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PubMed2001

Cell-wall composition and polysaccharide synthase activity changes following photoinduction in Trichoderma viride.

The differentiation and metabolism in the soil-borne saprophytic deuteromycete Trichoderma viride are subject to control by light. We have investigated the effect of illumination of mycelia on the activities of cell-wall synthesizing enzymes beta-1,3-glucan synthase and chitin synthase and on the composition of cell alls. After 10 min illumination of dark-grown mycelia with white fluorescent light at 600 1x, a gradual rise in specific activity of membrane-bound enzymes beta-1,3-glucan synthase by about 130% and a decrease in specific activity of chitin synthase by about 50% in relation to the dark control were observed. The changes in enzyme activities were caused by de novo synthesis of corresponding polysaccharide synthases(s) and/or their regulatory components since they were not observed when protein synthesis was blocked with 50 microg/ml cycloheximide. The content of beta-1,3-glucan in the cell walls of illuminated mycelia has increased by 35-50% in comparison to the dark control while the content of chitin remained practically unchanged.

Cell Wall↗
PubMed2001

Characterization of the extracellular enzyme systems of Trichoderma viride AH124.

A mycoparasitic Trichoderma viride strain was investigated for the production of extracellular enzymes important in antagonism, by using natural and chromogenic substrates. Some of these enzymes, such as beta-1,3-glucanases, and low levels of proteases were produced constitutively. Under inductive conditions, the measurable activities of beta-1,3-glucanase, protease and aspecific chitinase increased, while for the proteases and beta-1,3-glucanases, the levels depended on both the nitrogen and the carbon source. Gel filtration chromatography revealed at least 4 beta-1,3-glucanases, 6 proteases, 2 beta-glucosidases and 1 beta-1,4-N-acetyl-glucosaminidase izoenzyme under inductive conditions.

Culture Media↗
PubMed2002

Selection of strain and optimization of mutanase production in submerged cultures of Trichoderma harzianum.

Nineteen fungal strains belonging to different genera were tested for extracellular mutanase production in shaken flasks. The optimal enzymatic activity was achieved by Trichoderma harzianum F-470, a strain for which the mutanase productivity has not yet been published. Some of factors affecting the enzyme production in shaken flasks and aerated fermenter cultures have been standardized. Mandels mineral medium with initial pH 5.3, containing 0.25% mutan and inoculated with 10% of the 48-h mycelium, was the best for enzyme production. A slight mutanolytic activity was also found when sucrose, raffinose, lactose and melibiose were carbon sources. Application of optimized medium and cultural conditions, as well as use of a fermenter with automatic pH control set at pH 6.0 enabled to obtain a high mutanase yield (0.33 U/ml, 2.5 U/mg protein) in a short time (2-3 days). The enzyme in crude state was stable over a pH range of 4.5-6.0, and at temperatures up to 35 degrees C; its maximum activity was at 40 degrees C and at pH 5.5.

Fermentation↗
PubMed2006

Enhancement of mutanase production in Trichoderma harzianumby mutagenesis.

Conidia of Trichoderma harzianum F-340, an active producer of fungal mutanase, were mutagenized with physical and chemical mutagens used separately or in combination. After mutagenesis, the drop in conidia viability ranged from 0.004% to 71%. Among the applied mutagens, nitrosoguanidine gave the highest frequency of cultures with enhanced mutanase activity (98%). In total, 400 clones were isolated, and preliminarily evaluated for mutanase activity in flask microcultures. Eight most productive mutants were then quantified for mutanase production in shake flask cultures. The obtained results fully confirmed a great propensity of all the tested mutants to synthesize mutanase, the activity of which increased from 59 to 107% in relation to the parental T. harzianum culture. The best mutanase-overproducing mutant (T. harzianumn F-340-48), obtained with nitrosoguanidine, produced the enzyme activity of 1.36 U/ml (4.5 U/mg protein) after 4 days of incubation in shake flask culture. This productivity was almost twices higher than that achieved by the initial strain F-340, and, at present, is the best reported in the literature. The potential application of mutanase in dentistry is also discussed.

Dental Caries↗
PubMed2002

Ecophysiology and breeding of mycoparasitic Trichoderma strains (a review).

Losses due to plant diseases may be as high as 10-20% of the total worldwide food production every year, resulting in economic losses amounting to many billions of dollars and diminished food supplies. Chemical control involves the use of chemical pesticides to eradicate or reduce the populations of pathogens or to protect the plants from infection by pathogens. For some diseases chemical control is very effective, but it is often non-specific in its effects, killing beneficial organisms as well as pathogens, and it may have undesirable health, safety, and environmental risks. Biological control involves the use of one or more biological organisms to control the pathogens or diseases. Biological control is more specialized and uses specific microorganisms that attack or interfere with the pathogens. The members of the genus Trichoderma are very promising against soil-born plant parasitic fungi. These filamentous fungi are very widespread in nature, with high population densities in soils and plant litters [1]. They are saprophytic, quickly growing and easy to culture and they can produce large amounts of conidia with long lifetime.

Antibiosis↗
PubMed2003

Regulation of Trichoderma cellulase formation: lessons in molecular biology from an industrial fungus. A review.

The present article reviews the current understanding of regulation of cellulase gene transcription in Hypocrea jecorina (= Trichoderma reesei). Special emphasis is put on the mechanism of action of low molecular weight inducers of cellulase formation, the presence and role of recently identified transactivating proteins (Ace1, Ace2, Hap2/3/5), and the role of the carbon catabolite repressor Cre1. We also report on some recent genomic approaches towards understanding how cellulase inducers signal their presence to the transcriptional apparatus.

Base Sequence↗
PubMed2005

Peptaibols and related peptaibiotics of Trichoderma. A review.

Peptaibols and the related peptaibiotics are linear, amphipathic polypeptides. More than 300 of these secondary metabolites have been described to date. These compounds are composed of 5-20 amino acids and are generally produced in microheterogeneous mixtures. Peptaibols and peptaibiotics with unusual amino acid content are the result of non-ribosomal biosynthesis. Large multifunctional enzymes known as peptide synthetases assemble these molecules by the multiple carrier thiotemplate mechanism from a remarkable range of precursors, which can be N-methylated, acylated or reduced. Peptaibols and peptaibiotics show interesting physico-chemical and biological properties including the formation of pores in bilayer lipid membranes, as well as antibacterial, antifungal, occasionally antiviral activities, and may elicit plant resistance. The three-dimensional structure of peptaibols and peptaibiotics is characterized predominantly by one type of the helical motifs alpha-helix, 3(10)-helix and beta-bend ribbon spiral. The aim of this review is to summarize the data available about the biosynthesis, biological activity and conformational properties of peptaibols and peptaibiotics described from Trichoderma species.

Alamethicin↗
PubMed2003

Oviposition response of the mosquito, Culex quinquefasciatus to the secondary metabolite(s) of the fungus, Trichoderma viride.

Secondary metabolites produced by Trichoderma viride, a deuteromycetes fungus, under submerged culture condition were formulated and evaluated for oviposition attractancy against gravid females of Culex quinquefasciatus mosquito. At a concentration of 10 g ml-1 the formulation showed remarkable attractancy with an oviposition active index (OAI) of +0.52. When the oviposition attractancy of the formulation was compared with a known oviposition attractant, p-cresol, both at 10 g ml-1, the former was found to be more attractive to result in 70% egg laying than the later with 30% egg laying. Thin layer chromatography fractions of the secondary metabolites showed that a fraction with Rf value of 0.88 was highly active as oviposition attractant with an OAI of +0.65. Further work on identification of the active principle(s) of the microbial formulation might lead to an oviposition attractant useful in mosquito vector management.

Animals↗
PubMed2006

Impact of transgenic Bt maize residues on the mycotoxigenic plant pathogen Fusarium graminearum and the biocontrol agent Trichoderma atroviride.

Transformation of maize with genes encoding for insecticidal crystal (Cry) proteins from Bacillus thuringiensis (Bt) could have an impact on the saprophytic survival of plant pathogens and their antagonists on crop residues. We assessed potential effects on the mycotoxin deoxynivalenol (DON)-producing wheat and maize pathogen Fusarium graminearum and on the biocontrol agent Trichoderma atroviride. Purified Cry1Ab protein caused no growth inhibition of these fungi on agar plates. Cry1Ab concentrations above levels common in Bt maize tissue stimulated the growth of F. graminearum. The fungi were also grown on gamma-radiation-sterilized leaf tissue of four Bt maize hybrids and their non transgenic isolines collected at maize maturity on a field trial in 2002 and 2003. Both fungi degraded the Cry1Ab protein in Bt maize tissue. Fungal biomass quantification with microsatellite-based polymerase chain reaction (PCR) assays revealed differential fungal growth on leaf tissue of different maize varieties but no consistent difference between corresponding Bt and non-Bt hybrids. Generally, year of maize tissue collection had a greater impact on biomass production than cultivar or Bt transformation. The mycotoxin DON levels observed in maize tissue experiments corresponded with patterns in F. graminearum biomass, indicating that Bt transformation has no impact on DON production. In addition to bioassays, maize leaf tissue was analyzed with a mass spectrometer-based electronic nose, generating fingerprints of volatile organic compounds. Chemical fingerprints of corresponding Bt and non-Bt leaf tissues differed only for those hybrid pairs that caused differential fungal biomass production in the bioassays. Our results suggest that Cry1Ab protein in maize residues has no direct effect on F. graminearum and T. atroviride but some corresponding Bt/non-Bt maize hybrids differ more in composition than Cry protein content alone, which can affect the saprophytic growth of fungi on crop residues.

Biodegradation, Environmental↗
PubMed2004

Direct comparison of the crystal and solution structure of xylanase from Trichoderma longibrachiatum.

The small angle X-ray scattering (SAXS) data of xylanase XYNII (endo-1,4-beta-xylan xylanohydrolase EC 3.2.1.8) from Trichoderma longibrachiatum, an enzyme catalysing the reaction of accidental hydrolysis of beta-1,4-D-xylosidic linkages of xylan, were recorded for protein solution using synchrotron radiation. The experimental data were compared with those of theoretical scattering calculated on the basis of the known crystal structure. The radius of gyration measured by SAXS (RG = 1.7 nm) was about 3.5% larger and the maximum dimension in the distance distribution function about 5 % larger than the corresponding values calculated on the basis of the crystal structure.

Crystallography, X-Ray↗
PubMed2000

Synergy between ruminal fibrolytic enzymes and enzymes from Trichoderma longibrachiatum.

The mechanism by which enzyme additives improve feed digestion in ruminants is not fully understood. Direct hydrolysis of feed in the rumen is a potential mode of action, but the importance of this mode needs to be quantified because of the relatively low exogenous hydrolase activity added compared with the total activity present in the rumen. We examined the interactions between ruminal and exogenous enzymes on fiber degradation using a completely randomized experimental design, with an 11 (enzyme preparations and their combinations) x 5 (assay pH) arrangement of treatments. Ruminal enzymes were extracted from cattle receiving high fiber or high concentrate diets and exogenous enzymes were Trichoderma longibrachiatum preparations containing different proportions of xylanase and cellulase activities. Ruminal and exogenous enzyme preparations and their combinations were tested for the ability to degrade soluble cellulose, xylan, and corn silage over a range of pH from 4.5 to 6.5 at 39 degrees C. T. longibrachiatum enzymes acted synergistically with enzymes from mixed rumen microorganisms in degrading soluble cellulose, xylan, and corn silage. Hydrolysis increased by up to 35, 100, and 40% for soluble cellulose, xylan, and corn silage, respectively, and was most evident at a pH range between 5.0 and 6.0. The synergistic effect between ruminal and exogenous enzymes increases the hydrolytic potential within the rumen environment and is likely a significant mechanism by which enzyme additives improve feed digestion.

Animal Feed↗
PubMed1981

The viscosity interaction of barley beta-glucan with Trichoderma viride cellulase in the chick intestine.

A culture filtrate from Trichoderma viride was used as a source of cellulose degrading enzymes. This filtrate, when added to a barley ration fed to chicks, improved growth and feed efficiency by 19% and 8%, respectively. The response was saturable since increasing amounts of filtrate above 50 mg/kg produced no further improvement. Barley eta-glucan was added to a corn-based diet (10 g/kg) to evaluate the effect of the culture filtrate on the viscosity of beta-glucan in the chick intestinal contents. Barley beta-glucan increased the viscosity of the supernatant of the chick intestinal contents threefold and the culture filtrate reduced the viscosity to near control levels when combined with the diet containing beta-glucan. The presence of beta-glucan in barley probably limits growth of chicks by increasing viscosity of intestinal contents.

Animals↗
PubMed1985

Effects of cellulase from Trichoderma viride on nutrient utilization by broilers.

Studies were conducted to evaluate the effects of cellulase from Trichoderma viride in a diet for broilers containing high levels of wheat bran. Broiler-type, mixed-sex chicks were fed from 3 to 8 weeks of age. Wheat bran was added at 0, 10, and 20% levels. A fourth group received the 20% wheat bran plus the cellulase enzyme added at the level of .008%. A portion of the chicks was used in a digestibility study with chromic oxide as an indicator. The summarized data showed that cellulose treatment had a significant effect on reducing feed consumption (P less than .01) and an apparent effect in improving feed-to-gain ratio. Cellulase supplementation significantly improved the digestibility of cell wall components (P less than .01). Calcium, phosphorus, iron, zinc, and copper associated with cell walls were solubilized by cellulase. Iron balance was negative in the groups without cellulase; however, iron, which is bound by the bran, apparently was made available for absorption by cellulase.

Animals↗
PubMed1996

Influence of Trichoderma viride enzyme complex on nutrient utilization and performance of laying hens in diets with and without antibiotic supplementation.

The experiment was designed to test possible interactions of an enzyme complex (product from Trichoderma viride) and a feed antibiotic (flavophospholipol) in a barley diet on metabolism variables and egg production performance of Warren Brown laying hens. The basal diet contained 40% winter barley (French cultivar "Express", six row). The four treatments were as follows: O, control (without supplement); E, enzyme complex, 600 ppm; A, flavophospholipol, 10 ppm; EA, enzyme complex, 600 ppm and flavophospholipol, 10 ppm. The enzyme complex contained the following main activities: cellulase (10,500 U/g), endo-beta-(1:3)(1:4)-glucanase (24,000 U/g), and xylanase (32,000 U/g). The enzyme positively influenced AME content of the feed, organic matter (OM) utilization, and neutral detergent fiber (NDF) degradability (P < or = 0.01). When supplemented alone, the antibiotic had no influence on energy and nutrient utilization. No significant differences in egg production due to dietary treatments were observed. A significant enzyme by antibiotic interaction for AME (P < or = 0.01) and OM utilization (P < or = 0.001) as well as NDF degradability (P < or = 0.01) indicated a reduced enzyme effect in the diet containing antibiotic. Negative enzyme by antibiotic interaction for energy utilization in laying hens suggested that the positive response to dietary enzyme supplementation in the mature laying hen (Treatment O vs E) was to great extent mediated by the activity of intestinal microbes.

Animal Feed↗
PubMed2002

The expression of extracellular fungal cell wall hydrolytic enzymes in different Trichoderma harzianum isolates correlates with their ability to control Pyrenochaeta lycopersici.

Four isolates of Trichoderma harzianum (ThN3, Th11, Th12 and Th16) were selected for their ability to control the in vitro development of the tomato root pathogen Pyrenochaeta lycopersici. Analysis of the mechanisms involved in biocontrol showed that the formation of non-volatile metabolites appears to be one of those involved in biocontrol of P. lycopersici by all T. harzianum isolates tested. Nevertheless, the higher secretion of chitinases, both in number of isoenzymes and activity by the Th11 strain, correlated well with its higher ability to control this agent in laboratory and greenhouse experiments as compared to the other T. harzianum isolates tested. The secretion of beta-1,3-endoglucanases and/or proteases appeared to have less significance than endochitinases in the biological control of P. lycopersici.

Cell Wall↗
PubMed1999

Trichoderma viride suppresses fumonisin B1 production by Fusarium moniliforme.

Biocontrol activity against Fusarium moniliforme was analyzed for a Trichoderma viride strain isolated from root segments of corn plants grown in Piedmont Georgia. The isolate suppressed radial extension of F. moniliforme colonies during cocultivation on potato dextrose agar and fumonisin B1 (FB1) production during incubation of both fungi on corn kernels. T. viride decreased radial extension of F. moniliforme by 46% after 6 days and by 90% after 14 days. Furthermore, the colony diameter of F. moniliforme was less at 14 days than at 5 days, suggesting that F. moniliforme mycelia were undergoing lysis. FB1 production by F. moniliforme on corn kernels decreased by 85% when both organisms were inoculated the same day onto corn kernels and by 72% when inoculation of T. viride was delayed by 7 days after F. moniliforme inoculation. These results are the first to demonstrate that T. viride can suppress FB1 production by F. moniliforme, thereby functioning to control mycotoxin production. Thus, this isolate may be useful in biological control to inhibit F. moniliforme growth as a preharvest agent to prevent disease during plant development and/or as a postharvest agent during seed storage to suppress FB1 accumulation when kernels are dried inadequately.

Carboxylic Acids↗
PubMed1994

Tricholin, a new antifungal agent from Trichoderma viride, and its action in biological control of Rhizoctonia solani.

Tricholin, a ribosome-inactivating protein isolated from the culture broth of Trichoderma viride, has been shown to exert fungicidal effects on Rhizoctonia solani through a multi-hit kinetic interaction. Tricholin causes a parallel cessation of growth, uptake of amino acids, and protein biosynthesis. The in vivo mode of action of tricholin on protein synthesis and cell growth appears to be attributed to the diminishing of the polysome formation in R. solani through damage to large ribosomal subunits. These results concur with previous data and prove that tricholin is an effective inhibitor of protein synthesis. The efficacy of tricholin as an antibiotic agent was estimated to have a duration of approximately 42 hours.

Antifungal Agents↗