山东农业科学Issue(3)：106-109,4.
应用real-time PCR 定量检测土壤中小麦纹枯病菌方法的建立
Quantitative Detection of Rhizoctonia cerealis in Soil by Real-Time PCR
摘要
Abstract
A pair of specific primers,WKF -8/WKR -8,were designed according to the DNA se-quence of the internal transcribed spacers (ITS)region of Rhizoctonia cerealis AG-D fusion group.Their spe-cificity and sensitivity were detected.A real-time polymerase chain reaction system was developed and opti-mized based on SYBR GreenⅠfluorescent staining method.And the standard curve was drawn.A good linear relationship between the template DNA amount and cycle threshold (Ct)value was observed in the range of 1.0 ×10 -3 ~10 ng/μl.The correlation coefficient was 0.995,and the amplification efficiency was 99.7%. The sensitivity was 100 times higher than that of conventional PCR.The results showed that this method had characteristics of speediness,high sensitivity and specificity.关键词
小麦纹枯病菌/SYBR GreenⅠ荧光染色法/real-time PCRKey words
Rhizoctonia cerealis/SYBR Green Ⅰ fluorescent staining method/Real-time PCR分类
农业科技引用本文复制引用
徐娜娜,李鹏昌,于金凤..应用real-time PCR 定量检测土壤中小麦纹枯病菌方法的建立[J].山东农业科学,2014,(3):106-109,4.基金项目
山东省现代产业技术体系建设经费资助。 （）
