高山杜鹃ISSR-PCR反应体系建立

Establishment of ISSR-PCR Reaction System for Rhododendron lapponicum

[目的]建立高山杜鹃的ISSR-PCR反应体系。[方法]运用正交试验分析模板DNA、TaqDNA聚合酶、Primer、Mg2+和dNTPs 5个因子对杜鹃ISSR-PCR反应影响,建立高山杜鹃ISSR-PCR反应体系。[结果]杜鹃ISSR-PCR 25μl反应体系中5个因子较适水平为:DNA模板浓度为1.6 ng/μl,TaqDNA聚合酶浓度为0.040 U/μl,Primer浓度为0.64 mmol/L,dNTPs浓度为0.68 mmol/L,Mg2+浓度为2.08mmol/L。[结论]研究结果为利用ISSR分子标记技术研究杜鹃提供了理论支持。

[ Objective] To establish ISSR-PCR reaction system for Rhododendron lapponicum. [ Method] Effects of five factors （DNA template, TngDNA, Primer, Mg^2＋ and dNTPs） on ISSR-PCR reaction of Rhododendron lapponicum were analyzed by orthogonal design for establishing and optimizing ISSR-PCR reaction system. [ Result] As a result, the optimal ISSR -PCR reaction system （25 μl） mixture contained 40. 013 ng DNA template, 1.0 U TagDNA, 0.64 mmol/L primer, 2.08 mmol/L Mg^2＋ and 0.68 mmol/L dNTPs. [ Conclusion] Theory sustaining for researching Rhododendron by ISSR molecular marker were afforded from this experiment.