【摘要】 对转基因抗病小麦根际土壤中镰刀菌含量进行绝对定量,建立了实时荧光定量PCR(Real-time QPCR)检测体系。应用该反应体系,对转基因小麦各个生长时期的镰刀菌拷贝数进行绝对定量。结果表明,镰刀菌通用引物具有较好的特异性;Real-time QPCR反应的扩增曲线中各梯度标准质粒的循环阈值间隔均匀,溶解曲线峰值较突出,该标准曲线的相关系数(r)=0.999 20,斜率为-3.203,计算其扩增效率为105%,符合荧光定量PCR方法中对各项指标的要求。应用试验结果表明,在转基因小麦的播种前期、灌浆期、成熟期镰刀菌数量较低,在苗期、返青期、拔节期镰刀菌数量较高。更多还原

【Abstract】 Fusarium sp.is the major group of soil fungi.The effect of transgenic crop on soil rhizosphere microorganism is an important aspect of environmental biosafety research.In this paper,a real-time quantitative PCR detection system for Fusarium was established.The primer showed good specificity,and all indexes met the demands of normal real-time QPCR method.The spacing of cycle threshold of standard substance at each concentration gradient was uniform.The melting curve showed a simple peak,with the correlation coefficient(r) and slope being 0.999 20 and-3.203,respectively.The amplification efficiency was 105%.Using this method,the Fusarium in transgenic wheat rhizosphere soil at various growth stages was quantified low at early seeding,grain filling,and maturity,and high at seedling,turning green and jointing stages of wheat.更多还原