D. M. Benson

Etiology of Phytophthora drechsleri and P. nicotianae (=P. parasitica) diseases affecting floriculture crops

Phytophthora nicotianae and P. drechsleri isolates (n = 413) recovered from eight floricultural hosts at 11 different production sites were described according to compatibility type, resistance to mefenoxam, and amplified fragment length polymorphism (AFLP) profiles. Sample sizes ranged from 2 to 120. In all cases, isolates recovered from a single facility had the same compatibility type and resistance to mefenoxam. AFLP analysis indicated that six clonal lineages of P. nicotianae and two clonal lineages of P. drechsleri were responsible for the 11 epidemics and that isolates recovered from the same facility were identical. A single clone of P. nicotianae was recovered from snapdragons at two field production sites in the southeastern United States receiving seedlings from the same source. This clone persisted at one site from 2000 to 2001. Another clone was recovered from verbena at three separate greenhouse facilities where one facility was supplying verbena to the other two. These results suggest that asexual reproduction of these pathogens plays an important role in epidemics and spread may occur between distant facilities via movement of plants.

Temporal and Spatial Epidemiology of Phytophthora Root Rot in Fraser Fir Plantations

In 1999, 19 plots of Fraser fir (Abies fraseri) with a disease focus were established in commercial plantings grown for Christmas tree production in the mountains of five western North Caro-lina counties. Progress of Phytophthora root rot caused by Phytophthora cinnamomi as estimated by mortality was followed in each plot over 3 to 4 years in an attempt to understand dispersal of inoculum. Slope, aspect, and field production age at the time plots were established were recorded. Rainfall estimated from National Weather Service stations each growing season also was recorded. The relationship of site parameters and rainfall to dispersal and disease was investigated. Disease incidence and mortality were assessed in June and September each year for 3 or 4 years depending on plot. Phytophthora root rot as estimated by mortality counts over time in a logistic regression model progressed in only five of 19 plots over 3 years. None of the site parameters correlated with mortality data, although slightly more disease was found in plots with a north aspect. Rainfall was below normal in the 3 years of the study and did not correlate with mortality in any year. Lack of disease progress in the majority of plots was attributed to drought conditions in the region. In the five plots where mortality increased over time, spatial analysis suggested an aggregated pattern of diseased plants. Aggregation was apparent but not very strong among nearest neighbors, but was considerably stronger among groups of trees within a local area. This aggregation within groups was stronger when larger group sizes were examined by beta-binomial analysis. A spatial analysis by distance indices method (SADIE) indicated the presence of secondary clusters occurring several meters away from the main focus. A stochastic model also was employed that indicated a combination of spatial processes were likely involved, specifically a tendency toward spread within a local area, but not necessarily to the nearest neighboring trees, combined with an influence of background inoculum that could not be accounted for within local areas and may have come from external sources. Thus, all sources of inoculum including infected planting stock, inoculum in soil, infected trees, and contaminated equipment were equally important in epidemics of Phytophthora root rot in Fraser fir and dispersal of P. cinnamomi.

Overwintering of Sclerotium rolfsii and S. rolfsii var. delphinii in Different Latitudes of the United States

Previously known only from the southern United States, hosta petiole rot recently appeared in the northern United States. Sclerotium rolfsii var. delphinii is believed to be the predominant petiole rot pathogen in the northern United States, whereas S. rolfsii is most prevalent in the southern United States. In order to test the hypothesis that different tolerance to climate extremes affects the geographic distribution of these fungi, the survival of S. rolfsii and S. rolfsii var. delphinii in the northern and southeastern United States was investigated. At each of four locations, nylon screen bags containing sclerotia were placed on the surface of bare soil and at 20-cm depth. Sclerotia were recovered six times from November 2005 to July 2006 in North Dakota and Iowa, and from December 2005 to August 2006 in North Carolina and Georgia. Survival was estimated by quantifying percentage of sclerotium survival on carrot agar. Sclerotia of S. rolfsii var. delphinii survived until at least late July in all four states. In contrast, no S. rolfsii sclerotia survived until June in North Dakota or Iowa, whereas 18.5% survived until August in North Carolina and 10.3% survived in Georgia. The results suggest that inability to tolerate low temperature extremes limits the northern range of S. rolfsii.

Cellulase activity as a mechanism for suppression of phytophthora root rot in mulches.

Wood-based mulches are used in avocado production and are being tested on Fraser fir for reduction of Phytophthora root rot, caused by Phytophthora cinnamomi. Research with avocado has suggested a role of microbial cellulase enzymes in pathogen suppression through effects on the cellulosic cell walls of Phytophthora. This work was conducted to determine whether cellulase activity could account for disease suppression in mulch systems. A standard curve was developed to correlate cellulase activity in mulches with concentrations of a cellulase product. Based on this curve, cellulase activity in mulch samples was equivalent to a cellulase enzyme concentration of 25 U ml(-1) or greater of product. Sustained exposure of P. cinnamomi to cellulase at 10 to 50 U ml(-1) significantly reduced sporangia production, but biomass was only reduced with concentrations over 100 U ml(-1). In a lupine bioassay, cellulase was applied to infested soil at 100 or 1,000 U ml(-1) with three timings. Cellulase activity diminished by 47% between 1 and 15 days after application. Cellulase applied at 100 U ml(-1) 2 weeks before planting yielded activity of 20.08 μmol glucose equivalents per gram of soil water (GE g(-1) aq) at planting, a level equivalent to mulch samples. Cellulase activity at planting ranged from 3.35 to 48.67 μmol GE g(-1) aq, but no treatment significantly affected disease progress. Based on in vitro assays, cellulase activity in mulch was sufficient to impair sporangia production of P. cinnamomi, but not always sufficient to impact vegetative biomass.

Microbial Profiling of Cultural Systems for Suppression of Phytophthora Root Rot in Fraser Fir

Phytophthora root rot of Fraser fir, caused by several Phytophthora spp., is a severe problem in Christmas tree production. Since fungicides are not economically viable for disease management in field plantings and host resistance is not available, cultural control methods were investigated. Mulches, dairy compost, and soil pH adjustment were tested at five field sites in North Carolina. Treatments included wood chips, wood chips plus compost, or pine bark as raised beds, and compost or sulfur tilled into soil. Soil and mulch microbial populations were characterized by dilution plating and calculation of a log series diversity index, and by enzyme analyses at 5, 12, 17, and 24 months after planting. Bacterial and fungal counts, microbial activity, and cellulase activity were higher in mulch than in soil at all sites and times (P < 0.01), and generally did not differ among mulch types or among soils. Treatments significantly affected disease ratings and tree survival at three of five sites, with one or more mulch treatments yielding lower disease ratings and greater survival than controls. Tree mortality at each time point varied significantly with cellulase activity in the upper root zone (P = 0.005). Other biological variables did not show significant relationships with disease ratings or mortality.