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PubMed1995

Antibiotic production and biocontrol activity by Bacillus subtilis CL27 and Bacillus pumilus CL45.

Bacillus subtilis CL27 and B. pumilus CL45 showed similar activity against Botrytis cinerea in in vitro plate assays. In a seedling bioassay, however, B. subtilis CL27 had activity similar to a commercial fungicide while B. pumilus CL45 failed completely to prevent seedling damping-off caused by Bot. cinerea. Antibiotic production by the two Bacillus strains was found to depend on the growth substrate and highest antibiotic production was found on media based on homogenized cabbage tissue. Antibiotic activity was found to depend on the pH and nutrient concentration in the assay medium. Antifungal antibiotics produced by B. subtilis CL27 and B. pumilus CL45 in different fermentation media were separated by thin layer chromatography. As suspected from the activity spectrum, three antibiotics (one with activity against Alternaria brassicicola, one with activity against Botrytis cinerea and one with activity against both fungi) could be detected in the fermentation broth of CL27, but only one in the fermentation broth of CL45. The two antibiotics produced by strain CL27 with activity against A. brassicicola were identified as peptides since their bands on the TLC plates developed a green to blue/green colour after treatment with 4,4'-tetramethyldiamino-diphenylmethane (TDM) reagent. The third antibiotics produced by strain CL27 and antibiotic produced by CL45 had a similar Rf-value and appeared not to be peptides based on the reaction with TDM. However, they showed a slightly different activity spectrum when tested against a range of different fungi. Antibiotic production was clearly indicated as the mode of action of in vivo biocontrol by strain CL27 against damping off caused by Bot. cinerea of Astilbe micro-plants, because a u.v.-induced antibiotic negative mutant strain CL27b showed no activity in seedling bioassays in vivo. Also the mutant strain CL27a which produced the two peptide antibiotics but had lost the ability to produce the non-peptide antibiotic, showed greatly reduced in vivo activity.

Antibiosis↗
PubMed2005

A pathogen-inducible patatin-like lipid acyl hydrolase facilitates fungal and bacterial host colonization in Arabidopsis.

Genes and proteins related to patatin, the major storage protein of potato tubers, have been identified in many plant species and shown to be induced by a variety of environmental stresses. The Arabidopsis patatin-like gene family (PLPs) comprises nine members, two of which (PLP2 and PLP7) are strongly induced in leaves challenged with fungal and bacterial pathogens. Here we show that accumulation of PLP2 protein in response to Botrytis cinerea or Pseudomonas syringae pv. tomato (avrRpt2) is dependent on jasmonic acid and ethylene signaling, but is not dependent on salicylic acid. Expression of a PLP2-green fluorescent protein (GFP) fusion protein and analysis of recombinant PLP2 indicates that PLP2 encodes a cytoplasmic lipid acyl hydrolase with wide substrate specificity. Transgenic plants with altered levels of PLP2 protein were generated and assayed for pathogen resistance. Plants silenced for PLP2 expression displayed enhanced resistance to B. cinerea, whereas plants overexpressing PLP2 were much more sensitive to this necrotrophic fungus. We also established a positive correlation between the level of PLP2 expression in transgenic plants and cell death or damage in response to paraquat treatment or infection by avirulent P. syringae. Interestingly, repression of PLP2 expression increased resistance to avirulent bacteria, while PLP2-overexpressing plants multiplied avirulent bacteria close to the titers reached by virulent bacteria. Collectively, the data indicate that PLP2-encoded lipolytic activity can be exploited by pathogens with different lifestyles to facilitate host colonization. In particular PLP2 potentiates plant cell death inflicted by Botrytis and reduces the efficiency of the hypersensitive response in restricting the multiplication of avirulent bacteria. Both effects are possibly mediated by providing fatty acid precursors of bioactive oxylipins.

Amino Acid Sequence↗
PubMed1991

Use of the extended Phadebas RAST panel in the diagnosis of mould allergy in asthmatic children.

The clinical significance of mould allergens in Phadebas RAST panel was investigated in 121 asthmatic children. They were selected from a total population of 1649 patients. The patients were distributed into four groups, based on the combination of positive or negative skin prick tests (SPT) together with symptoms suggestive or not of mould sensitivity. Mould-specific IgE antibodies were investigated using the original RAST panel (Alternaria, Aspergillus, Candida, Cladosporium, Mucor and Penicillium) and a set of 10 additional mould-allergen discs (Aureobasidium, Botrytis, Curvularia, Epicoccum, Fusarium, Helminthosporium, Phoma, Rhizopus, Stemphylium and Trichoderma). The set of additional RAST discs revealed patients with mould-reaginic antibodies not found with the original RAST panel. This occurred in four of 49 (8.2%) RAST positive (class greater than or equal to 2) patients. The allergens most frequently positive were Cladosporium (in 28% of the patients), Candida (28%) and Helminthosporium (26%). A remarkable degree of simultaneous reactivity to almost all moulds tested was observed. Patients with multiple (greater than or equal to 7) mould sensitization were effectively pinpointed using any duplicate combination of Aureobasidium, Botrytis, Candida, Cladosporium, Helminthosporium, Penicillium and Stemphylium.

Adolescent↗
PubMed1977

The fungal flora of loganberries in relation to storage and spoilage.

Botrytis cinerea, Cladosporium spp., Penicillium spp., Aureobasidium pullulans, Mucor mucedo, Phoma state of Didymella applanata, Cryptococcus laurentii var. laurentii, C. albidus var. albidus, Candida sake and Sporobolomyces roseus were consistently present on freshly harvested loganberries. Botrytis cinerea was the main spoilage organism with Mucor mucedo of minor importance. The importance of benomyl-resistant strains of B. cinerea is discussed.

Ascomycota↗
PubMed2003

Purification and characterization of chitinase from Streptomyces sp. M-20.

Chitinase (EC 3.2.1.14) was isolated from the culture filtrate of Streptomyces sp. M-20 and purified by ammonium sulfate precipitation, DEAE-cellulose ion-exchange chromatography, and Sephadex G-100 gel filtration. No exochitinase activity was found in the culture filtrate. The molecular mass of the purified chitinase was 20 kDa, estimated by a sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was confirmed by activity staining with Calcofluor White M2R. Chitinase was optimally active at pH of 5.0 and at 30 degrees C. The enzyme was stable from pH 4 to 8, and up to 40 degrees C. Among the metals and inhibitors that were tested, the Hg(+), Hg(2+), and p-chloromercuribenzoic acid completely inhibited the enzyme activity. The chitinase activity was high on colloidal chitin, chitotriose, and chitooligosaccharide. The purified chitinase showed antifungal activity against Botrytis cinerea, and lysozyme activity against the cell wall of Botrytis cinerea.

Animals↗
PubMed2005

Identification of an antifungal chitinase from a potential biocontrol agent, Bacillus cereus 28-9.

Bacillus cereus 28-9 is a chitinolytic bacterium isolated from lily plant in Taiwan. This bacterium exhibited biocontrol potential on Botrytis leaf blight of lily as demonstrated by a detached leaf assay and dual culture assay. At least two chitinases (ChiCW and ChiCH) were excreted by B. cereus 28-9. The ChiCW-encoding gene was cloned and moderately expressed in Escherichia coli DH5alpha. Near homogenous ChiCW was obtained from the periplasmic fraction of E. coli cells harboring chiCW by a purification procedure. An in vitro assay showed that the purified ChiCW had inhibitory activity on conidial germination of Botrytis elliptica, a major fungal pathogen of lily leaf blight.

Amino Acid Sequence↗
PubMed2000

Extrinsic allergic alveolitis in domestic environments (Domestic allergic alveolitis) caused by mouldy tapestry

Extrinsic allergic alveolitis often occures as bird fancier's lung or is caused by occupational treatment with antigenic materials. In housing environments antigens of allergic alveolitis are also found, especially mould fungi. lf a source of antigens is absent in the anamnesis and the clinical picture as well as the clinical findings are ambiguous, the diagnosis of extrinsic allergic alveolits is delayed or unobtained. The following example shows that in spite of a detailed allergic anamnesis the source of antigens may remain occult and only an inspection of the dwelling rooms leads to an elucidation. - A sixty-one year old non-smoking women was twice admitted to hospital with a temperature of more than 39 degrees C, intense dyspnea on exertion and a strong dry cough under the persumed diagnosis pneumonia. The laboratory values showed nonspecific signs of inflammation, the blood gas analysis was changed to a heavy hypoxemia (pO subset2 49.2 mmHg) and in the chest x-ray there were seen miliary and partly reticular alterations. The chest computed tomography showed extensive densities in both upper and lower parts of the lungs and the pulmonary function test corresponded with a low-grade to middle-grade restriction (VC subsetin = 67%, TLC = 69%). A high dose of corticosteroids produced an improvement of the radiological findings and of the pulmonary function. Also the specialized diagnostic in a hospital for pulmonary diseases yielded no new knowledge, and an interstitial pneumonia was diagnosed. Only the new formation of an acute pneumonic clinical picture on the day of returning to the patients own habitation suggested a noxious substance in the domestic environment. The inspection of the rooms finally showed the source of antigen to be from a condensation water soaked, moulded tapestry on the embrasure of a small unopenable window in the bedroom just on the head of the bed. The nutritive mediums left in the rooms furnished evidence of the mould-species Penicillium, Cladosporium and Botrytis, in the bedroom however mostly Penicillium. The RAST of specific IgG proved a positive result of all three mould species (Penicillium sp. 1 : 100, Cladosporium sp. 1 : 200 and Botrytis sp. 1 : 200). The diagnosis of an extrinsic allergic alveolitis caused by mould was confirmed by a controlled re-exposure test, which promptly gave rise to a relapse and forced the patient to change habitation with the result of no further ailments. - Characteristic for the extrinsic allergic alveolitis in the described case is the inefficiency of antibiotic therapy, the immediate recovery by corticosteroids and furthermore the outbreak of relapses caused by re-exposure. If the source of antigens is unclear, the inspection of the habitation by an experienced allergologist may lead to success.

Journal Article↗
PubMed2004

Fungal spores in the atmosphere of Rzeszów (South-East Poland).

Continuous aerobiological monitoring has been conducted in Rzeszów from 2000-2002, using the volumetric method. On each microscope slide 1 horizontal band was analyzed, divided into 24 fields corresponding to hours. For the analysis, 10 easy to identify fungal spores were chosen from ones regarded as allergenic: Alternaria, Botrytis, Cladosporium, Drechslera (type), Epiccocum, Ganoderma, Pithomyces, Polythrincium, Stemphylium, and Torula. The results were statistically tested using the chi2 test as well as the Kruskal-Wallis test and ANOVA. The results were used to develop a calendar of the occurrence of fungal spores in Rzeszów. The spores occurred in the air throughout the whole year, but maximum concentrations were usually reached in July and August. Two groups of taxa were distinguished. Alternaria, Cladosporium, Botrytis, Epicoccum, Ganoderma spp. and Drechslera belong to the first group, and their spores and conidia were characterised by high frequency and abundance in the air. Low SFI values (Seasonal Fungal Index) and frequency of below 50% occurred in the second group of taxa, i.e. Pithomyces, Polythrincium, Stemphylium and Torula spp. Conidia of Cladosporium spp. were the most frequent, SFI values were very high and average annual concentrations did not differ significantly throughout the 3 years of study. The research confirmed the overlap of the period of maximum concentration of allergenic spores and the period of the domination of Poaceae and Artemisia pollen in the air.

Classification↗
PubMed1990

[Research of antifungal substances secreted by higher fungi in culture].

A screening in vitro of antifungic activity of 24 strains of Basidiomycetes was realized with their culture filtrate. Lycoperdon perlatum Pers. = Pers., Oudemansiella platyphylla (Pers. ex Fr.) Mos., Agrocybe dura (Bolt) Singer have shown an activity against Candida albicans, Candida tropicalis and Aspergillus fumigatus; Pholiota spumosa (Fr.) Singer towards Botrytis cinerea and Lycoperdon perlatum Pers. = Pers. towards Alternaria solani, Botrytis cinerea and Verticillium dahliae. More extensive studies of the kinetics of antifungic substances production from Lycoperdon perlatum Pers. = Pers. were effectued with 2 different media and 2 different seeding technics on the strains which were sensible to the inhibitory character of this Gasteromycete.

Antifungal Agents↗
PubMed1989

Specific IgE antibodies to 16 widespread mold genera in patients with suspected mold allergy.

A panel of 55 sera from patients with suspected mold allergy from two different geographic areas, A and B, was investigated for specific IgE antibodies to 16 individual molds using Phadebas RAST technique. The molds investigated were Alternaria, Cladosporium, Aspergillus, Penicillium, Mucor, and Candida from the former RAST panel and ten additional new mold genera. The RAST screening revealed that 73% of patients had specific IgE antibodies to at least one of the 16 molds. Eighteen percent were negative to the six molds of the previous RAST panel but had specific IgE antibodies to at least one of the ten new molds. The highest frequency of IgE antibodies in the A group could be assigned to Cladosporium, followed by Botrytis and Helminthosporium. In group B, sensitivity to Botrytis and Phoma showed the highest frequencies. This study revealed that the relative importance of mold genera may vary greatly between patient groups from different geographic areas. Some of the species of the expanded RAST panel seem to be at least as important as Alternaria and Cladosporium. Multiple sensitivities seem to be caused by sensitization by many mold species rather than by cross-reactivity.

Alabama↗
PubMed1996

Generation of superoxide anion and induction of superoxide dismutase and peroxidase in bean leaves infected with pathogenic fungi.

Generation of superoxide anion (O2-.) and peroxidase activity were significantly increased in bean leaves infected with incompatible and compatible pathogens: Botrytis fabae and Botrytis cinerea, respectively, but the induction was greater on direct inoculation with B. fabae, than with B. cinerea. A slightly higher O2-. level was also detected in the parts of leaves surrounding the inoculation side. Overproduction of O2-. was observed earlier than the increase in peroxidase activity. Pretreatment of the leaves with methyl jasmonate enhanced both O2-. production and peroxidase activity following inoculation with B. cinerea. Induction of superoxide dismutase activity after the infection was less pronounced than changes in O2-. level. The differences in the rate of NADH oxidation in the extracts from control and inoculated leaves, correlated with the differences in the rate of O2-. production. The results indicate that O2-. level is one of the essential factors responsible for the difference in the interactions between bean plant and compatible and incompatible pathogens.

Fabaceae↗
PubMed2000

Synthesis and antifungal activity of some new quinazoline and benzoxazinone derivatives.

The hitherto unknown 2-isopropyl-6,8-dibromo-4H-3,1-benzoxazin-4-one (2) was subjected to condensation with either primary or secondary amines affording the benzamide derivatives (3-7), while with alcohols in presence of the base, corresponding esters were obtained (8 and 9). Acylation of the hydrazide (12) or its cyclized form (13) gave (14-17). The quinazolinone derivative (18) was obtained either when (12) was reacted with nitrous acid or via fusion of (2) with ammonium acetate. The thione (20) which was obtained via reaction of (18) with Lawesson's reagent, was subjected to either alkylation yielding (21-25) or desulphurization with primary amines affording (26 and 27). Treatment of (18) as well as (20) with a chlorinating agent provided (29, 30) and (28, 29) mixtures, respectively. Ten of our compounds were examined against Sclerotium cepivorum as well as Botrytis allii on PDA media. These compounds showed a significant reduction of mycelial growth and scleratia number of these fungi which cause the white rot and neck rot diseases of onion.

Basidiomycota↗
PubMed2004

Synthesis and determination of the absolute configuration of fugomycin and desoxyfugomycin: CD spectroscopy and fungicidal activity of butenolides.

The dibromoalkenes (S)-3 and (R)- and (S)-4 are intermediates in the syntheses of the naturally occurring fungicidal butenolides fugomycin (1) and desoxyfugomycin (2), respectively. The stereoselective bromine-lithium exchange that leads to the carbenoid 12 and the vinyllithium reagent 17 a on the one hand, and palladium-catalyzed coupling reactions of the dibromoalkene 3 and the bromolactone 22 on the other are key steps en route to the butenolides 1 and 2. The chiral building blocks (S)-3, (R)-4, and (S)-4 are readily available from (R)-isopropylideneglyceraldehyde 5, isobutyl (R)-lactate 6 a, and ethyl (S)-lactate 6 b, respectively. The synthetic procedure adopted here permits the absolute configuration of the natural products fugomycin (1) and desoxyfugomycin (2) to be assigned by comparison of their chiroptical properties with those of the synthetic products. The CD spectra of the bromolactone 22, calculated by two different density functional methods (TDDFT, DFT/MRCI), are found to be in good agreement with the measured spectra. On the basis of these calculations, the two CD bands observed could be assigned to n-pi* and pi-pi* transitions, respectively. Fugomycin (1) and the synthetic butenolide 20 displayed high fungicidal activity against botrytis in greenhouse experiments, whereas the saturated lactone 21 was practically inactive.

4-Butyrolactone↗
PubMed2005

Isolation and antifungal activity of kakuol, a propiophenone derivative from Asarum sieboldii rhizome.

An antifungal substance active against Colletotrichum orbiculare (Berk & Mont) Arx was isolated from the methanol extracts of Asarum sieboldii (Miq) Maek rhizomes. High-resolution MS, NMR and UV spectral data confirmed that the antifungal substance is kakuol, 2-hydroxy-4,5-methylenedioxypropiophenone. Colletotrichum orbiculare was most sensitive to kakuol, with MIC of 10 microg ml(-1). Kakuol also completely inhibited the mycelial growth of Botrytis cinerea Pers ex Fr and Cladosporium cucumerinum Ellis & Arthur at 50 microg ml(-1) and 30 microg ml(-1), respectively. However, no antimicrobial activity was found against yeast and bacteria even at 100 microg ml(-1). Kakuol exhibited a protective activity against the development of anthracnose disease on cucumber plants. The control efficacy of kakuol against the anthracnose disease was in general somewhat less than that of the commercial fungicide chlorothalonil. This is the first report to demonstrate in vitro and in vivo antifungal activity of kakuol against C. orbiculare infection.

Asarum↗
PubMed2006

Impact of fungal drug transporters on fungicide sensitivity, multidrug resistance and virulence.

Drug transporters are membrane proteins that provide protection for organisms against natural toxic products and fungicides. In plant pathogens, drug transporters function in baseline sensitivity to fungicides, multidrug resistance (MDR) and virulence on host plants. This paper describes drug transporters of the filamentous fungi Aspergillus nidulans (Eidam) Winter, Botrytis cinerea Pers and Mycosphaerella graminicola (Fückel) Schroter that function in fungicide sensitivity and resistance. The fungi possess ATP-binding cassette (ABC) drug transporters that mediate MDR to fungicides in laboratory mutants. Similar mutants are not pronounced in field resistance to most classes of fungicide but may play a role in resistance to azoles. MDR may also explain historical cases of resistance to aromatic hydrocarbon fungicides and dodine. In clinical situations, MDR development in Candida albicans (Robin) Berkhout mediated by ABC transporters in patients suffering from candidiasis is common after prolonged treatment with azoles. Factors that can explain this striking difference between agricultural and clinical situations are discussed. Attention is also paid to the risk of MDR development in plant pathogens in the future. Finally, the paper describes the impact of fungal drug transporters on drug discovery.

Agriculture↗
PubMed2006

Enhancement of fungicidal and insecticidal activity by reductive alkylation of chitosan.

A series of N-alkyl chitosan (NAC) derivatives were synthesized using a reductive alkylation reaction to examine their fungicidal and insecticidal activity. The chemical structures were characterized by IR and (1)H NMR spectroscopy, and the degree of substitution (DS) ranged from 0.02 to 0.37. Their fungicidal activity was evaluated against the grey mould Botrytis cinerea Pers ex Fr (Leotiales: Sclerotiniaceae) and the rice leaf blast pathogen Pyricularia grisea Sacc [Teleomorph: Magnaporthe grisea (Hebert) Barr] by a radial growth bioassay. It was of interest that most of the NAC derivatives were more active against both fungi than chitosan itself. The most active derivative was N-(2,2-diphenylethyl)chitosan with EC50 values of 0.031 and 0.23 g L(-1) against B. cinerea and P. grisea respectively. In addition, some derivatives, at higher concentrations up to 1.0 g L(-1), inhibited the mycelial growth and spore formation of P. grisea. Bioassays against larvae of the cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) with the NAC derivatives at a rate of 5.0 g kg(-1) in artificial diet demonstrated that N-(3-phenylbutyl)chitosan was the most active compound. In addition, N-propylchitosan, N-undecanylchitosan and N-(3-phenylpropyl)chitosan derivatives strongly inhibited larval weight gain in S. littoralis, with respective reductions of 76, 66 and 65% after 4 days of feeding on treated diet.

Alkylation↗
PubMed2001

Control of post-harvest decay of apples by pre-harvest and post-harvest application of ammonium molybdate.

Ammonium molybdate was tested as a potential fungicide for use in apples (cv Golden Delicious) against blue and grey mould, important post-harvest diseases of pome fruits. In tests in vivo at 20 degrees C, ammonium molybdate (15 mM) reduced lesion diameters of Penicillium expansum, Botrytis cinerea and Rhizopus stolonifer by 84%, 88% and 100% respectively. When apples treated with ammonium molybdate were stored at 1 degree C for three months, a significant reduction in severity and incidence of P expansum and B cinerea was observed in both years of study (1998 and 1999). In the second year of the experiment the reduction in disease severity was greater than 88% for both pathogens, and the level of control was similar to, or greater than, that observed with the fungicide imazalil. When ammonium molybdate was applied as a pre-harvest treatment, a significant reduction in blue mould decay was observed after three months in cold storage. In vitro, ammonium molybdate greatly inhibited spore germination of P expansum and B cinerea, although better inhibition was obtained against grey mould. Ammonium dimolybdate, sodium molybdate and potassium molybdate were also tested in vitro in comparison with ammonium molybdate as inhibitors of spore germination, but only ammonium molybdate inhibited spore germination by more than 50%.

Agriculture↗
PubMed2002

Molluscicidal and antifungal activity of Erigeron speciosus steam distillate.

The steam-distilled fraction of the aerial parts of Erigeron speciosus (Lindl) DC was tested for activity against strawberry plant pathogenic fungi Botrytis cinerea Pers ex Fr, Colletotrichum acutatum Simmonds, C fragariae Brooks, C gloeosporioides (Penz) Penz & Sacc, and the intermediate host snail Planobdella trivolvis that harbors the trematode, Bolbophorus confusus, that infests and causes severe infections in pond-raised catfish in the Mississippi Delta region of the USA. Bioautography on silica TLC plates demonstrated antifungal activity in the steam distillate. Preliminary bioassays of the steam distillate indicated the presence of phytochemicals toxic to P trivolvis. The bioactive compounds methyl 2Z, 8Z-deca-2,8-diene-4,6-diynoate and its 2E, 8E isomer were isolated by bioassay-guided fractionation and chromatographic techniques and identified by 1H NMR spectroscopy.

Alkynes↗