Ralstonia solanacearum (Smith 1896) Yabuuchi et al. 1996 is ranked second among the top 10 most economically important plant pathogenic bacteria. The soil-borne bacterium affects over 200 plant species worldwide, including economically and nutritionally important crops, such as potato (Solanum tuberosum), tomato (Solanum lycopersicum), and bananas (Musa spp.). R. solanacearum is a species complex, meaning that the species is composed of strains with differential characteristics, including different metabolic requirements, centers of origin, host range, and ideal environmental conditions for infection. Its nature and the fact that it is a species complex can make R. solanacearum a difficult bacterium to work with, especially when lacking experience. Inappropriate isolation or storage of the pathogen can lead to inaccurate diagnostics or misleading conclusions. Thus, the objectives of this diagnostic guide are to provide adequate methods for isolation, storage, and identification and to discuss other relevant aspects related to this important plant pathogenic bacterium.
Take-all root rot is a disease of ultradwarf bermudagrass putting greens caused by Gaeumannomyces graminis (Gg), Gaeumannomyces sp. (Gx), Gaeumannomyces graminicola (Ggram), Candidacolonium cynodontis (Cc), and Magnaporthiopsis cynodontis (Mc). Many etiological and epidemiological components of this disease remain unknown. Improving pathogen identification and our understanding of the aggressiveness of these pathogens along with growth at different temperatures will advance our knowledge of disease development to optimize management strategies. Take-all root rot pathogens were isolated from symptomatic bermudagrass root and stolon pieces from 16 different golf courses. Isolates of Gg, Gx, Ggram, Cc, and Mc were used to inoculate ‘Champion’ bermudagrass in an in planta aggressiveness assay. Each pathogen was also evaluated at 10, 15, 20, 25, 30, and 35C to determine growth temperature optima. Infected plant tissue was used to develop a real-time PCR high resolution melt assay for pathogen detection. This assay was able to differentiate each pathogen directly from infected plant tissue using a single primer pair. In general, Ggram, Gg, and Gx were the most aggressive while Cc and Mc exhibited moderate aggressiveness. Pathogens were more aggressive when incubated at 30C compared to 20C. While they grew optimally between 24.4 and 27.8C, pathogens exhibited limited growth at 35C and no growth at 10C. These data provide important information on this disease and its causal agents that may improve take-all root rot management.
Spring dead spot (SDS) (Ophiosphaerella spp.) is a soilborne disease of warm-season turfgrasses grown where winter dormancy occurs. The edaphic factors that influence where SDS epidemics occur are not well defined. A study was conducted spring of 2020 and repeated spring of 2021 on four ‘TifSport’ hybrid bermudagrass (Cynodon dactylon (L.) Pers. x transvaalensis Burtt Davy) golf course fairways expressing SDS symptoms in Cape Charles, VA, USA. Spring dead spot within each fairway was mapped from aerial imagery collected spring of 2019 with a 20 MP CMOS 4k true color sensor mounted on a DJI Phantom 4 Pro drone. Three disease intensity zones were designated from the maps (low, moderate, high) based on the density of SDS patches in an area. Disease incidence and severity, soil samples, surface firmness, thatch depth, and organic matter measurements were taken from ten plots within each disease intensity zone from each of the four fairways (n=120). Multivariate pairwise correlation analyses (P < 0.1) and best subset stepwise regression analyses were conducted to determine which edaphic factors most influenced the SDS epidemic within each fairway and each year. Edaphic factors that correlated with an increase in SDS or were selected for the best fitting model varied across holes and years. However, in certain cases, soil pH and thatch depth were predictors for an increase in SDS. No factors were consistently associated with SDS occurrence, but results from this foundational study of SDS epidemics can guide future research on correlating factors that may drive disease development.
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