Submitted to: Plant and Animal Genome VX Conference Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 10/11/2006
Publication Date: 1/13/2007
Citation: Okubara, P.A., Li, C., Lawrence, N. 2007. Pressure Cycling Technology Sample Preparation System (PCT SPS) Improves Quantification of Pathogen DNA in Plants and Soil. Plant and Animal Genome VX Conference Abstracts.
Technical Abstract: Wheat and barley are susceptible to Rhizoctonia root rot, bare patch, and damping-off in the dryland cereal production regions of the Pacific Northwest and throughout the world. Detection and quantification of Rhizoctonia spp., the causal agents of this yield-limiting disease, are inconsistent at low population densities and in hard-to-extract samples. The Pressure Cycling Technology Sample Preparation System (PCT SPS) from Pressure BioSciences, Inc. is a novel, efficient, and reproducible extraction system in which samples are subjected to cycles of hydrostatic pressure (from ambient to as high as 35,000 psi) under controlled conditions. Using the PCT SPS, we obtained detectable amounts of Rhizoctonia DNA from wheat roots that were previously recalcitrant to homogenization. In 80% of the root samples, pathogen DNA was extracted in amounts high enough to be quantified using real-time PCR. The PCT SPS improved the extraction of Rhizoctonia DNA from agricultural soils up to 30-fold, compared to samples without the pressure cycling treatment. Furthermore, as the PCT SPS is a closed system, samples were free from contamination that can occur during conventional extraction procedures. The PCT SPS confers a significant advantage in germplasm screening, food security assessment, and characterization of host-microbe interactions.