锈菌夏孢子DNA的微量快速提取方法  

A Minim and Rapid Method for DNA Extraction from Uredinium of Rusts Fungi

作　　者：余仲东[1] 高爱琴[1] 曹支敏[1] 

机构地区：[1]西北农林科技大学林学院,陕西杨凌712100

出　　处：《生物技术通讯》2005年第1期48-50,共3页Letters in Biotechnology

基　　金：国家自然科学基金项目(39970616)

摘　　要：采用4种方法对毛白杨锈病单个夏孢子堆DNA提取和ITS-PCR扩增表明,钢珠法、玻片法均是锈菌基因组DNA微量快速提取的合适方法,以钢珠法最优,整个过程仅需30min,并且可以获得与CTAB法、氯化苄法相同的PCR扩增结果。钢珠法是在提取液中加入2～3颗钢珠,靠钢珠在涡旋中的相互碰撞将夏孢子破壁,以同时加入NaOH(终浓度3.3%)和Chelex-100(终浓度1.6%)效果最好,ITS-PCR扩增能稳定得到700bp片段。玻片法也能得到相同的结果。分析比较了4种方法的优缺点。The genomic DNA of single uredinium on Melampsora rostrupii was extracted via four methods, and ITS-PCR was carried with the DNA substrates. The results showed that steel ball method and glass slides method are both the good way for rust fungi DNA minim and rapid extraction. Especially, the steel ball method for DNA extraction can be accomplished within 30 min, it was the best one among these methods. Cell wall was crashed by two bouncing steeL balls when the tube was put on the vortex machine. For this method, adding NaOH(3.3%) and Chelex-100(1.6%) was the best way for DNA extraction. One 700 bp ITS-PCR fragment can be achieved repeatedly through both steel balls method and glass slides method.

关 键 词：锈菌 夏孢子 DNA 微量快速提取方法 植物病原菌 钢珠法 玻片法 

分 类 号：S432.42[农业科学—植物病理学]