Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: July 31, 2004
Publication Date: August 1, 2004
Citation: Barnes, C.W., Szabo, L.J., Nguyen, K.P. 2004. Detection of four rust pathogens of cereals and grasses using real-time PCR. Phytopathology. 94:S6. Technical Abstract: Puccinia rust species are widespread pathogens of cereals and grasses that annually cause significant crop losses worldwide. Duplex real-time PCR assays were developed for rapid detection of four rust pathogens, P. graminis, P. recondita, P. striiformis, and P. triticina, using TaqMan chemistry. Nuclear rDNA provided a variable region (ITS1) to distinguish between species, and a conserved region (28S) as an internal control. Specific ITS1 primer/probe sets exhibited high specificity and sensitivity with detection limits below 1 pg of rust DNA. Specificity of the assays was tested across multiple isolates representing a range of races and forma specialis within a species. Additionally, assay specificity was evaluated by testing a range of other grass rusts, as well as other fungi. The 28s primer/probe combination was successful in detecting all rust species tested within the duplex assays, validating the integrity of each assay. Rust species were identified within 30 min form the beginning of the assay, demonstrating the advantages of real-time PCR for detection of cereal rust pathogens.