【摘要】 作者对在小麦DNA的RAPD扩增中 ,Mg2 + 浓度、TaqDNA聚合酶和模板DNA的用量对扩增结果的影响进行了探讨 ,研究结果发现 ,在Mg2 + 浓度为 2 .0mmol/L ,TaqDNA聚合酶为 2U/2 5μL ,模板DNA为 15～ 2 0ng/2 5μL时 ,对小麦DNA的RAPD扩增结果较好 .实验共使用了 10 0个随机引物 ,对抗禾谷孢囊线虫基因供体亲本易变山羊草、抗性近等基因系 (near isogenicline,NILs)E- 10 和回交亲本寄主品系Lutin进行RAPD扩增 ,发现有 3个引物 :OPB12 ,OPB14和OPB11的扩增产物 ,在抗感NILs材料间揭示了多态性 ,但只有引物OPB12在 4次以上重复中 ,获得稳定相同的结果 ,其多态标记为OPB12 - 1350 ,它只在NILs和抗性供体亲本中出现 ,而回交亲本中未出现 .初步认为该多态性分子标记OPB12 - 1350 与抗禾谷孢囊线虫基因可能相关更多还原

【Abstract】 The factors that influencing RAPD (Random Amplified polymorphic DNA) reaction, including the concentration of Mg 2+ ,Taq DNA polymerase and template DNA were examined in the study. The results indicated that the optimum RAPD condition for wheat were:Mg 2+ concentration 2.0mmol/L,Taq DNA polymerase 2U,template DNA concentration 15～20ng in 25μL reaction system. The RAPD analysis was made in CNN (cereal cyst nematode) resistance gene near isogenic kines (NILs) with 100 random primers. Three primers the NILs.Only OPB12 showed the same discriminating results in more than four replications.The polymorphic DNA marker was OPB12 -1350 . It was amplified only in NILs E -10 and the parent carrying the CNN resistance gene Ae.variabilis,but the back cross parent Kutin wasn’t found. The authors are studing the polymorphic DNA molecular marker OPB12 -1350 to confirm the relation of the CNN resistance gene in wheat.更多还原