Factors Associated with Leguminous Green Manure Incorporation and Fusarium Wilt Suppression in Watermelon.

Fall-planted Vicia villosa or Trifolium incarnatum cover crops, incorporated in spring as a green manure, can suppress Fusarium wilt (Fusarium oxysporum f. sp. niveum) of watermelon. During cover crop growth, termination, and incorporation into the soil, many factors such as arbuscular mycorrhizae colonization, leachate, and soil respiration differ. How these cover-crop-associated factors affect Fusarium wilt suppression is not fully understood. Experiments were conducted to evaluate how leachate, soil respiration, and other green-manure-associated changes affected Fusarium wilt suppression, and to evaluate the efficacy of the biocontrol product Actinovate AG (Streptomyces lydicus WYEC 108). General and specific suppression was examined in the field by assessing the effects of cover crop green manures (V. villosa, T. incarnatum, Secale cereale, and Brassica juncea) on soil respiration, presence of F. oxysporum spp., and arbuscular mycorrhizal colonization of watermelon. Cover crop treatments V. villosa, T. incarnatum, and S. cereale and no cover crop were evaluated both alone and in combination with Actinovate AG in the greenhouse. Additionally, in vitro experiments were conducted to measure the effects of cover crop leachate on the mycelial growth rates of F. oxysporum f. sp. niveum race 1 and Trichoderma harzianum. Soil microbial respiration was significantly elevated in V. villosa and Trifolium incarnatum treatments both preceding and following green manure incorporation, and was significantly negatively correlated with Fusarium wilt, suggesting that microbial activity was higher under the legumes, indicative of general suppression. Parallel to this, in vitro growth rates of F. oxysporum f. sp. niveum and Trichoderma harzianum on V. villosa leachate amended media were 66 and 213% greater, respectively, than on nonamended plates. The F. oxysporum spp. population (based on CFU and not differentiated into formae specialis or races) significantly increased in V. villosa-amended field plots. Additionally, the percentage of watermelon roots colonized by arbuscular mycorrhizae following V. villosa and Trifolium incarnatum green manures was significantly higher than in watermelon following bare ground (58 and 44% higher, respectively). In greenhouse trials where cover crops were amended to soil, Actinovate AG did not consistently reduce Fusarium wilt. Both general and specific disease suppression play a role in reducing Fusarium wilt on watermelon.

Five Reasons to Consider Phytophthora infestans a Reemerging Pathogen.

Phytophthora infestans has been a named pathogen for well over 150 years and yet it continues to "emerge", with thousands of articles published each year on it and the late blight disease that it causes. This review explores five attributes of this oomycete pathogen that maintain this constant attention. First, the historical tragedy associated with this disease (Irish potato famine) causes many people to be fascinated with the pathogen. Current technology now enables investigators to answer some questions of historical significance. Second, the devastation caused by the pathogen continues to appear in surprising new locations or with surprising new intensity. Third, populations of P. infestans worldwide are in flux, with changes that have major implications to disease management. Fourth, the genomics revolution has enabled investigators to make tremendous progress in terms of understanding the molecular biology (especially the pathogenicity) of P. infestans. Fifth, there remain many compelling unanswered questions.

Impact of Five Cover Crop Green Manures and Actinovate on Fusarium Wilt of Watermelon.

Triploid watermelon cultivars are grown on more than 2,023 ha in Maryland and in Delaware. Triploid watermelon cultivars have little host resistance to Fusarium wilt of watermelon (Fusarium oxysporum f. sp. niveum). The effects of four different fall-planted cover crops (Vicia villosa, Trifolium incarnatum, Secale cereale, and Brassica juncea) that were tilled in the spring as green manures, and bare ground, were evaluated alone and in combination with the biocontrol product Actinovate (Streptomyces lydicus) on Fusarium wilt severity and watermelon fruit yield and quality. Six field experiments were conducted over 3 years in Beltsville and Salisbury, MD and Georgetown, DE. Both V. villosa and T. incarnatum significantly suppressed Fusarium wilt of watermelon as much as 21% compared with watermelon in nonamended plots. However, no suppression of Fusarium wilt occurred at low disease levels or where low cover crop biomass was present. In general, Beltsville, MD had lower disease levels than Salisbury, MD and Georgetown, DE. T. incarnatum was the only cover crop that yielded significantly more fruit than nonamended treatments (129% more fruit per hectare) but only for one field trial. The Actinovate product either did not reduce Fusarium wilt or the magnitude of the reduction was nominal. Actinovate significantly reduced Fusarium wilt by 2% in 2009 and as much as 7% in 2010, and increased Fusarium wilt severity by 2.5% in 2011. Actinovate significantly increased yield for one field trial but only when applied to nonamended or Secale cereal-amended plots. This is the first report of a reduction in Fusarium wilt following a T. incarnatum cover crop incorporated as a green manure.

The 2009 Late Blight Pandemic in the Eastern United States - Causes and Results.

The tomato late blight pandemic of 2009 made late blight into a household term in much of the eastern United States. Many home gardeners and many organic producers lost most if not all of their tomato crop, and their experiences were reported in the mainstream press. Some CSAs (Community Supported Agriculture) could not provide tomatoes to their members. In response, many questions emerged: How did it happen? What was unusual about this event compared to previous late blight epidemics? What is the current situation in 2012 and what can be done? It's easiest to answer these questions, and to understand the recent epidemics of late blight, if one knows a bit of the history of the disease and the biology of the causal agent, Phytophthora infestans.

Extension plant pathology: strengthening resources to continue serving the public interest.

Extension plant pathologists deliver science-based information that protects the economic value of agricultural and horticultural crops in the United States by educating growers and the general public about plant diseases. Extension plant pathologists diagnose plant diseases and disorders, provide advice, and conduct applied research on local and regional plant disease problems. During the last century, extension plant pathology programs have adjusted to demographic shifts in the U.S. population and to changes in program funding. Extension programs are now more collaborative and more specialized in response to a highly educated clientele. Changes in federal and state budgets and policies have also reduced funding and shifted the source of funding of extension plant pathologists from formula funds towards specialized competitive grants. These competitive grants often favor national over local and regional plant disease issues and typically require a long lead time to secure funding. These changes coupled with a reduction in personnel pose a threat to extension plant pathology programs. Increasing demand for high-quality, unbiased information and the continued reduction in local, state, and federal funds is unsustainable and, if not abated, will lead to a delay in response to emerging diseases, reduce crop yields, increase economic losses, and place U.S. agriculture at a global competitive disadvantage. In this letter, we outline four recommendations to strengthen the role and resources of extension plant pathologists as they guide our nation's food, feed, fuel, fiber, and ornamental producers into an era of increasing technological complexity and global competitiveness.

Anthracnose and Gummy Stem Blight Are Reduced on Watermelon Grown on a No-Till Hairy Vetch Cover Crop.

Multiple applications of fungicides are used to manage anthracnose caused by Colletotrichum orbiculare and gummy stem blight caused by Didymella bryoniae, the two most common and destructive diseases on watermelon (Citrullus lanatus) in the mid-Atlantic region of the United States. To develop a sustainable, nonchemical management option, a split-plot experiment was conducted over 3 years to evaluate the effects of a no-till hairy vetch (Vicia villosa) cover crop on disease severity, plant growth, and fruit yield compared with two conventional bedding systems and fungicide application. The main plots were bedding strategies consisting of bare ground, polyethylene covering, or a hairy vetch cover crop that was planted in the fall, killed the following spring, and left on the soil surface as an organic mulch. The subplots were a nonfungicide control or a weekly application of a standard fungicide program. Hairy vetch mulch provided greater than a 65% reduction in the area under the disease progress curves of anthracnose and gummy stem blight and greater than an 88% decrease in diseased fruit compared with bare ground or polyethylene mulch. The reductions were comparable with those achieved by fungicide applications. Watermelon vine lengths in plots with hairy vetch were similar to or greater than those in plots with polyethylene or bare ground that were treated with fungicides. Marketable fruit in plots with hairy vetch was higher compared with bare ground in 2 of 3 years and was similar to that in plots treated with fungicides in all 3 years. Addition of fungicide application to hairy vetch treatment further reduced anthracnose in 1 year and gummy stem blight in 2 years but did not significantly increase fruit yield in all 3 years. This is the first demonstration that a no-till hairy vetch production system can reduce anthracnose and gummy stem blight on watermelon and that the production system has the potential to mitigate damage caused by these diseases.

Race 3, a New and Highly Virulent Race of Fusarium oxysporum f. sp. niveum Causing Fusarium Wilt in Watermelon.

Three races (0, 1, and 2) of Fusarium oxysporum f. sp. niveum have been previously described in watermelon (Citrullus lanatus) based on their ability to cause disease on differential watermelon genotypes. Four isolates of F. oxysporum f. sp. niveum collected from wilted watermelon plants or infested soil in Maryland, along with reference isolates of races 0, 1, and 2, were compared for virulence, host range, and vegetative compatibility. Race identification was made on the watermelon differentials Sugar Baby, Charleston Gray, Dixielee, Calhoun Gray, and PI-296341-FR using a root-dip, tray-dip, or pipette inoculation method. All four Maryland isolates were highly virulent, causing 78 to 100% wilt on all differentials, one of which was PI-296341-FR, considered highly resistant to race 2. The isolates also produced significantly greater colonization in the lower stems of PI-296341-FR than a standard race 2 reference isolate. In field microplots, two of the isolates caused over 90% wilt on PI-296341-FR, whereas no disease was caused by a race 2 isolate. All four isolates were nonpathogenic on muskmelon, cucumber, pumpkin, and squash, confirming their host specific pathogenicity to watermelon. The Maryland isolates were vegetatively compatible to each other but not compatible with the race 2 isolates evaluated, indicating their genetic difference from race 2. This study proposes that the Maryland isolates belong to a new race, race 3, the most virulent race of F. oxysporum f. sp. niveum described to date.

First Report of Alternaria alternata f. sp. cucurbitae Causing Alternaria Leaf Spot of Melon in the Mid-Atlantic Region of the United States.

Alternaria alternata f. sp. cucurbitae, the casual agent of Alternaria leaf spot, was first described in Greece where it caused severe losses to greenhouse-grown cucumbers (Cucumis sativus) (3,4). The fungus also attacks melon (C. melo) and watermelon (Citrullus lanatus) (1-3). In late June of 2006, following a period of windy and rainy days, numerous dark brown, circular lesions, 0.5 to 1 mm in diameter, were observed on leaves of melons in a field in Wicomico County, Maryland. The lesions gradually enlarged and coalesced into large, nearly circular, or irregularly shaped lesions that could be as long as 3 cm. The center of the lesions was light tan, surrounded by a dark brown ring and a chlorotic halo, and tended to split in the later development stages. Most of the lesions appeared on the edge of the leaves and no lesions developed on the stems and fruit. Lesions first started on old leaves and then developed on leaves in the middle part of the canopy. Leaf lesions were observed on melon cvs. Ananas, Honeydew Greenflesh, and Israeli. Disease severity ranged from 3 to 20% of the leaf area affected. Small pieces (3 × 3 mm) of tissue removed from the margin between healthy and diseased tissue were surface disinfected in 0.5% NaOCl for 2 min and plated on acidified, »-strength potato dextrose agar. Isolations made from diseased tissue frequently (61%) yielded fungal colonies with morphological features and spore dimensions that were consistent with the description of A. alternata f. sp. cucurbitae (1,3). Fungal isolates were characterized by small, short-beaked, multicellular conidia. Conidia were ovoid, obclavate, and sometimes ellipsoidal with the average overall body length of 39 µm (range, 17 to 80 µm) and width of 14 µm (range, 7 to 20 µm). Conidia were produced on short conidiophores in chains. The beaks were short (often less than one-third the body length) and conical or cylindrical. Pathogenicity of six single-spore isolates was determined on four melon cultivars (Honeydew Greenflesh, Israeli, Tam Dew, and Topmark) and one watermelon cultivar (Sugar Baby) in a greenhouse. Twenty plants of each cultivar at the one-true-leaf stage were sprayed with a conidial suspension (106 conidia/ml) of each isolate amended with 0.1% (vol/vol) of Tween 20 until runoff (1.5 to 2 ml per plant). Inoculation with sterile distilled water amended with 0.1% Tween 20 served as controls. The plants were placed in a dew growth chamber for 48 h at 24°C and subsequently maintained in a greenhouse at 21 to 29°C. At 4 to 5 days after inoculation, each isolate induced leaf lesions on each inoculated cultivar similar to typical lesions observed in the field. There was no significant difference in disease severity among the cultivars tested or between melon and watermelon. Control plants remained symptomless. The fungus was readily reisolated from symptomatic tissues. To our knowledge, this is the first report of A. alternata f. sp. cucurbitae causing Alternaria leaf spot of melon in the Mid-Atlantic United States and the only report outside Georgia in the southern region of the United States (D. B. Langston, personal communication) and Greece. References: (1) D. L. Vakalounakis. Plant Dis. 74:227, 1990. (2) D. L. Vakalounakis. Ann. Appl. Biol. 117:507, 1990. (3) D. L. Vakalounakis. Alternaria leaf spot. Page 24 in: Compendium of Cucurbit Diseases. T. A. Zitter et al., eds. The American Phytopathological Society, St. Paul, MN, 1996. (4) D. L. Vakalounakis and N. E. Malathrakis. J. Phytopathol. 121:325, 1988.

Characterization of a Regional Population of Fusarium oxysporum f. sp. niveum by Race, Cross Pathogenicity, and Vegetative Compatibility.

ABSTRACT Eighty-eight isolates of Fusarium oxysporum f. sp. niveum, collected from wilted watermelon plants and infested soil in Maryland and Dela-ware, were characterized by cross pathogenicity to muskmelon, race, and vegetative compatibility. Four isolates (4.5%) were moderately pathogenic to >/=2 of 18 muskmelon cultivars in a greenhouse test, and one representative isolate also was slightly pathogenic in field microplots. The four isolates all were designated as race 2, and were in vegetative compatibility group (VCG) 0082. Of the 74 isolates to which a VCG could be assigned, 41 were in VCG 0080, the VCG distributed most widely; 27 were in VCG 0082, and were distributed in half of the 20 watermelon fields surveyed; and 6 were in the newly described VCG 0083, and were restricted to three fields. Among the isolates in VCG 0080, 8 were designated as race 0, 21 as race 1, and 12 as race 2. Of the isolates in VCG 0082, 6 were designated as race 0, 11 as race 1, and 10 as race 2. All isolates in VCG 0083 were designated as race 2. Isolates from more than one race within the same VCG or isolates from more than one VCG were recovered from single plants and fields. No differences in aggressiveness on differential watermelon cultivars were observed among isolates from different VCGs of the same race. A diverse association between virulence and VCG throughout the Mid-Atlantic region suggests that the pathotypes of F. oxysporum f. sp. niveum may be of local origin or at least long existent in the region.

Effects of Host Resistance and Inoculum Density on the Suppression of Fusarium Wilt of Watermelon Induced by Hairy Vetch.

Hairy vetch (Vicia villosa) used as a soil amendment is a newly described potential management tool for the suppression of Fusarium wilt of watermelon (Citrullus lanatus). However, the effect of inoculum density and the level of resistance in the host on the level of suppression are not understood. In this study, hairy vetch-induced wilt suppression was evaluated in the greenhouse on 12 watermelon cultivars with different levels of wilt resistance and in 16 naturally infested soil samples collected from commercial watermelon fields. Wilt suppression occurred in all but two cultivars and with the trend that suppression increased as the level of resistance in cultivars increased. Fusarium wilt suppression was 22, 53, and 63% in hairy vetch-amended soil compared with nonamended soil on cultivars ranked as susceptible, moderately resistant, and highly resistant, respectively. Suppression also occurred in nine of the soils that contained populations of Fusarium oxysporum f. sp. niveum below 1,100 CFU/g of soil. However, at this level or higher, significant wilt suppression was not observed. The magnitude of disease suppression decreased with the increase of inoculum in the soils. The induced wilt suppression appeared to be correlated with an increase in bacterial populations in soil. Hairy vetch-induced suppression to Fusarium wilt in watermelon is dependent on the resistance level of cultivars and is overcome by high inoculum level of F. oxysporum f. sp. niveum in soil.

Root-Knot and Root-Lesion Nematode Suppression by Cover Crops, Poultry Litter, and Poultry Litter Compost.

Experiments using soil-incorporated cover crops and amendments of poultry litter (PL) and PL compost to suppress root-knot (RKN) and root-lesion nematodes were conducted in response to increasing nematode populations in Maryland's potato production areas. Identical experiments were established in microplots infested with Meloidogyne incognita or Pratylenchus penetrans. Treatments consisted of 12 3-year rotational sequences comprising potato (year 1) and cucumber (year 2) followed by a moderately RKN-resistant or susceptible soybean cultivar, castor bean, grain sorghum, or sorghum sudangrass; PL or PL compost were amended to some of the RKN-susceptible soybean and sorghum sudangrass plots. In the third year of the rotation, potato followed by soybean was planted in all 12 treatments. The RKN-resistant soybean, castor bean, sorghum sudangrass, and fallow or tillage decreased the populations of M. incognita compared with microplots where RKN-susceptible soybean had been grown. However, RKN populations quickly recovered. Root-lesion nematode was reduced in the spring of 2001 following application of high rates of PL and PL compost in 2000. In the fall of 2001, sorghum sudangrass alone or in combination with PL or PL compost, grain sorghum, or fallow or tillage reduced root-lesion nematodes compared with either soybean cultivar. No treatment affected root-lesion nematode the following year. The use of cover crops and PL compost is an effective method to reduce nematode populations only if successively incorporated into rotational cropping sequences.

Quantification of Root and Stem Colonization of Watermelon by Fusarium oxysporum f. sp. niveum and Its Use in Evaluating Resistance.

ABSTRACT Colonization of watermelon root and stem tissues by Fusarium oxysporum f. sp. niveum race 1 and its relationship to the apparent resistance to Fusarium wilt was investigated. In each of 2 years, 17 differentially susceptible watermelon cultivars and one accession were tested in the greenhouse, and 7 cultivars also were tested in the field. Colonization by a chlorate-resistant marked isolate of the fungus was assayed by plating homogenized tissue samples on a selective medium. Six days after inoculation, seedlings of highly resistant, moderately resistant, and susceptible cultivars had F. oxysporum f. sp. niveum race 1 CFU counts in the lower stems of 10(2), 10(3), and 10(4) CFU/g of fresh tissue, respectively. Percent wilt (Y) of the seedlings was positively correlated with colonization (X) by F. oxysporum f. sp. niveum race 1 in roots (Y = 21.2 ln [X + 1] - 140.7, R(2) = 0.85) or lower stems (Y = 17.3 ln [X + 1] - 78.6, R(2) = 0.86). Percent wilt (Y) also was correlated with the ratio (X(r), 0 to 1 values) of lower stem to root colonization (Y = 34 ln X(r) + 112, R(2) = 0.36). Field evaluations confirmed these relationships, and a link between cultivar resistance and a reduced rate of spread of the fungus in primary stems during a season was observed. Fruit yield decreased with increased tissue colonization at linear rates of 9.9 to 12.7 t/ha per ln (CFU/g + 1) (R(2) >/= 0.58). The greenhouse seedling stem colonization assay described may be utilized as a collaborative method to quantify Fusarium wilt resistance in watermelon.

First Report of Benomyl Resistance in Didymella bryoniae in Delaware and Maryland.

Gummy stem blight, caused by Didymella bryoniae (Auersw.) Rehm, is the most severe foliar disease of watermelon, Citrullus lanatus (Thunb.) Matsum. & Nakai, in eastern Maryland and southern Delaware. The fungicide benomyl is used in combination with chlorothalonil to manage gummy stem blight. Under conducive environmental conditions, yield losses are high even when fields are sprayed weekly. Resistance of D. bryoniae to benomyl has been reported in New York State and South Carolina (1). Gummy stem blight-infected leaves and stems were collected from nine and three fields in Wicomico County, MD, and Sussex County, DE, respectively, in 1996. Infected tissue was also collected from two Wicomico County fields in 1997. One single-spore subculture was obtained to represent each field. Agar plugs were taken from actively growing subcultures and inverted on a 25% (quarter strength) potato dextrose agar medium amended with 0 and 33.1 mg of benomyl per liter, the concentration of benomyl that reduced relative colony diameter of four resistant isolates in New York and South Carolina by 50% (1). Two replicate plates were used per experiment and each experiment was repeated once. After 6 days of growth at 21°C in the dark, the colony diameter was measured. Isolates were classified as sensitive if they were unable to grow, moderately sensitive if colony diameter was reduced 40 to 60%, and resistant if colony diameter was reduced less than 10% on the benomyl-amended media, compared with unamended media. Isolates that had previously been tested were used as sensitive (W03) and moderately sensitive (NY1) standards (1). In 1996, two isolates were sensitive, four isolates were moderately sensitive, and six isolates were resistant to benomyl. One isolate from 1997 was resistant and the other was moderately sensitive. This is the first report of resistance to benomyl within the D. bryoniae population in eastern Maryland and southern Delaware. Reference: (1) A. P. Keinath and T. A. Zitter. Plant Dis. 82:479, 1998.

Suppression of Fusarium Wilt of Watermelon by Soil Amendment with Hairy Vetch.

Hairy vetch (Vicia villosa Roth) as a soil amendment was evaluated for suppression of Fusarium wilt of watermelon and soil populations of Fusarium oxysporum f. sp. niveum in greenhouse, microplot, and field studies. When mixed at 1 or 5% (wt/wt) in a loamy sand soil that was artificially or naturally infested with race 2 of F. oxysporum f. sp. niveum, pulverized dry hairy vetch, crab shell, and urea provided the best suppression (53 to 87% reduction) of Fusarium wilt on watermelon seedlings among 13 plant and animal residues screened. Soil amended with hairy vetch at 0.25 or 0.5% (wt/wt) in microplots resulted in 54 to 69% decreased wilt incidence and 100 to 220% increase of watermelon plant biomass. Hairy vetch winter cover crop incorporated into field plots under black plastic provided 42 to 48% reduction of wilt incidence, 64 to 100% increase of plant biomass, and a 34 to 68% increase in weight of fruit, comparable to improvements achieved by the soil fumigants methyl bromide or 1,3-dichloropropene plus 35% chloropicrin. Soil amendment with hairy vetch also increased the sugar content of watermelon fruit 10 to 15%. Significant reductions in the populations of F. oxysporum f. sp. niveum were not observed in hairy vetch-amended soil in microplots and field plots, but were observed in greenhouse pot soil amended with 5% (wt/wt) hairy vetch, which was attributed primarily to increased levels of fungicidal ammonia produced during decomposition. Incorporating hairy vetch into mulched soil can be an alternative or supplement to cultivar resistance and crop rotation for management of Fusarium wilt of watermelon.

Races and Inoculum Density of Fusarium oxysporum f. sp. niveum in Commercial Watermelon Fields in Maryland and Delaware.

A survey was conducted to determine races and inoculum density of Fusarium oxysporum f. sp. niveum, the causal agent of Fusarium wilt of watermelon in Maryland and Delaware. Virulence on six differential cultivars was tested for each of 63 isolates of F. oxysporum f. sp. niveum, obtained from 25 commercial watermelon fields. Thirteen isolates (21%) were identified as race 0, 36 isolates (57%) as race 1, and 14 isolates (22%) as race 2. Races 0 and 1 were present in 12 (48%) and 10 (40%) of the fields, respectively. The highly aggressive race 2 was identified from five fields in two counties in Maryland and from one field in Delaware, representing 24% of the fields. Race 2 was copresent with one or two other races. Race 2 (19 isolates) predominated among the 25 isolates obtained from a research field in Maryland. Nineteen commercial fields had inoculum densities of F. oxysporum f. sp. niveum ranging from 100 to 1,200 CFU/g of soil at harvest. Within this range of inoculum densities, >20% incidence of wilt was observed when the susceptible watermelon cv. Sugar Baby was planted in samples of soil collected from these fields. The relationship (P < 0.0001) between inoculum density of F. oxysporum f. sp. niveum (X) and incidence of Fusarium wilt (Y) on Sugar Baby was best described using the monomolecular equation, Y = 1 - exp[-0.0013 (X + 166)]. The ratio of pathogenic to total population of F. oxysporum in the fields linearly increased with increasing inoculum density of F. oxysporum f. sp. niveum (R 2 = 0.4; P < 0.0009).

Report of Leaf Spot of Spinach Caused by Stemphylium botryosum in Maryland and Delaware.

In October 2000, leaf spot symptoms were observed on spinach (Spinacia oleracea L. 'Seven R') at the University of Maryland Lower Eastern Shore Research and Education Center in Salisbury. In April 2001, a similar leaf spot disease was observed in two commercial spinach fields (cv. Vancouver) in Dorchester County, MD, and Sussex County, DE. Symptomatic plants occurred in foci, and overall disease incidence in the research and commercial fields was <10% of plants with lesions. However, low disease incidence may reduce the value of a spinach crop by requiring additional hand-sorting (fresh market) or lowering the grade (processing). Leaf spot lesions were small (0.2 to 0.7 cm), circular, tan, and papery and lacked visual signs of fungal infestation. Lesions resembled a new leaf spot of spinach reported in California (1) caused by Stemphylium botryosum Wallr. Plating surface-disinfested lesion margins on 0.25-strength potato dextrose agar consistently yielded S. botryosum. Single conidial cultures of three isolates were grown on V8 agar in a growth chamber with a 12 h light/dark regime at 21°C and were used for the pathogenicity test. Conidia were collected from 7-day-old colonies to test pathogenicity. Conidia were suspended in distilled water (1.1 × 105 conidia per milliliter), and sprayed on 4-week-old spinach plants (with four to six true leaves) of cvs. Seven R, Vancouver, and Melody. Noninoculated control plants were sprayed with deionized water. Plants were incubated for 72 h in a dew chamber (18°C, 9 to 15 h light/dark regime where dew formed during the dark periods) and then placed on a greenhouse bench (23°C) for 2 weeks. Plants that had been inoculated with any of the three isolates developed the aforementioned leaf spot lesions after 4 days in the greenhouse. Plants sprayed with deionized water were symptomless. One week after inoculation, more lesions were observed on 'Seven R' and 'Vancouver' than on 'Melody' (41, 39, and 1 lesion per plant, respectively; P< 0.0030), and the lesions were 1.5, 1.2, and 0.5 mm in diameter, respectively (P< 0.0001). S. botryosum was consistently reisolated from leaf spot lesions. The pathogenicity test was repeated with similar results. Isolates grown on V8 agar and incubated for ≈10 days produced conidia with mean dimensions of 31 × 19 µm. To our knowledge, this is the first report of leaf spot of spinach caused by S. botryosum in Maryland and Delaware. Reference: (1) S. T. Koike et al. Plant Dis. 85:126, 2001.

First Report of the Occurrence of Fusarium oxysporum f. sp. niveum Race 2 in Commercial Watermelon Production Areas of Maryland and Delaware.

Resistance to race 1 of Fusarium oxysporum f. sp. niveum (E.F. Sm.) W.C. Snyder & H.N. Hans) is the most widely used tool for management of Fusarium wilt of watermelon (Citrullus lanatus (Thunb.) Matsum. and Nakai). However, this resistance is ineffective against the more aggressive F. oxysporum f. sp. niveum race 2. Race 2 was first identified in Israel in 1973 (2) and was subsequently reported in Texas (1981), Oklahoma (1988), and Florida (1989) (1). From July through September 2000 in Maryland and Delaware, 123 isolates of F. oxysporum f. sp. niveum were collected from wilted plants or plants with vascular discoloration from commercial production fields with moderate to severe wilt severity. Race determination was performed in the greenhouse on five race-differential cultivars. Differential cvs. Sugar Baby, Crimson Sweet, Charleston Gray, Allsweet, and Dixielee range from susceptible to highly resistant (in the order listed) to race 1, but all are susceptible to race 2. Seeds were planted in pots containing pasteurized vermiculite and peat moss (2:3 mixture) growth medium infested with 1 × 105 conidia per g of growth medium. Conidia were harvested from 5- to 6-day-old cultures of F. oxysporum f. sp. niveum grown in a liquid mineral salts medium (2). Control pots were treated with water or diluted liquid medium. Pots were maintained in a greenhouse at 19 to 28°C. Three replicate pots containing seven plants each were used for each isolate-cultivar combination. Race 0 obtained from B. D. Bruton (USDA-ARS, Lane, OK), and races 1 and 2 obtained from the American Type Culture Collection (Manassas, VA) were included in the tests for comparative purposes. Eleven isolates from Maryland and three isolates from Delaware consistently caused 60 to 100% wilt or mortality of all cultivars, with a mean of more than 75% wilt. Percent wilt of cvs. Sugar Baby and Dixielee to races 0, 1, and 2, and the 14 isolates from Maryland and Delaware was 78, 76, 100, and 95 to 100% and 0, 14, 59, and 63 to 93%, respectively. All control plants remained healthy. In each of the three replicated tests, these 14 isolates caused an equal or greater incidence of wilt as the reference race 2 isolate, and cvs. Dixielee and Allsweet were susceptible to these isolates. Therefore, these isolates were identified as F. oxysporum f. sp. niveum race 2. This is the first report of F. oxsporum f. sp. niveum race 2 occurring in Maryland and Delaware. References: (1) R. D. Martyn and B. D. Bruton. HortScience 24:696, 1989. (2) D. Netzer. Phytoparasitica 4:131, 1976.

HSP27 elevated in mild allergic inflammation protects airway epithelium from H2SO4 effects.

Inflammation in allergic individuals is hypothesized to elevate stress proteins [heat shock proteins (HSP)] in airway epithelium, which may protect cells from further adverse conditions. Allergic, either asthmatic or not, and normal volunteers participated in a 2-day segmental allergen challenge bronchoscopic procedure. Bronchial epithelium was obtained before and after challenge. Epithelium was exposed to medium with H2SO4 (pH5), returned to medium at pH 7.4, and finally harvested for Western blotting with anti-27-kDa HSP (HSP27) antibody. Prechallenge epithelium of all subjects had significantly inhibited ciliary function by H2SO4 (pH 5) conditions (P < 0.001); only epithelium of normals recovered (P = 0.02). Allergic subjects with mild inflammation (< 50 micrograms/ml increase in albumin in bronchoalveolar lavage) had significantly increased HSP27 postchallenge (P = 0.01) and little ciliary dysfunction at pH 5, whereas subjects with severe inflammation (> 50 micrograms/ml increase in albumin) had little change in HSP27 and significant ciliary inhibition (P = 0.02). Normal epithelium had similar trends in HSP27 and equivalent inhibition of ciliary activity at pH 5 before and after allergen challenge. These data indicate that mild inflammation to allergen elevates HSP27 stress protein levels, thereby potentially protecting epithelial function from additional adverse conditions.

IL-1 beta release from cultured bronchial epithelial cells and bronchoalveolar lavage cells from allergic and normal humans following segmental challenge with ragweed.

This work investigated whether interleukin 1 beta (IL-1 beta) release from epithelial cells is modulated by antigen challenge in vivo, and inflammatory cells in vitro. Bronchial epithelial cells were obtained before and after segmental allergen challenge in allergic and normal individuals, and were cultured with and without autologous bronchoalveolar lavage (BAL) cells. IL-1 beta in culture medium was quantitated by enzyme-linked immunoabsorbent assay (ELISA). Pre-challenge IL-1 beta levels from epithelial cells were similar in allergic (4.4 +/- 0.8 pg/ml, n = 32) and normal (6.8 +/- 1.7 pg/ml, n = 17) subjects. IL-1 beta levels were significantly elevated from epithelium with BAL cell co-culture vs without co-culture in both subject groups (allergic, 13.2 +/- 2.3 pg/ml, P = 0.006; normal: 16.4 +/- 4.0 pg/ml, P = 0.007). Post-challenge IL-1 beta from epithelial cells without BAL cells was increased in both groups, but significant only for allergic subjects (P = 0.003). Post-challenge IL-1 beta from epithelial with BAL cells changed little for allergic subjects (13.8 +/- 2.4 pg/ml), and increased for normal subjects (20.0 +/- 4.8 pg/ml). Decreased production of tumour necrosis factor alpha (TNF-alpha) was observed in allergic subjects (day 1: 447 pg/ml vs day 2: 258 pg/ml). Moreover, pre-challenge release of TNF-alpha and IL-1 beta levels from epithelial + BAL cells correlated well for allergic (r = 0.84) and normal subjects (r = 0.6), but post-challenge release correlated only in normal subjects (r = 0.90). These results indicate that bronchial epithelial cells and BAL cells interact, modulating release of these inflammatory cytokines.