Full TGIF Record # 233619
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Web URL(s):https://web.archive.org/web/20160212131534/http://www.turfgrasssociety.eu/home/articles/code/433?headline=Molecular%20Identification%2C%20Diagnosis%2C%20and%20Infectivity%20of%20Puccinia%20Species%20Causing%20Rust%20Diseases%20of%20Turfgrass
    Last checked: 04/15/2016
Publication Type:
Author(s):Beirn, L. A.; Clarke, B. B.; Meyer, W. A.; Crouch, J. A.
Title:Molecular identification, diagnosis, and infectivity of Puccinia species causing rust diseases of turfgrass
Meeting Info.:Kristiansand, Norway: June 24-26, 2012
Source:3rd European Turfgrass Society Conference Proceedings. Vol. 3, 2012, p. Unknown.
# of Pages:0
Publishing Information:Angers, France: European Turfgrass Society
Keywords:TIC Keywords: Breeding program; Cultivar improvement; Disease evaluation; Disease resistance; Inoculation; Poa pratensis; Puccinia coronata var. coronata; Puccinia graminis subsp. graminis; Puccinia striiformis
Abstract/Contents:"Rust (Puccinia spp.) is a common fungal disease of cool-season turfgrasses that can result in severe thinning, reduced seed yield, and diminished aesthetic quality. Turfgrass breeders have been developing rust resistant varieties since the 1950s; however, breeding programs are still lacking the necessary tools to effectively evaluate the performance of newly developed cultivars and selections against rust disease. Currently, turfgrass rust species are identified by ambiguous and sometimes misleading morphological characteristics, often resulting in the misidentification of the rust species and inaccurate assessment of cultivar resistance. Furthermore, there remains no reliable method for evaluating cultivar performance in the greenhouse, forcing turfgrass breeders to rely on natural rust infections in the field to evaluate new varieties and selections, a situation that can result in wasted time and resources if conditions are not conducive to disease development. In this research, our objective was to develop more efficient and effective tools to screen and evaluate rust disease resistance in turfgrass. Specifically, we aimed to (1) design an accurate and reliable, sequence-based diagnostic protocol and (2) develop a reproducible inoculation method for three turfgrass rust species (P. coronata, P. graminis, and P. striiformis) commonly found in temperate regions. The internal transcribed spacer (ITS) region was amplified and sequenced from 66 turfgrass samples exhibiting rust disease collected from across the United States and four countries (Australia, Canada, Chile, and the United Kingdom). Phylogenetic analysis revealed sufficient nucleotide variation within the ITS region, thus this region was used to develop primers and probes for real-time PCR. Fluorescence labeled hydrolysis probes and primer pairs were designed from the ITS-1 region for detection of P. graminis and P. striiformis, while the ITS-2 region was used for the P. coronata assay. The P. graminis and P. striiformis probes were designed within the same ITS-1 region and relied upon the same primer set, thus allowing for multiplexing of the two probes for simultaneous detection of these pathogens. The resulting assays were 97% accurate for the identification of P. graminis, P. coronata, and P. striiformis directly from diseased turfgrass tissue and could detect the presence of multiple species in mixed infections. Species-specific identifications were made using as few as 50-150 urediniospores (1-9 pg DNA), and were completed from DNA samples in approximately 1 hour. A reproducible inoculation protocol was developed using a modified inoculation procedure adapted from the cereal rust system. Viable urediniospores were sprayed onto susceptible plants in a mineral oil suspension and placed in a dew chamber over night. Uredinia of P. coronata and P. graminis were produced on Kentucky bluegrass (Poa pratensis) approximately 10-14 days following inoculation. However, even after repeat attempts, uredinia of P. striiformis could not be induced. Infection and species identification was confirmed by morphological and real-time PCR methods. The techniques reported here represent significant improvements for turfgrass breeding programs focusing on the development of germplasm with enhanced resistance to rust disease. Turfgrass breeders can now rapidly screen new germplasm for susceptibility/resistance to particular rust species prior to planting in the field, and confirm pathogen identity with confidence, thus greatly expediting the breeding process. Moreover, this diagnostic and infectivity research will serve as a foundation for developing additional inoculation protocols for other turfgrass rust pathogens, as well as aiding in the design and implementation of a series of race-differentials for turfgrass rust fungi that can be used to identify resistant genes and new fungal races."
Note:Summary appears as abstract
ASA/CSSA/SSSA Citation (Crop Science-Like - may be incomplete):
Beirn, L. A., B. B. Clarke, W. A. Meyer, and J. A. Crouch. 2012. Molecular identification, diagnosis, and infectivity of Puccinia species causing rust diseases of turfgrass. Eur. Turfgrass Soc. Conf. Proc. 3:p. Unknown.
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    Last checked: 04/15/2016
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