珍稀濒危植物五小叶槭ISSR-PCR反应体系的建立和优化

Establishment and Optimization of ISSR-PCR Reaction System for Acer pentaphyllum

摘要: 五小叶槭为珍稀濒危植物，在研究被子植物的系统发育和古地理、古气候等方面具有重要的科学价值。本研究采用2XTaqMaster Mix，综合考虑Taq酶、Mg²⁺和dNTPs，对影响五小叶槭ISSR-PCR反应的3个因素（DNA模板、引物和Master Mix）进行3水平正交试验，从而优化出五小叶槭ISSR-PCR反应最佳体系。结果表明，引物用量对反应结果影响最大，其次为DNA模板，2XTaqMaster Mix对反应结果影响最小。最佳反应体系（25μL）为:DNA模板（50ng·μL^-1）1μL，引物2μL，2XTaqMaster Mix 13.5μL。

Abstract: Acer pentaphyllum is a kind of rare and endangered plants and it has important scientific value in studying angiosperm's phylogeny,paleogeography and paleoclimate.It this study,2XFaq Master Mix was adopted and the effect of Taq polymerase,dNTPs and Mg²⁺ were considered aiming to establish and optimize the best ISSR-PCR reaction system for Acer pentaphyllum.The results showed that the most remarkable factor was primers,followed by DNA template,and 2X Master Mix had the least effect on the reaction result.The optimum reaction system (25 μL) was 1 μL of DNA template (50 ng·μL^-1),2 μL of primer and 13.5 μL of 2XTaq Master Mix.