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出 处:《时珍国医国药》2009年第7期1834-1835,共2页Lishizhen Medicine and Materia Medica Research
摘 要: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25μl,内含10×PCR buffer2.5μl,dNTPs2.0μl,MgCl22.0μl,引物1.0μl,模板2ng,Taq酶0.3μl。扩增程序为:94℃预变性4min,然后进行45循环:94℃变性15s,36℃复性1min,72℃延伸1.3min,循环结束后72℃延伸4min。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。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.
关 键 词:远志 卵叶远志 遗传多样性 随机扩增多态性DNA
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