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PubMed2005

Trichoderma mitogen-activated protein kinase signaling is involved in induction of plant systemic resistance.

The role of a mitogen-activated protein kinase (MAPK) TmkA in inducing systemic resistance in cucumber against the bacterial pathogen Pseudomonas syringae pv. lacrymans was investigated by using tmkA loss-of-function mutants of Trichoderma virens. In an assay where Trichoderma spores were germinated in proximity to cucumber roots, the mutants were able to colonize the plant roots as effectively as the wild-type strain but failed to induce full systemic resistance against the leaf pathogen. Interactions with the plant roots enhanced the level of tmkA transcript in T. virens and its homologue in Trichoderma asperellum. At the protein level, we could detect the activation of two forms reacting to the phospho-p44/42 MAPK antibody. Biocontrol experiments demonstrated that the tmkA mutants retain their biocontrol potential against Rhizoctonia solani in soil but are not effective against Sclerotium rolfsii in reducing disease incidence. Our results show that, unlike in many plant-pathogen interactions, Trichoderma TmkA MAPK is not involved in limited root colonization. Trichoderma, however, needs MAPK signaling in order to induce full systemic resistance in the plant.

Cucumis sativus↗

PubMed1976

Competition between Phytophthora cinnamomi and Trichoderma spp. in autoclaved soil.

Results from analyses of beta-glucosidase (EC 3.2.1.21) and phosphatase (EC 3.1.3.1;EC 3.1.3.2) activities indicated that presence of a Trichoderma isolate reduced development of Phytophthora cinnamomi. It was also observed that P. cinnamomi was more competitive in coinoculated cultures than in cultures where Trichoderma was added on day 3. Analysis of trehalase (EC 3.2.1.28) activity indicated that Trichoderma either utilized portions of the P. cinnamomi mycelium as substrate or the action of P. cinnamomi released additional nutrients not normally available to Trichoderma. Ther stronger Trichoderma isolate was T. harzianum.

Acid Phosphatase↗

PubMed1999

Synthesis of a Trichoderma chitinase which affects the Sclerotium rolfsii and Rhizoctonia solani cell walls.

A Trichoderma sp. isolate, hereafter called T6, produces a 46-kDa endochitinase (CHIT 46) which had been shown to drastically affect in vitro the cell walls of the phytopathogens Sclerotium rolfsii and Rhizoctonia solani. We attempted to gain insight into its properties. The CHIT 46 N-terminal amino acid sequence shares a very high homology with other fungal chitinases. Western blot analysis using polyclonal antibodies anti-CHIT 46 revealed that this enzyme is immunologically distinct from other proteins produced by the same Trichoderma isolate T6, but is immunologically identical with proteins having equivalent molar mass, probably chitinases, produced by other Trichoderma spp. isolates. In addition, the antibodies revealed also that a substantial amount of this enzyme is secreted into the culture medium 2 d after the Trichoderma isolate T6 comes into contact with chitin.

Animals↗

PubMed2002

Expression regulation of the endochitinase chit36 from Trichoderma asperellum (T. harzianum T-203).

The presence of the endochitinase CHIT36 from Trichoderma harzianum TM was assessed in several antagonistic Trichoderma strains belonging to different molecular taxonomic groups. CHIT37 from T. harzianum CECT 2413 was sequenced and found to display 89% homology with CHIT36 at the amino acid level. Northern analysis showed that chit36Y from T. asperellum is regulated both by glucose and nitrogen levels. Stress conditions, colloidal chitin and N-acetyl-glucosamine are effective inducers of this gene. The promoter of chit36Y was cloned and was used to direct expression of a gfp reporter gene in Trichoderma transformants. Confrontation experiments with the plant pathogen Rhizoctonia solani revealed that direct contact between the fungi is not necessary for gfp expression. The R. solani-inducing factor appears to be a soluble molecule capable of diffusing through a dialysis membrane (<12 kDa). CHIT36 recombinant protein from the yeast Pichia pastoris was active against different phytopathogens, confirming the importance of this endochitinase in the mycoparasitic activity of Trichoderma antagonistic strains.

Alternaria↗

PubMed2004

Cell wall-degrading isoenzyme profiles of Trichoderma biocontrol strains show correlation with rDNA taxonomic species.

Trichoderma is known for being the most frequently used biocontrol agent in agriculture. A fundamental part of the Trichoderma antifungal system relies on a series of genes coding for a variety of extracellular lytic enzymes. Characterization of the polymorphism between five putative isoenzymatic activities [beta-1,3-glucanase (EC 3.2.1.39, EC 3.2.1.58), beta-1,6-glucanase (EC 3.2.1.75), cellulase (EC 3.2.1.4; EC 3.2.1.21, EC 3.2.1.91), chitinase (EC 3.2.1.30, EC 3.2.1.52), protease (EC 3.4.11; EC 3.4.13-19; EC 3.4.21-24, EC 3.4.99)] was carried out using 18 strains from three sections of Trichoderma. Of these, seven strains were from T. sect. Pachybasium, nine from T. sect. Trichoderma and two from T. sect. Longibrachiatum. Thirty-seven different alleles in total were identified: 13 for beta-1,3-glucanase, four for beta-1,6-glucanase, three for cellulase, eight for chitinase and nine for protease activity. A dendrogram (constructed by the unweighted pair group method with arithmetic averages) based on isoenzymatic data separated the 18 strains into three main enzymatic groups: T. harzianum, T. atroviride/T. viride/T. koningii and T. asperellum/T. hamatum/T. longibrachiatum. Isoenzymatic groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location, based on their sequence of internal transcribed spacer 1 in ribosomal DNA and their antifungal activities.

Antifungal Agents↗

PubMed1998

The expression of genes involved in parasitism by Trichoderma harzianum is triggered by a diffusible factor.

The mycoparasite Trichoderma harzianum has been extensively used in the biocontrol of a wide range of phytopathogenic fungi. Hydrolytic enzymes secreted by the parasite have been directly implicated in the lysis of the host. Dual cultures of Trichoderma and a host, with and without contact, were used as means to study the mycoparasitic response in Trichoderma. Northern analysis showed high-level expression of genes encoding a proteinase (prb1) and an endochitinase (ech42) in dual cultures even if contact with the host was prevented by using cellophane membranes. Neither gene was induced during the interaction of Trichoderma with lectin-coated nylon fibres, which are known to induce hyphal coiling and appressorium formation. Thus, the signal involved in triggering the production of these hydrolytic enzymes by T. harzianum during the parasitic response is independent of the recognition mediated by this lectin-carbohydrate interaction. The results showed that induction of prb1 and ech42 is contact-independent, and a diffusible molecule produced by the host is the signal that triggers expression of both genes in vivo. Furthermore, a molecule that is resistant to heat and protease treatment, obtained from Rhizoctonia solani cell walls induces expression of both genes. Thus, this molecule is involved in the regulation of the expression of hydrolytic enzymes during mycoparasitism by T. harzianum.

Base Sequence↗

PubMed2006

Trichoderma theobromicola and T. paucisporum: two new species isolated from cacao in South America.

Trichoderma theobromicola and T. paucisporum spp. nov. are described. Trichoderma theobromicola was isolated as an endophyte from the trunk of a healthy cacao tree (Theobroma cacao, Malvaceae) in Amazonian Peru; it sporulates profusely on common mycological media. Trichoderma paucisporum is represented by two cultures that were obtained in Ecuador from cacao pods partially infected with frosty pod rot, Moniliophthora roreri; it sporulates sporadically and most cultures remain sterile on common media and autoclaved rice. It sporulates more reliably on synthetic low-nutrient agar (SNA) but produces few conidia. Trichoderma theobromicola was reintroduced into cacao seedlings through shoot inoculation and was recovered from stems but not from leaves, indicating that it is an endophytic species. Both produced a volatile/diffusable antibiotic that inhibited development of M. roreri in vitro and on-pod trials. Neither species demonstrated significant direct in vitro mycoparasitic activity against M. roreri.

Base Sequence↗

PubMed2002

Release of ferulic acid from oat hulls by Aspergillus ferulic acid esterase and trichoderma xylanase.

Oat hulls, an agricultural byproduct, contain a relatively high amount of ferulic acid (FA; 4-hydroxy-3-methoxycinnamic acid), which is believed to be inhibitory to oat hull biodegradability by rumen microorganisms. In this paper, Aspergillus ferulic acid esterase (FAE) was investigated for its ability to release FA from oat hulls. The objectives were to determine the effects of particle size of oat hulls (ground to pass through 1 mm and 250 microm screens and a 100 microm sieve) on release of FA by FAE both in the presence and in the absence of Trichoderma xylanase. The results show that the release of FA by FAE was dependent upon the particle size of oat hulls (< or = 250 microm). In the absence of Trichoderma xylanase, little FA was released by FAE. In the presence of Trichoderma xylanase, there was a significant release of FA by FAE, indicating a synergistic interaction between FAE and Trichoderma xylanase on release of FA from oat hulls. These results indicate that FAE is able to break the ester linkage between FA and the attached sugar, releasing FA from oat hulls. This may leave the remainder of the polysaccharides open for further hydrolytic attack by rumen microorganisms. It is likely that removing FA from oat hulls could improve rumen biodegradability, thus improving the nutritional value of oat hulls.

Aspergillus↗

PubMed1997

In vitro studies on the potential for biological control of Aspergillus flavus and Fusarium moniliforme by Trichoderma species. 1. Macroscopical and microscopical observations of fungal interactions.

The possible biological control of the seed-associated fungi, Aspergillus flavus and Fusarium moniliforme by Trichoderma species was investigated. A study of the fungal growth in dual cultures revealed that four of nine isolates of two Trichoderma spp. (T. harzianum and T. viride) inhibited fungal growth. A scanning electron microscopical investigation of fungal interactions demonstrated no obvious hyphae penetration of A. flavus or F. moniliforme by Trichoderma spp. Morphological alteration of Fusarium hyphae with pronounced collapse and the production of aberrant conidial heads and microheads by A. flavus were observed. The results suggest that mycoparasitism is not the mechanism involved in the inhibitory interaction of either A. flavus or F. moniliforme with Trichoderma spp.

Antibiosis↗

PubMed2001

Antimicrobial activity of ergokonin A from Trichoderma longibrachiatum.

AIMS: Natural fungal products were screened for antifungal compounds. The mode of action of one of the hits found and the taxonomy of the producing organism were analysed. METHODS AND RESULTS: An extract from a Trichoderma species showed a more potent activity in an agar-based assay against the null mutant fks1::HIS strain than against the wild-type strain, suggesting that it could contain a glucan synthesis inhibitor. The active component was identified as the known compound ergokonin A. The compound exhibited activity against Candida and Aspergillus species, but was inactive against Cryptococcus species. It induced alterations in the hyphal morphology of Aspergillus fumigatus. The identification of the producing isolate was confirmed by sequencing of the rDNA internal transcribed spacers and comparison with the sequences of other Trichoderma species. The analysis showed that the producing fungus had a high homology with other strains classified as Trichoderma longibrachiatum and its teleomorph Hypocrea schweinitzii. CONCLUSIONS: The antifungal activity spectrum of ergokonin A and the morphology alterations induced on A. fumigatus are consistent with glucan synthesis as the target for ergokonin A. The production of ergokonin A is not uncommon, but is probably restricted to Trichoderma species. SIGNIFICANCE AND IMPACT OF THE STUDY: The discovery that ergokonin A could be an inhibitor of glucan synthesis, having a structure very different to other inhibitors, increases the likelihood that orally active agents with this fungal-specific mode of action may be developed.

Animals↗

PubMed1995

The role of recognition in the induction of specific chitinases during mycoparasitism by Trichoderma harzianum.

The induction of chitinolytic enzymes in the biocontrol agent Trichoderma harzianum during parasitism on Sclerotium rolfsii and the role of fungal-fungal recognition in this process were studied. A change in the chitinolytic enzyme profile was detected during the interaction between the fungi, grown in dual culture on synthetic medium. Before coming into contact with each other, both fungi contained a protein with constitutive 1,4-beta-N-acetylglucosaminidase activity. As early as 12 h after contact, the chitinolytic activity in S. rolfsii disappeared, while that of T. harzianum (a protein with a molecular mass of 102 kDa, CHIT 102) greatly increased. After 24 h of interaction, the activity of CHIT 102 diminished concomitantly with the appearance of a 73 kDa 1,4-beta-N-acetylglucosaminidase, which became clear and strong at 48 h. This phenomenon did not occur if the S. rolfsii mycelium was autoclaved prior to incubation with T. harzianum, suggesting its dependence on vital elements from the host. Cycloheximide inhibited this phenomenon, indicating that de novo synthesis of enzymes is taking place in Trichoderma during these stages of the parasitism. A biomimetic system based on the binding of a purified surface lectin from the host S. rolfsii to nylon fibres was used to dissect the effect of recognition. An increase in CHIT 102 activity was detected, suggesting that the induction of chitinolytic enzymes in Trichoderma is an early event which is elicited by the recognition signal (i.e. lectin-carbohydrate interactions). It is postulated that recognition is the first step in a cascade of antagonistic events which triggers the parasitic response in Trichoderma.

Acetylglucosaminidase↗

PubMed1999

PCR-based genotyping of epidemic and preepidemic Trichoderma isolates associated with green mold of Agaricus bisporus.

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.

Agaricus↗

PubMed2000

Molecular characterization and identification of biocontrol isolates of Trichoderma spp.

The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as "Trichoderma harzianum" Rifai and one biocontrol strain recognized as T. viride, was carried out using several molecular techniques. A certain degree of polymorphism was detected in hybridizations using a probe of mitochondrial DNA. Sequencing of internal transcribed spacers 1 and 2 (ITS1 and ITS2) revealed three different ITS lengths and four different sequence types. Phylogenetic analysis based on ITS1 sequences, including type strains of different species, clustered the 17 biocontrol strains into four groups: T. harzianum-T. inhamatum complex, T. longibrachiatum, T. asperellum, and T. atroviride-T. koningii complex. ITS2 sequences were also useful for locating the biocontrol strains in T. atroviride within the complex T. atroviride-T. koningii. None of the biocontrol strains studied corresponded to biotypes Th2 or Th4 of T. harzianum, which cause mushroom green mold. Correlation between different genotypes and potential biocontrol activity was studied under dual culturing of 17 BCAs in the presence of the phytopathogenic fungi Phoma betae, Rosellinia necatrix, Botrytis cinerea, and Fusarium oxysporum f. sp. dianthi in three different media.

DNA, Mitochondrial↗

PubMed1999

Fatal disseminated Trichoderma longibrachiatum infection in an adult bone marrow transplant patient: species identification and review of the literature.

Trichoderma longibrachiatum was recovered from stool surveillance cultures and a perirectal ulcer biopsy specimen from a 29-year-old male who had received an allogeneic bone marrow transplant for acute lymphoblastic leukemia. The amphotericin B (2.0 microgram/ml) and itraconazole (1.0 microgram/ml) MICs for the organism were elevated. Therapy with these agents was unsuccessful, and the patient died on day 58 posttransplantation. At autopsy, histologic sections from the lungs, liver, brain, and intestinal wall showed infiltration by branching septate hyphae. Cultures were positive for Trichoderma longibrachiatum. While Trichoderma species have been recognized to be pathogenic in profoundly immunosuppressed hosts with increasing frequency, this is the first report of probable acquisition through the gastrointestinal tract. Salient features regarding the identification of molds in the Trichoderma longibrachiatum species aggregate are presented.

Adult↗

PubMed1983

Beta-glucosidase excretion in Trichoderma strains with different cell wall bound beta-1,3-glucanase activities.

Two different strains of Trichoderma pseudokoningii (SE1 A8 and SE1 D81) and Trichoderma viride QM 9123 release into the medium different proportions of the total beta-glucosidase activity produced. This observation correlates with the degree of beta-1,3-glucanase binding to the cell wall found for each strain. DEAE-Sephadex ion-exchange chromatography revealed three peaks of beta-1,3-glucanase activity. These three enzymes (enzyme I, enzyme II, and enzyme III) differ in their extent of binding to the cell walls, their activity on isolated cell walls and Trichoderma beta-glucan, and their affinity for beta-glucan. Of these enzymes, enzyme II shows the largest variation in relative importance among the three strains and is located predominantly within the mural compartment. Enzyme II has the highest activity on and affinity for Trichoderma beta-glucan. Enzyme II is also the most active in releasing beta-glucosidase from cell walls of strain SE1 A8 (the strain excreting a high proportion of its beta-glucosidase into the culture fluid) as well as from strain SE1 D81 (little beta-glucosidase activity in the culture fluid). It is concluded that the action of beta-1,3-glucanase II on cell wall beta-glucan may be responsible for the in vivo release of cell wall bound beta-glucosidase into the culture fluid.

Cell Wall↗

PubMed1996

Mycoparasitic species of Trichoderma produce lectins.

Culture filtrates and mycelial extracts of two mycoparasitic Trichoderma species were tested for the presence of lectins, by haemagglutination with human and marsupial erythrocytes. In Trichoderma viride, haemagglutinating activity was present in both mycelial extracts and culture filtrate. While secreted lectins were only detected after 6 days of growth, the presence of mycelium-associated lectins was first noted in 3-day-old cultures. Agglutinating activity was also demonstrated in the mycelium of 6-, 9- and 13-day-old cultures of Trichoderma harzianum. In this species, however, lectins were not secreted. In all instances, haemagglutination was inhibited by N-acetylgalactosamine and related sugars. This is the first report on the occurrence of lectins in Trichoderma spp.

ABO Blood-Group System↗

PubMed2003

Clinical importance of the genus Trichoderma. A review.

Opportunistic fungal infections have been observed with increasing frequency in recent years in immunocompromised patients. Several data were published in the last decade about the clinical importance of the filamentous fungal genus Trichoderma, indicating that Trichoderma strains--besides their agricultural and biotechnological importance--may be potential opportunistic pathogens in immunocompromised hosts as well. This review is going to summarize the clinical case reports about Trichoderma infections, and to discuss the information available on the antifungal susceptibility and on the ecophysiological, enzymological and systematic aspects of clinical Trichoderma isolates.

Drug Resistance, Fungal↗

PubMed2004

Production of extracellular proteases by human pathogenic Trichoderma longibrachiatum strains.

Species belonging to the filamentous fungal genus Trichoderma are well known as potential candidates for the biological control of plant pathogenic fungi and as cellulase producers of biotechnological importance. Several data were published in the last decade also about the clinical importance of this genus, indicating that Trichoderma strains may be potential opportunistic pathogens in immunocompromised patients. However, there is a lack of information about the potential virulence factors of clinical Trichoderma strains. This study was designed to examine the extracellular proteolytic enzymes of six clinical T. longibrachiatum isolates. Supernatants from induced liquid cultures of the examined strains were screened for proteolytic enzyme activities with 11 different chromogenic p-nitroaniline substrates. The production of trypsin-like, chymotrypsin-like and chymoelastase-like protease activities cleaving N-Benzoyl-L-Phe-L-Val-L-Arg-p-nitroanilide, N-Succinyl-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide, and N-Succinyl-L-Ala-L-Ala-L-Pro-L-Leu-p-nitroanilide, respectively, was common among the strains examined. Separation of trypsin- and chymotrypsin-like activities by column chromatography revealed, that both systems are complex consisting of several isoenzymes. The pH-dependence of these two protease systems was also studied. Based on the results, the different isoenzymes seem to have different optimal pH values. Extracellular proteolytic enzymes may be involved in the pathogenecity of Trichoderma strains as facultative human pathogens.

Endopeptidases↗