彩色马蹄莲ISSR体系的建立及初步分析

Optimization of ISSR Reaction System and Preliminary Study on Zantedeschia

选取白花马蹄莲和10个颜色有代表性的彩色马蹄莲栽培品种为材料,采用改进的SDS法提取叶片基因组DNA,通过正交设计研究Mg2+、Taq DNA聚合酶和模板DNA、引物的影响,确定彩色马蹄莲最佳ISSR反应体系为:每20μL中含1×PCR Buffer、1.5mmol/L MgCl2、0.6μmol/L引物、20ng模板DNA、1UTaqDNA聚合酶。从100个引物中筛选出的15条进行分析,共获得了109条带,其中多态性为99条,占总数的90.8%。选用NTSYS软件进行聚类分析,绘出了供试材料的亲缘关系图,为系统进行马蹄莲属植物的遗传多样性分析和品种选育提供了参考。

The total genome DNA of a white calla lily （Zantedeschia aethiopica） and 10 representative varieties of colorful calla lily （Zantedeschia hybrid ） have been obtained through the method of modified SDS. The concentration of Mg^2＋, Taq DNA polymerase, primer and template DNA, greatly influencing ISSR-PCR, were optimized by orthogonal design in this study. The results showed that optimum ISSR reaction system of calla lily was every 20 μL reaction system containing 1 ×PCR Buffer, 1.5 mmol/L MgCI2, 0.6 μmol/L primer, 20 ng DNA template and 1 U Taq DNA polymerase. In this study, 109 pattern bands results from ISSR-PCR by amplifying 15 primers, 99 bands （90.8%） were polymorphic. Based on the soft NTSYS, the genetic relationship were analyzed. This study will be helpful to the breeding work and research on the genetic diversity of the genus Zantedeschia.