RESUMEN
Gray leaf spot caused by Magnaporthe oryzae is a serious disease of perennial ryegrass (Lolium perenne) turf in golf course fairways in the United States and Japan. Genetic relationships among M. oryzae isolates from perennial ryegrass (prg) isolates within and between the two countries were examined using the repetitive DNA elements MGR586, Pot2, and MAGGY as DNA fingerprinting probes. In all, 82 isolates of M. oryzae, including 57 prg isolates from the United States collected from 1995 to 2001, 1 annual ryegrass (Lolium multiflorum) isolate from the United States collected in 1972, and 24 prg isolates from Japan collected from 1996 to 1999 were analyzed in this study. Hybridization with the MGR586 probe resulted in approximately 30 DNA fragments in 75 isolates (designated major MGR586 group) and less than 15 fragments in the remaining 7 isolates (designated minor MGR586 group). Both groups were represented among the 24 isolates from Japan. All isolates from the United States, with the exception of one isolate from Maryland, belonged to the major MGR586 group. Some isolates from Japan exhibited MGR586 fingerprints that were identical to several isolates collected in Pennsylvania. Similarly, fingerprinting analysis with the Pot2 probe also indicated the presence of two distinct groups: isolates in the major MGR586 group showed fingerprinting profiles comprising 20 to 25 bands, whereas the isolates in the minor MGR586 group had less than 10 fragments. When MAGGY was used as a probe, two distinct fingerprint types, one exhibiting more than 30 hybridizing bands (type I) and the other with only 2 to 4 bands (type II), were identified. Although isolates of both types were present in the major MGR586 group, only the type II isolates were identified in the minor MGR586 group. The parsimony tree obtained from combined MGR586 and Pot2 data showed that 71 of the 82 isolates belonged to a single lineage, 5 isolates formed four different lineages, and the remaining 6 (from Japan) formed a separate lineage. This study indicates that the predominant groups of M. oryzae associated with the recent outbreaks of gray leaf spot in Japan and the United States belong to the same genetic lineage.
RESUMEN
Sixty-seven isolates of Sclerotinia homoeocarpa, causing dollar spot disease in creeping bentgrass, annual bluegrass, Bermudagrass, and perennial ryegrass turf, collected from 23 golf courses in various geographical regions of the United States and Canada between 1972 and 2001, were characterized by vegetative compatibility, genetic diversity, and pathogenicity. Eleven vegetative compatibility groups (VCGs A to K) were identified among the isolates tested in this study, and five of them (VCGs G to K) were new. VCG B was the most predominant group, typifying 33 isolates (51%) tested. S. homoeocarpa isolates collected from golf courses in Pennsylvania belonged to seven VCGs (A, B, E, F, G, I, and K), whereas three groups were observed in those collected from New York (B, E, and G) and New Jersey (E, H, and I). Two isolates, one each from Pennsylvania and Canada, were incompatible when paired with the tester isolates in all possible combinations, and did not fall into any known VCG. An isolate collected from Canada was compatible with tester isolates from two VCGs (C and D). Genetic analyses using amplified fragment length polymorphism (AFLP) showed the presence of two genetically distinct groups, designated as major group and the minor group. The major group included 36 isolates collected from various golf courses in the United States and Canada. Two isolates collected from bermudagrass in Florida formed a separate cluster, the minor group. Isolates that belonged to the major group were further divided into two subgroups (1 and 2). Subgroup 1 consisted of all the isolates that belonged to VCGs A, E, G, H, and I. Three of the four isolates that belonged to VCG K also were clustered with isolates of subgroup 1. Subgroup 2 consisted of all the isolates from VCG B, and one each from VCGs F and K. Pathogenicity assays on Penncross creeping bentgrass showed significant differences (P = 0.05) in virulence among the isolates. Overall, a relationship between virulence and VCGs was observed, in which certain virulence groups corresponded to specific VCGs; however, such a relationship was not observed between virulence and AFLPs. Close similarity among isolates of S. homoeocarpa collected from different locations in the United States and Canada suggests that isolates of the same genotype could be involved in outbreaks of dollar spot epidemics at multiple locations.
RESUMEN
ABSTRACT Gray leaf spot is a serious disease of perennial ryegrass (Lolium perenne), causing severe epidemics in golf course fairways. The effects of temperature and leaf wetness duration on the development of gray leaf spot of perennial ryegrass turf were evaluated in controlled environment chambers. Six-week-old Legacy II ryegrass plants were inoculated with an aqueous conidial suspension of Pyricularia grisea (approximately 8 x 10(4) conidia per ml of water) and subjected to four different temperatures (20, 24, 28, and 32 degrees C) and 12 leaf wetness durations (3 to 36 h at 3-h intervals). Three days after inoculation, gray leaf spot developed on plants at all temperatures and leaf wetness durations. Disease incidence (percent leaf blades symptomatic) and severity (index 0 to 10; 0 = leaf blades asymptomatic, 10 = >90% leaf area necrotic) were assessed 7 days after inoculation. There were significant effects ( alpha = 0.0001) of temperature and leaf wetness duration on disease incidence and severity, and there were significant interactions ( alpha = 0.0001) between them. Among the four temperatures tested, 28 degrees C was most favorable to gray leaf spot development. Disease incidence and severity increased with increased leaf wetness duration at all temperatures. A shorter leaf wetness duration was required for disease development under warmer temperatures. Analysis of variance with orthogonal polynomial contrasts and regression analyses were used to determine the functional relationships among temperature and leaf wetness duration and gray leaf spot incidence and severity. Significant effects were included in a regression model that described the relationship. The polynomial model included linear, quadratic, and cubic terms for temperature and leaf wetness duration effects. The adjusted coefficients of determination for the fitted model for disease incidence and severity were 0.84 and 0.87, respectively. The predictive model may be used as part of an integrated gray leaf spot forecasting system for perennial ryegrass turf.
RESUMEN
An immuno-recognition assay using a monoclonal antibody was developed to detect Pyricularia grisea, the causal agent of gray leaf spot of perennial ryegrass (Lolium perenne). In vitro assays with isolates of P. grisea from perennial ryegrass, tall fescue (Festuca arundinacea), St. Augustinegrass (Stenotaphrum secundatum), crabgrass (Digitaria sanguinalis), finger millet (Eleusine coracana), wheat (Triticum aestivum), triticale (× Triticosecale rimpaui), and rice (Oryza sativa) showed positive reactions; however, the strength of the reactions differed among isolates. Reactions were more intense with isolates from perennial ryegrass, wheat, and triticale. All P. grisea isolates from perennial ryegrass collected from various regions of the United States showed positive reactions. P. grisea was detected at antigen dilution rates of 0.5×, 0.25×, 0.13×, 0.06×, and 0.03×. Dot-blot assays with Bipolaris sorokiniana, Colletotrichum graminicola, Curvularia lunata, Microdochium nivale, Pythium aphanidermatum, Rhizoctonia solani, or Sclerotinia homoeocarpa isolated from turfgrasses were negative. In vivo assays of symptomatic leaves of perennial ryegrass plants inoculated with P. grisea also showed positive reactions, and those inoculated with B. sorokiniana, P. aphanidermatum, R. solani, or S. homoeocarpa were negative. Intensity of reaction between the monoclonal antibody and P. grisea was proportional to disease severity in perennial ryegrass inoculated with P. grisea; however, there was no reaction in dot blots of leaf tissue collected during the latent period. P. grisea was detected in perennial ryegrass samples from golf course fairways affected by gray leaf spot in Connecticut, Massachusetts, Maine, New Jersey, Pennsylvania, and Rhode Island using this procedure. The monoclonal antibody recognition system is highly sensitive to P. grisea and can be used effectively for the rapid diagnosis of gray leaf spot of perennial ryegrass turf.
RESUMEN
Isolates of Magnaporthe grisea causing gray leaf spot of perennial ryegrass (PR) (Lolium perenne) and St. Augustinegrass (SA) (Stenotaphrum secundatum) were analyzed for mating compatibility and fertility. A total of 312 isolates of M. grisea from PR and 62 isolates from SA were paired with hermaphroditic tester strains from finger millet (Eleusine coracana), rice (Oryza sativa), and wheat (Triticum aestivum). All the PR isolates belonged to a single mating type, MAT1-2. Male fertility was observed in all these isolates. Asci and ascospores were not produced regardless of their developmental stage. Of the 139 (44.6%) isolates from PR that formed perithecia with the fertile tester strains, 83 (59.7%) were highly fertile, 33 (23.7%) were intermediately fertile, and 23 (16.5%) were low in fertility. Both mating types were observed among the isolates of SA, where MAT1-1 predominated the MAT1-2 type. An equal number of male and female fertile isolates were detected among these isolates obtained from a single location; however, none of the isolates behaved as hermaphrodites. Few ascospores were produced in crosses between two isolates of SA and a finger millet tester. Of the 62 monoconidial isolates of SA tested, 19 (30.6%) isolates formed perithecia, of which 5 (26.3%) were highly fertile, 7 (36.8%) were intermediately fertile, 7 (36.8%) were low in fertility, and 43 (69.4%) were infertile. The results of this study indicate that the sexual stage may not be a significant factor contributing to the genetic variation the gray leaf spot pathogen population.
RESUMEN
Gray leaf spot of perennial ryegrass (Lolium perenne L.) turf was first reported in the United States in 1991. The disease epidemic was primarily confined to golf course fairways in southeastern Pennsylvania (1). Subsequently, moderate to severe outbreaks of gray leaf spot occurred in perennial ryegrass fairways and roughs in numerous locations throughout the eastern and midwestern United States. In August 2001, a serious decline of perennial ryegrass turf was observed in a bermudagrass (Cynodon dactylon (L.) Pers) baseball field in Dodger Stadium in Los Angeles, CA, that had been overseeded with perennial ryegrass. The bermudagrass turf was not affected. The perennial ryegrass turf developed necrotic lesions that resulted in blighting of leaf blades. In laboratory assays, Pyricularia grisea (Cooke) Sacc., was consistently isolated from symptomatic ryegrass blades from turf samples collected from the site. Of the 12 P. grisea isolates collected from the assayed leaf blades, five isolates were selected for a pathogenicity assay. Twenty-five 'Legacy II' perennial ryegrass plants were grown from seeds in 4 × 4 in.-plastic pots, (10 × 10 cm) which were filled to 1 cm below the rim with granular calcine clay medium (Turface MVP, Allied Industrial Material Corp., Buffalo Grove, IL). Three weeks after seeding, plants were fertilized with a water-soluble 20-20-20 N-P-K fertilizer (1.3 g/liter of water) once per week. Treatments (isolates of P. grisea and a control) were arranged as a randomized complete block design with five replications. Five-week-old plants were sprayed with an aqueous suspension of P. grisea conidia (≈5 × 104 conidia per ml of sterilized distilled water with 0.1% Tween 20) using an atomizer until the leaves were completely wet. Plants sprayed with sterilized distilled water served as the control. After inoculation, individual pots were covered with clear polyethylene bags and placed in a controlled environment chamber maintained at 28°C and continuous fluorescent light (88 µE m-2 s-1). Four days after inoculation, necrotic lesions (<2 mm diameter) developed on ryegrass blades inoculated with each isolate of P. grisea. Lesions did not develop on leaves of control plants. Seven days after inoculation, the polyethylene bags were removed, and 50 symptomatic blades from each pot were collected, and disease incidence (percent infected leaves) and severity (index 0 to 10; 0 = none, 10 = >90% of the leaf blade necrotic ) were assessed. P. grisea was isolated from symptomatic leaves of plants inoculated with the fungus. Disease incidence and severity on inoculated plants were 92 to 96% and 8.8 to 10, respectively. There were no significant differences in disease incidence and severity (P = 0.05) among the isolates of P. grisea included in the test. To our knowledge, this is the first report of gray leaf spot of perennial ryegrass turf in California. Reference: (1) P. J. Landschoot and B. F. Hoyland. Plant Dis. 76:1280, 1992.
RESUMEN
Gray leaf spot is a serious disease of perennial ryegrass (Lolium perenne) turf in the United States. Isolates of Pyricularia grisea causing the disease in perennial ryegrass were characterized using molecular markers and pathogenicity assays on various gramineous hosts. Genetic relationships among perennial ryegrass isolates were determined using different types of trans-posons as probes. Phylogenetic analysis using Pot2 and MGR586 probes, analyzed with AMOVA (analysis of molecular variance), showed that these isolates from perennial ryegrass consist of three closely related lineages. All the isolates belonged to a single mating type, MAT1-2. Among 20 isolates from 16 host species other than perennial ryegrass, only the isolates from wheat (Triticum aestivum) and triticale (× Triticosecale), showed notable similarity to the perennial ryegrass isolates based on their Pot2 fingerprints. The copy number and fingerprints of Pot2 and MGR586 in isolates of P. grisea from perennial ryegrass indicate that they are genetically distinct from the isolates derived from rice (Oryza sativa) in the United States. The perennial ryegrass isolates also had the same sequence in the internal transcribed spacer (ITS) region of the genes encoding ribosomal RNA as that of the wheat and triticale isolates, and exhibited rice isolate sequence polymorphisms. In pathogenicity assays, all the isolates of P. grisea from Legacy II perennial ryegrass caused characteristic blast symptoms on Marilee soft white winter wheat, Bennett hard red winter wheat, Era soft white spring wheat, and Presto triticale, and they were highly virulent on these hosts. An isolate from wheat and one from triticale (from Brazil) were also highly virulent on perennial ryegrass and Rebel III tall fescue (Festuca arundinacea). None of the isolates from perennial ryegrass caused the disease on Lagrue rice, and vice versa. Understanding the population structure of P. grisea isolates infecting perennial ryegrass and their relatedness to isolates from other gramineous hosts may aid in identifying alternate hosts for this pathogen.
RESUMEN
Blast disease (gray leaf spot) of perennial ryegrass (Lolium perenne L.), caused by Pyricularia grisea (Cooke) Sacc., in golf course fairways in Pennsylvania, was first reported in 1991 (1). The disease was primarily confined to the southeastern region of the state. In August and September 1998, severe outbreaks of blast disease occurred in perennial ryegrass fairways in numerous golf courses in the same region in Pennsylvania. Additionally, in 1998, the disease was diagnosed in ryegrass fairways for the first time in the western region of the state, where serious disease outbreaks occurred. The disease develops on leaf blades as small, watersoaked lesions that become necrotic spots. The spots expand rapidly, and develop into gray, grayish-brown, or light brown, circular spots with purple to dark brown borders often surrounded by a yellow halo on the leaf blades. As the disease progresses, the circular necrotic spots coalesce, become irregular in shape, and cause partial (tip blight) or complete blighting of the leaf blades. The blighted blades may often appear twisted. Turfgrass loss due to the disease was over 90% in several golf courses. During the 1998 epidemics, a survey was conducted in the affected regions, which included three golf courses in western Pennsylvania and 15 golf courses in southeastern Pennsylvania. P. grisea was consistently isolated from the symptomatic ryegrass leaf blades of turf samples (12 to 28 samples per location) collected from the blighted fairways. Of the 122 isolates of P. grisea collected from the 18 golf courses, seven isolates (two from western Pennsylvania and five from southeastern Pennsylvania) were selected for pathogenicity assays. Five Pennfine perennial ryegrass plants were grown in each Cone-Tainer (4 mm in diameter) that was filled to 1 cm below the rim with granular calcine clay medium (Turface MVP; Allied Industrial Material, Buffalo Grove, IL). Three weeks after seeding, grasses were fertilized with water-soluble 20-20-20 fertilizer (1.3 g per liter of water) once a week to field capacity of the growing medium. Treatments (the isolates) were arranged as a randomized complete block with four replications (a Cone-Tainer per replication). Six-week-old ryegrass plants were atomized with an aqueous suspension of P. grisea conidia (approximately 8 × 104 conidia per ml of sterilized, distilled water) until the leaves were completely wet. Individual Cone-Tainers were placed in clear polyethylene bags, enclosed, and were placed in an incubator that was maintained at continuous 28°C and 12-h-day fluorescent light (72 µE s-1 m-2). Three days after inoculation, water-soaked lesions (<2 mm in diameter) developed on leaves of the ryegrass inoculated with each isolate of P. grisea. No lesions developed on leaves of the control plants. Seven days after inoculation, the polyethylene bags were removed, and disease incidence (percent infected leaves) was assessed. P. grisea was isolated from the necrotic lesions or the blighted leaf blades of every plant inoculated with the fungus. This is the first report of blast disease outbreak in golf course fairways in western Pennsylvania. The impact of blast disease epidemics on golf courses in the northeastern United States in 1998 was significant, and caused serious concern to turf managers. The survey indicates that blast disease of perennial ryegrass may be emerging as a new problem for the turfgrass industry in the northeastern United States. Reference: (1) P. J. Landschoot and B. F. Hoyland. Plant Dis. 76:1280, 1992.
RESUMEN
Production of perithecia, asci, and ascospores by Indian isolates of Magnaporthe grisea is rare and has not been found among the Southern Indian isolates of the blast pathogen. From among 138 monoconidial isolates that infect rice and other hosts, we now report the distribution of mating types (MAT1-1 and MAT1-2) of M. grisea in finger millet and paragrass (Brachiaria mutica)-infecting isolates. Twenty-eight of the 96 finger millet isolates, 5 of the 16 paragrass isolates, and none of the 26 rice isolates produced fertile perithecia in laboratory matings with fertile testers. Backcrosses of ascospore progenies to the parental M. grisea isolate but not to the tester strain resulted in fertile perithecial formation, and a further backcrossing scheme indicated definite fertility patterns of Mendelian inheritance in M. grisea.
