苍耳柄锈菌三裂叶豚草专化型冬孢子基因组DNA快速提取方法的比较研究

The methods for rapid DNA extraction from teliospores of Puccinia xanthii f. sp. ambrosiae-trifidae

以苍耳柄锈菌三裂叶豚草专化型Puccinia xanthii f.sp.ambrosiae-trifidae为研究试材,比较研究了其冬孢子DNA提取的9种方法,其中CTAB-钢珠法、改良的微型电钻法以及EZ-Kit改良法获得的基因组DNA经检测质量较好。在此基础上,利用ITS-PCR和ISSR引物UBC#835将待用DNA进行PCR扩增检测。结果表明,上述3种方法提取的DNA适合于ISSR反应。研究结果为专性寄生锈菌分子遗传变异的研究提供了保障。

The genomic DNA of Puccinia xanthii f. sp. ambrosiae-trifidae was extracted by nine methods from the teliospores. The results show that the methods of CTAB-steel balls,modified mini-drill and modified EZ-Kit were better than the other six methods due to their higher quality of DNA extraction with less impurity. The template DNA samples generated from these methods were detected by methods of ITS-PCR and ISSR-PCR with the primer UBC#835. They all produced expected band patterns. The research confirms that the DNA samples generated from these three methods could be used for further molecular study of the rust fungus.