Specific polymerase chain reaction primers for the detection of Plasmodiophora brassicae in soil and water
Specific polymerase chain reaction primers for the detection of Plasmodiophora brassicae in soil and water
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DOI：
10.1094/phyto.1999.89.5.392
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发表时间：
1999-05-01
期刊：
影响因子：
3.2
通讯作者：
Porter, IJ
中科院分区：
文献类型：
--
作者：
Faggian, R;Bulman, SR;Porter, IJ
针对芸苔根肿菌的特异性寡核苷酸引物的开发导致了土壤和水中芸苔根肿菌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，并将数据用于设计针对核糖体RNA基因和内部转录间隔区的引物。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芸苔疫霉分离株进行了特异性测试。对于纯模板，灵敏度被确定为0.1毫微克（fg; 10（-15）g），并且低至1，000个孢子/g灌封混合物。在土壤中，在接种物足以导致根肿病症状的所有土壤中检测到芸苔疫霉。本文还介绍了一种从土壤中提取DNA的简便方法。
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.