Search National Agricultural Library (NAL) Digital Collections
Showing item 0 of
from your search.
Detection and Identification of Four Common Rust Pathogens of Cereals and Grasses Using Real-Time Polymerase Chain Reaction
- Puccinia spp. are widespread pathogens of cereals and grasses that annually cause significant yield losses worldwide, especially in barley, oat, and wheat. Urediniospore morphology and early symptom development have limited usefulness for distinguishing Puccinia spp. Therefore, we developed real-time polymerase chain reaction assays for rapid detection of the four rust pathogen species, Puccinia graminis (Pers.:Pers.), P. striiformis (Westend.), P. triticina (Eriks.), and P. recondita (Roberge ex Desmaz.). Duplex assays were constructed for the nuclear rDNA gene, using the variable internal transcribed spacer 1 (ITS1) region to distinguish between species, and the conserved 28S region as an internal control. Species-specific ITS1 primer/probe sets were highly specific and could detect <1 pg of DNA. The species-specific primer/probe sets showed positive results over a linear range of DNA five orders of magnitude or greater. Specificity of the assays was tested using multiple collections representing a range of races and formae speciales within a species. Additionally, assay specificity was evaluated by testing a range of other grass rust pathogens, as well as other fungi. The 28S primer/probe combination was successful in detecting all Puccinia spp. tested within the duplex assays, validating the integrity of each assay. Finally, the assays were used to identify unknown rust fungi infecting pasture grasses.
Barnes, C.W. , Szabo, L.J.
Poaceae , rust diseases , Hordeum vulgare , barley , Avena sativa , oats , Triticum aestivum , wheat , grasses , grain crops , Puccinia graminis , Puccinia striiformis , Puccinia recondita , plant pathogenic fungi , pathogenicity , disease detection , DNA profiling , polymerase chain reaction , ribosomal DNA , internal transcribed spacers , genetic markers
- Includes references
- Phytopathology 2007 June, v. 97, no. 6
Journal Articles, USDA Authors, Peer-Reviewed
- Works produced by employees of the U.S. Government as part of their official duties are not copyrighted within the U.S. The content of this document is not copyrighted.