scholarly journals A Temperature and Leaf Wetness Duration-Based Model for Prediction of Gray Leaf Spot of Perennial Ryegrass Turf

2003 ◽  
Vol 93 (3) ◽  
pp. 336-343 ◽  
Author(s):  
W. Uddin ◽  
K. Serlemitsos ◽  
G. Viji

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 × 104 conidia per ml of water) and subjected to four different temperatures (20, 24, 28, and 32°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 ( α = 0.0001) of temperature and leaf wetness duration on disease incidence and severity, and there were significant interactions ( α = 0.0001) between them. Among the four temperatures tested, 28°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.

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 689-689
Author(s):  
A. Nasehi ◽  
J. B. Kadir ◽  
M. Nasr Esfahani ◽  
F. Mahmodi ◽  
H. Ghadirian ◽  
...  

In 2011, a severe gray leaf spot was observed on eggplant (Solanum melongena) in major eggplant growing areas in Malaysia, including the Pahang, Johor, and Selangor states. Disease incidence was >70% in severely infected areas of about 150 ha of eggplant greenhouses and fields examined. Symptoms initially appeared as small (1 to 5 mm diameter), brownish-black specks with concentric circles on the lower leaves. The specks then coalesced and developed into greyish-brown, necrotic lesions, which also appeared on the upper leaves. Eventually, the leaves senesced and were shed. Tissue cut from the edges of leaf spots were surface-sterilized in 1% NaOCl for 2 min, rinsed in sterilized water, dried, and incubated on potato dextrose agar (PDA). Fungal colonies were greyish green to light brown, and produced a yellow pigment. Single, muriform, brown, oblong conidia formed at the terminal end of each conidiophore, were each 21.6 to 45.6 μm long and 11.5 to 21.6 μm wide, and contained 2 to 7 transverse and 1 to 4 longitudinal septa. The conidiophores were tan to light brown and ≤220 μm long. Based on these morphological criteria, 25 isolates of the fungus were identified as Stemphylium solani (1). To produce conidia in culture, 7-day-old single-conidial cultures were established on potato carrot agar (PCA) and V8 juice agar media under an 8-h/16-h light/dark photoperiod at 25°C (4). Further confirmation of the identification was obtained by molecular characterization in which fungal DNA was extracted and the internal transcribed spacer (ITS) region of ribosomal DNA amplified using primers ITS5 and ITS4 (2), followed by direct sequencing. A BLAST search in the NCBI database revealed that the sequence was 99% identical with published ITS sequences for two isolates of S. solani (Accession Nos. AF203451 and HQ840713). The amplified ITS region was deposited in GenBank (JQ736023). Pathogenicity testing of a representative isolate was performed on detached, 45-day-old eggplant leaves of the cv. 125066-X under laboratory conditions. Four fully expanded leaves (one wounded and two non-wounded leaflets/leaf) were placed on moist filter paper in petri dishes, and each leaflet inoculated with a 20-μl drop of a conidial suspension containing 1 × 105 conidia/ml in sterilized, distilled water (3). The leaves were wounded by applying pressure to leaf blades with the serrated edge of forceps. Four control leaves were inoculated similarly with sterilized, distilled water. Inoculated leaves were incubated in humid chambers at 25°C with 95% RH and a 12-h photoperiod. After 7 days, symptoms similar to those observed in the original fields developed on both wounded and non-wounded inoculated leaves, but not on control leaves, and S. solani was reisolated consistently from the symptoms using the same method as the original isolations. Control leaves remained asymptomatic and the fungus was not isolated from these leaves. The pathogenicity testing was repeated with similar results. To our knowledge, this is the first report of S. solani on eggplant in Malaysia. References: (1) B. S. Kim et al. Plant Pathol. J. 20:85, 2004. (2) Y. R. Mehta et al. Curr. Microbiol. 44:323, 2002. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiv. Series 6:775, 2007.


Plant Disease ◽  
2012 ◽  
Vol 96 (8) ◽  
pp. 1226-1226
Author(s):  
A. Nasehi ◽  
J. B. Kadir ◽  
M. A. Zainal Abidin ◽  
M. Y. Wong ◽  
F. Mahmodi

In June 2011, tomatoes (Solanum lycopersicum) in major growing areas of the Cameron Highlands and the Johor state in Malaysia were affected by a leaf spot disease. Disease incidence exceeded 80% in some severely infected regions. Symptoms on 50 observed plants initially appeared on leaves as small, brownish black specks, which later became grayish brown, angular lesions surrounded by a yellow border. As the lesions matured, the affected leaves dried up and became brittle and later developed cracks in the center of the lesions. A survey was performed in these growing areas and 27 isolates of the pathogen were isolated from the tomato leaves on potato carrot agar (PCA). The isolates were purified by the single spore technique and were transferred onto PCA and V8 agar media for conidiophore and conidia production under alternating light (8 hours per day) and darkness (16 hours per day) (4). Colonies on PCA and V8 agar exhibited grey mycelium and numerous conidia were formed at the terminal end of conidiophores. The conidiophores were up to 240 μm long. Conidia were oblong with 2 to 11 transverse and 1 to 6 longitudinal septa and were 24 to 69.6 μm long × 9.6 to 14.4 μm wide. The pathogen was identified as Stemphylium solani on the basis of morphological criteria (2). In addition, DNA was extracted and the internal transcribed spacer region (ITS) was amplified by universal primers ITS5 and ITS4 (1). The PCR product was purified by the commercial PCR purification kit and the purified PCR product sequenced. The resulting sequences were 100% identical to published S. solani sequences (GenBank Accestion Nos. AF203451 and HQ840713). The amplified ITS region was deposited with NCBI GenBank under Accession No. JQ657726. A representative isolate of the pathogen was inoculated on detached 45-day-old tomato leaves of Malaysian cultivar 152177-A for pathogenicity testing. One wounded and two nonwounded leaflets per leaf were used in this experiment. The leaves were wounded by applying pressure to leaf blades with the serrated edge of a forceps. A 20-μl drop of conidial suspension containing 105 conidia/ml was used to inoculate these leaves (3). The inoculated leaves were placed on moist filter paper in petri dishes and incubated for 48 h at 25°C. Control leaves were inoculated with sterilized distilled water. After 7 days, typical symptoms for S. solani similar to those observed in the farmers' fields developed on both wounded and nonwounded inoculated leaves, but not on noninoculated controls, and S. solani was consistently reisolated. To our knowledge, this is the first report of S. solani causing gray leaf spot of tomato in Malaysia. References: (1) M. P. S. Camara et al. Mycologia 94:660, 2002. (2) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiversity Series 6:775, 2007.


Plant Disease ◽  
2012 ◽  
Vol 96 (8) ◽  
pp. 1227-1227 ◽  
Author(s):  
A. Nasehi ◽  
J. B. Kadir ◽  
M. A. Zainal Abidin ◽  
M. Y. Wong ◽  
F. Abed Ashtiani

Symptoms of gray leaf spot were first observed in June 2011 on pepper (Capsicum annuum) plants cultivated in the Cameron Highlands and Johor State, the two main regions of pepper production in Malaysia (about 1,000 ha). Disease incidence exceeded 70% in severely infected fields and greenhouses. Symptoms initially appeared as tiny (average 1.3 mm in diameter), round, orange-brown spots on the leaves, with the center of each spot turning gray to white as the disease developed, and the margin of each spot remaining dark brown. A fungus was isolated consistently from the lesions using sections of symptomatic leaf tissue surface-sterilized in 1% NaOCl for 2 min, rinsed in sterile water, dried, and plated onto PDA and V8 agar media (3). After 7 days, the fungal colonies were gray, dematiaceous conidia had formed at the end of long conidiophores (19.2 to 33.6 × 12.0 to 21.6 μm), and the conidia typically had two to six transverse and one to four longitudinal septa. Fifteen isolates were identified as Stemphylium solani on the basis of morphological criteria described by Kim et al. (3). The universal primers ITS5 and ITS4 were used to amplify the internal transcribed spacer region (ITS1, 5.8, and ITS2) of ribosomal DNA (rDNA) of a representative isolate (2). A 570 bp fragment was amplified, purified, sequenced, and identified as S. solani using a BLAST search with 100% identity to the published ITS sequence of an S. solani isolate in GenBank (1). The sequence was deposited in GenBank (Accession No. JQ736024). Pathogenicity of the fungal isolate was tested by inoculating healthy pepper leaves of cv. 152177-A. A 20-μl drop of conidial suspension (105 spores/ml) was used to inoculate each of four detached, 45-day-old pepper leaves placed on moist filter papers in petri dishes (4). Four control leaves were inoculated similarly with sterilized, distilled water. The leaves were incubated at 25°C at 95% relative humidity for 7 days. Gray leaf spot symptoms similar to those observed on the original pepper plants began to develop on leaves inoculated with the fungus after 3 days, and S. solani was consistently reisolated from the leaves. Control leaves did not develop symptoms and the fungus was not reisolated from these leaves. Pathogenicity testing was repeated with the same results. To our knowledge, this is the first report of S. solani causing gray leaf spot on pepper in Malaysia. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. P. S. Camara et al. Mycologia 94:660, 2002. (3) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (4) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002.


Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 689-689
Author(s):  
A. Nasehi ◽  
J. B. Kadir ◽  
M. Nasr Esfahani ◽  
F. Mahmodi ◽  
H. Ghadirian ◽  
...  

In June 2011, lettuce (Lactuca sativa) plants cultivated in major lettuce growing areas in Malaysia, including the Pahang and Johor states, had extensive leaf spots. In severe cases, disease incidence was recorded more than 80%. Symptoms on 50 observed plants initially were as water soaked spots (1 to 2 mm in diameter) on leaves, and then became circular spots spreading over much of the leaves. In this research, main lettuce growing areas infected by the pathogen in the mentioned states were investigated and the pathogen was isolated onto potato dextrose agar (PDA). Colonies observed were greyish green to light brown. Single conidia were formed at the terminal end of conidiophores that were 28.8 to 40.8 μm long and 11.0 to 19.2 μm wide, and 2 to 7 transverse and 1 to 4 longitudinal septa. To produce conidia, the fungus was grown on potato carrot agar (PCA) and V8 juice agar media under 8-h/16-h light/dark photoperiod. Fourteen isolates were identified Stemphylium solani based on morphological criteria described by Kim et al. (1). To confirm morphological characterization, DNA of the fungus was extracted from mycelium and PCR was done using universal primers ITS5 (5′-GGAAGTAAAAGTCGTAACAAGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′), which amplified the internal transcribed spacer (ITS) region of rDNA (2). The sequencing result was subjected to BLAST analysis which was 99% identical to the other published sequences in the GenBank database (GenBank Accession Nos. AF203451 and HQ840713). The nucleotide sequence was deposited in GenBank under Accession No. JQ736022. Pathogenicity testing of representative isolate was done using 20 μl of conidial suspension with a concentration of 1 × 105/ml in droplets (three drops on each leaf) on four detached 45-day-old lettuce leaves cv. BBS012 (3). Fully expended leaves were placed on moist filter paper in petri dishes and were incubated in humid chambers at 25°C. The leaves inoculated with sterile water served as control. After 7 days, disease symptoms were observed, which were similar to those symptoms collected in infected fields and the fungus was reisolated and confirmed as S. solani based on morphological criteria (1) and molecular characterization (2). Control leaves remained healthy. Pathogenicity testing was completed twice. To our knowledge, this is the first report of S. solani on lettuce in Malaysia and it may become a serious problem because of its broad host range, variability in pathogenic isolates, and prolonged active phase of the disease cycle. Previous research has shown that S. solani is a causal agent of gray leaf spot on lettuce in China (4). References: (1) B. S. Kim et al. Plant Pathol. J. 20:85, 2004. (2) Y. R. Mehta et al. Current Microbiol. 44:323, 2002. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) F. L. Tai. Sylloge Fungorum Sinicorum, Sci. Press, Acad. Sin., Peking, 1979.


Plant Disease ◽  
2008 ◽  
Vol 92 (6) ◽  
pp. 870-877 ◽  
Author(s):  
U. N. Nanayakkara ◽  
W. Uddin ◽  
L. E. Datnoff

Silicon amendments have been proven effective in controlling fungal diseases of various crops. However, effects of silicon amendments on gray leaf spot (Magnaporthe oryzae) of perennial ryegrass are not known. Studies were conducted in controlled-environment chambers and microplots where perennial ryegrass pots were buried among perennial ryegrass turf to determine the effects of silicon amendments on gray leaf spot development. Plants were grown in two soil types: peat:sand mix (soil Si = 5.2 mg/liter) and Hagerstown silt loam (soil Si = 70 mg/liter). Both soil types were amended with two sources of silicon—wollastonite and calcium silicate slag—at 0, 0.5, 1, 2, 5, and 10 metric tons/ha and 0, 0.6, 1.2, 2.4, 6, and 12 metric tons/ha, respectively. Nine-week-old perennial ryegrass was inoculated with M. oryzae. Gray leaf spot incidence and severity were assessed 2 weeks after inoculation. Gray leaf spot incidence and severity of perennial ryegrass significantly decreased by different rates of wollastonite and calcium silicate slag applied to both soils under both experimental conditions. Tissue silicon content increased consistently with increasing amount of silicon in the soils, while disease incidence decreased consistently with increasing tissue silicon content in all four soil and source combinations under both experimental conditions. These findings suggest that silicon amendments may be utilized in integrated gray leaf spot management programs on perennial ryegrass.


2015 ◽  
Vol 105 (6) ◽  
pp. 748-757 ◽  
Author(s):  
Alamgir Rahman ◽  
Christopher M. Wallis ◽  
Wakar Uddin

Sustainable integrated disease management for gray leaf spot of perennial ryegrass may involve use of plant defense elicitors with compatible traditional fungicides to reduce disease incidence and severity. Silicon (Si) has previously been identified as a potential inducer or modulator of plant defenses against different fungal pathogens. To this end, perennial ryegrass was inoculated with the causal agent of gray leaf spot, Magnaporthe oryzae, when grown in soil that was nonamended or amended with three different levels of calcium silicate (1, 5, or 10 metric tons [t]/ha). When applied at a rate of 5 t/ha, calcium silicate was found to significantly suppress gray leaf spot in perennial ryegrass, including a significant reduction of disease incidence (39.5%) and disease severity (47.3%). Additional studies observed nonpenetrated papillae or cell-wall appositions harboring callose, phenolic autofluorogens, and lignin-associated polyphenolic compounds in grass grown in the Si-amended soil. Regarding defense-associated enzyme levels, only following infection did grass grown in Si-amended soil exhibit greater activities of peroxidase and polyphenol oxidase than equivalent inoculated control plants. Also following infection with M. oryzae, grass levels of several phenolic acids, including chlorogenic acid and flavonoids, and relative expression levels of genes encoding phenylalanine ammonia lyase (PALa and PALb) and lipoxygenase (LOXa) significantly increased in Si-amended plants compared with that of nonamended control plants. These results suggest that Si-mediated increase of host defense responses to fungal pathogens in perennial ryegrass has a great potential to be part of an effective integrated disease management strategy against gray leaf spot development.


Plant Disease ◽  
2002 ◽  
Vol 86 (1) ◽  
pp. 75-75 ◽  
Author(s):  
W. Uddin ◽  
G. Viji ◽  
L. Stowell

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.


Plant Disease ◽  
2000 ◽  
Vol 84 (4) ◽  
pp. 492-492 ◽  
Author(s):  
P. Harmon ◽  
K. Rane ◽  
G. Ruhl ◽  
R. Latin

Pyricularia grisea, the causal agent of gray leaf spot on turfgrass, was isolated from symptomatic perennial ryegrass (Lolium perenne) leaves collected from a golf course in north-central Indiana in August 1999. Gray leaf spot is an emerging threat to stands of perennial ryegrass in the mid-Atlantic and Midwestern United States (1). Symptoms were first evident in taller (6 cm) mown, rough areas surrounding golf course fairways. Field symptoms included diffuse patches (1 to 4 m in diameter) of thin, yellow-tan turf. Within larger affected areas, some of the turf was dead and matted. Close inspection revealed the presence of typical tan-gray lesions with brown margins and fish hook-like distortion of infected leaf blade tips. Incubation of affected turf in a saturated environment at 23°C for 16 h resulted in production of numerous three-celled, pear-shaped conidia characteristic of those produced by P. grisea. A pure culture of the isolate was grown on V8-juice agar in darkness at 29°C. After 10 days, the culture was exposed to continuous light for 4 days at 23°C to induce sporulation. Conidia were washed from the colony surface with sterile distilled water. Two-week-old perennial ryegrass plants in 8-cm-diameter pots were inoculated with the conidial suspension. Typical gray leaf spot symptoms resulted after incubation of inoculated plants at 27°C for 72 h in a saturated atmosphere. Uninoculated control plants exposed to the same environmental conditions remained healthy. This is the first report of gray leaf spot on perennial ryegrass in Indiana. Reference: (1) P. J. Landschoot and B. F. Hoyland. Plant Dis. 76:1280, 1992.


Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1151-1151 ◽  
Author(s):  
D. K. Pedersen ◽  
R. T. Kane ◽  
H. T. Wilkinson

Each year from 1991 to 1999, a disease matching the description of gray leaf spot (1) was observed in the central and north central regions of Illinois. Disease severity was low (<10% blight) from 1991 to 1994 and 1999 and was severe (>50% blight in some areas) from 1995 to 1998. The disease was observed on Lolium perenne (perennial ryegrass) golf course fairways and sports fields. Isolations of Pyricularia grisea were made from L. perenne collected from golf courses in Bloomington, Decatur, Kankakee, Pekin, Urbana, and Moline, IL. All isolates were collected from surface-sterilized, symptomatic leaves. Cultures were maintained on one-fifth strength potato-dextrose agar (PDA) and induced to sporulate on full-strength oatmeal agar. All isolates in culture displayed vegetative and conidial characteristics similar to those previously described for P. grisea (1). Twenty-five different L. perenne germ plasms were inoculated with isolate WF9826 (Kankakee) using a suspension of 1 × 105 conidia per milliliter. The 4-week-old lawns (100 plants per 3-cm-diameter cone-tainer) of each ryegrass germ plasm were inoculated by spraying foliage with the conidial suspension until runoff. Inoculated and uninoculated lawns were enclosed in plastic bags and placed in an incubator (16 h light; 28°C) for 7 days. Disease severity was rated using a scale of 0 to 10 (10 = 100% blight). Each treatment was replicated three times, and all experiments were repeated four times. Small blue-gray, water-soaked lesions with dark brown borders were observed on leaves of all inoculated ryegrass germ plasms. Advanced symptoms included blighting of much of the leaves. The mean disease severity rating was 3.8 (range 2 to 7) for all experimental units and all 25 germ plasms. P. grisea was isolated from leaves that were inoculated with WF9826. This is the first report of gray leaf spot of perennial ryegrass caused by P. grisea in Illinois. Reference: (1) P. J. Landschoot et al. Plant Dis. 76:1280, 1992.


Plant Disease ◽  
2005 ◽  
Vol 89 (2) ◽  
pp. 207-207 ◽  
Author(s):  
S. Gaetán ◽  
M. Madia

Canola (Brassica napus) is a developing oleaginous crop grown commercially in Argentina. During 2003, typical symptoms of a foliar disease were observed on canola plants in experimental field plots in Buenos Aires. Average disease incidence across 14 6-month-old canola cultivars was 27% (range 12 to 42%). Climatic conditions in Buenos Aires during August 2003 included moderate temperatures and periods with high humidity, which were apparently favorable for disease development. Symptoms were observed on leaves, stems, and pods. Leaf symptoms were randomly distributed on the adaxial surfaces and consisted of zonate lesions of alternating light gray and dark brown areas that were 6 to 10 mm in diameter. Remaining leaf tissue was chlorotic and affected leaves abscised. Stem infections appeared as irregular and elongated black lesions, 0.7 to 1.2 cm long. Pods lesions were circular, 6 to 8 mm in diameter, gray in the center, and surrounded by a diffuse dark brown margin. The disease developed progressively from the lower leaves to the pods, resulting in premature senescence of the tissues, chlorosis, and defoliation. Conidiophores bearing conidia colonized the lesions as a dark gray growth of spore masses. Segments (0.5 cm long) taken from leaves, stems, and pods of diseased plants were dipped in 70% ethanol, surface sterilized with NaOCl (1%) for 2 min, and rinsed in sterile water. Each segment was blotted dry and placed on potato dextrose agar. Plates were incubated in the dark at 25°C for 2 to 3 days, followed by incubation under NUV light and a 12-h light/dark photoperiod for 6 to 8 days. Six fungal isolates were obtained. Fungal colonies were pale gray with dark concentric rings. Conidia were yellow to pale brown, ellipsoid to ovoid, produced singly or in short chains, with 8 to 10 transverse septa and 2 to 6 longitudinal septa. The spore body measured 13 to 22 × 68 to 135 µm with a beak cell 42 to 101 µm long. On the basis of conidial and cultural characteristics, the fungus was identified as Alternaria brassicae (Berk.) Sacc (1). Koch's postulates were completed for three isolates by spray-inoculating foliage of 6-week-old canola plants of cvs. Caviar, Dunkeld, Eclipse, Impulse, Mistral, and Sponsor with a conidial suspension (1 × 105 conidia per ml). The experiment, which included four inoculated plants and two noninoculated control plants for each cultivar per isolate, was conducted in the greenhouse at 22 to 24°C and maintained at 75% relative humidity with no supplemental light. Inoculated and control plants were covered with polyethylene bags for 48 h after inoculation. Within 12 days, inoculated plants developed small, brown lesions on leaves and stems for all three isolates; the pathogen was successfully reisolated in all instances. Control plants, inoculated only with sterile distilled water, remained symptomless. The experiment was repeated with similar results. The results suggest that A. brassicae may be a threat to the main cultivars being grown in Argentina. To our knowledge, this is the first report of A. brassicae causing gray leaf spot of canola in Argentina. Reference: (1) J. Joly. Le genre Alternaria. Recherches Physiologiques, Biologiques, et Systématiques. Paul Lechevalier, ed. Paris, France, 1964.


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