Showing papers on "Trichoderma harzianum published in 1999"
TL;DR: This is the first report of the ability of a Trichoderma strain to solubilize some insoluble or sparingly soluble minerals via three possible mechanisms: acidification of the medium, production of chelating metabolites, and redox activity.
Abstract: We investigated the capability of the plant-growth-promoting and biocontrol fungus Trichoderma harzianum Rifai 1295-22 (T-22) to solubilize in vitro some insoluble or sparingly soluble minerals via three possible mechanisms: acidification of the medium, production of chelating metabolites, and redox activity. T-22 was able to solubilize MnO2, metallic zinc, and rock phosphate (mostly calcium phosphate) in a liquid sucrose-yeast extract medium, as determined by inductively coupled plasma emission spectroscopy. Acidification was not the major mechanism of solubilization since the pH of cultures never fell below 5.0 and in cultures containing MnO2 the pH rose from 6.8 to 7.4. Organic acids were not detected by high-performance thin-layer chromatography in the culture filtrates. Fe2O3, MnO2, Zn, and rock phosphate were also solubilized by cell-free culture filtrates. The chelating activity of T-22 culture filtrates was determined by a method based on measurement of the equilibrium concentration of the chrome azurol S complex in the presence of other chelating substances. A size exclusion chromatographic separation of the components of the culture filtrates indicated the presence of a complexed form of Fe but no chelation of Mn. In liquid culture, T. harzianum T-22 also produced diffusible metabolites capable of reducing Fe(III) and Cu(II), as determined by the formation of Fe(II)-Na2-bathophenanthrolinedisulfonic acid and Cu(I)-Na2-2, 9-dimethyl-4,7-diphenyl-1,10-phenanthrolinedisulfonic acid complexes. This is the first report of the ability of a Trichoderma strain to solubilize insoluble or sparingly soluble minerals. This activity may explain, at least partially, the ability of T-22 to increase plant growth. Solubilization of metal oxides by Trichoderma involves both chelation and reduction. Both of these mechanisms also play a role in biocontrol of plant pathogens, and they may be part of a multiple-component action exerted by T-22 to achieve effective biocontrol under a variety of environmental conditions.
785 citations
TL;DR: Biochemical analyses revealed that inoculation with Trichodermainitiated increased peroxidase and chitinase activities within 48 and 72 h, respectively, providing evidence that T. harzianum may induce systemic resistance mechanisms in cucumber plants.
Abstract: The potential of the biocontrol agent Trichoderma harzianum T-203 to trigger plant defense responses was investigated by inoculating roots of cucumber seedlings with Trichoderma in an aseptic, hydroponic system. Trichoderma-treated plants were more developed than nontreated plants throughout the experiment. Electron microscopy of ultrathin sections from Trichoderma-treated roots revealed penetration of Trichoderma into the roots, restricted mainly to the epidermis and outer cortex. Strengthening of the epidermal and cortical cell walls was observed, as was the deposition of newly formed barriers. These typical host reactions were found beyond the sites of potential fungal penetration. Wall appositions contained large amounts of callose and infiltrations of cellulose. The wall-bound chitin in Trichoderma hyphae was preserved, even when the hyphae had undergone substantial disorganization. Biochemical analyses revealed that inoculation with Trichoderma initiated increased peroxidase and chitinase activities within 48 and 72 h, respectively. These results were observed for both the roots and the leaves of treated seedlings, providing evidence that T. harzianum may induce systemic resistance mechanisms in cucumber plants.
768 citations
TL;DR: It is confirmed that substances released by the AM fungus in the growth medium is the main factor explaining differential growth of the microorganisms tested and suggested that direct interactions exist between AM fungi and soil microorganisms, which might lead to changes in microbial equilibrium detrimental to pathogens.
Abstract: Arbuscular mycorrhizal (AM) fungi can reduce the incidence and importance of plant root diseases caused by pathogens. The mechanisms involved are not well characterized. We used an in vitro experimental system to test the hypothesis that the extraradical mycelium of AM fungi can interfere directly with microorganisms in the mycosphere and directly or indirectly reduce the population of plant pathogens. This system permitted the isolation of soluble substances released by the extraradical mycelium of Glomus intraradices. The AM fungus was grown on Daucus carota transformed roots in one compartment, while only the extraradical mycelium was allowed to grow in a second compartment. A freezing and centrifugation technique was developed for the extraction and concentration of substances present in the compartment containing only the AM fungal mycelium. Four soil-inhabiting microorganisms were selected, and conidial germination (fungi) or growth (bacteria) of these was studied in the presence and absence (control) of the extract. In comparison with the control, the results indicated that both the growth of Pseudomonas chlororaphis and the conidial germination of Trichoderma harzianum were stimulated in the presence of the AM fungal extract. In contrast, conidial germination of Fusarium oxysporum f. sp. chrysanthemi was reduced while the growth of Clavibacter michiganensis subsp. michiganensis was not affected. The measured effects in general were directly correlated with extract concentration. Differences in pH were noted between the extract containing substances released by the AM fungus and the non-AM control, but no significant influence of the pH on growth or conidial germination was noted, confirming that substances released by the AM fungus in the growth medium is the main factor explaining differential growth of the microorganisms tested. The results suggest that direct interactions exist between AM fungi and soil microorganisms, which might lead to changes in microbial equilibrium detrimental to pathogens.
313 citations
TL;DR: The results indicate that ech42 is expressed before contact of T. harzianum with R. solani and its induction is triggered by soluble chitooligosaccharides produced by constitutive activity of CHIT42 and/or other chitinolytic enzymes.
256 citations
TL;DR: Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39.
Abstract: The role of protease of Trichoderma harzianum in the biocontrol of Botrytis cinerea was examined. Two isolates of T. harzianum were compared for their ability to produce protease in liquid culture medium and on the surface of bean leaves. The biocontrol agent T. harziaum T39 produced 58 mU/ml of protease and T. harzianum NCIM1185 produced 54 mU/ml on the 5th day of growth in liquid culture medium. On bean leaves, combinations of B. cinerea and T. harzianum isolates were examined for the synthesis of protease. The protease activities were 0.9 and 0.6 mU/ml for T. harzianum T39 and NCIM1185, respectively, and 0.5 mU/ml for B. cinerea alone after 48 h of incubation. In the presence of T. harzianum T39 culture liquid containing protease, a 55% reduction in B. cinerea germination and a 80% reduction in the germ tube length were observed after 17 h of incubation in vitro. When T. harzianum isolates were added to B. cinerea on bean leaves, increased synthesis of protease was observed (1.0 and 1.2 mU/ml for T39 and NCIM1185, respectively). In the presence of T. harzianum NCIM1185 protease, although the rate of germination was reduced, B. cinerea attained 98% germination after 17 h of incubation. The hydrolytic enzymes produced by B. cinerea, endo-polygalacturonase (PG) and exoPG were partially deactivated by protease from the T. harzianum isolates. Carboxymethyl cellulase was deactivated only by protease of NCIM1185. On the surface of bean leaves, the protease (obtained from liquid culture medium of T. harzianum isolates) resulted in 56–100% reduction of disease severity. The culture liquid containing protease synthesized on the surface of bean leaves treated with B. cinerea and with T. harzianum was collected and added to fresh leaves infected by B. cinerea. There was 56–100% and 30–75% reduction of disease severity with liquid droplet collected from the leaves treated with T. harzianum T39 and NCIM1185, respectively. Increased control of disease was obtained by combining the conidia of T. harzianum isolates with protease obtained from culture media. Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39. T39 was found to be a poor producer of chitinase and β-1,3-glucanase in vitro. These enzymes were not detected on leaves treated with T39. Involvement of protease in biocontrol of B. cinerea is suggested.
248 citations
TL;DR: The level of extracellular endochitinase activity when T. harzianum was grown under inducing conditions increased up to 42-fold in multicopy strains as compared with the wild type, whereas gene disruptants exhibited practically no activity.
Abstract: The role of the Trichoderma harzianum endochitinase (Ech42) in mycoparasitism was studied by genetically manipulating the gene that encodes Ech42, ech42. We constructed several transgenic T. harzianum strains carrying multiple copies of ech42 and the corresponding gene disruptants. The level of extracellular endochitinase activity when T. harzianum was grown under inducing conditions increased up to 42-fold in multicopy strains as compared with the wild type, whereas gene disruptants exhibited practically no activity. The densities of chitin labeling of Rhizoctonia solani cell walls, after interactions with gene disruptants were not statistically significantly different than the density of chitin labeling after interactions with the wild type. Finally, no major differences in the efficacies of the strains generated as biocontrol agents against R. solani or Sclerotium rolfsii were observed in greenhouse experiments.
176 citations
TL;DR: It seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum, and the use of a compartmented growth system with root-free soil compartments suggests that nutrient competition is a likely means of interaction.
Abstract: Trichoderma harzianum is an effective biocontrol agent against several fungal soilborne plant pathogens. However, possible adverse effects of this fungus on arbuscular mycorrhizal fungi might be a drawback in its use in plant protection. The objective of the present work was to examine the interaction between Glomus intraradices and T. harzianum in soil. The use of a compartmented growth system with root-free soil compartments enabled us to study fungal interactions without the interfering effects of roots. Growth of the fungi was monitored by measuring hyphal length and population densities, while specific fatty acid signatures were used as indicators of living fungal biomass. Hyphal 33P transport and beta-glucuronidase (GUS) activity were used to monitor activity of G. intraradices and a GUS-transformed strain of T. harzianum, respectively. As growth and metabolism of T. harzianum are requirements for antagonism, the impact of wheat bran, added as an organic nutrient source for T. harzianum, was investigated. The presence of T. harzianum in root-free soil reduced root colonization by G. intraradices. The external hyphal length density of G. intraradices was reduced by the presence of T. harzianum in combination with wheat bran, but the living hyphal biomass, measured as the content of a membrane fatty acid, was not reduced. Hyphal 33P transport by G. intraradices also was not affected by T. harzianum. This suggests that T. harzianum exploited the dead mycelium but not the living biomass of G. intraradices. The presence of external mycelium of G. intraradices suppressed T. harzianum population development and GUS activity. Stimulation of the hyphal biomass of G. intraradices by organic amendment suggests that nutrient competition is a likely means of interaction. In conclusion, it seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum; in contrast, T. harzianum was adversely affected by G. intraradices.
168 citations
TL;DR: In this article, the expression of the two major chitinase genes, ech42 and nag1, was investigated by using a reporter system based on the Aspergillus niger glucose oxidase.
Abstract: Regulation of the expression of the two major chitinase genes, ech42 (encoding the CHIT42 endochitinase) and nag1 (encoding the CHIT73 N-acetyl-β-d-glucosaminidase), of the chitinolytic system of the mycoparasitic biocontrol fungus Trichoderma atroviride (= Trichoderma harzianum P1) was investigated by using a reporter system based on the Aspergillus niger glucose oxidase. Strains harboring fusions of the ech42 or nag1 5′ upstream noncoding sequences with the A. niger goxA gene displayed a glucose oxidase activity pattern that was consistent under various conditions with expression of the native ech42 and nag1 genes, as assayed by Northern analysis. The expression product of goxA in the mutants was completely secreted into the medium, detectable on Western blots, and quantifiable by enzyme-linked immunosorbent assay. nag1 gene expression was triggered during growth on fungal (Botrytis cinerea) cell walls and on the chitin degradation product N-acetylglucosamine. N-Acetylglucosamine, di-N-acetylchitobiose, or tri-N-acetylchitotriose also induced nag1 gene expression when added to mycelia pregrown on different carbon sources. ech42 expression was also observed during growth on fungal cell walls but, in contrast, was not triggered by addition of chitooligomers to pregrown mycelia. Significant ech42 expression was observed after prolonged carbon starvation, independent of the use of glucose or glycerol as a carbon source, suggesting that relief of carbon catabolite repression was not involved in induction during starvation. In addition, ech42 gene transcription was triggered by physiological stress, such as low temperature, high osmotic pressure, or the addition of ethanol. Four copies of a putative stress response element (CCCCT) were found in the ech42 promoter.
163 citations
TL;DR: Transformants of the biocontrol agent Trichoderma harzianum strain CECT 2413 that overexpressed a 33-kDa chitinase (Chit33) were obtained and characterized and indicated that the chit33 gene was integrated ectopically, mostly in tandem.
Abstract: Transformants of the biocontrol agent Trichoderma harzianum strain CECT 2413 that overexpressed a 33-kDa chitinase (Chit33) were obtained and characterized. Strain CECT 2413 was cotransformed with the amdS gene and its own chit33 gene under the control of the pki constitutive promoter from T. reesei. Southern blotting indicated that the chit33 gene was integrated ectopically, mostly in tandem. Some transformants showed the same restriction pattern, indicating preferable sites of integration. There was no correlation between the number of integrated copies and the level of expression of the chit33 gene in the transformants. When grown in glucose, the extracellular chitinase activity of the transformants was up to 200-fold greater than that of the wild type, whereas in chitin, the activity of both the transformants and the wild type was similar. Under both conditions, the transformants were more effective in inhibiting the growth of Rhizoctonia solani as compared with the wild type. Similar results were obtained when culture supernatants from the transformants and the wild type were tested against R. solani.
135 citations
TL;DR: In vitro antifungal activity of the ech42 disruptant culture filtrates against Botrytis cinerea and Rhizoctonia solani was reduced about 40%, compared with wild type; antif fungus activity was fully restored by adding an equivalent amount of CHIT42 as secreted by P1.
Abstract: The biocontrol strain P1 of Trichoderma harzianum was genetically modified by targeted disruption of the single-copy ech42 gene encoding for the secreted 42-kDa endochitinase (CHIT42). Stable mutants in which ech42 was interrupted, and unable to produce CHIT42, were obtained and characterized. These mutants lacked the ech42 transcript, the protein, and endochitinase activity in culture filtrates, and they were unable to clear a medium containing colloidal chitin. Other chitinolytic and glucanolytic enzymes expressed during mycoparasitism were not affected by the disruption of ech42. The disrupted mutant D11 grew and sporulated similarly to the wild type. In vitro antifungal activity of the ech42 disruptant culture filtrates against Botrytis cinerea and Rhizoctonia solani was reduced about 40%, compared with wild type; antifungal activity was fully restored by adding an equivalent amount of CHIT42 as secreted by P1. The mutant exhibited the same biocontrol effect against Pythium ultimum as strain P1, but t...
134 citations
TL;DR: A 110kDa novel extracellular beta-1,3-exoglucanase from T. harzianum is purified and contains a unique C-terminal embodying cysteine motifs, enabling Trichoderma to utilize both their cell walls and cellular contents for nutrition.
TL;DR: Two novel type I ribosome-inactivating proteins (RIPs) were found in the storage roots of Mirabilis expansa, an underutilized Andean root crop, and their antimicrobial activity was tested against root-rot microorganisms, among others.
Abstract: Two novel type I ribosome-inactivating proteins (RIPs) were found in the storage roots of Mirabilis expansa, an underutilized Andean root crop. The two RIPs, named ME1 and ME2, were purified to homogeneity by ammonium sulfate precipitation, cation-exchange perfusion chromatography, and C4 reverse-phase chromatography. The two proteins were found to be similar in size (27 and 27.5 kD) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their isoelectric points were determined to be greater than pH 10.0. Amino acid N-terminal sequencing revealed that both ME1 and ME2 had conserved residues characteristic of RIPs. Amino acid composition and western-blot analysis further suggested a structural similarity between ME1 and ME2. ME2 showed high similarity to the Mirabilis jalapa antiviral protein, a type I RIP. Depurination of yeast 26S rRNA by ME1 and ME2 demonstrated their ribosome-inactivating activity. Because these two proteins were isolated from roots, their antimicrobial activity was tested against root-rot microorganisms, among others. ME1 and ME2 were active against several fungi, including Pythium irregulare, Fusarium oxysporum solani, Alternaria solani, Trichoderma reesei, and Trichoderma harzianum, and an additive antifungal effect of ME1 and ME2 was observed. Antibacterial activity of both ME1 and ME2 was observed against Pseudomonas syringae, Agrobacterium tumefaciens, Agrobacterium radiobacter, and others.
TL;DR: Analysis of the fungal populations in the plant growth substrate showed that T. harzianum consistently reduced that of Phytophthora capsici over time, and this reduction in the pathogen population was associated with a reduction in root rot.
Abstract: The ability of Trichoderma harzianum to control the rotting of pepper (Capsicum annuum) plant roots caused by Phytophthora capsici was studied. Interactions between the fungi were assessed in vitro on three culture media (V8c, Czapek and 2% water agar) and in vivo in plants grown in a substrate inoculated with P. capsici and T. harzianum. Studies on mutual antagonism in vitro showed that P. capsici was inhibited by T. harzianum; however, the intensity of inhibition differed according to the medium used, being greatest on Czapek. Analysis of the fungal populations in the plant growth substrate showed that T. harzianum consistently reduced that of P. capsici over time. This reduction in the pathogen population was associated with a reduction in root rot of between 24 and 76%, although plant growth (dry weight) was still reduced by 21.2–24.7%, compared with the uninoculated control. In the absence of T. harzianum with the same pathogen inoculum levels, the reduction in dry weight was 59.8–68.6%, suggesting that T. harzianum reduced the damage.
TL;DR: Prospects for biocontrol of B. cinerea in greenhouse vegetables appear good under a range of conditions, but regression analysis showed that high temperature during the day and high vapour pressure deficit during the night reduced biOControl efficacy.
Abstract: The efficacy of Trichoderma harzianum T39 and the yeasts Aureobasidium pullulans and Cryptococcus albidus against Botrytis cinerea in cucumber and tomato was compared with chemical control. Four experiments were conducted in cucumber grown under different climatic conditions in The Netherlands, and two experiments were done in tomato both in the Netherlands and in Israel. T. harzianum and A. pullulans showed the most consistent control of B. cinerea, reducing stem lesions and death of plants by 40–100% in most cases. Control of stem lesions and subsequent wilting was generally better than control of symptoms on fruits. In some cases, the biocontrol agents were more effective than the broad-spectrum fungicide tolylfluanid and the selective fungicide iprodione. The climatic conditions did not strongly influence the efficacy of the biocontrol agents, but regression analysis showed that high temperature during the day and high vapour pressure deficit during the night reduced biocontrol efficacy. From the results, prospects for biocontrol of B. cinerea in greenhouse vegetables appear good under a range of conditions.
TL;DR: Light induction ofphr1 in non-sporulating mutants shows that a complete sporulation pathway is not required for photoregulation, suggesting that photoinduction of sporulation and of photolyase expression is distinct in their photoreceptor system or in the transduction of the blue light signal.
TL;DR: A polymerase chain reaction-amplified DNA containing the internal transcribed spacer (ITS)-1, 5.8S, and ITS-2 regions of the nuclear ribosomal DNA transcriptional unit was sequenced for 81 isolates of Trichoderma spp.
Abstract: A polymerase chain reaction-amplified DNA containing the internal transcribed spacer (ITS)-1, 5.8S, and ITS-2 regions of the nuclear ribosomal DNA transcriptional unit was sequenced for 81 isolates of Trichoderma spp. associated with mushroom culture or used for biological control of plant pathogens. Phylogenetic analyses revealed that the biocontrol isolates were more closely related to an isolate of T. harzianum biotype 1 (Th1) than to the aggressive biotypes 2 and 4. Th1 has been isolated from mushroom compost but is not the cause of widespread green mold epidemics that have occurred during the last 12 years in Europe and North America. Three isolates of T. harzianum obtained from shiitake (Lentinula edodes; Shi1B and S3-96) and maitake (Grifola frondosa; Mai1) substrates were placed within the biocontrol group. We also found evidence suggesting that some isolates of T. harzianum originally identified as Th4 from Pennsylvania are more closely related to Th2 from Europe. Finally, considering the wide range in sequence distribution of our samples, we propose that the consensus sequence found in this investigation be used as the reference sequence for further studies involving the identification and taxonomy of T. harzianum.
TL;DR: Of fifteen isolates of yeasts, filamentous fungi and bacteria and a commercial product, tested in a bioassay with stem segments, eleven isolates consistently reduced incidence of disease and sporulation of Botrytis cinerea Pers; Fr in tomato and seven isolates in cucumber.
Abstract: Of fifteen isolates of yeasts, filamentous fungi and bacteria and a commercial product, tested in a bioassay with stem segments, eleven isolates consistently reduced incidence of disease and sporulation of Botrytis cinerea Pers; Fr in tomato and seven isolates in cucumber. Several isolates reduced disease by more than 75% in all experiments. Six antagonists that performed well in the bioassays and that were fairly easy to produce in vitro, were selected for further testing in two glasshouse experiments with cucumbers. After application of spores of B. cinerea and the antagonists or the fungicide tolylfluanid to pruning wounds, disease incidence was reduced by 50–100% by all antagonists in both experiments and only in one experiment by tolylfluanid.
TL;DR: Among these, Phoma sp.
Abstract: A number of fungal cultures were screened to select an organism suitable to be used in the detoxification of aflatoxin B1. They were co-cultured in Czapek-Dox-Casamino acid medium with aflatoxin B1 producing Aspergillus flavus. Several fungal cultures were found to prevent synthesis of aflatoxin B1 in liquid culture medium. Among these Phoma sp., Mucor sp., Trichoderma harzianum, Trichoderma sp. 639, Rhizopus sp. 663, Rhizopus sp. 710, Rhizopus sp. 668, Alternaria sp. and some strains belonging to the Sporotrichum group (ADA IV B14(a), ADA SF VI BF (9), strain 720) could inhibit aflatoxin synthesis by ≥90 %. A few fungi, namely ADA IV B1, ADA F1, ADA F8, also belonging to the Sporotrichum group, were less efficient than the Phoma sp. The Cladosporium sp. and A. terreus sp. were by far the least efficient, registering <10 % inhibition. The cultures which prevent aflatoxin biosynthesis are also capable of degrading the preformed toxin. Among these, Phoma sp. was the most efficient destroying about 99 % of aflatoxin B1. The cell free extract of Phoma sp. destroyed nearly 50 µg aflatoxin B1 100 ml−1 culture medium (90 % of the added toxin), and this was more effective than its own culture filtrate over 5 days incubation at 28 ± 2 °C. The degradation was gradual: 35 % at 24 h, 58 % at 48 h, 65 % at 72 h, 85 % at 96 h and 90 % at 120 h. The possibility of a heat stable enzymatic activity in the cell free extract of Phoma is proposed. Copyright © 1999 John Wiley & Sons, Ltd.
TL;DR: Formulations of P fluorescens were effective in reducing the F moniliforme infection and also in increasing the seed germination, vigour index and field emergence, followed by T harzianum and C globosum treatments in comparison with control.
Abstract: Five different cultivars of sorghum seeds infected with a varied degree of Fusarium moniliforme were treated with biocontrol agents. Pure cultures of Pseudomonas fluorescens, Trichoderma harzianum and Chaetomium globosum at the rate of 1 x 10(8) cfu g(-1) and talcum based formulations of (28 x 10(7) cfu g(-1)), (19 x 10(7) cfu g(-1)) and (4 x 10(6) cfu g(-1)) at the rate of 6 g kg(-1) and 10 g kg(-1) of seeds were used, respectively. The treated seeds were evaluated for per cent reduction of F moniliforme, seed germination, vigour index and field emergence. It was found that the pure culture of P fluorescens was more effective in reducing the F moniliforme infection followed by T harzianum and C globosum than the Bavistin treated and untreated seeds. Formulations of P fluorescens were effective in reducing the F moniliforme infection and also in increasing the seed germination, vigour index and field emergence, followed by T harzianum and C globosum treatments in comparison with control. (C) 1999 Society of Chemical Industry.
TL;DR: The purification to electrophoretical homogeneity of BGN16.1, the second beta-1, 6-glucanase enzyme, shows an endo-hydrolytic mode of action and the probable role of this enzyme in the antagonistic action of T. harzianum is discussed.
Abstract: The antagonistic fungus Trichoderma harzianum CECT 2413 produces at least two extracellular beta-1,6-glucanases, among other hydrolases acting on polysaccharides from fungal cell walls, when grown in chitin as the sole carbon source We have previously reported on the purification and biochemical characterization of the major activity, which corresponds to an acidic enzyme named BGN162 [de la Cruz, J, Pintor-Toro, JA, Benitez, T & Llobell, A (1995) J Bacteriol 177, 1864-1871] In this paper, we report on the purification to electrophoretical homogeneity of BGN161, the second beta-1, 6-glucanase enzyme BGN161 was purified by ammonium sulfate precipitation followed by adsorption and digestion of pustulan (a beta-1,6-glucan), chromatofocusing and gel-filtration chromatography BGN161 is a non-glycosylated protein with an apparent molecular mass of 51 kDa and a basic isoelectric point (pI 74-77) The enzyme was active toward substrates containing beta-1,6-glycosidic linkages, including yeast cell walls The Km was 08 mg x mL-1 with pustulan as the substrate Reaction product analysis by HPLC clearly indicated that BGN161 has an endo-hydrolytic mode of action The probable role of this enzyme in the antagonistic action of T harzianum is also discussed
TL;DR: Five microbial products claimed by their manufacturers to protect against soil-borne plant pathogens were tested in the greenhouse for efficacy against Pythium ultimum on cucumber and Rhizoctonia solani on peas, and SOILGARD was the only biocontrol product that gave significant control of P. ultimum.
TL;DR: Two strains with known biocontrol capabilities were found to relate to the genotypic group containing Th1 and Th2 forms and, based on variation within this group, to belong to a homogeneous group of form Th1 strains.
TL;DR: Treatment of banana fruits with T. viride 4 h prior to inoculation with L. theobromae provided better protection than simultaneous application or treatment 4 h after inoculation, and Trichoderma harzianum exhibited the greatest inhibition in dual culture.
TL;DR: This case illustrates the widening spectrum of opportunistic Trichoderma spp.
Abstract: We describe the second known case of human infection by Trichoderma harzianum. A disseminated fungal infection was detected in the postmortem examination of a renal transplant recipient and confirmed in culture. The only other reported infection by this fungus caused peritonitis in a diabetic patient. The in vitro antifungal susceptibilities of the clinical strain and three other strains of Trichoderma species to six antifungal drugs are provided. This case illustrates the widening spectrum of opportunistic Trichoderma spp. in immunocompromised patients and emphasizes the problems in diagnosing invasive fungal diseases.
TL;DR: It is proposed that a signal for mycoparasitic behaviour from the host cell surface is transduced by heterotrimeric G protein(s) and mediated by cAMP, and will be a valuable assay to aid in the genetic manipulation of this pathway.
TL;DR: The self-directing optimization or the rotating simplex method of optimization was employed to determine the best suitable combination of parameters, pH (controlled), aeration rate and agitation rate for maximal production of chitinase by Trichoderma harzianum in batch culture.
TL;DR: Ampelomyces quisqualis, a mycoparasite of fungi causing powdery mildews, exhibited high levels of extracellular exo-beta-1,3-glucanase activity in culture compared with Neurospora crassa and Gliocladium roseum, and transcription was induced by fungal cell wall components.
Abstract: Ampelomyces quisqualis, a mycoparasite of fungi causing powdery mildews, exhibited high levels of extracellular exo-β-1,3-glucanase activity in culture compared with Neurospora crassa and Gliocladium roseum. A. quisqualis culture filtrates affected powdery mildew caused by Sphaerotheca fusca in a manner indicative of cell wall degradation, as determined by microscopic examination. A gene encoding an exo-β-1,3-glucanase in A. quisqualis, designated exgA, was isolated and sequenced. The predicted polypeptide deduced from exgA had 46, 42, and 30% identity with amino acid sequences of Trichoderma harzianum exo-β-1,3-glucanase and Cochliobolus carbonum EXG1 (both encoding exo-β-1,3-glucanase) and T. harzianum bng13.1 (encoding an endo-β-1,3-glucanase), respectively. The exgA gene had a predicted molecular mass of 84 kDa and a pI of 4.79. The gene was expressed during the late stages of growth in culture, and transcription was induced by fungal cell wall components. Transcript levels for exgA were pres...
TL;DR: The findings suggest that the onset of the green mold epidemic in North America resulted from the recent introduction of a highly virulent genotype of the pathogen into cultivated mushrooms.
Abstract: We used randomly amplified polymorphic DNA (RAPD)-PCR to estimate genetic variation among isolates of Trichoderma associated with green mold on the cultivated mushroom Agaricus bisporus. Of 83 isolates examined, 66 were sampled during the recent green mold epidemic, while the remaining 17 isolates were collected just prior to the epidemic and date back to the 1950s. Trichoderma harzianum biotype 4 was identified by RAPD analysis as the cause of almost 90% of the epidemic-related episodes of green mold occurring in the major commercial mushroom-growing region in North America. Biotype 4 was more closely allied to T. harzianum biotype 2, the predominant pathogenic genotype in Europe, than to the less pathogenic biotype 1 and Trichoderma atroviride (formerly T. harzianum biotype 3). No variation in the RAPD patterns was observed among the isolates within biotype 2 or 4, suggesting that the two pathogenic biotypes were populations containing single clones. Considerable genetic variation, however, was noted among isolates of biotype 1 and T. atroviride from Europe. Biotype 4 was not represented by the preepidemic isolates of Trichoderma as determined by RAPD markers and PCR amplification of an arbitrary DNA sequence unique to the genomes of biotypes 2 and 4. Our findings suggest that the onset of the green mold epidemic in North America resulted from the recent introduction of a highly virulent genotype of the pathogen into cultivated mushrooms.
01 Jan 1999
TL;DR: Factors that favour microbial growth, such as physiological senescence of fruits, mechanical injuries, as well as physiological disorders due to undesirable storage conditions can promote and explain these postharvest decays.
Abstract: Since early 1970’s, postharvest diseases of apple annually cause losses of 15–25 % despite modern storage facilities including controlled atmosphere (CA) or Ultra Low Oxygen (ULO) facilities (Bondoux, 1992). Factors that favour microbial growth, such as physiological senescence of fruits, mechanical injuries, as well as physiological disorders due to undesirable storage conditions can promote and explain these postharvest decays.