小麦根际土壤中禾谷多黏菌实时荧光定量 PCR检测体系的建立及应用

摘要

为了研究转基因小麦N12-1对其根际土壤中禾谷多黏菌( Polymyxa graminis)数量的影响,建立了基于实时荧光定量PCR( Quantitative Real-Time PCR)方法检测多黏菌数量的体系,并将该体系用于转基因小麦N12-1及其受体扬麦158不同生长期根际土壤中禾谷多黏菌数量的检测。结果显示,标准曲线的扩增效率为95%,相关系数为0.999,熔解曲线呈现单一峰,符合荧光定量扩增的各项指标；在各个生长期,转基因小麦N12-1与其受体扬麦158根际土壤中的禾谷多黏菌数量没有显著差异。由此推测,转基因小麦N12-1的种植对其根际土壤中禾谷多黏菌的数量没有显著影响。%A quantitative real-time PCR system for the detection of the number of Polymyxa graminis was established in this study. In the system, the standard curve showed a linear correlation between the input DNA and the CT values, with PCR efficiency of 95% and the correlation coefficient of 0. 999, and the melting curve presented a single peak, indicative of the feasibility of the system. The quantity of P. graminis in the rhizosphere soil of transgenic wheat N12-1 and its paren-tal material Yangmai158 during growth stages was detected using the system. No significant difference was detected in the quantity of P. graminis between transgenic wheat N12-1 and Yangmai158. The results suggested that the planting of trans-genic wheat N12-1 had few adverse effects on the quantity of P. graminis in rhizosphere soil.