中国李RAPD的优化反应体系及其在品种鉴定中的应用

Optimal Reaction System of RAPD in Chinese Plum and Its Preliminary Use for Cul tivar Identification

以5'-GACCGCTTGT-3'为随机引物,以奉化李(Prunussalicinacv.Fenghuali)为试材,对中国李RAPD反应体系进行了优化研究,结果表明:25μL反应体系中,TaqDNA聚合酶、Mg2+、引物、模板DNA和dNTPs5种主要成分的适宜浓度或用量分别是:1.0U、2.5mmol/L、20ng、40ng和0.2mmol/L。利用该优化反应体系,用10个随机引物,构建了5个中国李品种的RAPD指纹图谱;并以共有谱带率对这些品种遗传关系进行了分析,5个中国李品种的共有谱带率平均为33.2%,变异幅度为20.1%～44.9%。

The optimal reaction of RAPD(Random Amplified Polymorphic DNA)in Chinese plum(Prunus salicina Lindl.)was studied with5'-GACCGCTTGT-3' primer for cv.Fenghuali.The results showed that the optimum concentration of five important components i.e.Taq DNA polymerase,Mg 2+ ,primer,template DNA and dNTPs in25μL RAPD reaction system were1.0U,2.5mmol/L,20ng,40ng and0.2mmol/L,respectively.The RAPD fingerprinting of five Chinese plum culti-vars were established by using optimum reaction system with ten random primers.The genetic relationship of these cultivars were analyzed with their Band Sharing(BS).The Band Sharing average of five plum cultivars was33.2%,and their variant range was20.1%～40.9%.