Mycoparasitism is a key feature of Trichoderma (Hypocreales, Ascomycota) biocontrol agents. Recent studies of intracellular signal transduction pathways of the potent mycoparasite Trichoderma atroviride revealed the involvement of Tmk1, a mitogen-activated protein kinase (MAPK), in triggering the mycoparasitic response. We previously showed that mutants missing Tmk1 exhibit reduced mycoparasitic activity against several plant pathogenic fungi. In this study, we identified the most robustly regulated targets that were governed by Tmk1 during mycoparasitism using transcriptome and proteome profiling. Tmk1 mainly exerts a stimulating function for T. atroviride during its mycoparasitic interaction with the fungal plant pathogen Rhizoctonia solani, as reflected by 89% of strongly differently responding genes in the ∆tmk1 mutant compared to the wild type. Specifically, 54% of these genes showed strong downregulation in the response with a deletion of the tmk1 gene, whereas in the wild type the same genes were strongly upregulated during the interaction with the fungal host. These included the gene encoding the mycoparasitism-related proteinase Prb1; genes involved in signal transduction pathways such as a candidate coding for a conserved 14-3-3 protein, and a gene coding for Tmk2, the T. atroviride cell-wall integrity MAP kinase; genes encoding a specific siderophore synthetase, and multiple FAD-dependent oxidoreductases and aminotransferases. Due to the phosphorylating activity of Tmk1, different (phospho-)proteomics approaches were applied and identified proteins associated with cellular metabolism, energy production, protein synthesis and fate, and cell organization. Members of FAD- and NAD/NADP-binding-domain proteins, vesicular trafficking of molecules between cellular organelles, fungal translational, as well as protein folding apparatus were among others found to be phosphorylated by Tmk1 during mycoparasitism. Outstanding downregulation in the response of the ∆tmk1 mutant to the fungal host compared to the wild type at both the transcriptome and the proteome levels was observed for nitrilase, indicating that its defense and detoxification functions might be greatly dependent on Tmk1 during T. atroviride mycoparasitism. An intersection network analysis between the identified transcripts and proteins revealed a strong involvement of Tmk1 in molecular functions with GTPase and oxidoreductase activity. These data suggest that during T. atroviride mycoparasitism this MAPK mainly governs processes regulating cell responses to extracellular signals and those involved in reactive oxygen stress.
- MeSH
- Hypocreales * metabolismus MeSH
- mitogenem aktivované proteinkinasy genetika metabolismus MeSH
- proteom metabolismus MeSH
- regulace genové exprese u hub MeSH
- signální transdukce MeSH
- Trichoderma * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- mitogenem aktivované proteinkinasy MeSH
- proteom MeSH
BACKGROUND: Trichoderma spp. are filamentous fungi causing invasive fungal diseases in patients with haematological malignancies and in peritoneal dialysis patients. OBJECTIVES: To analyse clinical presentation, predisposing factors, treatment and outcome of Trichoderma infections. METHODS: A systematic literature review was conducted for published cases of invasive Trichoderma infection in PubMed until December 2021 and by reviewing the included studies' references. Cases from the FungiScope® registry were added to a combined analysis. RESULTS: We identified 50 invasive infections due to Trichoderma species, including 11 in the FungiScope® registry. The main underlying conditions were haematological malignancies in 19 and continuous ambulatory peritoneal dialysis (CAPD) in 10 cases. The most prevalent infection sites were lung (42%) and peritoneum (22%). Systemic antifungal therapy was administered in 42 cases (84%), mostly amphotericin B (n = 27, lipid-based formulation 13/27) and voriconazole in 15 cases (30%). Surgical interventions were performed in 13 cases (26%). Overall mortality was 48% (n = 24) and highest for allogeneic HSCT and solid organ transplantation (SOT) recipients [80% (4/5) and 77% (7/9), respectively]. In patients treated with amphotericin B, voriconazole and caspofungin, mortality was 55% (15/27), 46% (7/15) and 28% (2/7), respectively. Three out of four patients treated with a combination therapy of voriconazole and caspofungin survived. CONCLUSIONS: Despite treatment with antifungal therapies and surgery, invasive Trichoderma infections are life-threatening complications in immunocompromised patients, especially after HSCT and SOT. In addition, Trichoderma spp. mainly affect the lungs in patients with haematological malignancies and the peritoneum in CAPD patients.
- MeSH
- amfotericin B terapeutické užití MeSH
- antifungální látky terapeutické užití MeSH
- hematologické nádory * komplikace MeSH
- kaspofungin MeSH
- lidé MeSH
- registrace MeSH
- Trichoderma * MeSH
- vorikonazol terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- systematický přehled MeSH
- Názvy látek
- amfotericin B MeSH
- antifungální látky MeSH
- kaspofungin MeSH
- vorikonazol MeSH
Fungal metabolic carbon acquisition and its subsequent partitioning between biomass production and respiration, i.e. the carbon-use efficiency (CUE), are central parameters in biogeochemical modeling. However, current available techniques for estimating these parameters are all associated with practical and theoretical shortcomings, making assessments unreliable. Gene expression analyses hold the prospect of phenotype prediction by indirect means, providing new opportunities to obtain information about metabolic priorities. We cultured four different fungal isolates (Chalara longipes, Laccaria bicolor, Serpula lacrymans and Trichoderma harzianum) in liquid media with contrasting nitrogen availability and measured growth rates and respiration to calculate CUE. By relating gene expression markers to measured carbon fluxes, we identified genes coding for 1,3-β-glucan synthase and 2-oxoglutarate dehydrogenase as suitable markers for growth and respiration, respectively, capturing both intraspecific variation as well as within-strain variation dependent on growth medium. A transcript index based on these markers correlated significantly with differences in CUE between the fungal isolates. Our study paves the way for the use of these markers to assess differences in growth, respiration and CUE in natural fungal communities, using metatranscriptomic or the RT-qPCR approach.
- Klíčová slova
- carbon-use efficiency, fungi, gene markers, growth, metatranscriptomics, respiration,
- MeSH
- Ascomycota genetika metabolismus MeSH
- Basidiomycota genetika MeSH
- biologické markery * analýza MeSH
- fungální proteiny * genetika metabolismus MeSH
- houby * genetika metabolismus MeSH
- Hypocreales genetika metabolismus MeSH
- Laccaria genetika metabolismus MeSH
- transkriptom * MeSH
- Trichoderma genetika metabolismus MeSH
- uhlík * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biologické markery * MeSH
- fungální proteiny * MeSH
- uhlík * MeSH
There are increasing efforts to identify biocontrol-active microbial metabolites in order to improve strategies for biocontrol of phytopathogens. In this work, Fusarium oxysporum f. sp. conglutinans was confronted with three different biocontrol agents: Trichoderma harzianum, Bacillus amyloliquefaciens, and Pseudomonas aeruginosa in dual culture bioassays. Metabolites produced during the microbial interactions were screened by a matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). T. harzianum exhibited the strongest inhibition of growth of F. oxysporum resulting in overlay of the pathogen colony with its mycelium. Recorded metabolite profiles suggested a direct attack of F. oxysporum mycelium by T. harzianum and B. amyloliquefaciens by means of membrane-attacking peptaibols and a set of antimicrobial lipopeptides and siderophores, respectively. The direct mode of the biocontrol activity of T. harzianum and B. amyloliquefaciens corresponded to their ability to suppress F. oxysporum production of mycotoxin beauvericin suggesting that this ability is not specific only for Trichoderma species. In the case of P. aeruginosa, siderophores pyoverdine E/D and two rhamnolipids were produced as major bacterial metabolites; the rhamnolipid production was blocked by F. oxysporum. The results showed that this type of biocontrol activity was the least effective against F. oxysporum. The effective application of MALDI-MS profiling to the screening of nonvolatile microbial metabolites produced during the interaction of the phytopathogen and the biocontrol microorganisms was demonstrated.
- MeSH
- Bacillus amyloliquefaciens metabolismus fyziologie MeSH
- biologická ochrana * MeSH
- druhová specificita MeSH
- Fusarium růst a vývoj metabolismus MeSH
- glykolipidy metabolismus MeSH
- kokultivační techniky MeSH
- metabolomika MeSH
- mikrobiální interakce MeSH
- mycelium růst a vývoj metabolismus MeSH
- mykotoxiny metabolismus MeSH
- nemoci rostlin mikrobiologie prevence a kontrola MeSH
- Pseudomonas aeruginosa metabolismus fyziologie MeSH
- siderofory metabolismus MeSH
- Trichoderma metabolismus fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologická ochrana * MeSH
- glykolipidy MeSH
- mykotoxiny MeSH
- rhamnolipid MeSH Prohlížeč
- siderofory MeSH
Sustainability and a more environment-friendly approach is an emerging issue relevant to crop production. Abiotic stresses like drought, salinity, heat, cold or heavy metal pollution can severely compromise yields, and in this respect, plant protection practices should be highly efficient as well as safe for the environment and people. Among the many ways to achieve high productivity of healthy, safe and tasty food, the use of beneficial micro-organisms as biostimulants is the most promising one. Two types of soil fungi can be considered efficient natural plants stimulants: arbuscular mycorrhizal fungi (AMF) and Trichoderma spp. (TR). Generally, most investigations indicated AMF and TR were effective, as well as safe, for use as natural biopreparations dedicated to horticultural crops, although some reports pointed to their negative impact on plants. This review focuses on the mutual interaction of AMF and TR, as well as complex relationships with plants analysed on a multidimensional level: biochemical, morphological, ecological and agrotechnical. AMF and TR were found to be effective elicitors of root system development, nutrient uptake, plant stress response and production of secondary metabolites. As natural plant stimulants, beneficial fungi are compatible with modern trends of crop management, environmental conservation and functional food production. Herein, we demonstrate the advantages and disadvantages of AMF and TR use in horticulture and their prospects, as well as the points that need further exploring.
- Klíčová slova
- abiotic stresses, arbuscular mycorrhizal symbiosis, biostimulants, plant-microbial interactions, sustainability,
- MeSH
- fyziologický stres MeSH
- mykorhiza fyziologie MeSH
- půdní mikrobiologie MeSH
- symbióza MeSH
- Trichoderma fyziologie MeSH
- zemědělské plodiny růst a vývoj metabolismus mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
The study of the soil microbial community represents an important step in better understanding the environmental context. Therefore, biological characterisation and physicochemical integration are keys when defining contaminated sites. Fungi play a fundamental role in the soil, by providing and supporting ecological services for ecosystems and human wellbeing. In this research, 52 soil fungal taxa were isolated from in situ pilot reactors installed to a contaminated site in Czech Republic with a high concentration of hexachlorocyclohexane (HCH). Among the identified isolates, 12 strains were selected to evaluate their tolerance to different isomers of HCH by using specific indices (Rt:Rc; T.I.) and to test their potential in xenobiotic biotransformation. Most of the selected taxa was not significantly affected by exposure to HCH, underlining the elevated tolerance of all the tested fungal taxa, and different metabolic intermediates of HCH dechlorination were observed. The oxidative stress responses to HCH for two selected species, Penicillium simplicissimum and Trichoderma harzianum, were investigated in order to explore their toxic responses and to evaluate their potential functioning in bioremediation of contaminated environments. This research suggests that the isolated fungal species may provide opportunities for new eco-friendly, integrated and cost-effective solutions for environmental management and remediation, considering their efficient adaptation to stressful conditions.
- Klíčová slova
- Bioremediation, Contaminated sites, HCH, Oxidative stress responses, Persistent organic pollutants, Soil fungi, Tolerance,
- MeSH
- biodegradace MeSH
- biotransformace * MeSH
- ekosystém MeSH
- hexachlorcyklohexan analýza metabolismus MeSH
- houby metabolismus MeSH
- isomerie MeSH
- látky znečišťující půdu analýza metabolismus MeSH
- oxidační stres MeSH
- Penicillium metabolismus MeSH
- půda MeSH
- půdní mikrobiologie * MeSH
- tolerance léku MeSH
- Trichoderma metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- hexachlorcyklohexan MeSH
- látky znečišťující půdu MeSH
- půda MeSH
Efficient hydrolysis of holocellulose depends on a proper balance between cellulase (endoglucanase, exoglucanase, β-glucosidase) and xylanase activities. The present study aimed to induce the production of cellulases and xylanases using liquid cultures (one, two, three, and four fungal strains on the same bioreactor) of wild strains of Trichoderma harzianum, Aspergillus niger, and Fusarium oxysporum. The strains were identified by amplification and analysis of the ITS rDNA region and the obtained sequences were deposited in Genbank. Enzymes (endoglucanase, exoglucansae, β-glucosidase, and xylanase activities) and the profile of extracellular protein isoforms (SDS-PAGE) produced by different fungal combinations (N = 14) were analyzed by Pearson's correlation matrix and principal component analysis (PCA). According to our results, induction of endoglucanase (19.02%) and β-glucosidase (6.35%) were obtained after 4 days when A. niger and F. oxysporum were cocultured. The combination of A. niger-T. harzianum produced higher endoglucanase in a shorter time than monocultures. On the contrary, when more than two strains were cultured in the same reactor, the relationships of competition were established, trending to diminish the amount of enzymes and the extracellular protein isoforms produced. The xylanase production was sensible to stress produced by mixed cultures, decreasing their activity. This is important when the aim is to produce cellulase-free xylanase. In addition, exoglucanase activity did not change in the combinations tested.
- Klíčová slova
- Aspergillus, Endoglucanase, Exoglucanase, Filamentous fungi, Fusarium, Trichoderma, β-Glucosidase,
- MeSH
- Ascomycota enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- Aspergillus niger enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- biomasa MeSH
- bioreaktory mikrobiologie MeSH
- celulasy biosyntéza metabolismus MeSH
- celulosa metabolismus MeSH
- fermentace MeSH
- fungální proteiny biosyntéza metabolismus MeSH
- Fusarium enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- kokultivační techniky * MeSH
- mikrobiální interakce fyziologie MeSH
- průmyslová mikrobiologie metody MeSH
- Trichoderma enzymologie růst a vývoj izolace a purifikace metabolismus MeSH
- xylosidasy biosyntéza metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- celulasy MeSH
- celulosa MeSH
- fungální proteiny MeSH
- xylosidasy MeSH
The study evaluates the survivability and storage stability of seven Trichoderma strains belonging to the species: T. harzianum (1), T. atroviride (4), and T. virens (2) after the lyophilization of their solid state cultures on wheat straw. Biomass of Trichoderma strains was freeze-dried with and without the addition of maltodextrin. Furthermore, in order to determine the ability of tested Trichoderma strains to preserve selected technological features, the biosynthesis of extracellular hydrolases (cellulases, xylanases, and polygalacturonases) after a 3-month storage of lyophilizates was investigated. Strains of T. atroviride (except TRS40) and T. harzianum TRS85 showed the highest viability after lyophilization process (up to 100%). After 3 months of storage, T. atroviride TRS14 exhibited the highest stability (95.23%); however, the number of active conidia remained at high level of 106-107 cfu/g for all tested T. atroviride strains and T. harzianum TRS85. Interestingly, after a 3-month storage of lyophilized formulations, most of the tested Trichoderma strains exhibited higher cellulolytic and xylanolytic activities compared to the control, i.e., before freeze-drying process. The highest activities of these enzymes exhibited the following: T. atroviride TRS14-2.37 U/g and T. atroviride TRS25-21.47 U/g, respectively, whereas pectinolytic activity was weak for all tested strains, with the highest value of 0.64 U/g registered for T. virens TRS109.
- MeSH
- biomasa MeSH
- časové faktory MeSH
- fermentace MeSH
- hydrolasy metabolismus MeSH
- lyofilizace * MeSH
- mikrobiální viabilita * MeSH
- pšenice metabolismus MeSH
- skladování léků MeSH
- spory hub růst a vývoj MeSH
- Trichoderma klasifikace růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- hydrolasy MeSH
Molecular markers that enable monitoring of fungi in their natural environment or assist in the identification of specific strains would facilitate Trichoderma utilization, particularly as an agricultural biocontrol agent (BCA). In this study, sequence analysis of internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of the ribosomal RNA (rRNA) gene cluster, a fragment of the translation elongation factor 1-alpha (tef1) gene, and random amplified polymorphic DNA (RAPD) markers were applied to determine the genetic diversity of Trichoderma atroviride strains collected in Poland, and also in order to identify loci and PCR-based molecular markers useful in genetic variation assessment of that fungus. Although tef1 and RAPD analysis showed limited genetic diversity among T. atroviride strains collected in Poland, it was possible to distinguish major groups that clustered most of the analyzed strains. Polymorphic RAPD amplicons were cloned and sequenced, yielding sequences representing 13 T. atroviride loci. Based on these sequences, a set of PCR-based markers specific to T. atroviride was developed and examined. Three cleaved amplified polymorphic sequence (CAPS) markers could assist in distinguishing T. atroviride strains. The genomic regions identified may be useful for further exploration and development of more precise markers suitable for T. atroviride identification and monitoring, especially in environmental samples.
- MeSH
- DNA fungální chemie genetika MeSH
- elongační faktor 1 genetika MeSH
- fylogeneze MeSH
- genetická variace * MeSH
- genetické lokusy * MeSH
- genetické markery MeSH
- mezerníky ribozomální DNA chemie genetika MeSH
- molekulární sekvence - údaje MeSH
- molekulární typizace MeSH
- mykologické určovací techniky MeSH
- sekvenční analýza DNA MeSH
- shluková analýza MeSH
- technika náhodné amplifikace polymorfní DNA MeSH
- Trichoderma klasifikace genetika izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Polsko MeSH
- Názvy látek
- DNA fungální MeSH
- elongační faktor 1 MeSH
- genetické markery MeSH
- mezerníky ribozomální DNA MeSH
Xylanase III (Xyn III), a specific endoxylanase that belongs to family 10 of the glycoside hydrolases, was overexpressed in Trichoderma reesei QM9414 using a constitutive strong promoter of the gene encoding pyruvate decarboxylase (pdc). The maximum recombinant xylanase activity achieved was 817.2 ± 65.2 U/mL in the transformant fermentation liquid. The productivities of Xyn III accounted for approximately 53% of the total protein secreted by the recombinant. The enzyme was optimally active at 60 °C and pH 6. The recombinant Xyn III was stable at pH 5-8. This is the first report on the homologous expression of xyn3 in T. reesei QM9414. The properties of Xyn III make it promising in a variety of industrial use.
- MeSH
- aktivace enzymů MeSH
- endo-1,4-beta-xylanasy genetika metabolismus MeSH
- exprese genu * MeSH
- klonování DNA MeSH
- koncentrace vodíkových iontů MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Trichoderma genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- endo-1,4-beta-xylanasy MeSH
