Utility of six fungicides for selective isolation of Evlachovaea sp. and Tolypocladium cylindrosporum

The effects of the fungicides dodine, benomyl, thiabendazole, mancozeb, cupric sulfate, and copper oxychloride were examined in vitro upon germination and further development of Evlachovaea sp. and Tolypocladium cylindrosporum. Fungicidal activity depended on concentrations and varied among products, fungi and the strains tested. Depending on the fungicidal concentration, germination of conidia was induced but germlings produced neither mycelium nor new conidia. There was a good recovery of both Evlachovaea sp. and T. cylindrosporum from previously sterilized soils with fungicide-supplemented medium. Fungi were resistant to copper oxychloride up to 30 g/l, and this fungicide was found to have no utility for a selective medium. Minimal fungicide concentrations for successful isolations were 1 mg/l for benomyl, 200 mg/l for cupric sulfate, 50 mg/l for dodine, 100 mg/l for mancozeb, and 4 mg/l for thiabendazole. Thiabendazole, which is easy to obtain and can be used in low quantities, showed the greatest utility for a selective medium with these entomopathogenic fungi. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The control of apple canker (Nectria galligena) in a young orchard with established infections

In a factorial experiment on canker control the efficiency of phenylmercurie nitrate (PMN) applied at leaf fall, before bud burst or both was compared with the application of dodine, dithianon, triforine, thiabendazole, benomyl or carbendazim in May and June. Of the total number of cankers which developed in unsprayed trees 76–78% resulted from infections in April to August of each year of the experiment. Infection was reduced significantly by all of the fungicides applied in summer. Of these carbendazim was outstanding, controlling both summer and autumn infections. Sporulation throughout the summer was suppressed by carbendazim and to a lesser extent benomyl, but whereas the suppressant effect of carbendazim persisted until long after leaf fall that of benomyl was evident only until August. Dodine, dithianon, triforine and thiabendazole had no significant effect on spore production. Dithianon and dodine showed highest toxicity to the germination of Nectria galligena spores of all fungicides used in summer. All of the fungicides controlled apple scab (Venturia inaequalis) although thiabendazole was the least effective. Autumn applications of PMN reduced canker incidence but their value alone was not as great as carbendazim, dithianon, benomyl or dodine applied in summer. PMN applied in spring reduced the number of cankers in trees receiving no other fungicides but tended to increase the incidence of infections, particularly in autumn, when used in conjunction with fungicides applied in May and June. PMN applied before bud burst reduced sporulation of N. galligena for a few weeks, after which production resumed and in late summer and autumn actually exceeded the controls. None of the fungicides had any direct effect on leaf fall. Infection of the crotch and basal leaf scars was more common in summer than in autumn whereas infection of leaf scars above the basal region was more common in autumn than in summer. The rootstocks of many of the trees became infected via callus tissue associated with adventitious root development and this was controlled by the carbendazim treatment.     

Effects of fungicides on the growth and conidial germination of Colletotrichum coccodes and on the development of black dot disease of potatoes

The effects of 10 fungicides on the growth of Colletotrichum coccodes in agar culture and on the germination of conidia was investigated. In field experiments in 1990 and 1991 the extent to which treating black dot-affected potato seed tubers with fungicides affected the development of the disease on stem bases, roots and tubers was assessed. Black dot was also assessed on plants from field trials in 1990 which were designed to investigate the efficacy of the soil sterilant 1,3-dichloropropene (Telone) and two nematicides, aldicarb (Temik) and ethoprophos (Mocap). Prochloraz and fenpiclonil were the most effective fungicides in decreasing the size of C. coccodes colonies on agar. Imazalil, propiconazole and dichlorophen were also effective, but at higher concentrations, whereas tolclofos-methyl, thiabendazole and benomyl were only moderately effective. Resistant sectors developed from inhibited colonies on agar containing fenpiclonil and tolclofos-methyl. Conidial germination was prevented at 1 mg/litre fenpiclonil and 5 mg/ litre dichlorophen; imazalil, benomyl and thiabendazole were also moderately effective. Fenpiclonil and propiconazole seed tuber treatments consistently decreased black dot infection on roots, stem bases and daughter tubers early in the season, but only fenpiclonil decreased disease on tubers at harvest in October. Propiconazole also delayed emergence and decreased stem numbers. Soil treatment with 1,3-dichloropropene, aldicarb or ethoprophos had no effect on black dot but Rhizoctonia solani tuber infection and black scurf were increased.     

Comparison of fungicides for the control of Rhizoctonia solani causing datnping-off of mung bean (Phaseohis aureus)

Of 41 fungicides tested in the laboratory, copper carbonate, copper sulphate, mercuric chloride, Agrosan GN, quintozene, kasugamycin, carboxin, pyracar-bolid, carbendazim, chloroneb, benomyl, Ohric, RH 893 (2-n-octyl-4-isothiazole-3-one) and Terrazole were most inhibitory to the mycelial growth of Rhizoctonia solani on Czapek's agar plates and had EC₅₀ values of less than 1 μg a.i./ml, while copper oxychloride, Udonkor, zineb, ziram, F 319 (3-hydroxy-5-methyl isoxazole) and anilazine were much less toxic, ziram being least inhibitory with an EC₅₀ of 214 μg a.i./ml. Of 17 fungicides tested in the greenhouse as seed treatments, thiabendazole, carbendazim, benomyl, thiophanate-methyl, dichlozoline and Ohric gave 80–90% control of damping-off of mung bean seedlings. A single soil drench with thiophanate-methyl and two drenches with benomyl gave about 90% disease control, More seedlings with R. solani infection survived when thiophanate-methyl was used as a post-inoculation soil drench than when benomyl or chloroneb were used.     

Growth regulatory effect of three benzimidazole fungicides on the germination of celery (Apium graveolens) seeds

Seeds of four celery cultivars did not germinate when incubated in buffered solutions in the dark at 22 °C. This high-temperature-induced dormancy was broken with either of the systemic fungicides benomyl or BAS 3460 F (mainly methyl-2-benzimidazole carbamate) when used in combination with a mixture of the gibberellins A₄ and A₇ (GA_4/7). The fungicide thiabendazole was ineffective in these tests.     

Efficacy of benzimidazole and related fungicides against Rhizoctonia solani and R. bataticola

Five formulations of four benzimidazole derived fungicides, carbendazim, benomyl, thiophanate methyl and methyl 4-[2-(2-dimethylamino acetamide) phenyl]-3-thioallophanate were compared for their toxicity towards two pathogenic isolates of Rhizoctonia solani and three of R. bataticola. The isolates of two fungi showed significant differences in mycelial growth inhibition by the five fungicides. Benomyl and carbendazim were most inhibitory to all isolates of both fungi while the sesame isolate of R. bataticola was least sensitive to all fungicides. Disease control (90%) was obtained with low concentrations of benomyl against root rot of cowpea caused by R. solani, and with thiophanate methyl against root rot of sesame and sunflower, and leaf blight of mung bean caused by R. bataticola. The spread of stalk-end rot of sunflower heads was best checked with a spray of thiophanate methyl. The results suggest that benzimidazole fungicides having similar toxophores act differently for disease control in different host-parasite combinations.     

EFFECTS OF BENLATE ON FERN GAMETOPHYTE DEVELOPMENT

The widely used fungicide, benlate, was tested for its effect(s) on gametophyte development in the fern Ceratopteris thalictroides (L.) Brongn. The active ingredient of benlate, benomyl, represents 50% of the fungicide by weight. Seven concentrations of benomyl were tested on C. thalictroides gametophytes: 0, 0.001, 0.01, 0.1, 1.0, 10.0 and 100 mg/l. Five developmental stages were observed for possible effects of benomyl. These were 1) germination, and the initiation of 2) antheridia, 3) notch meristems, 4) archegonia, and 5) sporophytes. Overall inhibition was greatest at 100 mg/l benomyl. At 10 mg/l and 100 mg/l, sporophyte initiation was completely blocked. This was probably the consequence of two characteristics found only on gametophytes growing on medium containing 10 mg/l and 100 mg/l benomyl. These characteristics were lack of sperm motility and production of callus growths in the areas proximal to the notch meristems, just proximal to the younger archegonia. Besides blocking the completion of sexual reproduction, the highest concentrations tested also produced smaller (cell number) and chlorotic gametophytes (especially at 100 mg/l). The bigametophyte population (made up of hermaphroditic and male gametophytes) was changed from 51% hermaphrodites (at 0 mg/l benomyl) to 26% hermaphrodites at 100 mg/l. This would, since only hermaphrodites possess archegonia, also decrease the potential for the production of sporophytes.     

Susceptibility of Metarhizium spp. and other entomopathogenic fungi to dodine-based selective media

The fungicide dodine has been widely used in selective media to isolate entomopathogenic fungi (EF) from contaminating microorganisms, primarily bacteria and non-entomopathogenic fungi. In order to isolate the fungus Metarhizium acridum from soil for grasshopper and Mormon cricket control in the western USA, the susceptibility of M. acridum was compared with two Metarhizium spp. and other EF species. The isolates were inoculated onto mycological media with concentrations of dodine ranging from 0.0001 to 0.03% active ingredient (A.I.). In addition, susceptibilities of five Metarhizium spp. isolates to two sources of dodine, Syllit® commercial fungicide (65% A.I.) and Sigma® reagent grade (99% A.I.), were compared using Czapek agar medium. Responses to the two dodine sources were virtually identical. Accordingly, subsequent experiments used the less expensive Syllit dodine. Three media [Czapek, potato dextrose agar plus yeast extract (PDAY) and oatmeal agar] were evaluated for appropriateness as the base in selective media. Germination of all three of the M. acridum isolates tested was almost completely inhibited by dodine concentrations of 0.002% A.I. in Czapek or 0.006% A.I. in PDAY. On the other hand, M. robertsii and M. anisopliae isolates were considerably more tolerant, with germination not being inhibited until 0.010% A.I. in Czapek or 0.030% A.I. in PDAY. The higher vulnerability of the isolates to low concentrations of dodine in Czapek medium suggests that this medium would be less effective than PDAY in a selective medium. Oatmeal agar greatly improved fungal growth, but the levels of inhibition were lower. Therefore, PDAY was selected as the best selective basal medium. The lowest concentration that inhibited a common soil-inhabiting fungus, Aspergillus nidulans, was 0.001% A.I. Dodine tolerances were highest with M. robertsii, M. anisopliae, and Beauveria bassiana, followed by Isaria fumosorosea and Lecanicillium spp. The least tolerant EF isolates were M. acridum.     

A field study using the fungicide benomyl to investigate the effect of mycorrhizal fungi on plant fitness

Summary The effect of vesicular-arbuscular mycorrhiza (VAM) on the fecundity ofVulpia ciliata ssp.ambigua was investigated at two field sites in eastern England by applying the fungicide benomyl to reduce VAM infection. The application of benomyl at the two sites produced very different results. At one site the application of the fungicide reduced the fecundity of plants whereas at the other fecundity was increased. At the first site the reduction in fecundity was linked to a significant reduction in VAM infection on the sprayed plants. The mechanism of the benefit associated with the VAM infection is however unclear: there was no treatment effect on morphology or on phosphorus inflow. At the second site, where fecundity was increased, there was only a negligible amount of VAM infection amongst the unsprayed plants and it is suggested that the increase in fecundity with the application of benomyl may have resulted from a reduction in infection by other, presumably pathogenic, fungi. The value of VAM fungi to the host plant may therefore not be restricted to physiological benefits. They may also provide protection to the plant by competing for space with other species of pathogenic fungi.     

Effect of five fungicides with different modes of action on cobweb disease (Cladobotryum mycophilum) and mushroom yield

The fungicides chlorothalonil, metrafenone, prochloraz‐Mn, thiabendazole and thiophanate‐methyl were tested in vitro and in vivo for their effect on Cladobotryum mycophilum, the mycoparasite that causes cobweb disease in white button mushroom. In vitro experiments showed that metrafenone (EC₅₀= 0.025 mg L^?1) and prochloraz‐Mn (EC₅₀= 0.045 mg L^?1) were the most effective fungicides for inhibiting the mycelial growth of C. mycophilum. Selectivity indexes of the tested fungicides on both C. mycophilum and Agaricus bisporus indicated that metrafenone was also the most selective fungicide, while chlorothalonil was the most toxic fungicide against A. bisporus mycelium. The in vivo efficacy of fungicides for controlling cobweb was evaluated in three mushroom cropping trials, which were artificially inoculated with C. mycophilum (10⁶ conidia m^?2). Prochloraz‐Mn provided good control, although the surface colonised by cobweb reached 12% by the end of the crop cycles. None of the inoculated cropping trials treated with metrafenone showed any cobweb disease symptoms, and neither were any significant phytotoxic effects on mushroom yield recorded. These results indicated that metrafenone can be used as an alternative to prochloraz‐Mn in the control of cobweb disease.     

The disappearance of benomyl from mushroom casing

The failure of Benlate (50% benomyl) to control Mycogone perniciosa on mushroom farms where isolates of the organism were benomyl sensitive was investigated. A comparison was made of carbendazim levels, the major fungitoxic breakdown product of benomyl, in unsterilised casing, autoclaved casing and autoclaved casing mixed with a small proportion (1.7%) of casing previously treated with benomyl but from which all detectable levels of carbendazim had disappeared. Added benomyl was effective in controlling M. perniciosa in autoclaved casing, less effective in unamended casing and ineffective in the amended casing mixture. The recovery of carbendazim from these treatments was directly related to disease occurrence. Studies on farms where benomyl or other carbendazim generating fungicides had been used indicated that a lack of control of M. perniciosa was associated with the disappearance of fungicide from the casing before cropping began. This was not so on a farm where carbendazim generating fungicides had not been used before these investigations. Thiabendazole, a fungicide closely related to the carbendazim generators showed only a slight decline in concentration in casing capable of degrading benomyl. Bacteria isolated from the casing were examined in vitro for their ability to degrade benomyl. Five groups of bacteria were compared and three bacteria, Pseudomonas putida, Pseudomonas aeruginosa and an unidentified member of the Entero-bacteriaceae were most able to degrade benomyl.     

Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion

The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T.␣atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 g/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in␣vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 g/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T.␣atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.This work was presented as an oral lecture in section ‘Agriculture, Soil, Forest Microbiology’ at the BioMicroWorld2005 conference.     

Fungicides and some biological controller agents effects on the growth of fusarium oxysporum causing paprika wilt: Wirkung von fungiziden und einigen biologischen kontrollwirkstoffen auf das wachstum des paprikawelke verursachenden fusarium oxysporum

Soil samples from both healthy and diseased paprika roots were tested to identify their mycoflora. Thirty-one species belonging to 16 genera were collected from rhizosphere and rhizoplane samples. The most frequently isolated fungi were Aspergillus flavus, A. niger, A. terreus, Fusarium oxysporum, Penicillium jensenii and Trichoderma harzianum. Fusarium oxysporum was the most common Fusarium species in the rhizoplane samples of diseased roots and identification was confirmed by RAPD-PCR technique. Trichoderma harzianum, T. pseudokoningii and Glioclaium roseum were chosen to study their biological control efficiency against Fusarium oxysporum. These fungal species reduced the percentage of seedling infection to 25, 40 and 50%, respectively. With the increasing of fungicide (Folicur and Ridomil) doses the dry weight of F. oxysporum decreased. Also, the increasing of fungicide dose lead to a slight decrease in the dry weight of T. harzianum, T. pseudokoningii and Glioclaium roseum.     

A Comparative Study of Caffeine Degradation by Four Different Fungi

The objective of the present study is to investigate the caffeine-degrading abilities of different fungi and to apply this knowledge to environmental remediation and industrial decaffeination process. Chrysosporium keratinophilum, Gliocladium roseum, Fusarium solani, and Aspergillus restrictus were isolated from the coffee pulp obtained from a coffee estate. Pure cultures of fungi were isolated on standard conventional potato dextrose broth (PDB) medium and authenticated. Pure cultures were subjected to a caffeine tolerance study at different concentrations of caffeine (1–8 g/L) in potato dextrose agar (PDA) and minimal media. On PDA, Fusarium solani could tolerate caffeine concentration up to 8 g/L, whereas Chrysosporium keratinophilum, Gliocladium roseum, and Aspergillus restrictus could tolerate up to 6 g/L. On minimal agar medium containing different concentrations of caffeine (1–8 g/L), Fusarium solani tolerated up to 8 g/L and the other fungi up to 2 g/L. A time-bound caffeine degradation study was undertaken at 1 g/L concentration of caffeine and glucose in nitrogen-containing and nitrogen-free liquid minimal media by subjecting the four fungi to shake flask culture at 120 rpm and 30°C. Degradation of caffeine up to 7 days at 24-h intervals was analyzed by high-performance liquid chromatography (HPLC). Gliocladium roseum followed by Aspergillus restrictus showed maximum degradation of caffeine at 0.47 and 0.3 mg/ml, respectively, by 96 h in nitrogen-containing minimal medium, whereas Fusarium solani showed maximum degradation of caffeine by 48 h (0.35 mg/ml) and Chrysosporium keratinophilum by 72 h (0.29 g/ml). In nitrogen-free minimal medium, Chrysosporium keratinophilum showed maximum degradation of caffeine at 72 h (0.45 mg/ml), followed by Gliocladium roseum, Fusarium solani (0.3 mg/ml), and Aspergillus restrictus (0.25 mg/ml) at 96 h. Overall, Chrysosporium keratinophilum showed a comparatively higher rate of caffeine degradation in minimal medium with or without a nitrogen source as compared with the other three fungi, indicating that nitrogen affects caffeine metabolism.     

In vitro compatibility of Beauveria bassiana strain ATCC 74040 with copper fungicides

Copper fungicides and mycoinsecticides based on entomopathogenic fungi Beauveria spp. are the most common pesticides used in organic crop production systems. The in vitro effects of the copper fungicides copper oxide, copper hydroxide, copper oxychloride, copper sulphate, dicopper chloride trihydroxide and tribasic copper sulphate were investigated for mycelial growth, sporulation and conidial germination of the ATCC 74040 commercial strain of Beauveria bassiana. Mycelial growth was evaluated on potato dextrose agar plates with 100%, 75%, 50%, 25%, 12.5%, 6.25% and 0% of the recommended application rates of each fungicide at 15 and 25°C. Sporulation and conidial germination were determined at the recommended field doses of each fungicide at 25°C. All copper fungicides had fungistatic or fungicidal effects on B. bassiana that varied according to the dose. Only in two cases, copper oxide at 15°C and copper hydroxide at 25°C, at the lowest concentration of 6.5%, was mycelial growth not statistically significantly inhibited. Inhibition of mycelial growth depended both on the fungicide and its concentration, and partly on temperature. Both sporulation and conidial germination of B. bassiana were significantly inhibited by all fungicides. All fungicides inhibited the sporulation in a similar way (99.8%–100%). With the exception of copper oxychloride (inhibition, 13.6%), the other fungicides showed high detrimental effects on conidial germination (inhibition, 91.7%–100%). The fungus was strongly affected by some fungicides even at the lowest doses. The biological index used for the B. bassiana with copper fungicides ranged from 0.6 (copper sulphate) to 18.1 (copper oxychloride). Therefore, the tested fungicides were classified into the upper half of the highly toxic (T) category and are considered incompatible with the entomopathogenic fungus B. bassiana strain ATCC 74040 under in vivo experimental conditions. These results need to be further verified in vitro under both greenhouse and open-field conditions.     

The effect of treating seed potato tubers with benzimidazole, imidazole and phenylpyrrole fungicides on the control of rot and skin blemish diseases

Over 6 yr the effectiveness of imazalil, prochloraz and fenpiclonil, applied either alone or in a mixture, in controlling gangrene, dry rot, skin spot and silver scurf on potato tubers in store was compared with that of 2-aminobutane and thiabendazole. An assessment was also made of their efficiency in controlling the development of these diseases on the daughter tubers of seed tubers treated at harvest or before planting. Overall, 2-aminobutane was more effective in controlling gangrene (Phoma foveata) in store than the spray-applied fungicides. Deposits of imazalil, thiabendazole and fenpiclonil were greater when sprays were applied with an electrostatic sprayer than with a hydraulic sprayer. The opposite was found with the mixture of prochloraz Mn and tolclofos-methyl. More effective gangrene control was associated with the highest deposits. Fenpiclonil and the mixture of thiabendazole and imazalil were more effective in controlling dry rot (Fusarium solani var. coeruleum) than imazalil alone. The development of dry rot was, however, increased by 2-aminobutane treatment on eight out of 14 stocks. 2-aminobutane gave the greatest reduction (83%) in the severity of skin spot during storage whereas thiabendazole alone, and the mixture of thiabendazole and imazalil, gave mean reductions of 70% and 65% respectively. This mixture and fenpiclonil gave the greatest reduction in the severity of silver scurf although, in general, reductions in silver scurf with fungicide treatment were less than with skin spot. The type of sprayer used to apply a fungicide did not affect the effectiveness of the fungicides in controlling either skin spot or silver scurf on tubers in store, or on the daughter tubers. The incidence of gangrene and dry rot on daughter tubers was not reduced consistently by fungicide treatment of seed tubers of the six stocks tested. However, the severity of skin spot and silver scurf was reduced by fungicide treatments of all eight stocks but the reduction in disease was greater for skin spot than for silver scurf. All fungicides gave reductions in the severity of skin spot, and fenpiclonil and the mixture of thiabendazole and imazalil were the most effective for silver scurf. The effectiveness of the fungicides in controlling these diseases was similar for seed treated at harvest and that treated before planting.     

Endophytic fungi from Musa acuminata and their reintroduction into axenic plants

Sixteen fungal taxa were isolated from 2400 leaf fragments of mature Musa acuminata plants collected from three different localities in São Paulo State, Brazil. The most frequently found endophytic fungi were Xylaria sp., Colletotrichum musae and Cordana musae. The standard distribution of endophytes was similar in the three localities. Spontaneous resistant mutants to the fungicides thiabendazole (thi-1) and benomyl (ben-5), were obtained from Colletotrichum musae wild-type isolates. Equal amounts of conidia mixtures of combinations of wild-types and mutants were reintroduced in axenic banana plantlets derived from tissue culture; thi-1 showed a selective advantage when compared to both wild-types used. For ben-5, no significant difference was found. Results showed that wild-types and spontaneous mutants of endophytic fungi as Colletotrichum musae, can be successfully reintroduced in plantlets derived from tissue culture.     

Benomyl: A broad spectrum fungicide for use in plant cell and protoplast culture

The systemic fungicide methyl-1-(butylcarbamoyl)-2-benzimidazole carbamate (benomyl), is a broad spectrum fungicide. Benomyl at concentrations up to 50 mg/l does not inhibit the growth of suspension cultures ofNicotiana tabacum, Datura innoxia, Daucus carota, Glycine canescens, andSolanum tuberosum nor growth ofN. tabacum orN. plumbaginifolia protoplasts if benomyl is dissolved by autoclaving or boiling. Addition of benomyl dissolved in dimethyl sulfoxide results in a visible toxicity. Benomyl, at 6.25–50 mg/l preventsPenicillium spp. growth in both protoplast and cell cultures and can be used to remove fungal contaminates after one to three transfers without visibly retarding plant cell growth. Due to the broad spectrum of fungicidal activity, and nontoxicity at high concentrations when dissolved by boiling or autoclaving, benomyl can be used effectively to control or prevent fungal contamination in plant cell and protoplast cultures.     

Influence of commercial fungicides on the germination,growth and virulence of four species of entomopathogenic fungi

The influence of 15 commercially available fungicides on the germination, growth and virulence of Metarhizium anisopliae, Beauveria bassiana, Isaria fumosorosea, and Lecanillium longisporum was evaluated. The influence of the fungicides on conidial germination was dependant on the fungicide type and dose. Most fungicides retarded conidial germination of all the fungi tested at 10× and at the recommended rate of application, however, their toxicity declined at lower concentrations. Most of the fungicides inhibited mycelial growth of B. bassiana, whereas L. longisporum growth was unaffected. Only two and eight fungicides influenced mycelial growth of I. fumosorosea and M. anisopliae, respectively. None of the fungicides influenced the virulence of B. bassiana and L. longisporum, however, tolylfluanid and azoxystrobin reduced the virulence of M. anisopliae and I. fumosorosea, respectively. These studies clearly show that certain fungicides have the potential to inhibit germination of entomopathogenic fungi in vitro but appear to have little or no effect on their virulence against target insects.