期贮藏可采用-20℃冷冻保存的方法；较长时间的保存建议采用液氮速冻的贮藏方式。 关键词：贮藏条件；棉叶；DNA提取；纯度；得率
中图分类号：S562+S188 文献标识号：A 文章编号：1001-4942（2015）08-0011-03 Abstract This study was conducted based on the problems that inappropriate storage could lead to poor extraction rate and purity of DNA from cotton leaves. With the fresh leaves as control， the DNA extraction effects of cotton leaves were compared after storage in five kinds of conditions to screen out a best storage method. The results showed that the DNA extraction rate of cotton leaves was the highest after -20℃ frozen storage for 3 days with better purity， which increased by 51.4% compared to that of the control. The DNA yield of 3-day frozen storage was obviously higher than that of 3-day 4℃ storage， and the 7-day liquid nitrogen frozen had clear advantages over the silica gel dry storage and 7-day frozen storage. The DNA purity of 7-day liquid nitrogen frozen was close to that of 7-day silica gel dry storage. In conclusion， -20℃ frozen storage could be used for short-term storage， while the liquid nitrogen frozen could be used for longer-time storage.
Key words Storage condition； Cotton leaves； DNA extraction； Purity； Extraction rate 近年来， 分子生物技术在棉花育种上的应用，使棉花在抗性、产量及品质改良等方面均取得了较大进展。但棉花组织中富含酚类及多糖类物质，棉叶总DNA的提取较为困难，若采集的鲜叶样品保存不当，将严重影响其DNA的提取纯度及得率，进而影响后续分子工作的开展[1～3]。因此，通过实验手段找出满足不同要求的植物叶片贮藏条件，对各项实验的开展具有重要意义。
本研究以棉花新鲜嫩叶为对照，比较了不同贮藏条件对棉叶DNA提取得率及纯度的影响，以期筛选出最佳的棉叶贮藏方法，为今后开展分子生物学和生物技术试验提供技术支持。