Data for designing two isothermal amplification assays for the detection of root-infecting fungi on cool-season turfgrasses.

Loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) are two rapid isothermal amplification methods for detecting three common fungal root pathogens of cool-season turfgrass: Gaeumannomyces avenae, Magnaporthiopsis poae and Ophiosphaerella korrae, "Detection of root-infecting fungi on cool-season turfgrasses using loop-mediated isothermal amplification and recombinase polymerase amplification" (Karakkat et al., 2018) [1]. The data provided here describe the information for designing primers and probes for LAMP and RPA, how specific they are for each of the three fungi, and the evaluation of RPA on field samples.

Detection of root-infecting fungi on cool-season turfgrasses using loop-mediated isothermal amplification and recombinase polymerase amplification.

Root-infecting fungal pathogens such as Gaeumannomyces avenae, Ophiosphaerella korrae, and Magnaporthiopsis poae cause extensive damage to amenity turfgrasses in temperate climates. The diseases they cause are difficult to diagnose by visual symptoms or microscopic inspection, and traditional polymerase chain reaction-based assays require large financial investments in equipment such as thermal cyclers and highly trained staff. The primary objective of this research was to develop fast and accurate detection assays for the three pathogens listed above that did not require the use of thermal cycling equipment. Loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) assays were developed for each pathogen based on known fungal cultures. The assays were tested on 27 samples received at the University of Wisconsin's Turfgrass Diagnostic Laboratory in 2016 and 2017 and both methods provided accurate diagnoses within about 30 min with minimal sample preparation. However, the RPA assays had lower levels of false positive contamination relative to the LAMP assays and are more likely to be effective in a field or diagnostic laboratory for improved turf root-pathogen detection.

Incidence and Distribution of Puccinia coronata and P. graminis on Turfgrass in the Midwestern United States.

Crown rust (caused by Puccinia coronata) and stem rust (caused by P. graminis) are two common and destructive diseases of turfgrass in the United States. Crown rust has been associated with perennial ryegrass and stem rust with Kentucky bluegrass when identified based solely on fungal morphology. However, recent studies using molecular identification methods have indicated the host-pathogen relationship of rusts on turf to be more complex. Our primary objective was to quickly and accurately identify P. coronata and P. graminis in symptomatic turfgrass leaves over 3 years on turfgrass samples from across the Midwestern United States. Between 2013 and 2015, 413 samples of symptomatic cool-season turfgrass from Wisconsin and surrounding states were screened using real-time polymerase chain reaction. Of these samples, 396 were Kentucky bluegrass and 17% of them contained P. coronata, 69% contained P. graminis, and 13% contained both P. coronata and P. graminis. In addition, both year and location effects were observed on the distribution of Puccinia spp. collected annually from two locations in southern Wisconsin. This research supports previous conclusions that have identified variability among P. graminis and P. coronata host relationships on turfgrass, and further demonstrates that rust fungal populations on Kentucky bluegrass may not be consistent between locations in the same year or over multiple years at the same location. The increasing evidence of variation in the turfgrass rust populations will likely affect future rust management and turfgrass breeding efforts.

Two members of the Ustilago maydis velvet family influence teliospore development and virulence on maize seedlings.

Members of the fungal-specific velvet protein family regulate sexual and asexual spore production in the Ascomycota. We predicted, therefore, that velvet homologs in the basidiomycetous plant pathogen Ustilago maydis would regulate sexual spore development, which is also associated with plant disease progression in this fungus. To test this hypothesis, we studied the function of three U. maydis velvet genes, umv1, umv2 and umv3. Using a gene replacement strategy, deletion mutants were made in all three genes in compatible haploid strains, and additionally for umv1 and umv2 in the solopathogenic strain, SG200. None of the mutants showed novel morphological phenotypes during yeast-like, in vitro growth. However, the Δumv1 mutants failed to induce galls or teliospores in maize. Chlorazol black E staining of leaves infected with Δumv1 dikaryons revealed that the Δumv1 hyphae did not proliferate normally and were blocked developmentally before teliospore formation. The Δumv2 mutants were able to induce galls and teliospores in maize, but were slow to do so and thus reduced in virulence. The Δumv3 mutants were not affected in teliospore formation or disease progression. Complementation of the Δumv1 and Δumv2 mutations in the SG200 background produced disease indices similar to those of SG200. These results indicate that two U. maydis velvet family members, umv1 and umv2, are important for normal teliospore development and disease progression in maize seedlings.