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Amplified fragment length polymorphisms (AFLPs) were used to estimate phylogenetic relationships within Magnaporthe grisea and determine the genetic structure of M. grisea populations associated with tall fescue and St. Augustinegrass in Georgia. Sixteen clonal lineages were identified in a sample population of 948 isolates. Five lineages were isolated from tall fescue (E, G1, G2, G4, and H), with lineage G4 comprising 90% of the population. Isolates from tall fescue were closely related to those from perennial ryegrass, weeping lovegrass, and wheat. Two M. grisea lineages were isolated from St. Augustinegrass (C and K), with lineage C comprising 99.8% of the population. Populations from crabgrass were dominated (98%) by lineage K, but also contained a single lineage C isolate. Haplotype diversity indices ranged from 0.00 to 0.29 in tall fescue populations and from 0.00 to 0.04 in St. Augustinegrass populations. Selection due to host species was the primary factor determining population structure according to analysis of molecular variance; host cultivar and geographical region had no significant effect. The host range of M. grisea lineages from turfgrasses was determined in growth chamber experiments and supports the prominent role of host species in determining the genetic structure of M. grisea populations from turfgrasses in Georgia.

Irrigation and an in vitro agitation assay were used to determine the percentage of the epiphytic yeast community (Cryptococcus, Pseudozyma, Rhodotorula, and Sporobolomyces) adhering to the phylloplane of creeping bentgrass (Agrostis palustris (Huds.) Pers.). Colony-forming units (cfu) of total epiphytic yeast populations (adherent and nonadherent cells) and of adherent populations (cells not removed by agitation) were determined by leaf washing and dilution plating. In an in vitro assay, 40.0% and 57.1% of the yeast adhered to the leaves, whereas, in initial field trials the percentage of adherent yeasts ranged from 40.0% to 71.9% of the total population. Adherent yeast cfu on leaves in the morning were significantly lower on bentgrass (8.0 × 10 3 to 3.1 × 10 4 cfu·cm -2 ) compared with total yeast cfu (1.4 × 10 4 to 4.7 × 10 4 cfu·cm -2 ) on the nonirrigated control. No differences in yeast populations were observed between irrigated and nonirrigated plots 2 h after the 0900 treatments. Yeast populations followed a diurnal pattern, with larger cfu recovered from bentgrass leaves in the morning and significantly lower populations recovered in the afternoon. At 1400 the adherent yeast were 83.1%-100% of the total yeast population recovered from the leaves. The relative adhesiveness of the epiphytic yeast community on bentgrass leaves is dynamic with nonadherent cells making up a larger percentage of the population in the mornings than the afternoons.Key words: adherence, Cryptococcus, leaf surface, Rhodotorula, turfgrass.

The efficacy of nine granular formulations of a bioherbicide containing the fungus Sclerotinia sclerotiorum (Lib.) de Bary was determined by assessing vegetative growth from tap roots of treated plants after incubation in a controlled environment. Plants treated with formulations 0788-03 or 0788-04 did not produce new petioles and leaves from tap roots after 96 h at 23 C and 100% relative humidity.

The effects of fungicides on population size and the development of fungicide resistance in the phylloplane yeast flora of bentgrass was investigated. In the spring of 2001, azoxystrobin, chlorothalonil, flutolanil, and propiconazole were applied separately over a 6-week period to creeping bentgrass (Agrostis palustris Huds.). Total and fungicide-resistant yeast populations were assessed by dilution plating onto either potato dextrose agar or potato dextrose agar amended with the test fungicides. Total yeast populations in the fungicide- treated plots were significantly lower than the check plots on three out of four sample dates. In the fall, azoxystrobin or propiconazole were applied twice to the bentgrass over 3 weeks. Significantly larger total yeast populations were observed compared with resistant or highly resistant populations for each treatment on every sample date. Total yeast populations were significantly higher in the check plots compared with either the propiconazole- or azoxystrobin-treated plots on the first three of five sample dates. A collection of yeasts (N = 114) with no prior exposure to fungicides were more sensitive to chlorothalonil, propiconazole, flutolanil, and iprodione than a second group (N = 115) isolated from fungicide-treated turfgrass. These results suggest that fungicide resistance among phylloplane yeasts is widespread and could be an important factor in the development of biological control agents for turfgrass diseases.Key words: yeast, biological control, fungicide, resistance, phylloplane.

The ability of yeasts to attach to hyphae or conidia of phytopathogenic fungi has been speculated to contribute to biocontrol activity on plant surfaces. Attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa was determined using in vitro attachment assays. Yeasts were incubated for 2 d on potato dextrose agar (PDA) prior to experimentation. A total of 292 yeasts cultured on PDA were screened for their ability to attach to conidia of B. cinerea; 260 isolates (89.1%) attached to conidia forming large aggregates of cells, and 22 isolates (7.5%) weakly attached to conidia with 1 or 2 yeast cells attached to a few conidia. Ten yeasts (3.4%), including 8 isolates of Cryptococcus laurentii, 1 isolate of Cryptococcus flavescens, and an unidentified species of Cryptococcus, failed to attach to conidia. All non-attaching yeasts produced copious extracellular polysaccharide (EPS) on PDA. Seventeen yeast isolates did not attach to hyphal fragments of B. cinerea, R. solani, and S. homoeocarpa after a 1 h incubation, but attachment was observed after 24 h. Culture medium, but not culture age, significantly affected the attachment of yeast cells to conidia of B. cinerea. The 10 yeast isolates that did not attach to conidia when grown on agar did attach to conidia (20%-57% of conidia with attached yeast cells) when cultured in liquid medium. Attachment of the biocontrol yeast Rhodotorula glutinis PM4 to conidia of B. cinerea was significantly greater at 1 × 10 7 yeast cells·mL -1 than at lower concentrations of yeast cells. The ability of yeast cells to attach to fungal conidia or hyphae appears to be a common phenotype among phylloplane yeasts.Key words: adhesion, biological control, Cryptococcus laurentii, Rhodotorula glutinis.

Populations of Magnaporthe grisea associated with tall fescue and St. Augustinegrass in Georgia were analyzed for mating type distribution and fertility status in 1999 and 2000. A polymerase chain reaction based assay for mating type was developed to facilitate population analysis. M. grisea populations from St. Augustinegrass in Georgia were dominated by the Mat1-1 mating type, whereas populations from tall fescue were dominated by Mat1-2. The opposite mating type was found in low frequency (0 to 5.7%) associated with each host. The fertility status of isolates from two populations was determined using controlled crosses in vitro. Seventy-eight Mat1-1 isolates from St. Augustinegrass were sterile in test crosses, but a single Mat1-2 isolate from St. Augustinegrass was male fertile. Of 87 Mat1-2 isolates from tall fescue, 47 were male fertile in test crosses, but 19 produced perithecia that were barren. All Mat1-1 isolates from tall fescue were sterile. Although both mating types exist in M. grisea populations from turfgrasses in Georgia, no female fertile isolates were identified in sample populations. The predominance of one mating type in eight sample populations and absence of female fertile isolates in two sample populations indicates that sexual reproduction may not occur with significant frequency in M. grisea populations associated with turfgrasses in Georgia.

The components of resistance in tall fescue to Magnaporthe grisea, the causal agent of gray leaf spot, were measured in growth chamber experiments. Cultivars ranging in susceptibility to M. grisea were selected: 'Kentucky 31' (susceptible), 'Rebel III' (moderately susceptible), 'Coronado' (resistant), and 'Coyote' (resistant). Plants were inoculated with nine M. grisea isolates representing five clonal lineages associated with tall fescue in Georgia. Compared to Kentucky 31, Coronado and Coyote exhibited longer incubation and latent periods, reduced rates of disease progress and lesion expansion, and lower final disease incidence, final foliar blight incidence, final mean lesion length, area under the lesion expansion curve, and area under the disease progress curve. No evidence of hypersensitive response was observed, all M. grisea isolates completed the disease cycle by producing secondary inoculum, and no differential response to isolates from different clonal lineages was detected in Coronado and Coyote. These results indicate that Coronado and Coyote have partial resistance to M. grisea. Measurement of resistance components using primary parameters and derived parameters yielded similar results. Foliar blight incidence data exhibited increased variation relative to other parameters and was less powerful for detection of M. grisea resistance. Measurements of incubation period, latent period, final disease incidence, and final mean lesion length were the most effective and efficient methods for detecting M. grisea resistance in tall fescue.

An unusual and undescribed foliar blight of tall fescue was observed in a home lawn and in turf grass research plots near Griffin, GA in May and June, 2000 and 2001. Isolation from lesions yielded mycelium of a basidiomycete with hyphal characteristics (binucleate cells, absence of clamp connections) associated with Laetisaria and Limonomyces spp. Isolates from blighted tall fescue and an isolate of Limonomyces roseipellis formed a clade distinct from isolates of Laetisaria fuciformis based on ribosomal DNA sequences. These data, in conjunction with cultural morphology, indicate that the basidiomycete from tall fescue represents a biotype of Limonomyces roseipellis that lacks clamp connections. In a controlled environment, isolates of the biotype induced foliar blight in the fescue cvs. Kentucky 31 and Rebel III. Histological observations revealed that the fungus colonized leaf surfaces as branched hyphae and aggregated hyphal strands. Penetration occurred via stomatal pores on the abaxial leaf surface. Colonization of leaf tissues was inter- and intracellular, with no evidence of papilla formation in response to invading hyphae. The name “cream leaf blight” is proposed for this new disease of tall fescue.

Hyphal anastomosis was studied among 85 isolates of binucleate Rhizoctonia solani-like fungi. On the basis of pairings, seven anastomosis groups (CAG) were discovered and designated CAG 1 through CAG 7. Seventy-one isolates were assigned to the seven CAG. No hyphal fusion occurred in pairings between 54 isolates, randomly selected from the seven CAG and isolates of R. solani AG 1, 2, 3, or 4. Within each CAG there was little homogeneity with respect to host, plant part invaded, and geographic origin, with the exception of CAG 1 isolates, which were found associated only with members of the Gramineae. CAG 2 included isolates of R. ramicola and R. fragariae. Four isolates in this group were induced to sporulate and were identified as Ceratobasidium cornigerum. CAG 6 included an isolate of R. muneratii. The absence of dark mycelial and sclerotial pigmentation among the majority of isolates in CAG 1, 2, and 7 was the significant cultural feature distinguishing isolates in these groups from isolates in other anastomosis groups and from isolates of R. solani. Isolates in CAG 3, 4, and 5 formed dark mycelial and sclerotial pigments similar to those produced by isolates of R. solani. The biologic and taxonomic implications of the anastomosis groups are discussed.

Sixty isolates of the plant pathogenic fungus Sclerotinia sclerotiorum (Lib.) de Bary and six isolates of Sclerotinia minor Jagger were evaluated in a controlled environment for virulence on leaves excised from 8-week-old dandelion plants. Significant negative correlations were obtained between the relative virulence of the isolates and the dry weights of dandelion plants inoculated in a controlled environment; and positive correlations were detected between the relative virulence of isolates and reduction in number of dandelion plants in turfgrass swards infested with inoculum of the isolates. In August 1988, an 80.7% reduction in the number of dandelion plants was detected in a turfgrass sward treated in 1987 with four applications of heat-killed seed of perennial ryegrass (100 g m−2 application–1) infested with isolate R30 of S. sclerotiorum, followed by six applications at the same rate in 1988. Populations of dandelions in untreated swards increased by 22.2% during the same period. Heat-killed seed of perennial ryegrass (100 g m−2) infested with isolate R30 applied simultaneously with dandelion seed (25 g m−2) onto a sward of Kentucky bluegrass reduced the establishment of dandelion seedlings by 85.5%. Necrosis or discoloration did not develop on Kentucky bluegrass, creeping bentgrass, annual bluegrass, or quackgrass treated with inoculum of Sclerotinia. Sclerotia of Sclerotinia spp. were found, on occasion, in crowns but not on the foliage of diseased dandelions.