Phytopathology. 1999 May;89(5):392-7. doi: 10.1094/PHYTO.1999.89.5.392.
ABSTRACT The development of specific oligonucleotide primers for Plasmodiophora brassicae has led to a nested polymerase chain reaction (PCR) detection method for P. brassicae in soil and water. Initially, the PCR was used to amplify a section of the rDNA repeat. The PCR products were sequenced and the data used to design primers that were directed at the ribosomal RNA genes and internal transcribed spacer regions. Specificity was tested against more than 40 common soil organisms, host plants, and spore suspension contaminants, as well as P. brassicae isolates from around Australia and the world. Sensitivity was determined to be 0.1 fentograms (fg; 10(-15) g) for pure template and as low as 1,000 spores per g of potting mix. In soil, P. brassicae was detected in all soils where the inoculum was sufficient to result in clubroot symptoms. Also outlined is a simple method of DNA extraction from soil.
摘要 针对根肿菌（Plasmodiophora brassicae）特异性寡核苷酸引物的开发，实现了根肿菌在土壤和水中的巢式聚合酶链反应（PCR）检测方法。最初，PCR 用于扩增 rDNA 重复序列的一部分。对 PCR 产物进行测序，并利用这些数据设计针对核糖体 RNA 基因和内部转录间隔区的引物。该方法针对 40 多种常见土壤生物、宿主植物和孢子悬浮污染物以及来自澳大利亚和世界各地的根肿菌分离株进行了特异性测试。其灵敏度为纯模板时为 0.1 飞克（fg；10(-15) g），盆栽混合物中每克低至 1000 个孢子。在土壤中，当接种足够导致根肿病症状的菌量时，根肿菌可在所有土壤中被检测到。本文还概述了一种从土壤中提取 DNA 的简单方法。
