【摘要】 为建立土壤中黑胫病菌的SYBR GreenⅠ实时荧光定量PCR快速检测方法,依据疫霉菌特异性par A1基因序列差异位点,设计1对特异性引物。通过构建含有目的片段的重组质粒,建立标准曲线,以SYBR GreenⅠ荧光染料法对烟草黑胫病菌进行荧光定量PCR检测,检测灵敏度达1×102个/g土壤,建立了基于SYBR GreenⅠ荧光染料法的荧光定量PCR的快速定量检测体系,并对云南、四川烟草主要栽培区土壤进行检测。表明该体系可以对烟草不同时期的田间土壤黑胫病菌的数量进行动态监测,可用于烟草黑胫病的预测和防治。更多还原

【Abstract】 The aim of this study was to establish the SYBR GreenⅠreal-time fluorescence quantitative PCR assay for rapid detection of Phytophthora nicotianae in soil.According to the sequence of the par A1 gene,the specific pair of primers were designed.By constructing a recombinant plasmid containing the desired fragment and establishing the standard curve,the real-time q PCR with SYBR GreenⅠdye fluorescence was used to detect Phytophthora nicotianae in soil.The sensitivity of detection was Up to 1 × 102 spores per gram of soil. A rapid quantitative detection system of q PCR based on SYBR GreenⅠfluorescence dye method was established,for the detection of the soil of the main tobacco cultivation area soil in Yunnan and Sichuan.These results indicated that the amount of Phytophthora nicotianae in different tobacco growth periods of field soil can be dynamically monitored by this system,and could be used in predicting the occurrence of tobacco black shank.更多还原