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PubMed2005

Influence of autoclaved fungal materials on spearmint (Mentha spicata L.) growth, morphogenesis, and secondary metabolism.

The influence of autoclaved fungal materials such as culture filtrate, freeze-dried mycelium (FDM), mycelium suspension, and spore suspension (SS) on the growth, morphogenesis, and carvone production of spearmint (Mentha spicata L.) plants was studied. Fungal materials were either applied as a drench or spray on the plants. Spearmint plants (cv. "294099") drenched with SS (1 x 10(8) spores/ml) of Trichoderma reesei showed no significant differences in leaf numbers, root numbers, or shoot numbers compared with nontreated controls. However, significantly higher fresh weights and carvone levels were observed in plants drenched with T. reesei SS compared with the untreated controls. Fungal materials derived from Aspergillus sp., Fusarium graminearum, F. sporotrichoides, Penicillium sp., P. acculeatum, Rhizopus oryzae, and T. reesei were sprayed on spearmint foliage. F. graminearum, F. sporotrichoides, or R. oryzae elicited no enhanced growth, morphogenesis, or secondary metabolism responses. The best growth and morphogenesis responses were obtained employing Aspergillus sp., Penicillium sp., or T. reesei foliar sprays. For example, spearmint cv. "557807" plants sprayed with 100 mg/l FDM T. reesei isolate NRRL 11460 C30 stimulated higher fresh weights (75%), shoot numbers (39%), leaf numbers (57%), and root numbers (108%) compared with untreated plants. This effect was not dose-dependent because similar growth and morphogenesis responses were obtained by testing 10, 100, or 1000 mg/l FDM concentrations. Carvone levels in fungal-treated foliar-sprayed plants were comparable to nontreated controls. However, total carvone levels per plant were higher in fungal-treated plants because of their increased fresh weight.

Animals↗

PubMed2005

Fungal contamination of some imported spices.

Seventeen imported raw spice samples obtained from retail outlets were examined for spoilage mould profile. A total of 665 fungal isolates, representing 14 species, were recovered and identified from dried and ground spice samples on several media using standard dilution plate method. Moisture content varied greatly among various samples and were generally high. The most heavily contaminated spice samples examined were observed in red chili and black pepper in order of magnitude of 1580 and 1120 cfu/g, respectively. The most predominant fungal genera encountered were Aspergillus, Penicillium, Rhizopus, Cladosporium and Trichoderma. Yeasts were also frequently recovered, but not identified. Relative occurrence values of taxa disclosed ranged between 36.4% for A. flavus and 0.6% for A. parasiticus and Absidia corymbifera. Samples obtained from gunny bags encounter higher colony counts and contamination frequency than other packing methods. The present magnitude of contamination and spectra of mycobiota approximate those reported for similar spice samples. Although several potentially mycotoxigenic fungi were isolated during the present study, neither the foodstuff nor the fungi were assayed for the presence of these toxins.

Colony Count, Microbial↗

PubMed2005

Non-dermatophytic molds as agents of onychomycosis in Izmir, Turkey - a prospective study.

The purpose of this study was to determine the prevalence of causative non-dermatophytic filamentous fungi in onychomycosis. Totally 1,222 (1,222 x 3 = 3,666) samples of nail scrapings from 1,146 patients (from 76 patients two specimens: both from finger- and toe-nails) with prediagnosis of onychomycosis sent to the Mycology Laboratory from the Clinic of Dermatology, Ege University Hospital, Izmir, Turkey, July 2001-December 2003, were prospectively studied with conventional mycological procedures. The set criteria for the diagnosis of onychomycosis due to non-dermatophytic molds were: (1) Observation of fungal elements in 15% KOH-preparations made from nail scrapings, (2) growth of the same mold in all three consecutive cultures of the specimens taken three times from the same patient with one-week intervals, (3) no growth of a dermatophyte or yeast in three consecutive cultures. As agents of onychomycosis molds were detected in 33 (9%), dermatophytes in 175 (48%), yeasts in 150 (41%), and mixed (two different fungi) in 8 (2%) patients. In cases of mold onychomycosis, 11 (33%) had finger-nail and 22 (67%) toe-nail infection; 25 (76%) were female and 8 (24%) male; and 27 (82%) were above 40 years of age. The agents of mold onychomycosis, in order of frequency, were Aspergillus niger (7), Acremonium spp. (6), Fusarium spp. (6), Ulocladium spp. (4), sterile mycelia (2), Alternaria sp. (1), Aspergillus flavus (1), Aspergillus fumigatus (1), Aspergillus terreus (1), Cladosporium sp. (1), Paecilomyces spp. (1), Scopulariopsis sp. (1) and Trichoderma sp. (1). In conclusion, this study showed that non-dermatophytic molds were responsible for nearly 10% of onychomycoses cases attending the dermatology outpatient clinic of a university hospital in Izmir, Turkey. Since molds are common contaminants in the laboratory, cultures from consecutively taken nail scrapings should be made and carefully evaluated in order to diagnose a "mold onychomycosis".

Acremonium↗

PubMed2006

Susceptibility of the ant-cultivated fungus Leucoagaricus gongylophorus (Agaricales: Basidiomycota) towards microfungi.

The aim of this study was to select virulent strains of microfungi against Leucoagaricus gongylophorus, a symbiotic fungus cultivated by leaf-cutting ants. The results from in vitro assays showed that microfungal strains had a variable and significant impact on the colony development of L. gongylophorus. Specifically, Trichoderma harzianum, Escovopsis weberi CBS 810.71 and E. weberi A088 were more effective, inhibiting the L. gongylophorus colonies by 75, 68 and 67%, respectively (P < 0.05) after 15 days. Strain E. weberi A086 and Acremonium kiliense were less effective: 43 and 26%, respectively (P < 0.05). In spite of the current negative perspective of a microbiological control approach for these ants, the present work discusses the possibility of using mycopathogenic fungi for the control of these insects, and points out the importance of encouraging more studies in this area.

Animals↗

PubMed2006

Carbon requirements of some nematophagous, entomopathogenic and mycoparasitic hyphomycetes as fungal biocontrol agents.

Thirty-three carbon sources were evaluated for their effects on spore germination, hyphal growth and sporulation of 11 fungal biocontrol agents, i.e. the nematophagous fungi Paecilomyces lilacinus, Pochonia chlamydosporia, Hirsutella rhossiliensis, H. minnesotensis and Arkansas Fungus 18, the entomopathogenic fungi Lecanicillium lecanii, Beauveria bassiana and Metarhizium anisopliae, and the mycoparasitic fungus Trichoderma viride. Variations in carbon requirements were found among the fungal species or strains tested. All strains studied except for T. viride grew on most carbon sources, although B. bassiana had more fastidious requirements for spore germination. Monosaccharides and disaccharides were suitable for fungal growth. For most isolates, D-glucose, D-mannose, sucrose and trehalose were superior to pectin and soluble starch among the polysaccharides and lactic acid among the organic acids. Both ethanol and methanol could accelerate growth of most isolates but not biomass. D-mannose, D-fructose and D-xylose were excellent carbon sources for sporulation, while D-glucose, sucrose, cellobiose, trehalose, chitin, dextrin, gelatin and lactic acid were better for some isolates. Neither sorbic acid nor linoleic acid could be utilized as a single carbon source. These findings provided a better understanding of the nutritional requirements of different fungal biocontrol agents that can benefit the mass production process.

Carbon↗

PubMed2025

Microbe-induced gene silencing of fungal gene confers efficient resistance against Fusarium graminearum in maize.

UNLABELLED: Small RNAs (sRNAs), the main effectors of RNA interference (or RNA silencing, RNAi), mediate cell-autonomous and non-cell-autonomous gene silencing. The discoveries of trans-kingdom RNAi and interspecies RNAi have accelerated the development of RNAi-based crop protection technologies. Recently, based on interspecies RNAi, a practical technology termed microbe-induced gene silencing (MIGS) without the need of host genetic modification is developed for crop protection against Verticillium dahliae and Fusarium oxysporum in cotton and rice plants. In this study, we utilized MIGS technology to protect maize against Fusarium graminearum, which is responsible for maize stalk rot. An RNAi-engineered Trichoderma harzianum strain, Th-FgPmt2i, was exploited to generate double-stranded RNAs (dsRNAs) to trigger the silencing of the FgPTM2 gene. Our data verify that sRNAs generated from Th-FgPmt2i can silence the FgPMT2 gene via translational inhibition in F. graminearum. We further demonstrated that Th-FgPmt2i has a stronger capacity than does the T. harzianum chassis for protection of maize against F. graminearum. Coupled with our studies on crop protection against V. dahliae and F. oxysporum, our findings reveal that MIGS can be exploited to protect various crops against distinct fungal pathogens and has extensive applicability. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s42994-025-00212-9.

Fusarium graminearum↗

PubMed1989

Isolation of DNA from yeasts.

Methods are described that allow DNA to be prepared from widely different yeasts (Candida utilis, Saccharomyces cerevisiae, and Schizosaccharomyces pombe). The methods are reliably reproducible, and the DNA obtained is of appropriate quality for the construction of gene libraries (upper limit of size range consistently 50-150 kbp). In method A, yeast cells are converted into spheroplasts by treatment with a highly purified mixture of enzymes from Trichoderma harzianum, the spheroplasts are lysed in a lauroylsarcosinate/EDTA buffer, and the lysate is incubated with proteinase K and then directly centrifuged through a cesium trifluoroacetate gradient. DNA is recovered from the appropriate fractions by ethanol precipitation, and the redissolved precipitate is incubated with ribonuclease. For the rest of the isolation, two protocols are given, one avoiding and one including phenol/chloroform extraction. In this way, DNA up to about 150 kbp in size can be obtained. In method B, spheroplasts are not made. Yeast cells are broken by grinding under liquid nitrogen and are then worked up in a manner similar to method A, protocol 2. Subsequent steps depend on the purpose for which the DNA is required. Traditional methods of sucrose or salt density gradient centrifugation or agarose gel electrophoresis are applicable for size selection. A sodium iodide/silica matrix technique allows fast and effective DNA recovery from agarose gels.

Centrifugation, Density Gradient↗

PubMed1989

A method for detection of cellulases in polyacrylamide gels using 5-bromoindoxyl-beta-D-cellobioside: high sensitivity and resolution.

The assay of endo-1,4-beta-glucanases (cellulases) from Trichoderma reesei, T. longibrachiatum, and Sporotrichum pulverulentum by 5-bromoindoxyl-beta-D-cellobioside is described. The substrate is enzymatically cleaved to afford 5-bromoindoxyl and latter undergoes immediate azo coupling with Fast Red or oxidation by nitroblue monotetrazolium chloride, various forms of endoglucanases which can thus be assayed in polyacrylamide gel.

Acrylic Resins↗

PubMed1991

Solubilization and properties of GDP-fucose: xyloglucan 1,2-alpha-L-fucosyltransferase from pea epicotyl membranes.

GDP-fucose:xyloglucan 1,2-alpha-L-fucosyltransferase from pea (Pisum sativum) epicotyl microsomal membranes was readily solubilized by extraction with the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (Chaps). When using GDP-[14C]fucose as fucosyl donor and tamarind xyloglucan (XG) as acceptor, maximum activation was observed at 0.3% (w/v) Chaps and the highest yield of solubilized activity at 0.4%. The reaction product was hydrolyzed by Trichoderma cellulase to yield labeled oligosaccharides that peaked on gel permeation chromatography at the same elution volume as pea XG nona- and decasaccharide subunits. The apparent Km for fucosyl transfer to tamarind XG by the membrane-bound or solubilized enzyme was about 80 microM GDP-fucose. This was 10 times the apparent Km for fucosyl transfer to endogenous pea nascent XG. Optimum activity was between pH 6 and 7, and the isoelectric point was close to pH 4.8. The solubilized enzyme showed no requirement for, or stimulation by, added cations or phospholipids, and was stable for several months at -70 degrees C. Solubilization and gel permeation chromatography on columns of Sepharose CL-6B enriched the specific activity of the enzyme by about 20-fold relative to microsomes. Activity fractionated on columns of CL-6B with an apparent molecular weight of 150 kDa. The solubilized fucosyltransferase was electrophoresed on nondenaturing polyacrylamide slab gels containing 0.02% (w/v) tamarind XG, and its activity located by incubation in GDP-[14C]fucose, washing, and autoradiographing the gel. A single band of labeled reaction product appeared with an apparent molecular weight of 150 kDa.

Chromatography, Gel↗

PubMed1983

High field NMR study of antibiotic peptides: 1H assignment of trichorzianine A1 spectra by 2D experiments.

A new antibiotic peptide, trichorzianine A1, was isolated from a culture of Trichoderma harzianum. It contains 19 residues, the N terminus is blocked by an acetyl group and the C terminus is tryptophanol. As a first part of the structural study of this new peptide, we here present the analysis of the 1H NMR spectrum accomplished by 2 DJ resolved and spin echo correlated spectroscopy.

Amides↗

PubMed1992

Apple-fruit xyloglucans: a comparative study of enzyme digests of whole cell walls and of alkali-extracted xyloglucans.

Apple cell walls or alkali-extracted xyloglucans were digested with an endo-glucanase from Trichoderma viride and the resulting oligosaccharides were isolated by chromatography on Bio-Gel P-4. Three main oligosaccharides were present in similar proportions, and their structures were shown to be [Xyl(Glc)]3-Glc, [Xyl(Glc)]2-(FucGalXyl)Glc-Glc, and XylGlc-(GalXyP)Glc-(FucGalXyl)Glc-Glc. Each non-reducing-end Glc was 6-linked, each reducing-end Glc was 4-substituted, and each other Glc was 4,6-disubstituted. The Xyl was either terminal or 2-substituted, the Fuc was terminal, and the Gal was either terminal or 2-substituted. The 1H-NMR spectra of the oligosaccharides extracted directly from the cell wall showed that they are not acetylated. Other oligosaccharides, notably GalXyl3Glc4, Xyl2Glc4, and Xyl2Glc3, were present in smaller proportions in the digest of the cell walls.

Carbohydrate Sequence↗

PubMed1995

Eleven newly characterized xyloglucan oligoglycosyl alditols: the specific effects of sidechain structure and location on 1H NMR chemical shifts.

Eleven previously uncharacterized oligosaccharides, each containing from seventeen to twenty glycosyl residues, were isolated from the xyloglucan produced by suspension-cultured Acer pseudo-platanus cells and characterized by 1H NMR spectroscopy, fast-atom bombardment mass spectrometry, and matrix-assisted laser-desorption mass spectrometry. The complex mixture of xyloglucan oligosaccharides released by endo-(1-->4)-beta-glucanase (Trichoderma reesei) treatment of cell walls was similar to that released by digestion of the soluble xyloglucan present in the culture medium. The oligosaccharides were converted to oligoglycosyl alditols by borohydride reduction and purified by a combination of gel-permeation (Bio-Gel P-2) chromatography, normal-phase HPLC, reversed-phase HPLC, and high-performance anion-exchange (HPAE) chromatography. Eleven new oligoglycosyl alditols (along with several others that had been previously characterized) were isolated and characterized, allowing additional correlations between xyloglucan structure and specific chemical shift effects in the 1H NMR spectra to be determined. The correlations between structural and spectral features deduced in this study will facilitate the structural determination of a wide range of xyloglucans and their subunit oligosaccharides.

Carbohydrate Sequence↗

PubMed1995

Characterisation of 4-deoxy-beta-L-threo-hex-4-enopyranosyluronic acid attached to xylan in pine kraft pulp and pulping liquor by 1H and 13C NMR spectroscopy.

A new acidic sidegroup in xylans, from both kraft pulp and pulping liquor, was identified by NMR spectroscopy. Unmodified oligosaccharides from kraft pulp xylan were obtained by enzymatic hydrolysis with xylanase (Trichoderma reesei). The acidic oligosaccharides were separated from the natural forms on an anion exchange resin. The new acidic sidegroup was identified as 4-deoxy-beta-L-threo-hex-4-enopyranosyluronic acid (hexenuronic acid) by 1H and 13C NMR spectroscopy. Hexenuronic acid is a beta-elimination product of 4-O-methylglucuronic acid and is formed during kraft pulping. HMBC and NOESY experiments showed that hexenuronic acid is attached beta-(1 --> 2) to xylose. The NOESY data further indicated that hexenuronic acid protrudes from the main xylan chain. The pKa values for hexenuronic acid (3.03) and 4-O-methylglucuronic acid (3.14) attached (1 --> 2) to xylose were determined from pH-dependent chemical shifts.

Carbohydrate Sequence↗

PubMed1994

Trichokonin VI, a new Ca2+ channel agonist in bullfrog cardiac myocytes.

The effects of trichokonin VI (= gliodeliquescin A), a peptaibol isolated from the culture broth of Trichoderma koningii Oudemans, on L-type Ca2+ channel currents in single bullfrog atrial cells were investigated. Our results showed that trichokonin VI is a new potent agonist of L-type Ca2+ channels in cardiac membranes.

Alamethicin↗

PubMed1991

Two antifungal thaumatin-like proteins from barley grain.

Antifungal activity has been associated with 2 immunochemically distinct proteins, protein R and S (Mr approximately 23 kDa; pI 9-10), which were isolated in pure form from barley grain. The proteins are homologous with thaumatin- and pathogenesis-related proteins of the PR5 family. The proteins inhibit growth of i.a. Trichoderma viride and Candida albicans in microtiter plate assays and act synergistically with barley grain chitinase C. Like maize zeamatin, protein R and S but not chitinase C retarded fungal growth in synergism with nikkomycin Z, a nucleoside-peptide inhibitor of fungal chitin synthesis. Although no inhibition of alpha-amylases or serine proteases could be associated with protein R or S the results indicate that the homologous maize grain bifunctional inhibitor of insect alpha-amylase and trypsin is very similar to or identical with maize zeamatin, which was proposed to have permeabilizing activity towards fungal membranes. Thus, in addition to the intensely sweet properties of thaumatin, multiple unrelated defense functions against insect and fungal pests can now be associated with the family of thaumatin-homologous proteins.

Amino Acid Sequence↗

PubMed1992

Cellobiose oxidase of Phanerochaete chrysosporium enhances crystalline cellulose degradation by cellulases.

The effect of Phanerochaete chrysosporium cellobiose oxidase (CBO) on microcrystalline cellulose hydrolysis by Trichoderma cellulases was determined. Addition of 10 micrograms.ml-1 CBO to a reaction mixture containing T. viride cellulase increased glucose and cellobiose production by 10% and 48%, respectively. Cellulose weight loss was also enhanced by 19%. At higher concentrations (20-80 micrograms.ml-1), CBO decreased glucose and cellobiose production. Cellulose weight loss at 60 micrograms.ml-1 CBO was 76% compared to control cellulase reactions. This decrease appears to be due to inactivation of cellulase by H2O2 produced via CBO reaction, because addition of catalase enhances sugar production and cellulose weight loss. These findings suggest that at low, perhaps physiologically relevant concentrations, CBO enhances crystalline cellulose degradation by cellulases.

Basidiomycota↗

PubMed1993

Amino acid sequence and thermostability of xylanase A from Schizophyllum commune.

The amino acid sequence (197 residues) of xylanase A from the fungus, Schizophyllum commune, was determined by automated analysis of peptides from proteolytic and acid cleavage. The sequence is similar to two Trichoderma xylanases (approximately 56% identical amino acids), but also shows at least 40% identities with xylanases from Bacillus subtilis, B. pumilus and B. circulans. The conserved regions of the enzyme contain only two glutamic acid residues which implicates their possible involvement in catalysis. The disulfide bond in xylanase A is not conserved in this family. In spite of this, the B. subtilis xylanase was found to be more thermostable than xylanase A.

Amino Acid Sequence↗

PubMed1995

Dynamic light scattering study of the two-domain structure of Humicola insolens endoglucanase V.

Endoglucanase V (EG V) of HUmicola insolens is composed of a catalytic domain and of a cellulose-binding domain linked by a 33 amino acid long peptide rich in Ser, Thr and Pro residues. This work describes the dynamic behavior of the two-domain structure of EG V as revealed by quasi-elastic light scattering experiments. For both the full-length and the isolated catalytic domain, the autocorrelation function is essentially described by a single relaxation mode. The equivalent hydrodynamic radius of the catalytic domain was found to correspond precisely to the dimensions measured from the previously determined three-dimensional structure. The results obtained with the full-length protein allow a description of the two domain structure of EG V similar to that resulting from earlier studies using small angle X-ray scattering on cellulases from Trichoderma reesei. The hydrodynamic dimensions of the entire enzyme can be approximated as an ellipsoid with dimensions of 42 x 133.6 A.

Binding Sites↗