Identification of a New Waitea circinata Variety Causing Basal Leaf Blight of Seashore Paspalum.

Seashore paspalum (Paspalum vaginatum) is a saline-tolerant, warm-season turfgrass species popular for golf course use in tropical and subtropical climates. A new variety of Waitea circinata (proposed as W. circinata var. prodigus) is described as the causal agent of basal leaf blight, a novel disease of seashore paspalum. Foliar necrosis and canopy thinning of seashore paspalum were observed on three different golf course fairways in Florida over an 18-month period. Five isolates with profuse, pink to yellow mycelia and small, salmon-colored or yellow to light-brown sclerotia were cultured from diseased turf foliage. Isolates grew rapidly over a temperature range of 25 to 35°C and were initially identified as an uncharacterized variety of W. circinata. Internal transcribed spacer sequences of rDNA from the isolates were compared with sequences from previously described W. circinata varieties. The paspalum isolates formed a phylogenetic clade that was distinct from the other W. circinata varieties. Pathogenicity was confirmed on 'SeaDwarf' and 'SeaIsle Supreme' seashore paspalum, 'Penncross' creeping bentgrass (Agrostis stolonifera), 'Senesta' bermudagrass (Cynodon dactylon), and 'Dark Horse' roughstalk bluegrass (Poa trivialis). The geographical distribution and potential impact of basal leaf blight is unknown. However, the range of potential turfgrass hosts and environmental conditions conducive for disease development suggest that the pathogen may infect other species in addition to seashore paspalum.

First Report of Root Rot Caused by Binucleate Rhizoctonia Anastomosis Group F on Musa spp.

A research program was initiated at the University of Georgia in 2003 to identify banana cultivars suitable for production in the coastal and southern areas of the state. During a root disease survey conducted in October 2007 on bananas (Musa spp.) grown at the University of Georgia Bamboo Farm and Coastal Gardens in Savannah, GA, root lesions and root rot were observed on banana cvs. Gold Finger, Kandarian, and Manzano. Root lesions were dark brown to black and irregular in shape, with partial or entire roots affected. Lateral roots and outer layers of cord roots (roots arising from interior layers of the corm) of infected plants were blackened and rotted. Diseased root samples were collected from three plants of each cultivar, surface sterilized with 0.6% sodium hypochlorite, and placed on tannic acid benomyl agar (TABA). Pure cultures of the fungus consistently associated with diseased tissue were obtained by subculturing hyphal tips on TABA. Mycelia of the fungus on potato dextrose agar (PDA) were light to deep brown and the hyphae tended to branch at right angles. A septum was present in each hyphal branch near the point of origin and a slight constriction at the branch was observed. The hyphae of two isolates were stained with 0.6% phenosafranin and 3% KOH and binucleate hyphal cells were observed. On the basis of these morphological features, the isolates appeared to be binucleate Rhizoctonia anamorphs (teleomorph Ceratobasidium Rogers). For molecular identification, the internal transcribed spacer (ITS) regions and the 5.8S gene from rDNA of the isolates were cloned and sequenced (GenBank Accession No. HQ168370). The ITS regions (775 bp) were 100% identical between the two isolates and 99% identical to Ceratobasidium sp. AG-F strain SIR-1 isolated from sweet potato in Japan (GenBank Accession No. AF354085). The anastomosis group of the isolates was confirmed by pairing with strain SIR-1 on PDA. On the basis of morphological and molecular characteristics and the anastomosis assay, the two isolates were identified as a Ceratobasidium sp. AG-F (1-3). Pathogenicity assays were conducted by inoculating banana plants (cv. Golden pillow, synonym = Manzano) grown in pots under greenhouse conditions (25 to 27°C). Twenty wheat seeds infested with each isolate were placed uniformly around each plant at a depth of 10 cm in the soil. The plants were incubated in the greenhouse and the roots were examined 2 months after inoculation. Brown-to-black lesions and root rot, identical to symptoms associated with field banana roots, were observed on all inoculated plants but not on the noninoculated control plants. The fungus was reisolated from affected root samples and the identity was confirmed by morphological and molecular characteristics and the anastomosis assay. To our knowledge, this is the first report of banana root rot caused by binucleate Rhizoctonia anastomosis group F. With the increased interest in producing bananas for food and ornamental purposes, the occurrence of Ceratobasidium root rot on bananas needs to be considered when designing disease management programs and searching for suitable cultivars for banana production. References: (1) L. L. Burpee et al. Mycologia 70:1281, 1978. (2) D. González et al. Mycologia 93:1138, 2001. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN. 1991.

Genetic Structure of Magnaporthe grisea Populations Associated with St. Augustinegrass and Tall Fescue in Georgia.

ABSTRACT 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.

Components of Resistance to Magnaporthe grisea in 'Coyote' and 'Coronado' Tall Fescue.

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.

Pathogenicity of a Novel Biotype of Limonomyces roseipellis 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.

Mating Type Distribution and Fertility Status in Magnaporthe grisea Populations from Turfgrasses in Georgia.

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.

Components of Resistance to Rhizoctonia solani Associated with Two Tall Fescue Cultivars.

Components of resistance to Rhizoctonia solani in the tall fescue cultivars Kentucky 31 (moderately resistant) and Mojave (susceptible) were evaluated under controlled environmental conditions. Size and expansion rate of foliar lesions were recorded on 100 individual plants of each cultivar. Lesions on the first fully expanded leaves of 6- to 10-month-old inoculated plants covered a significantly greater proportion of the leaf width on cv. Mojave compared to Kentucky 31. Rate of lesion expansion was also greater on cv. Mojave than on Kentucky 31. Lesion size and rate of expansion were greater on the second compared to the first fully expanded leaf of both cultivars. Wider leaves and slower lesion expansion are two components responsible for the greater level of resistance to R. solani in cv. Kentucky 31 compared to Mojave tall fescue.

Use of multispectral radiometry for assessment of rhizoctonia blight in creeping bentgrass.

ABSTRACT The ability to identify diseases early and quantify severity accurately is crucial in plant disease assessment and management. This study was conducted to assess changes in the spectral reflectance of sunlight from plots of creeping bentgrass during infection by Rhizoctonia solani, the cause of Rhizoctonia blight, and to evaluate multispectral radiometry as a tool to quantify Rhizoctonia blight severity. After inoculation of 6-year-old creeping bentgrass turf with R. solani anastomosis group 2-2, reflectance of sunlight from the foliar canopy was measured at light wavelengths of 460 nm (blue) to 810 nm (near infrared [NIR]), at 50-nm intervals. Visual estimates of disease severity and percentage of canopy reflectance were made daily throughout each of three epidemics of Rhizoctonia blight from the onset of visible symptoms until maximum disease severity was reached. In each experiment, linear regression analysis revealed a significant reduction in the percentage of NIR (760 and 810 nm) reflectance as disease severity increased. However, in the majority of analyses, regression models explained <50% of the variability between components. Multispectrum radiometry appears to function best when used to assess differences in disease severity at discrete points in time rather than over an entire epidemic.

Control of Dollar Spot of Creeping Bentgrass Caused by an Isolate of Sclerotinia homoeocarpa Resistant to Benzimidazole and Demethylation-Inhibitor Fungicides.

The effects of fungicides were studied on two isolates of Sclerotinia homoeocarpa that differed in fungicide sensitivity. Concentrations of benzimidazole (benomyl and thiophanate-methyl), demethylation inhibitor (fenarimol, propiconazole, and triadimefon), and nitrile (chlorothalonil) fungicides required to inhibit mycelial growth in vitro by 50 and 90% (effective concentration; EC50,90) were significantly greater for isolate S088 than for isolate S084. No differences were observed in the EC50,90 values of a pyridylaniline (fluazinam) or dicarboximide (iprodione) fungicide. In field tests conducted on creeping bentgrass, S088 had a significantly shorter incubation period than S084 in plots treated with propiconazole applied at 0.2 or 0.8 kg a.i. ha-1 in 1994 or 0.4 kg a.i. ha-1 in 1995, thiophanate-methyl applied at 1.5 or 3.0 kg a.i. ha-1 in 1994 and 1995, or a tank-mix of propiconazole and iprodione applied at 1.6 + 0.8 kg a.i. ha-1 in 1994. No differences in incubation periods occurred in plots treated with chlorothalonil or iprodione in either year, or with fluazinam in 1995 and 1996. Fewer days were required to reach 5% disease severity in turf inoculated with isolate S088, compared to isolate S084, in plots treated with propiconazole, thiophanate-methyl, or certain tank-mixes containing propiconazole, in 1994 and 1995. Several treatments, including propiconazole, thiophanate-methyl, and fluazinam, resulted in suppression of disease caused by isolate S084 to less than 5% severity for more than 21 days, while only fluazinam applied at 1.6 or 3.2 kg a.i. ha-1 provided more than 21 days of disease suppression in turf inoculated with either of the isolates tested. Slope coefficients from linear regressions between incubation period or days to 5% disease and concentrations of fungicide applied in 1996 were significantly different for isolate S084 compared to isolate S088 in plots treated with propiconazole, but not in plots treated with fluazinam.

The influence of alachlor, trifluralin, and diazinon on the development of endogenous mycorrhizae in soybeans.

Preplant incorporated treatments of 2 and 4 kg/ha of trifluralin and diazinon had no significant effect on growth, P accumulation or root colonization by mycorrhizal fungi in soybeans planted in an Andover clay loam. At 4 kg/ha, alachlor and trifluralin inhibited root development of 25 day-old plants. The 4 kg/ha alachlor treatment reduced shoot weight of 25 day old plants significantly and suppressed mycorrhizal development of 25 to 60 day old plants. At currently used commercial rates neither alachlor, trifluralin, nor diazinon affected mycorrhizal development under the conditions of the experiment.

The influence of Pratylenchus penetrans on the incidence and severity of verticillium wilt of potato.

The influence of Pratylenchus penetrans on the incidence and severity of Verticillium wilt was examined in the potato cultivars 'Kennebec', 'Katahdin', and 'Abnaki'. Single-stem plants were grown in soil maintained at a temperature of 22 +/- 1 C. Axenically cultured nematodes were suspended in water and introduced to the soil, at a rate of ca 5,000/25.4-cm pot, through holes made around each stem. Ten days after infestation with nematodes, conidial suspensions of Verticillium albo-atrum were introduced into the soil at a rate of ca 1,000,000/pot. Among Katahdin plants, the severity of foliar symptoms was increased in the presence of both pathogens 2 and 3 weeks after soil intestation. During the remaining 5 weeks, severity of foliar symptoms was not different between plants infected by both pathogens and those infected by Verticillium alone. Within the wilt-susceptible cultivar Kennebec and the resistant eultivar Abnaki, no effects on foliar symptom severity were observed. When plant heights, shoot weights, and tuber yields were analyzed, a Pratylenchus-Verticillium interaction was not evident within any of the cultivars tested. Nematode populations in roots and rhizosphere were suppressed in Kennebec and Katahdin plants in the presence of Verticillium.