Adhesion of Nongerminated Botrytis cinerea Conidia to Several Substrata

Conidia of the plant pathogenic fungus Botrytis cinerea adhered to tomato cuticle and to certain other substrata immediately upon hydration. This immediate adhesion occurred with both living and nonliving conidia. Adhesion was not consistently influenced by several lectins, sugars, or salts or by protease treatment, but it was strongly inhibited by ionic or nonionic detergents. With glass and oxidized polyethylene, substrata whose surface hydrophobicities could be conveniently varied, there was a direct relationship between water contact angle and percent adhesion. Immediate adhesion did not involve specific conidial attachment structures, although the surfaces of attached conidia were altered by contact with a substratum. Freshly harvested conidia were very hydrophobic, with more than 97% partitioning into the organic layer when subjected to a phase distribution test. Percent adhesion of germinated conidia was larger than that of nongerminated conidia. Evidence suggests that immediate adhesion of conidia of B. cinerea depends, at least in part, on hydrophobic interactions between the conidia and substratum.     

Growth, zearalenone production and some metabolic activities of Fusarium moniliforme under salt stress

With a increasing salinity, a decrease in the growth rate of Fusarium moniliforme was observed. The percentage of germinated conidia decreased with increasing salinity (% germination ranged from 80.3% at 0.0% NaCl to 0.0% at 15% NaCl). Concentration of 12.5% NaCl produced the highest chlamydospore-like structures. Concentration of 5% NaCl increased zearalenone production which decreased with increasing salt stress. Escherichia coli was more tolerant to the toxin than Bacillus subtilis. The total amount of lipids produced by F. moniliforme changed with increasing the concentration of NaCl in the growth medium. The genomic DNA of the control and treated samples showed a common band of more than 20 kilobases. Similar RAPD-PCR patterns were also produced.     

Fungicidal properties of two saponins from Capsicum frutescens and the relationship of structure and fungicidal activity

Two steroidal saponins have been purified from cayenne pepper (Capsicum frutescens). Both have the same steroidal moiety but differ in the number of glucose moieties: the first saponin has four glucose moieties (molecular mass 1081 Da) and the second contains three glucose moieties (molecular mass 919 Da). Solubility in aqueous solution is less for the saponin containing three glucose moieties than for the one containing four glucose moieties. The larger saponin was slightly fungicidal against the nongerminated and germinating conidia of Aspergillus flavus, A. niger, A. parasiticus, A. fumigatus, Fusarium oxysporum, F. moniliforme, and F. graminearum, whereas, the second saponin (molecular mass 919 Da) was inactive against these fungi. Results indicate that the absence of one glucose molecule affects the fungicidal and aqueous solubility properties of these similar molecules.     

Involvement of Hydrogen Peroxide, Peroxidase and Superoxide Dismutase in Response of Medicago truncatula Lines Differing in Susceptibility to Phoma medicaginis Infection

This study was undertaken to assess the involvement of hydrogen peroxide (H₂O₂), peroxidase (POX; EC 1.11.1.7) and superoxide dismutase (SOD; EC 1.15.1.1) in Medicago truncatula in relation with susceptibility to Phoma medicaginis infection. Several M. truncatula lines were studied in terms of their response to P. medicaginis infection. Fifteen days after inoculation (dai), differences in susceptibility were found. DZA45.5 was the least susceptible line and F83005.5 was the most susceptible line. Microscopic analysis of fungal development was performed in inoculated detached leaves of the DZA45.5 and F83005.5 lines. No significant difference was observed in events from conidia germination to penetration. Differences became apparent during the colonization process as the pathogen was able to sporulate rapidly increasing its concentration on the tissue of F83005.5 in comparison with DZA45.5. To characterize the susceptibility of the two lines, histochemical detection of H₂O₂ was made in detached leaves. H₂O₂ detection showed an early accumulation of this component in cells of DZA45.5 at 1 dai. However, H₂O₂ was detected in few, if any, cells in the tissues of the most susceptible line, F83005.5. The activity of POX and SOD were determined spectrophotometrically in leaves of intact inoculated plants of both lines. Phoma medicaginis inoculation of DZA45.5 and F83005.5 did not affect POX activity level in leaves when compared with control uninoculated plants. SOD activity showed a significant decrease in F83005.5 and DZA45.5 leaves at 4 dai and 9 dai, respectively, in comparison with control plants. In control plants POX activity was significantly higher in the least susceptible line DZA45.5 in comparison with F83005.5. Early and higher production of H₂O₂ and elevated basal POX activity in cells of the least susceptible line, DZA45.5 could explain its ability to be less favourable to the colonization and reproduction of P. medicaginis in comparison with the most susceptible line, F83005.5.     

Influence of pea-root exudates on germination of conidia and chlamydospores of physiologic races of Fusarium oxysporum f. pisi

Sterile root exudates from wilt susceptible and wilt resistant pea cultivars showed no differential effects on spore germination of Fusarium oxysporum Schl. f.pisi (Linf.) Snyd. & Hans, races 1 and 2 which could be correlated with the pathogenicity of a particular isolate to a given cultivar. Uniformly high percentages of germination were obtained with conidia of the two races in aseptic shake culture with exudates collected from resistant or susceptible plants of various ages. Chlamydospores of the two races incubated with exudates under sterile conditions germinated to uniformly high levels irrespective of exudate origin. Conidia and chlamydospores of Fusarium solani (Mart.) Sacc. f. pisi (Jones) Snyd. & Hans., used for comparative purposes, also germinated to high levels in the presence of exudate solutions of all cultivars. Non-specific germination of the two races of F. oxysporum f. pisi occurred in soil when the exudates were supplied to populations of chlamydospores via diffusion units. Germination was lower than that recorded under sterile conditions and was rapidly followed by germling lysis.     

Occurrence and toxigenicity of shape Fusarium moniliforme from freshly harvested maize ears with special references to fumonisin production in Egypt

Using the seed-plate technique, 18 different isolates of Fusarium moniliforme were isolated on pentachloronitrobenzene (PCNB) agar medium from 18 samples of a local variety of corn collected from locations in Minia Governorate. The isolates of F moniliforme were screened for their ability to produce fumonisins on polished rice grains using the solid state fermentation technique. Based on thin layer chromatographic (TLC) analyses using silica gel plates, 14 of the 18 isolates tested produced FB₁ and FB₂ with R _f (0.17) and (0.24), respectively. Concentration of FB₁ was estimated using high performance liquid chromatography (HPLC). Production of FB₁ by the 14 isolates of F. moniliforme tested ranged from 69 to 4495 ppm indicating that mouldy corn may represent a health hazard to consumers. This revised version was published online in August 2006 with corrections to the Cover Date.     

Evidence for Resistance-inducing Compounds in Conidia of Erysiphe graminis f.sp. hordei

Preparations of Erysiphe graminis f.sp. hordei conidia were spray-applied to the first leaf of barley plants which were subsequently challenge inoculated with virulent powdery mildew. The powdery mildew reducing effect of the preparations was assessed by scoring the outcome of the challenge inoculation. Homogenates of ungerminated conidia, germinated conidia, and methanol-water extracts of germinated conidia reduced the number of powdery mildew colonies. Cell wall fragments from ungerminated conidia, germinated conidia, and conidial germination fluid obtained from conidia germinated in aqueous suspension did not reduce the number of powdery mildew colonies. Microsconical analysis of the infection course following challenge inoculation indicated that the powdery mildew reducing effect is due partly to induced resistance.     

In vitro studies on the potential for biological control of Aspergillus flavus and Fusarium moniliforme by Trichoderma species

Culture filtrates of Trichoderma viride and Trichoderma harzianum were inhibitory of Fusarium moniliforme and, to a lesser extent, Aspergillus flavus. The degree of inhibition was, however, dependent on the carbon or nitrogen source incorporated into the medium. Scanning electron microscopy revealed the development of abnormal fruiting structures on exposure to some Trichoderma culture filtrate, while macroscopically, growth restriction and, in the case of A. flavus, altered colony colouration were observed. Based on the results of inverted colony culture, it would appear that some isolates of Trichoderma produce inhibitory volatile compounds. The production of possible antibiotics was also demonstrated. The aggressive behaviour (towards A. flavus and F. moniliforme) demonstrated by Trichoderma spp. may be partly explained by the liberation of extracellular enzymes by these fungi. An isolate of T. viride exhibited amylolytic, pectinolytic, proteolytic and cellulolytic activity. Based on the results of the present investigation, Trichoderma spp. are potential candidates for biocontrol of some mycotoxin-producing fungi, but there exists some doubt as to their osmotolerance within the air-dry seed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pathogenicity of the basidiospores of Filobasidiella neoformans

The chronology of Phomopsis ganjae conidia germination and infection of Cannabis sativa leaves was observed with the scanning electron microscope. A-conidia germination approached 100% after 24 h, appresoria initiation began after 36 h; B-conidia germinated by 52 h but were not infective. Four-week-old C. sativa seedlings were more susceptible than 16-week-old plants, males more than females. THC and CBD, extracted and separated via TLC, inhibited P. ganjae conidia germination and hyphal growth.     

Inhibition of plant-pathogenic fungi by a corn trypsin inhibitor overexpressed in Escherichia coli

The cDNA of a 14-kDa trypsin inhibitor (TI) from corn was subcloned into an Escherichia coli overexpression vector. The overexpressed TI was purified based on its insolubility in urea and then refolded into the active form in vitro. This recombinant TI inhibited both conidium germination and hyphal growth of all nine plant pathogenic fungi studied, including Aspergillus flavus, Aspergillus parasiticus, and Fusarium moniliforme. The calculated 50% inhibitory concentration of TI for conidium germination ranged from 70 to more than 300 microgram/ml, and that for fungal growth ranged from 33 to 124 microgram/ml depending on the fungal species. It also inhibited A. flavus and F. moniliforme simultaneously when they were tested together. The results suggest that the corn 14-kDa TI may function in host resistance against a variety of fungal pathogens of crops.     

Comparative efficacy and pathogenicity of keratinophilic soil fungi against Culex quinquefasciatus larvae

Out of seven fungal species belonging to four genera isolated from pond and wallow soils using feathers of Pavo cristatus as bait, four species viz., Aspergillus flavus, Aspergillus niger, Chrysosporium pseudomerdarium and Trichophyton ajelloi were most frequent. Chrysosporium and Trichophyton spp. were more pathogenic on Culex quinquefasciatus larvae than Aspergillus and Penicillium. The bioefficacy tests conducted as per the protocol of World Health Organization and the LC₅₀ values calculated by the Probit analysis showed that 3^rd-instar C. quinquefasciatus were more susceptible to the conidia of above fungi. Highest mortality was observed in the larvae of C. quinquefasciatus when exposed to T. ajelloi. The density of fungal conidia was greatest on the ventral brush, palmate hair and anal region of the mosquito larvae after exposing for 72 hours. The potentiality of these fungi for use in the control of C. quinquefasciatus is discussed which can be exploited as a suitable biocontrol agent in the tropics.     

Effects of Conidia of Various Aspergillus Species on Apoptosis of Human Pneumocytes and Bronchial Epithelial Cells

Aspergillus species can cause mycoses in human and animals. Previously, we demonstrated that A. fumigatus conidia from a human isolate inhibited apoptosis in human pneumocytes and bronchial epithelial cells. In the current study, we studied the effects of A. fumigatus conidia non-human origin and A. flavus, A. nidulans, A. niger and A. oryzae conidia on human cells apoptosis. Human pneumocytes or bronchial epithelial cells were simultaneously exposed to apoptotic inductors and aspergilli conidia. The cell cultures were analyzed by flow cytometry, immunoblotting, and examination of nuclear morphology. Similar to A. fumigatus conidia, A. flavus conidia inhibited cellular apoptosis while A. nidulans, A. niger and A. oryzae conidia did not affect apoptosis. We further studied the species specificity of conidia: there were no differences in the inhibition of apoptosis by A. fumigatus conidia from either human or bird isolates. In order to determine whether the inhibition of apoptosis by conidia is limited to certain strains, the effect on human cell apoptosis of different A. fumigatus human clinical isolates and A. fumigatus of environmental origin was evaluated. All A. fumigatus isolates inhibited apoptosis; an anti-apoptotic factor was released by conidia. For TNF-induced apoptosis, the anti-apoptotic effect of conidia of all isolates was found to be associated with a reduction of caspase-3 in human cells. The results suggest that suppression of apoptosis may play a role in reducing the efficacy of host defense mechanisms during infection with Aspergillus species. F. Féménia and D. Huet made an equal contribution to this work.     

Taka-Amylase A in the Conidia of Aspergillus oryzae RIB40

A study of Taka-amylase A of conidia from Aspergillus oryzae RIB40 was done. During the research, proteins from conidia and germinated conidia were analyzed using SDS–PAGE, 2-D gel electrophoresis, Western blot analysis, MALDI-TOF Mass spectrometry, and native-PAGE combined with activity staining of TAA. The results showed that TAA exists not only in germinated conidia but also in conidia. Some bands representing degraded products of TAA were detected. Conidia, which formed on starch (SCYA), glucose (DCYA), and glycerol (GCYA) plates, contained mature TAA. Only one active band of TAA was detected after native-PAGE activity staining. In addition, TAA activity was detected in cell extracts of conidia using 0.5 M acetate buffer, pH 5.2, as extraction buffer, but was not detected in whole conidia or cell debris. The results indicate that TAA exists in conidia in active form even when starch, glucose, or glycerol is used as carbon source. TAA might belong to a set of basal proteins inside conidia, which helps in imbibition and germination of conidia.     

Rhizosphere and rhizoplane fungi of Triticum vulgare cultivated in Saudi Arabia

131 species in addition to 7 varieties which belong to 40 genera were collected from the rhizosphere (36 genera and 120 species + 7 varieties) and rhizoplane (27 genera and 56 species + 2 varieties) of Triticum vulgare. More than 22 species and 3 varieties were not encountered previously from soils or other sources in Saudi Arabia.In the rhizosphere, Aspergillus and Penicillium contributed the broadest spectrum of species (26 species + 5 varieties; and 24 species, respectively). The most frequent fungi were Aspergillus niger, A. clavatus, A. flavus, A. terreus, A. carneus, Penicillium citrinum, P. notatum, P. chrysogenum and Fusarium solani. In the rhizoplane which is a more selective substratum for fungi than the rhizosphere, the picture of dominance became different and the most common fungi were Fusarium solani, F. oxysporum, F. moniliforme, Drechslera spicifera, Cephalosporium roseo-griseum, Stemphylium botryosum, Acremonium strictum and Rhizoctonia solani.     

Inactivation of aflatoxigenic Aspergilli by treatment with ozone

The D-values of conidia of aflatoxigenic Aspergillus flavus and Aspergillus parasiticus exposed to 1.74 ppm. ozone in 1 mM potassium phosphate buffer (pH 7.0 and 5.5) at 25 degrees C were determined. D-values of A. flavus conidia were 1.72 and 1.54 min at pH 5.5 and 7.0, respectively; D-values of A. parasiticus were 2.08 and 1.71 min, respectively. None of these D-values was significantly (P < or = 0.05) different from each other.     

Regularities in the Germination of Conidia of Phytopathogenic Fungi

The autoregulation of conidium germination in phytopathogenic micromycetes of the genera Fusarium, Botrytis, and Bipolaris was studied. It was shown that Trichoderma longibrachiatum was less competitive than Fusarium oxysporum after their simultaneous inoculation but inhibited the phytopathogen growth in the case of earlier introduction. In the latter case, no autoinhibition of the germination of F. oxysporum conidia occurred; moreover, a cooperative effect was observed, i.e., the number of germinated F. oxysporum conidia increased with an increase in their density.     

Dimorphic fungus characteristic of fumonisin-producing strains of Fusarium moniliforme from Zhejiang

Fusarium moniliforme and its fumonisins have been shown to be carcinogenic in lab animals and have been linked to high incidences of human esophageal cancer. In this study we report the dimorphic fungus characteristic of fumonisin-producing strains of F. moniliforme from foodstuffs in Zhejiang, China. All of the twenty strains of F. moniliforme shown produce fumonisin B₁ 475.9–6322.2 μg/g in corn medium. These strains of F. moniliforme form yeast-like colonies in Sabouraud's agar plates contained 9% NaCl at 37 °C incubator and shows mostly budding reproduction. In blood agar plates these strains of F. moniliforme appear grass-green haemolytic reactions. This is the first report that yeast-like growth, dimorphic pathogenic fungus feature is found in F. moniliforme. These results suggest that it is also important to program epidemiological surveys of F. moniliforme as a primary pathogenic fungus, while proceeding to produce mycotoxins of F. moniliforme in food hygiene. This revised version was published online in August 2006 with corrections to the Cover Date.     

努利虫疠霉感染桃蚜过程的扫描电镜观察

利用扫描电镜观察了努利虫疠霉Pandora nouryi (Remaudière & Hennebert) Humber初生分生孢子接种桃蚜Myzus persicae (Sulzer)后孢子萌发、入侵以及菌体突破虫体的整个侵染过程。结果表明：附着于虫体表面的初生分生孢子在3～5 h后即有60%以上的萌发率,萌发的孢子形成芽管或产生球形或棍棒状的附着胞;12 h后大部分孢子均已萌发,并成功入侵寄主虫体;接种60 h后,呈掌状分枝的假根首先从桃蚜胸部的腹面突破虫体长出体外,明显区别于新蚜虫疠霉Pandora neoaphidis (Remaudière & Hennebert) Humber假根突破虫体的位置;假囊状体不多见,且仅分布于蚜虫身体两侧,这可在一定程度上解释努利虫疠霉产孢对湿度条件要求较高的生物学现象。     

THE VIRULENCE OF THE ENTOMOPHTHORALEAN FUNGUS PANDORA DELPHACZS TO THE BROWN PLANTHOPPER NILAPARVATA LUGENS1)

Abstract The entomophthoralean fungus, Pandwa delphacis (Hori) Humber, is a pathogen frequently causing epizootics of the brown planthopper, Nilaparvata lugens Stal, in rice growing areas of China. Two isolates (F95127 and F95129) of the fungus obtained from mycosis-killed planthoppers collected in Zhejiang Province were bioassayed for their virulence to N. lugens. For inoculation, botches of 4– or 5-in-star-old nymphs from a laboratory population were exposed to time-varying spore shower from sporulating plates of fungal mass produced in Sabouraud liquid medium, resulting in 8 and 6 dosages for F95129 and F95127, respectively. The nymphs treated at each dosage were maintained on rice plants at 25°C and a photophase of 12L: 12D in growth chamber (nearly 100% RH) and were examined daily for mortality. The resulting data were well fitted to time-dose-mortality model, generating parameters of time and dose effects for estimation of virulence indices. The LD₅₀ values 6–8 d after exposure were 327, 122 and 46 conidia/mm² for F95129 and 409, 133 and 74 conidia/mm² for F95127, respectively. The LT₅₀ estimated for F95127 was 7. 6 d at 95 conidia/mm² whereas that for F95129 decreased from 6. 6 d at 172 conidia/mm² to 5. 6 d at 525 conidia/mm². Based on these virulence indices and comparison of the slopes for dose effects from the model, the two isolates of P. delphacis had moderate virulence to N. lugens with insignificant difference to each other.     

Intrafungal distribution of aflatoxins among conidia and sclerotia of Aspergillus flavus and Aspergillus parasiticus

This research examines the distribution of aflatoxins among conidia and sclerotia of toxigenic strains of Aspergillus flavus Link and Aspergillus parasiticus Speare cultured on Czapek agar (21 days, 28 degrees C). Total aflatoxin levels in conidia and sclerotia varied considerably both within (intrafungal) and among strains. Aspergillus flavus NRRL 6554 accumulated the highest levels of aflatoxin (conidia: B1, 84000 ppb; G1, 566000 ppb; sclerotia: B1, 135000 ppb; G1, 968000 ppb). Substantial aflatoxin levels in conidia could place at risk those agricultural workers exposed to dust containing large numbers of A. flavus conidia. Cellular ratios of aflatoxin B1 to aflatoxin G1 were nearly identical in conidia and sclerotia even though levels of total aflatoxins in these propagule types may have differed greatly. Aflatoxin G1 was detected in sclerotia of all A. flavus strains but in the conidia of only one strain. Each of the A. parasiticus strains examined accumulated aflatoxin G1 in both sclerotia and conidia. These results are examined in the context of current evolutionary theory predicting an increase in the chemical defense systems of fungal sclerotia, propagules critical to the survival of these organisms.