利用ISSR揭示不同类型月季遗传多样性

本研究应用正交设计法对ISSR反应体系中的各个主要影响因子进行了优化筛选，确立了适合月季ISSR-PCR反应的最佳体系。结果表明，25μL的ISSR反应体系中各组分的最适浓度分别为：1×PCR缓冲液、1U Taq DNA聚合酶、800pmol／L 引物、0.16mmol／L dNTPs、Mg^2＋ 1.5mmol／L。筛选了33个ISSR引物，共得到了11个多态性比较高的ISSR引物，占所筛引物的33.33％。利用筛选出的11条ISSR引物对3种月季类型的23份月季材料进行遗传多样性分析，共扩增出477条DNA带，其中多态性位点有14个，平均每条引物可以检测到4.5个多态性位点。用NTSYS软件对样品进行了UPGMA聚类分析，聚类结果表明丰花月季基本能聚为一类，切花月季与藤本月季交叉聚在一起。这表明月季种质的遗传差异与其应用分类的相关性不紧密。

Genetic Diversity of Different Roses Revealed by ISSR

Orthogonal design was used to optimize ISSR-PCR amplific reaction system. An optimal ISSR-PCR system for rose was obtained as 1×PCR buffer, 1 U Taq DNA polymerase, 800 pmol/L primer, 0.16 mmol/L dNTPs, 1.5 mmol/L Mg^2＋ for 25μL reaction system. We screen 33 ISSR primers. We have got 11 ISSR primers with high polymorphism, which represent 33.33% of the total number oflSSR. We selected 11 ISSR primers to do the genetic diversity analysis of three types roses. The result of PCR amplification showed that 477 DNA segments were obtained in the tested materials, of which, 14 were polymorphic bands and the average number of polymor-phism bands per ISSR primer was 4.5. UPGMA cluster analysis showed that all materials used in this study could be classified into 2 groups. All materials collected from the florbunda rose could be clustered together. The acces-sions from cut rose and vine rose were mixed by cluster analysis. All results indicated that the relationship between genetic variation of the germplasms and their different types were not so close.