Selection of fungal biological control agents of Sclerotium cepivorum for control of white rot by sclerotial degradation in a UK soil

A range of fungal isolates was tested in a three-stage screening system for their ability to degrade sclerotia of Sclerotium cepivorum on agar and in soil, and to reduce white rot disease on onion seedlings. Biological control agents (BCAs) were identified that could degrade up to 60% of sclerotia in soil and significantly reduce white rot disease on onion seedlings. The efficacy of the BCAs was enhanced when applied as wheat bran cultures compared with spore suspensions, and two of the best BCAs from the screening procedures were both identified as Trichoderma viride (L4, S17A). When L4 and S17A were fluid-drilled in guar gum with bulb onion seed in the field white rot symptoms were significantly reduced, but stem base applications applied mid-season had little effect. The strategy of selecting and using BCAs that degrade sclerotia of S. cepivorum and integration with other control methods is discussed.     

The effect of fungicides on fungal antagonists of onion white rot and selection of dicarboximide-resistant biotypes

The sensitivity of four fungal antagonists (Chaetomium globosum, Trichoderma harzianum, T. viride, Trichoderma sp.) to six fungicides was evaluated. The fungi were insensitive to captan, mancozeb and thiram but were sensitive to benomyl (EC₅₀ < 0 3/μg/ml) and the two dicarboximides iprodione and procymidone (EC₅₀ < 3 3/μg/ml). Conidia and mycelia of the four fungi were exposed to a series of ultraviolet irradiations in order to select biotypes resistant to benomyl and iprodione. Resistance to benomyl was not induced but a number of biotypes of each fungus were isolated from irradiated cultures that showed resistance to iprodione (EC₅₀ values of up to 56·4, 177·8, 171·5 and 216·4/μg/ml for Chaetomium globosum, Trichoderma harzianum, T. viride and Trichoderma sp., respectively). With three exceptions, all selected biotypes retained their fungicide-resistance after being extensively cultured on fungicide-free medium. In general, growth of the fungicide resistant biotypes on MEA plates and in MYE liquid medium was equal to that of the corresponding wild-type strains. One resistant biotype of Trichoderma sp. (C62UV3) grew significantly (P < 0·05) better both on solid and in liquid medium. This resistant biotype was also observed to have greatly enhanced conidial production on agar plates. All fungicide resistant-biotypes retained the ability to antagonize the pathogen Sclerotium cepivorum in dual culture and, in two instances (A53UV1, A53UV5), exhibited greater antagonism as evidenced by the production of larger inhibition zones. Similarly, with two exceptions, the resistant biotypes retained the ability to control onion white rot disease caused by S. cepivorum in the glasshouse. In particular, two fungicide-resistant biotypes (Trichoderma sp. C62UV3 and Chaetomium globosum A53UV1) reduced white rot of onion more effectively than did their respective wild-type parents.     

枯草芽孢杆菌（Bacillus subtilis）S9对植物病原真菌的溶菌作用

本研究共分离了976株细菌分离物，发现来自甘蔗根围的1株枯草芽孢杆菌(Bacillus subtilis)S9对立枯丝核菌(Rhizoctonia solani)、终极腐霉(Pythium ultimum)和西瓜枯萎病菌(Fusarium oxysporum f.sp.niveum)在PDA平板的对峙培养过程中不形成抑菌圈，但4d后可使上述植物病原真菌的菌丝溶解。扫描电镜观察发现S9菌株在待测的立枯丝核菌表面形成了溶菌斑。S9菌株对立枯丝核菌的作用过程是通过吸附在病原真菌的菌丝上，并随菌丝生长而生长，而后产生溶菌物质消解菌丝体。液体共培养测定也证明了S9菌株具有上述作用。本研究还发现，S9菌株对植物病原真菌的拮抗真菌绿色木霉(Trichoderma viride)、鲜红毛壳菌(Chaetomium cupreum)和球毛壳菌（Chaetomium globosum）的生长没有影响。盆栽试验证明了S9菌株可有效地控制立枯丝核菌(R．solani)引起的番茄苗期病害。S9菌株与其它拮抗真菌混合具有促进防治植物病原真菌引起的植物病害的潜力。     

Effect of environmental factors and Sclerotium cepivorum isolate on sclerotial degradation and biological control of white rot by Trichoderma

Laboratory assays demonstrated that two isolates of Trichoderma viride and one isolate of Trichoderma pseudokoningii degraded up to 80% of sclerotia of four isolates of Sclerotium cepivorum in a silty clay soil, and also degraded up to 60% of sclerotia in three other soil types. Relationships were defined between the degree of sclerotial degradation by the two T. viride isolates in the silty clay soil and both temperature and soil water potential. Sclerotia were degraded between 10 and 25°C at −0·00012 MPa, but there was little activity of T. viride at 5°C or at −4 MPa. Degradation of S. cepivorum sclerotia also occurred in the absence of Trichoderma at soil water potentials approaching saturation . Experiments using onion seedling bioassays showed that the efficacy of Trichoderma isolates for the control of white rot using the same selection of soils and S. cepivorum isolates was variable, but that there was significant disease control overall. The importance of environmental factors and pathogen isolate in relation to effective biological control of white rot is discussed.     

哈茨木霉拌种对冬小麦生长、土传病害及根际真菌群落的影响

为了明确哈茨木霉LTR-2拌种处理冬小麦的田间效果,为木霉拌种剂的推广应用提供依据,本试验从2016年－2018年连续3年,研究了哈茨木霉LTR-2拌种对小麦出苗率、幼苗生长、小麦纹枯病和茎基腐病发生情况和产量的影响,通过高通量测序和FUNGuild预测分析了木霉拌种对小麦根际土壤中真菌群落组成的影响。结果表明,哈茨木霉LTR-2拌种可以提高小麦的出苗率和冬前分蘖数;对小麦纹枯病的平均防效60%以上;对小麦茎基腐病的平均防效65%以上,优于6%戊唑醇悬浮种衣剂;与不拌种对照相比,哈茨木霉LTR-2拌种处理增产4.3%~6.34%,增产效果略高于6%戊唑醇悬浮种衣剂;木霉拌种可以降低小麦根际土壤中病原真菌的相对丰度,特别是土壤中镰孢属真菌的相对丰度。因此,哈茨木霉LTR-2可以作为化学拌种剂的绿色替代产品用于小麦生产。     

Characteristics of alginate pellets formulated with Trichoderma and Gliocladium and their effect on the proliferation of the fungi in soil

Pelletized formulations of wheat bran or kaolin clay in an alginate gel containing conidia, chlamydospores, or fermentor biomass (FB) of several isolates of the biocontrol fungi Trichoderma spp. and Gliocladium virens were prepared. The ability of fungal propagules within the pellets to proliferate in soil was determined. Higher population densities were obtained when alginate pellets added to soil contained chlamydospores rather than condia, and bran rather than kaolin as the bulking agent. The active ingredient in pellets prepared from FB was approximately 5% biomass by weight and contained many chlamydospores. Colony-forming units (cfu) ranged from 10⁶'to 10¹⁰/g of soil after soil amendment with FB pellets of 12 Trichoderma and G. virens isolates. Population densities were high during the first 3 weeks of incubation and declined only gradually during 9 weeks. Propagules in FB pellets were more viable at 5° than at 25°C. Viability at 25°C remained high (> 70%) after 1 week, but declined to less than 10% after 24 weeks. Despite reduction in propagule viability in stored pellets, numbers of cfu formed after adding these pellets to soil were comparable with those formed from freshly prepared pellets.     

Genetic diversity and identification of race 3 of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lactucae</Emphasis> in Taiwan

Sorghum is an important drought tolerant crop cultivated for food and fodder purposes. Anthracnose disease caused by Colletotrichum graminicola is a major constraint in sorghum productivity in India. Certain antagonistic fungi, that were isolated in the previous study from the rhizosphere and rhizoplane of perennial grasses in India, were studied for their antagonism in vitro to C. graminicola, root colonization ability and rhizosphere competence. Out of 138 isolates tested, 89 were antagonistic. Fifteen fungal isolates with greater than 70 % in vitro inhibition zone to the pathogen tested positive for root and rhizosphere colonization abilities. Three isolates – Chaetomium globosum isolate 57, Trichoderma harzianum isolate 184 and Fusarium oxysporum (NSF isolate 9) with prominent biocontrol potentials were tested for the control of sorghum anthracnose in greenhouse and field. Chaetomium globosum, Trichoderma harzianum and Fusarium oxysporum isolates decreased seedling mortality, and incidence and severity of disease at different growing stages. They promoted plant growth (dry biomass- 45.3, 40.0 and 46.7 %) and increased yield (grain biomass- 33.3, 23.8 and 49.2 %) respectively, over control in field. The population of the above fungi in soil was moderately high at harvest stage. The present investigation revealed that fungal isolates from rhizosphere and rhizoplane of perennial grasses could be employed to manage anthracnose and enhance plant growth and yield potentialities in sorghum, at the same time.     

Suppression of Diaporthe phaseolorum f. sp. meridionalis in soybean stems by Chaetomium globosum

The effect of two isolates of Chaetomium globosum (CgA-1, a producer and CgNA, a non-producer of antibiotic in vitro ) on the sporulation of pycnidia and survival of Diaporthe phaseolorum f. sp. meridionalis ( Dpm ) in soybean stems was determined under conditions of intermittent wetting and drying. Dpm -colonized stems placed on the surface of field soil or of quartz gravel were exposed to a 12- or 24-h wet period followed by a 48-h drying period until a cumulative wetness duration of 120, 240 or 480 h was attained. Both isolates reduced the pycnidia sporulation index to zero within 120 h of cumulative wetness duration if the wet period was 24 h. With a 12-h wet period the sporulation index decreased gradually, to between 4 and 8% after 480 h of cumulative wetness. In general, on the soil surface and with a 12-h wet period, CgA-1 reduced sporulation more efficiently than CgNA whereas with a 24-h wet period there was no difference between the isolates. Recovery of Dpm from C. globosum -treated stems decreased gradually with increase in wet period and cumulative wetness duration. The recovery rate from stems treated with isolate CgA-1 was always significantly less than that from stems treated with CgNA. In glasshouse experiments, disease incidence and the proportion of the stem length colonized by Dpm at the V₆ growth stage was reduced significantly when soybean seeds treated with ascospores of C. globosum were sown in Dpm -infested soil. Incorporation of bran inoculum of either of the isolates in the upper soil layer dramatically reduced disease incidence. Isolate CgA-1 reduced the disease more efficiently than CgNA.     

Effects of annual applications of sodium azide on soil fungal populations with emphasis on Trichoderma species

The effect of a granular formulation of sodium azide, applied annually to pine nursery beds at rates of 0, 67.2, and 134.5 kg a. i. ha^?1 under water seal or plastic seal, on soil fungal populations was determined over a 3-year period. Populations of fungi in the soil decreased following application of the sodium azide each year; the greatest decrease occurred at the highest rate of application. Populations of fungi in soil treated with the azide generally remained lower than in the controls throughout each of the 3 years; however, the population disparity between treated and control plots decreased in magnitude with each succeeding year. Populations of Trichoderma spp., in plots treated with 134.5 kg sodium azide ha^?1, increased 2 weeks after treatment each year, and the population peaks increased in magnitude each year. In addition the effect of sodium azide (technical grade > 99%) at concentrations of 0, 2, 5, 10, 20 and 50 μg ml^?1 in potato dextrose agar and blackstrap molasses agar media was determined in vitro for 14 isolates of Trichoderma harzianum. Growth and sporulation differed among the isolates and between the two media tested. Generally, the azide temporarily inhibited growth of the fungi, but the majority of the isolates were able to grow on either medium containing 50 μg sodium azide ml^?1, although sporulation was more profuse on the molasses than on the potato dextrose agar medium.     

Application of Trichoderma and Gliocladium in alginate pellets for control of Rhizoctonia damping-off

Alginate pellets were prepared from wet fermentor biomass of 11 isolates of Trichoderma spp. and Gliocladium virens , with wheat bran as a food base carrier. Pellets with eight of the isolates reduced survival (34-78%) of Rhizoctonia solani in infested beet seed in soil. Pellets containing a T. harzianum isolate (Th-58) and a T. hamatum isolate (TRI-4) were the most effective. All isolates significantly reduced growth of the pathogen from infested beet seed into natural soil. Populations of isolates proliferated in soil to 10⁶−10¹¹ colony-forming units/g (cfu) from propagules within the pellets. Pellets with TRI-4 reduced pathogen survival and growth (>70%) in six different soils and were effective against six R. solani isolates in a natural loamy sand. Survival of R. solani in infested beet seed was not reduced when TRI-4 pellets were added to soil 1-6 weeks before the pathogen; however, saprophytic growth was prevented. Small amounts of biomass (3.0–7.5 g wet weight) in pellets were as effective as a large amount (300 g) in suppressing the pathogen. The storage of pellets for more than 6 weeks at 5 or 25C reduced their effectiveness against R. solani. Pellets prepared with four and three of the 11 isolates prevented damping-off of cotton and sugar beet in the greenhouse, respectively.     

Biological control of Botrytis cinerea on chickpea seed with Trichoderma spp. and Gliocladium roseum: indigenous versus non-indigenous isolates

Establishment of chickpea seedlings from seed inoculated with Botrytis cinerea was increased under controlled environmental conditions by treatment of seed with Gliocladium roseum or Trichoderma virens . An isolate of G. roseum (GrH) from Horsham in the chickpea-growing region of western Victoria outperformed T. virens in three soil types, including one to which T. virens was indigenous. There was no effect of soil moisture on seedling establishment from seed treated with GrH, but recovery of G. roseum from seed in continuously wet soil was lower than from wet/dry soil. A factorial assay with three isolates of G. roseum and the soils from which they were isolated showed no interaction between isolate and soil type and no difference between isolates for establishment or recovery. In a field trial at Horsham sown in winter (July), seed treatment with GrH increased establishment from 26.0% (control) to 37.5%. Biocontrol by GrH was ineffective in a winter-sown field trial at Bundoora, but all three isolates increased establishment from 47.5% (control) to up to 87.5% in a trial at the same location sown in spring (October). The commercial implications of these observations are discussed.     

Comparison of Physical, Chemical and Biological Methods of Controlling Garlic White Rot

Treatment of garlic cloves with tebuconazole (at 1ml of Folicur 25% l^–1) achieved a significant reduction in the rate of disease progress and the final incidence of plant death by Sclerotium cepivorum: garlic yields were improved. Although soil solarization provided the best control of garlic white rot, bringing soil populations of S. cepivorum to negligible levels, similar levels of disease control and garlic yields were achieved when tebuconazole was sprayed to stem bases of plants grown from cloves also treated with tebuconazole. This double treatment almost doubled the yield compared with untreated plants and significantly increased bulb quality under high disease pressure conditions. Soil solarization was also highly effective in a second consecutive crop of garlic, with significant improvements in yield and garlic quality. In contrast, lower levels of disease control were obtained when selected isolates of Trichoderma harzianum and Bacillus subtilis were applied to the soil and cloves respectively.     

Evaluation of a polyclonal antibody immunoassay for detection and quantification of Macrophomina phaseolina

Detection and quantification of Macrophomina phaseolina , causal agent of charcoal/dry root rot disease in many crop plants, was carried out using the ELISA serological technique. Polyclonal antisera raised to water soluble extracts of mycelium, the residual water insoluble mycelial materials or ribosomal proteins were evaluated for specificity and cross-reactivity with 16 common soil fungi by ODD and DAS–ELISA. Soluble and cell wall antisera exhibited strong cross-reactivity with most of the fungal isolates. Ribosomal antibodies were less reactive to common soil fungi except Fusarium oxysporum f.sp. ciceri . Mycelial antigens of M. phaseolina on chickpea roots were detectable with DAS–ELISA at a minimum concentration of 10 ng g⁻¹ at 1 : 100 root : buffer dilution. Quantitative estimation of M. phaseolina on roots was evaluated by ELISA under different temperatures and moisture conditions, and in soil amended with a potential antagonist ( Trichoderma harzianum 25-92). A significant reduction in ELISA values was observed in T. harzianum -amended treatments. This method may be useful for detection and rapid screening of M. phaseolina under different environmental conditions.     

Integration of soil application and seed treatment formulations of Trichoderma species for management of wet root rot of mungbean caused by Rhizoctonia solani

BACKGROUND: The efficacy of seed dressing and soil application formulations from the isolates of Trichoderma viride (IARI P1; MTCC 5369), T. virens (IARI P3; MTCC 5370) and T. harzianum (IARI P4; MTCC 5371) were evaluated individually and in combination in pot and field experiments during the rainy seasons of 2005, 2006 and 2007 for the management of wet root rot (Rhizoctonia solani) and improvement in the yield of mungbean. RESULTS: A seed dressing formulation, Pusa 5SD, and soil application formulations, Pusa Biogranule 6 (PBG 6) and Pusa Biopellet 16G (PBP 16G), based on Trichoderma virens, were found to be superior to other formulations in reducing disease incidence and increasing seed germination and shoot and root lengths in mungbean. In field experiments, a combination of soil application with PBP 16G (T. virens) and seed treatment with Pusa 5SD (T. virens) + carboxin was superior to any of these formulations individually in increasing seed germination, shoot and root lengths and grain yield and reducing wet root rot incidence in mungbean. Seed treatment was more effective than soil application for all the evaluated parameters. The combined application of Pusa 5SD and carboxin was also superior to individual treatment. CONCLUSION: The efficacy of the evaluated formulations against wet root rot of mungbean proved that the integration of soil application of PBP 16G and seed treatment with Pusa 5SD + carboxin is highly effective for the management of wet root rot, increasing plant growth and grain yield of mungbean Copyright © 2011 Society of Chemical Industry     

Management of phytophthora root rot in radiata pine seedlings

Chemical and biological agents were evaluated for their ability to suppress root rot, caused by Phytophthora cactorum , in field-grown radiata pine seedlings in New Zealand. Trials were conducted over two seasons in an area of a forest nursery with a natural infestation of P. cactorum , and a history of root rot. In each season, symptoms of root rot developed during April, one month after root pruning, when seedlings were approximately six months old. In trial one, root rot incidence by mid July 2007 was 9·1% in untreated plots and 8·4% in plots that had been treated with metalaxyl-M/mancozeb (14 kg ha⁻¹) at seedling emergence. Disease incidence was lowest (2·1%) in plots that received seven monthly applications of phosphorous acid (6·5 L ha⁻¹). Other treatments, including seed coating with thiram or Trichoderma spp., and foliar applications of methyl jasmonate, did not control disease. In trial two, effects of treatment timing relative to root pruning were investigated. By late June 2008, three months after root pruning, root rot incidence was 22·2% in the untreated plots. Phosphorous acid was the most effective treatment and almost completely suppressed disease (0·1% incidence) when applied fortnightly from February until May (seven applications). Metalaxyl-M/mancozeb (15 kg ha⁻¹) was not effective (21·4% incidence) when applied five months before root pruning. However, disease incidence was reduced when the chemical was applied one week after root pruning (14·9% incidence) and greater control was achieved (8·2% incidence) when the application rate was increased to 50 kg ha⁻¹.     

Viability and stability of biological control agents on cotton and snap bean seeds

Cotton and snap bean were selected for a multi-year, multi-state regional (south-eastern USA) research project to evaluate the efficacy of both commercial and experimental bacterial and fungal biological control agents for the management of damping-off diseases. The goal for this portion of the project was to determine the viability and stability of biological agents after application to seed. The biological seed treatments used included: (1) Bacillaceae bacteria, (2) non-Bacillaceae bacteria, (3) the fungus Trichoderma and (4) the fungus Beauveria bassiana. Seed assays were conducted to evaluate the following application factors: short-term (< or = 3 months) stability after seed treatment; quality (i.e. isolate purity); compatibility with chemical pesticides and other biocontrol agents; application uniformity between years and plant species. For the bacterial treatments, the Bacillaceae genera (Bacillus and Paenibacillus) maintained the greatest population of bacteria per seed, the best viability over time and the best application uniformity across years and seed type. The non-Bacillaceae genera Burkholderia and Pseudomonas had the least viability and uniformity. Although Beauveria bassiana was only evaluated one year, the seed fungal populations were high and uniform. The seed fungal populations and uniformity for the Trichoderma isolates were more variable, except for the commercial product T-22. However, this product was contaminated with a Streptomyces isolate in both the years that it was evaluated. The study demonstrated that Bacillaceae can be mixed with Trichoderma isolates or with numerous pesticides to provide an integrated pest control/growth enhancement package.     

Biological Control of Plant Pathogens: Present Status and Perspectives in Italy

Interest in the biological control of plant pathogens has been increasing in Italy in the last few years. Current or completed research programmes at some University and/or CNR (National Research Council) Institutions concern the following topics: a) biological control of crown gall with Agrobacterium radlobacter strain 84; b) biocontrol of some forestry diseases caused by fungi; c) integrated biological control of grey mould on grapevine; d) biocontrol of Fusarium dry rot on maize and sorghum and of Rhizoctonia rot on eggplant and tomato with Trichoderma viride; e) seed and root≪ bacterization ≫ as a means of biological control of some fungal diseases of horticultural plants; f)≪ suppressive ≫ effect of some Italian soils to Fusarium oxysporum f.sp. dianthi : g) biologically induced resistance against bacterial, fungal and viral diseases. In this contribution, the results obtained are reported, together with comments on the prospects for practical application of each type of biological control. Evaluation of the reduction in the use of pesticides against the same pathogens is not currently possible, owing to lack of data (except for programmes b) and c) above). Biological control will only become a common and widespread agricultural practice if a number of conditions are satisfied, including the establishment of registration guidelines for microbial and viral pesticides. The criteria for the evaluation of biological control organisms in the agricultural environment must be revised and the technical-legislative problems must be solved. A Commission in the European Community is currently examining the question.     

Control of infection and sporulation ofBotrytis cinerea on bean and tomato by saprophytic bacteria and fungi

Sixty isolates of saprophytic microorganisms were screened for their ability to reduce the severity of grey mould (Botrytis cinerea) infection and sporulation. Isolates of the bacteriaXanthomonas maltophilia, Bacillus pumilus, Lactobacillus sp., andPseudomonas sp. and the fungusGliocladium catenulatum reduced germination of conidia of the pathogen and controlled disease on bean and tomato plants. Their activity under growth room conditions was good, consistent, and similar to the activity of the known biocontrol agent,Trichoderma harzianum T39 (non-formulated). Although the tested isolates may for nutrients with the germinating conidia ofB. cinerea, resistance induced in the host by live or dead cells were also found to be involved. Inhibitory compounds were not detected on treated leaves. Sporulation ofB. cinerea after its establishment on leaves was also reduced by the above mentioned isolates and byPenicillium sp.,Arthrinium montagnei, Ar. phaeospermum, Sesquicillium candelabrum, Chaetomium globosum, Alternaria alternata, Ulocladium atrum, andT. viride. These sporulation-inhibiting fungi did not reduce the infection of leaves byB. cinerea. Most of these selected fungi and bacteria were capable of reducing lesion expansion.     

The potential of nonpathogenic Fusarium oxysporum and other biological control organisms for suppressing fusarium wilt of banana

The aim of this study was to evaluate the ability of nonpathogenic F. oxysporum and Trichoderma isolates from suppressive soils in South Africa to suppress fusarium wilt of banana in the glasshouse. Several biological control agents and commercial biological control products were included in the study. The isolates were first screened in vitro on potato dextrose agar. In glasshouse evaluations, the fungal and bacterial isolates were established on banana roots before they were replanted in pathogen-infested soil, while the commercial biocontrol agents were applied as directed by the supplier. Banana plantlets were evaluated for disease development after 7 weeks. In vitro tests showed none of the nonpathogenic isolates suppressed Fusarium oxysporum f.sp. cubense ( Foc ), while slight suppression was observed with the two Trichoderma isolates. Results of the glasshouse evaluations revealed that two of the nonpathogenic F. oxysporum isolates, CAV 255 and CAV 241, reduced fusarium wilt incidence by 87·4 and 75·0%, respectively. The known biological control agent Fo47 did not suppress Foc significantly. Pseudomonas fluorescens strain WCS 417, known for its ability to suppress other fusarium wilt diseases (WCS 417), reduced disease incidence by 87·4%. These isolates should be further evaluated for potential application in the field, independently and in combination.     

Interaction of Four Antagonistic Fungi with Botrytis aclada in Dead Onion Leaves: A Comparative Microscopic and Ultrastructural Study

ABSTRACT The colonization of dead onion leaves by Botrytis aclada and the fungal antagonists Aureobasidium pullulans, Chaetomium globosum, Glio-cladium catenulatum, and Ulocladium atrum and the interactions between B. aclada and each of the four antagonists were studied at the microscopic and ultrastructural level. This approach was used in an attempt to understand the colonization pattern of these fungi and the nature of the biocontrol activity of the antagonists that have shown a potential to suppress spore production of Botrytis spp. on necrotic plant tissues. When applied alone, B. aclada and U. atrum were found throughout the leaf tissues in high densities after an incubation period of 6 days at 18 degrees C in a moist chamber. C. globosum and G. catenulatum colonized only the outer portions of the leaf, whereas A. pullulans appeared to be concentrated in the leaf stomata. When pathogen and antagonists were applied together, ultrastructural observations revealed that cells of B. aclada were plasmolyzed in the presence of G. catenulatum, suggesting a reaction to antifungal molecules. Antibiosis also seemed to be involved, albeit to a lesser extent, in the antagonistic interactions between B. aclada and A. pullulans or C. globosum. No evidence of direct parasitism was recorded. On the other hand, U. atrum appeared to completely exclude B. aclada from dead onion tissues when both fungi competed for the substrate. Ultrastructural observations of the in vitro interaction between the two fungi did not reveal parasitism or antibiosis by either fungus. Based on previous records of its biocontrol potential and observations of its colonizing properties, it appears that U. atrum can compete for and utilize necrotic tissues rapidly and extensively, thus, excluding competitors without any other antagonistic action.