采用CTAB(十六烷基三甲基溴化铵)法从10个玉米品种种子胚中提取全基因组DNA，用正交实验法优化ISSR-PCR反应体系，筛选扩增条带清晰的引物，然后用所筛引物对10个玉米品种DNA扩增，分析其扩增带纹差异，找出能够鉴别10个玉米品种的合适引物。研究结果表明：①优化的ISSR-PCR反应参数为：1.5 mmol/L Mg²⁺、0.375 mmol/L dNTP、0.5 μmol/L引物、2.5U Taq DNA聚合酶；②从40条ISSR引物筛选出11条扩增条带清晰、多态性较高的引物，它们共扩增出101条条带；③利用引物UBC808能将10个供试玉米样品区分开。

Genomic DNA of 10 maize varieties were extracted from the embryos using CTAB method, and suitable ISSR(Inter-simple sequence repeats) reaction system was established with multifactor orthogonal experiment. The feasible ISSR primers were screened which could be used to distinguish 10 varieties based on the ISSR fingerprints. The results showed that: ①The optimized ISSR reaction system, namely 20 μL reaction system contained 1×PCR buffer, 1.5 mmol/L Mg²⁺, 0.375 mmol/L dNTP, 0.5 μmol/L primer and 2.5U Taq DNA polymerase. ②101 polymorphism bands were obtained steadily in 10 maize varieties by 11 ISSR primers chosen from 40. ③All of the 10 varieties could be differentiated from each other based on their specific bands established by primer UBC808.

河南省青年骨干教师资助项目(教高2005-461-110)