远志种质资源遗传多样性随机扩增多态性DNA分析

目的 用随机扩增多态性DNA(RAPD)技术分析远志Polygala tenuifolia Willd.及卵叶远志P.sibirica L.的遗传多样性.方法 反应总体积25 μl,内含10×PCR buffer 2.5 μl,dNTPs 2.0 μl,MgCl2 2.0 μl,引物1.0 μl,模板2 ng,Taq酶0.3 μl.扩增程序为:94℃预变性4 min,然后进行45循环:94℃变性15 s,36℃复性1 min,72℃延伸1.3 min,循环结束后72℃延伸4 min.结果 10个引物共检测到154个位点,平均每个引物扩增9.24条带,其中148条带表现为多态性,占总带数的94.3%;距离系数为24时,11个样品明显聚为远志居群和卵叶远志居群两类.结论 远志与卵叶远志具有丰富的遗传多样性,遗传聚类与其地理分布有一定的相关性.

Genetic Diversity of Polygala tenuifolia Willd and P.sibirica L.with RAPD-PCR

Objective To analyze genetic diversity of Polygala tenuifolia Willd and P.sibirica L.with RAPD-PCR.Methods The RAPD-PCR reaction system was constructed with 2.5 μl 10×PCR buffer,2.0 μl dNTPs,2.0 μl MgCl2,1.0 μl primer,2 ng template and 0.3 μl Taq with total reaction volume of 25 μl.PCR amplifying sequences were as follows:Initial denaturating for 4 min at 94℃,then denaturating for 15 s at 94℃,annealing for 1 min at 36℃ and extension for 1.3 min at 72℃.After 45 times cycle,keeping extension for 4 min.Results 154 polymorphism strands were detected with 10 primers,among which 148 strands were of polymorphism.Cluster analysis indicated that all samples could be clustered into two populations with distance coefficient of 24.Conclusion Polygala tenuifolia Willd and P.sibirica L.have rich genetic diversity.The genetic clustering has some relevance with geographical distribution of P.tenuifolia Willd and P.sibirica L.