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Sample records for anamorph trichoderma reesei

  1. Evolution and ecophysiology of the industrial producer Hypocrea jecorina (Anamorph Trichoderma reesei and a new sympatric agamospecies related to it.

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    Irina S Druzhinina

    Full Text Available BACKGROUND: Trichoderma reesei, a mitosporic green mould, was recognized during the WW II based on a single isolate from the Solomon Islands and since then used in industry for production of cellulases. It is believed to be an anamorph (asexual stage of the common pantropical ascomycete Hypocrea jecorina. METHODOLOGY/PRINCIPAL FINDINGS: We combined molecular evolutionary analysis and multiple methods of phenotype profiling in order to reveal the genetic relationship of T. reesei to H. jecorina. The resulting data show that the isolates which were previously identified as H. jecorina by means of morphophysiology and ITS1 and 2 (rRNA gene cluster barcode in fact comprise several species: i H. jecorina/T. reesei sensu stricto which contains most of the teleomorphs (sexual stages found on dead wood and the wild-type strain of T. reesei QM 6a; ii T. parareesei nom. prov., which contains all strains isolated as anamorphs from soil; iii and two other hypothetical new species for which only one or two isolates are available. In silico tests for recombination and in vitro mating experiments revealed a history of sexual reproduction for H. jecorina and confirmed clonality for T. parareesei nom. prov. Isolates of both species were consistently found worldwide in pantropical climatic zone. Ecophysiological comparison of H. jecorina and T. parareesei nom. prov. revealed striking differences in carbon source utilization, conidiation intensity, photosensitivity and mycoparasitism, thus suggesting adaptation to different ecological niches with the high opportunistic potential for T. parareesei nom. prov. CONCLUSIONS: Our data prove that T. reesei belongs to a holomorph H. jecorina and displays a history of worldwide gene flow. We also show that its nearest genetic neighbour--T. parareesei nom. prov., is a cryptic phylogenetic agamospecies which inhabits the same biogeographic zone. These two species thus provide a so far rare example of sympatric speciation

  2. Light-dependent roles of the G-protein α subunit GNA1 of Hypocrea jecorina (anamorph Trichoderma reesei

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    Kubicek Christian P

    2009-09-01

    Full Text Available Abstract Background The filamentous ascomycete Hypocrea jecorina (anamorph Trichoderma reesei is primarily known for its efficient enzymatic machinery that it utilizes to decompose cellulosic substrates. Nevertheless, the nature and transmission of the signals initiating and modulating this machinery are largely unknown. Heterotrimeric G-protein signaling represents one of the best studied signal transduction pathways in fungi. Results Analysis of the regulatory targets of the G-protein α subunit GNA1 in H. jecorina revealed a carbon source and light-dependent role in signal transduction. Deletion of gna1 led to significantly decreased biomass formation in darkness in submersed culture but had only minor effects on morphology and hyphal apical extension rates on solid medium. Cellulase gene transcription was abolished in Δgna1 on cellulose in light and enhanced in darkness. However, analysis of strains expressing a constitutively activated GNA1 revealed that GNA1 does not transmit the essential inducing signal. Instead, it relates a modulating signal with light-dependent significance, since induction still required the presence of an inducer. We show that regulation of transcription and activity of GNA1 involves a carbon source-dependent feedback cycle. Additionally we found a function of GNA1 in hydrophobin regulation as well as effects on conidiation and tolerance of osmotic and oxidative stress. Conclusion We conclude that GNA1 transmits a signal the physiological relevance of which is dependent on both the carbon source as well as the light status. The widespread consequences of mutations in GNA1 indicate a broad function of this Gα subunit in appropriation of intracellular resources to environmental (especially nutritional conditions.

  3. Use of Hypocrea jecorina (anamorph Trichoderma reesei) as a model system for Trichoderma biocontrol of Pythium blight identifies new targets for genetic strain improvement

    Institute of Scientific and Technical Information of China (English)

    Seidl V; Schmoll M; Scherm B; Balmas V; Seiboth B; Migheli Q; Kubicek C P

    2004-01-01

    @@ Biocontrol by Trichoderma has been studied mainly with selected isolates of T. harzianum, T.atroviride and T. asperellum, which are members of sections Pachybasium and Trichoderma. In contrast, species from section Longibrachiatum have only rarely been studied. On the other hand, one taxon from this section-Hypocrea jecorina (anamorph: Trichoderma reesei)-has been widely used for the production of cellulolytic and hemicellulolytic enzymes and recombinant proteins. As far as Trichoderma is concerned, molecular genetic methods and tools are most advanced in H. jecorina,and its genome has recently been fully sequenced, thus making this taxon a model organism for the genus. Here we will demonstrate that H. jecorina is able to antagonize plant pathogenic fungi in plate confrontation tests, and can protect tomato and cucumber plants against Pythium ultimum blight.Using this as a model case, we made use of available H. jecorina mutants to investigate (a) whether carbon catabolite repression via the Cre1-regulator protein has an impact on biocontrol, and (b)whether cellulase gene expression is necessary for biocontrol of P. ultimum. In the first case, plate confrontation tests and in planta experiments yielded opposite results, i.e. while a Cre1 mutant was more active in antagonization of fungi on plates, the survival rates of P. ultimum-inoculated cucumber plants was lower than with the H. jecorina wild-type strain. Mutants of H. jecorina,unable to form cellulases, were still able to antagonize fungi on plates and provided similar protection of tomatos against P. ultimum as the wild type, indicating that the pronounced biocontrol ability of H.jecorina against fungi with cellulose-containing cell-walls is not due to its high cellulolytic activity. A strain disrupted in the light-modulator gene envoy (Schmoll et al., ms submitted) exhibited in planta biocontrol activity strongly exceeding that of the wild-type strain, thereby providing a first link between Trichoderma

  4. A versatile toolkit for high throughput functional genomics with Trichoderma reesei

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    Schuster, Andre; Bruno, Kenneth S.; Collett, James R.; Baker, Scott E.; Seiboth, Bernhard; Kubicek, Christian P.; Schmoll, Monika

    2012-01-02

    The ascomycete fungus, Trichoderma reesei (anamorph of Hypocrea jecorina), represents a biotechnological workhorse and is currently one of the most proficient cellulase producers. While strain improvement was traditionally accomplished by random mutagenesis, a detailed understanding of cellulase regulation can only be gained using recombinant technologies. RESULTS: Aiming at high efficiency and high throughput methods, we present here a construction kit for gene knock out in T. reesei. We provide a primer database for gene deletion using the pyr4, amdS and hph selection markers. For high throughput generation of gene knock outs, we constructed vectors using yeast mediated recombination and then transformed a T. reesei strain deficient in non-homologous end joining (NHEJ) by spore electroporation. This NHEJ-defect was subsequently removed by crossing of mutants with a sexually competent strain derived from the parental strain, QM9414.CONCLUSIONS:Using this strategy and the materials provided, high throughput gene deletion in T. reesei becomes feasible. Moreover, with the application of sexual development, the NHEJ-defect can be removed efficiently and without the need for additional selection markers. The same advantages apply for the construction of multiple mutants by crossing of strains with different gene deletions, which is now possible with considerably less hands-on time and minimal screening effort compared to a transformation approach. Consequently this toolkit can considerably boost research towards efficient exploitation of the resources of T. reesei for cellulase expression and hence second generation biofuel production.

  5. Small Angle Neutron Scattering Reveals pH-dependent Conformational Changes in Trichoderma reesei Cellobiohydrolase I: IMPLICATIONS FOR ENZYMATIC ACTIVITY*

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    Pingali, Sai Venkatesh; O'Neill, Hugh M.; McGaughey, Joseph; Urban, Volker S.; Rempe, Caroline S.; Petridis, Loukas; Jeremy C Smith; Evans, Barbara R.; Heller, William T.

    2011-01-01

    Cellobiohydrolase I (Cel7A) of the fungus Trichoderma reesei (now classified as an anamorph of Hypocrea jecorina) hydrolyzes crystalline cellulose to soluble sugars, making it of key interest for producing fermentable sugars from biomass for biofuel production. The activity of the enzyme is pH-dependent, with its highest activity occurring at pH 4–5. To probe the response of the solution structure of Cel7A to changes in pH, we measured small angle neutron scattering of it in a series of solut...

  6. A rapid, one step molecular identification of Trichoderma citrinoviride and Trichoderma reesei.

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    Saroj, Dina B; Dengeti, Shrinivas N; Aher, Supriya; Gupta, Anil K

    2015-06-01

    Trichoderma species are widely used as production hosts for industrial enzymes. Identification of Trichoderma species requires a complex molecular biology based identification involving amplification and sequencing of multiple genes. Industrial laboratories are required to run identification tests repeatedly in cell banking procedures and also to prove absence of production host in the product. Such demands can be fulfilled by a brief method which enables confirmation of strain identity. This communication describes one step identification method for two common Trichoderma species; T. citrinoviride and T. reesei, based on identification of polymorphic region in the nucleotide sequence of translation elongation factor 1 alpha. A unique forward primer and common reverse primer resulted in 153 and 139 bp amplicon for T. citrinoviride and T. reesei, respectively. Simplification was further introduced by using mycelium as template for PCR amplification. Method described in this communication allows rapid, one step identification of two Trichoderma species.

  7. Enhanced Expression of Endochitinase in Trichoderma harzianum with the cbh1 Promoter of Trichoderma reesei

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    Margolles-Clark, E.; Harman, G. E.; Penttila, M.

    1996-01-01

    Production of extracellular endochitinase could be increased 5-fold in the mycoparasite fungus Trichoderma harzianum by using the cellulase promoter cbh1 of Trichoderma reesei, whereas the total endochitinase activity increased 10-fold. The cbh1 promoter was not expressed on glucose and sucrose in T. harzianum and was induced by sophorose and on cellulase-inducing medium. The endogenous endochitinase gene was expressed at a low basal level on glucose and sucrose. No specific induction by crab...

  8. Examining the Potential of Plasma-Assisted Pretreated Wheat Straw for Enzyme Production by Trichoderma reesei

    DEFF Research Database (Denmark)

    Rodríguez Gómez, Divanery; Lehmann, Linda Olkjær; Schultz-Jensen, Nadja;

    2012-01-01

    Plasma-assisted pretreated wheat straw was investigated for cellulase and xylanase production by Trichoderma reesei fermentation. Fermentations were conducted with media containing washed and unwashed plasma-assisted pretreated wheat straw as carbon source which was sterilized by autoclavation...

  9. The influence of some factors on β-1,4-xylanase activity of the filamentous fungus Trichoderma reesei QM9414

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    Alexandru Manoliu

    2012-03-01

    Full Text Available The mesophyllic fungus Trichoderma reesei (anamorph to Hypocrea jecorina is an important biotechnological tool, known for its ability to secrete large quantities of hydrolytic enzymes. Renewable biomass, such as agricultural and forest wastes are used to produce microbial enzymes in various industrial processes such as food, feed and bioethanol industries. In raw biomass materials, such as wheat straws, barley straws and maize stalks, the main polysaccharide is cellulose which is closely associated with hemicelluloses like xylan, manan and xyloguclan. In consequence, the hydrolysis of these materials requires the concerted action of several enzymes, namely cellulases and xylanases. Endo-xylanase (endo-1,4--xylanase, EC 3.2.1.8 is the key enzyme involved in xylan hydrolysis, the mainhemicellulosic component of plant cell walls. The metabolic activity and enzyme productivity of Trichoderma reesei isinfluenced by various environmental conditions. In this context, we analysed the effect of pH, cultivation period, thenature of the substrate used and the nitrogen source on enzymatic activity. The maximum xylanase yield was recorded at a initial pH of 4 (116.189 IU/ml for barley and 5 for wheat (88.578 IU/ml, respectively maize (116.583 IU/ml. The bestsubstrate for endo-xylanase activity was maize stalks (90.446 IU/ml at a a concentration of 30g/L.

  10. A novel platform for heterologous gene expression in Trichoderma reesei (Teleomorph Hypocrea jecorina)

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    Jørgensen, Mikael Skaanning; Skovlund, Dominique Aubert; Johannesen, Pia Francke;

    2014-01-01

    ABSTRACT: BACKGROUND: The industrially applied filamentous fungus Trichoderma reesei has received substantial interest due to its highly efficient synthesis apparatus of cellulytic enzymes. However, the production of heterologous enzymes in T. reesei still remains low mainly due to lack of tools...

  11. Quantitative site-specific phosphoproteomics of Trichoderma reesei signaling pathways upon induction of hydrolytic enzyme production

    NARCIS (Netherlands)

    E.V. Nguyen; S.Y. Imanishi; P. Haapaniemi; A. Yadav; M. Saloheimo; G.L. Corthals; T.M. Pakula

    2016-01-01

    The filamentous fungus Trichoderma reesei is used for industrial production of secreted enzymes including carbohydrate active enzymes, such as cellulases and hemicellulases. The production of many of these enzymes by T. reesei is influenced by the carbon source it grows on, where the regulation syst

  12. Reduction of aflatoxins by Rhizopus oryzae and Trichoderma reesei.

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    Hackbart, H C S; Machado, A R; Christ-Ribeiro, A; Prietto, L; Badiale-Furlong, E

    2014-08-01

    This study evaluated the ability of the microorganisms Rhizopus oryzae (CCT7560) and Trichoderma reesei (QM9414), producers of generally recognized as safe (GRAS) enzymes, to reduce the level of aflatoxins B1, B2, G1, G2, and M1. The variables considered to the screening were the initial number of spores in the inoculum and the culture time. The culture was conducted in contaminated 4 % potato dextrose agar (PDA) medium, and the residual mycotoxins were determined every 24 h by HPLC-FL. The fungus R. oryzae has reduced aflatoxins B1, B2, and G1 in the 96 h and aflatoxins M1 and G2 in the range of 120 h of culture by approximately 100 %. The fungus T. reesei has reduced aflatoxins B1, B2, and M1 in the 96 h and aflatoxin G1 in the range of 120 h of culture by approximately 100 %. The highest reduction occurred in the middle of R. oryzae culture.

  13. Reduction of aflatoxins by Rhizopus oryzae and Trichoderma reesei.

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    Hackbart, H C S; Machado, A R; Christ-Ribeiro, A; Prietto, L; Badiale-Furlong, E

    2014-08-01

    This study evaluated the ability of the microorganisms Rhizopus oryzae (CCT7560) and Trichoderma reesei (QM9414), producers of generally recognized as safe (GRAS) enzymes, to reduce the level of aflatoxins B1, B2, G1, G2, and M1. The variables considered to the screening were the initial number of spores in the inoculum and the culture time. The culture was conducted in contaminated 4 % potato dextrose agar (PDA) medium, and the residual mycotoxins were determined every 24 h by HPLC-FL. The fungus R. oryzae has reduced aflatoxins B1, B2, and G1 in the 96 h and aflatoxins M1 and G2 in the range of 120 h of culture by approximately 100 %. The fungus T. reesei has reduced aflatoxins B1, B2, and M1 in the 96 h and aflatoxin G1 in the range of 120 h of culture by approximately 100 %. The highest reduction occurred in the middle of R. oryzae culture. PMID:24925827

  14. Cellulases and beyond: the first 70 years of the enzyme producer Trichoderma reesei.

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    Bischof, Robert H; Ramoni, Jonas; Seiboth, Bernhard

    2016-01-01

    More than 70 years ago, the filamentous ascomycete Trichoderma reesei was isolated on the Solomon Islands due to its ability to degrade and thrive on cellulose containing fabrics. This trait that relies on its secreted cellulases is nowadays exploited by several industries. Most prominently in biorefineries which use T. reesei enzymes to saccharify lignocellulose from renewable plant biomass in order to produce biobased fuels and chemicals. In this review we summarize important milestones of the development of T. reesei as the leading production host for biorefinery enzymes, and discuss emerging trends in strain engineering. Trichoderma reesei has very recently also been proposed as a consolidated bioprocessing organism capable of direct conversion of biopolymeric substrates to desired products. We therefore cover this topic by reviewing novel approaches in metabolic engineering of T. reesei. PMID:27287427

  15. Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization.

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    de Oliveira, Gabriel Stephani; Adriani, Patricia Pereira; Borges, Flavia Garcia; Lopes, Adriana Rios; Campana, Patricia T; Chambergo, Felipe S

    2016-08-01

    Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have biotechnological potential in chiral chemistry. We report the cloning, purification, enzymatic activity, and conformational analysis of the TrEH gene from Trichoderma reesei strain QM9414 using circular dichroism spectroscopy. The EH gene has an open reading frame encoding a protein of 343 amino acid residues, resulting in a molecular mass of 38.2kDa. The enzyme presents an optimum pH of 7.2, and it is highly active at temperatures ranging from 23 to 50°C and thermally inactivated at 70°C (t1/2=7.4min). The Michaelis constants (Km) were 4.6mM for racemic substrate, 21.7mM for (R)-(+)-styrene oxide and 3.0mM for (S)-(-)-styrene oxide. The kcat/Km analysis indicated that TrEH is enantioselective and preferentially hydrolyzes (S)-(-)-styrene oxide. The conformational stability studies suggested that, despite the extreme conditions (high temperatures and extremely acid and basic pHs), TrEH is able to maintain a considerable part of its regular structures, including the preservation of the native cores in some cases. The recombinant protein showed enantioselectivity that was distinct from other fungus EHs, making this protein a potential biotechnological tool. PMID:27177457

  16. Linking Hydrolysis Performance to Trichoderma reesei Cellulolytic Enzyme Profile

    DEFF Research Database (Denmark)

    Lehmann, Linda Olkjær; Petersen, Nanna; I. Jørgensen, Christian;

    2016-01-01

    Trichoderma reesei expresses a large number of enzymes involved in lignocellulose hydrolysis and the mechanism of how these enzymes work together is too complex to study by traditional methods, e.g. by spiking with single enzymes and monitoring hydrolysis performance. In this study a multivariate...... approach, partial least squares regression, was used to see if it could help explain the correlation between enzyme profile and hydrolysis performance. Diverse enzyme mixtures were produced by Trichoderma reesei Rut-C30 by exploiting various fermentation conditions and used for hydrolysis of washed...

  17. Xylanase XYN IV from Trichoderma reesei showing exo- and endo-xylanase activity

    Science.gov (United States)

    A novel xylanase from Trichoderma reesei Rut C30, named XYN IV, was purified from the cellulolytic system of the fungus. The enzyme was discovered on its ability to attack aldotetraohexenuronic acid (HexA-2Xyl-4Xyl-4Xyl, HexA3Xyl3), releasing the reducing-end xylose residue. XYN IV exhibited catalyt...

  18. Unraveling the Secondary Metabolism of the Biotechnological Important Filamentous Fungus Trichoderma reesei ( Teleomorph Hypocrea jecorina)

    DEFF Research Database (Denmark)

    Jørgensen, Mikael Skaanning

    The filamentous fungus Trichoderma reesei (teleomorph Hypocrea jecorina) is one of the most important industrial production organisms, owing to its highly efficient (hemi-)cellulase synthesis and secretion machineries. These enzymes, which in nature allow the fungus to utilize energy bound...

  19. Evaluation of Minimal Trichoderma reesei Cellulase Mixtures on Differently Pretreated Barley Straw Substrate

    DEFF Research Database (Denmark)

    Rosgaard, Lisa; Pedersen, Sven; Langston, J;

    2007-01-01

    The commercial cellulase product Celluclast 1.5, derived from Trichoderma reesei (Novozymes A/S, Bagsv ae rd, Denmark), is widely employed for hydrolysis of lignocellulosic biomass feedstocks. This enzyme preparation contains a broad spectrum of cellulolytic enzyme activities, most notably...

  20. Light-mediated control of gene expression in filamentous fungus Trichoderma reesei.

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    Wang, Wei; Shi, Xiang-Yu; Wei, Dong-Zhi

    2014-08-01

    We developed a light-mediated system based on synthetic light-switchable transactivators. The transactivators bind promoter upon blue-light exposure and rapidly initiate transcription of target transgenes in filamentous fungus Trichoderma reesei. Light is inexpensive to apply, easily delivered, and instantly removed, and thus has significant advantages over chemical inducers.

  1. Evaluation of minimal Trichoderma reesei cellulase mixtures on differently pretreated barley straw substrates

    DEFF Research Database (Denmark)

    Rosgaard, Lisa; Pedersen, Sven; Langston, Jim;

    2007-01-01

    The commercial cellulase product Celluclast 1.5, derived from Trichoderma reesei (Novozymes A/S, Bagsv ae rd, Denmark), is widely employed for hydrolysis of lignocellulosic biomass feedstocks. This enzyme preparation contains a broad spectrum of cellulolytic enzyme activities, most notably...

  2. Peningkatan Kadar Protein Putak melalui Fermentasi oleh Kapang Trichoderma reesei (THE INCREASE OF PROTEIN LEVEL FROM PUTAK THROUGH FERMENTATION OF FUNGI TRICHODERMA REESEI

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    Maritje Aleonor Hilakore

    2013-09-01

    Full Text Available A study was conducted was to increasing the protein level in putak by fermentation using fungiTrichoderma reesei. A laboratoryum experimental study was conducted using  factorial CompletelyRandomized Design 3 x 4 x 3. The main factor is were  inoculant levels of fungi T. reesei (T: 5,0; 7,5 and 10,0% (w/w,the level  and the second factor were of incubation time (W:  2; 3; and 4 days. Variables tested werecrude protein (CP, true protein (TP and crude fiber (CF. The result showed that  treatment with 7.5% ofT.reesei  and incubation time for 4 days gawe the highest of crude and true protein level (20,60%  from14,17% and 13,25% from 3,25%, and  lowest crude fiber 9,08% from 9,70%. Through fermentation of  fungiT.reesei can be increase the protein and decrease the fiber level of putak.

  3. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M.; Negro, M. J.; Saez, R.; Martin, C.

    1986-07-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  4. Secretome data from Trichoderma reesei and Aspergillus niger cultivated in submerged and sequential fermentation methods.

    Science.gov (United States)

    Florencio, Camila; Cunha, Fernanda M; Badino, Alberto C; Farinas, Cristiane S; Ximenes, Eduardo; Ladisch, Michael R

    2016-09-01

    The cultivation procedure and the fungal strain applied for enzyme production may influence levels and profile of the proteins produced. The proteomic analysis data presented here provide critical information to compare proteins secreted by Trichoderma reesei and Aspergillus niger when cultivated through submerged and sequential fermentation processes, using steam-explosion sugarcane bagasse as inducer for enzyme production. The proteins were organized according to the families described in CAZy database as cellulases, hemicellulases, proteases/peptidases, cell-wall-protein, lipases, others (catalase, esterase, etc.), glycoside hydrolases families, predicted and hypothetical proteins. Further detailed analysis of this data is provided in "Secretome analysis of Trichoderma reesei and Aspergillus niger cultivated by submerged and sequential fermentation process: enzyme production for sugarcane bagasse hydrolysis" C. Florencio, F.M. Cunha, A.C Badino, C.S. Farinas, E. Ximenes, M.R. Ladisch (2016) [1]. PMID:27419196

  5. Secretome data from Trichoderma reesei and Aspergillus niger cultivated in submerged and sequential fermentation methods

    Directory of Open Access Journals (Sweden)

    Camila Florencio

    2016-09-01

    Full Text Available The cultivation procedure and the fungal strain applied for enzyme production may influence levels and profile of the proteins produced. The proteomic analysis data presented here provide critical information to compare proteins secreted by Trichoderma reesei and Aspergillus niger when cultivated through submerged and sequential fermentation processes, using steam-explosion sugarcane bagasse as inducer for enzyme production. The proteins were organized according to the families described in CAZy database as cellulases, hemicellulases, proteases/peptidases, cell-wall-protein, lipases, others (catalase, esterase, etc., glycoside hydrolases families, predicted and hypothetical proteins. Further detailed analysis of this data is provided in “Secretome analysis of Trichoderma reesei and Aspergillus niger cultivated by submerged and sequential fermentation process: enzyme production for sugarcane bagasse hydrolysis” C. Florencio, F.M. Cunha, A.C Badino, C.S. Farinas, E. Ximenes, M.R. Ladisch (2016 [1].

  6. Enhanced cellulase production from Trichoderma reesei QM 9414 on physically treated wheat straw

    Energy Technology Data Exchange (ETDEWEB)

    Acebal, C.; Castillon, M.P.; Estrada, P.; Mata, I.; Costa, E.; Aguado, J.; Romero, D.; Jimenez, F.

    1986-06-01

    Trichoderma reesei QM 9414 was grown on wheat straw as the sole carbon source. The straw was pretreated by physical and chemical methods. The particle size of straw was less than 0.177 mm. Growth of T. reesei QM 9414 was maximal with alkali-pretreated straw whereas cellulase production was optimal when physically pretreated straw was used as substrate. Cellulase yields expressed as IU enzyme activity/g cellulose present in the cultures were considerably higher when alkali pretreatment of wheat straw was omitted. Cellulase yields of 666 IU/g cellulose for filter paper activity (FPA) are the highest described for cultures of T. reesei QM 9414 carried out in analogous conditions. Crystallinity index of the cellulose contained in wheat straw increased slightly after alkali pretreatment. This increase did not decrease cellulose accessibility to the fungus. Delignification of wheat straw was not necessary to achieve the best cellulase production.

  7. Transcriptional Profiling of the Trichoderma reesei Recombinant Strain HJ48 by RNA-Seq.

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    Huang, Jun; Wu, Renzhi; Chen, Dong; Wang, Qingyan; Huang, Ribo

    2016-07-28

    The ethanol production of Trichoderma reesei was improved by genome shuffling in our previous work. Using RNA-Seq, the transcriptomes of T. reesei wild-type CICC40360 and recombinant strain HJ48 were compared under fermentation conditions. Based on this analysis, we defined a set of T. reesei genes involved in ethanol production. Further expression analysis identified a series of glycolysis enzymes, which are upregulated in the recombinant strain HJ48 under fermentation conditions. The differentially expressed genes were further validated by qPCR. The present study will be helpful for future studies on ethanol fermentation as well as the roles of the involved genes. This research reveals several major differences in metabolic pathways between recombinant strain HJ48 and wild-type CICC40360, which relates to the higher ethanol production on the former, and their further research could promote the development of techniques for increasing ethanol production.

  8. Transcriptional Profiling of the Trichoderma reesei Recombinant Strain HJ48 by RNA-Seq.

    Science.gov (United States)

    Huang, Jun; Wu, Renzhi; Chen, Dong; Wang, Qingyan; Huang, Ribo

    2016-07-28

    The ethanol production of Trichoderma reesei was improved by genome shuffling in our previous work. Using RNA-Seq, the transcriptomes of T. reesei wild-type CICC40360 and recombinant strain HJ48 were compared under fermentation conditions. Based on this analysis, we defined a set of T. reesei genes involved in ethanol production. Further expression analysis identified a series of glycolysis enzymes, which are upregulated in the recombinant strain HJ48 under fermentation conditions. The differentially expressed genes were further validated by qPCR. The present study will be helpful for future studies on ethanol fermentation as well as the roles of the involved genes. This research reveals several major differences in metabolic pathways between recombinant strain HJ48 and wild-type CICC40360, which relates to the higher ethanol production on the former, and their further research could promote the development of techniques for increasing ethanol production. PMID:27056474

  9. Deciphering the molecular mechanisms behind cellulase production in Trichoderma reesei, the hyper-cellulolytic filamentous fungus.

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    Shida, Yosuke; Furukawa, Takanori; Ogasawara, Wataru

    2016-09-01

    The filamentous fungus Trichoderma reesei is a potent cellulase producer and the best-studied cellulolytic fungus. A lot of investigations not only on glycoside hydrolases produced by T. reesei, but also on the machinery controlling gene expression of these enzyme have made this fungus a model organism for cellulolytic fungi. We have investigated the T. reesei strain including mutants developed in Japan in detail to understand the molecular mechanisms that control the cellulase gene expression, the biochemical and morphological aspects that could favor this phenotype, and have attempted to generate novel strains that may be appropriate for industrial use. Subsequently, we developed recombinant strains by combination of these insights and the heterologous-efficient saccharifing enzymes. Resulting enzyme preparations were highly effective for saccharification of various biomass. In this review, we present some of the salient findings from the recent biochemical, morphological, and molecular analyses of this remarkable cellulase hyper-producing fungus. PMID:27075508

  10. Origin of initial burst in activity for Trichoderma reesei endo-glucanases hydrolyzing insoluble cellulose

    DEFF Research Database (Denmark)

    Murphy, Leigh; Cruys-Bagger, Nicolaj; Baumann, Martin J.;

    2012-01-01

    by the three main EGs from Trichoderma reesei (Tr): TrCel7B (formerly EG I), TrCel5A (EG II), and TrCel12A (EG III). These endo-glucanases show a distinctive initial burst with a maximal rate that is about 5-fold higher than the rate after 5 min of hydrolysis. The burst is particularly conspicuous for TrCel7B...

  11. Cost-effective lignocellulolytic enzyme production by Trichoderma reesei on a cane molasses medium

    OpenAIRE

    He, Jun; Wu, Ai-Min; Chen, Daiwen; Yu, Bing; Mao, Xiangbing; Zheng, Ping; Yu, Jie; Tian, Gang

    2014-01-01

    Background Cane molasses, an important residue of the sugar industry, have the potential as a cost-effective carbon source that could serve as nutrients for industrial enzyme-producing microorganisms, especially filamentous fungi. However, the enzyme mixtures produced in such a complex medium are poorly characterized. In this study, the secretome of Trichoderma reesei grown on a cane molasses medium (CMM) as well as on a lactose-based conventional medium (LCM) were compared and analyzed by us...

  12. Surface adhesion of fusion proteins containing the hydrophobins HFBI and HFBII from Trichoderma reesei

    OpenAIRE

    Linder, Markus; Szilvay, Geza R.; Nakari-Setälä, Tiina; Söderlund, Hans; Penttilä, Merja

    2002-01-01

    Hydrophobins are surface-active proteins produced by filamentous fungi, where they seem to be ubiquitous. They have a variety of roles in fungal physiology related to surface phenomena, such as adhesion, formation of surface layers, and lowering of surface tension. Hydrophobins can be divided into two classes based on the hydropathy profile of their primary sequence. We have studied the adhesion behavior of two Trichoderma reesei class II hydrophobins, HFBI and HFBII, as isolated proteins and...

  13. Protein disulfide isomerase homolog TrPDI2 contributing to cellobiohydrolase production in Trichoderma reesei.

    Science.gov (United States)

    Wang, Guokun; Lv, Pin; He, Ronglin; Wang, Haijun; Wang, Lixian; Zhang, Dongyuan; Chen, Shulin

    2015-09-01

    The majority of the cysteine residues in the secreted proteins form disulfide bonds via protein disulfide isomerase (PDI)-mediated catalysis, stabilizing the enzyme activity. The role of PDI in cellulase production is speculative, as well as the possibility of PDI as a target for improving enzyme production efficiency of Trichoderma reesei, a widely used producer of enzyme for the production of lignocellulose-based biofuels and biochemicals. Here, we report that a PDI homolog, TrPDI2 in T. reesei exhibited a 36.94% and an 11.81% similarity to Aspergillus niger TIGA and T. reesei PDI1, respectively. The capability of TrPDI2 to recover the activity of reduced and denatured RNase by promoting refolding verified its protein disulfide isomerase activity. The overexpression of Trpdi2 increased the secretion and the activity of CBH1 at the early stage of cellulase induction. In addition, both the expression level and redox state of TrPDI2 responded to cellulase induction in T. reesei, providing sustainable oxidative power to ensure cellobiohydrolase maturation and production. The results suggest that TrPDI2 may contribute to cellobiohydrolase secretion by enhancing the capability of disulfide bond formation, which is essential for protein folding and maturation. PMID:26138396

  14. Overexpression of bacterial ethylene-forming enzyme gene in Trichoderma reesei enhanced the production of ethylene

    Directory of Open Access Journals (Sweden)

    Xi Chen, Yong Liang, Jing Hua, Li Tao, Wensheng Qin, Sanfeng Chen

    2010-01-01

    Full Text Available In order to efficiently utilize natural cellulose materials to produce ethylene, three expression vectors containing the ethylene-forming enzyme (efe gene from Pseudomonas syringae pv. glycinea were constructed. The target gene was respectively controlled by different promoters: cbh I promoter from Trichoderma reesei cellobiohydrolases I gene, gpd promoter from Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase gene and pgk I promoter from T. reesei 3-phosphoglycerate kinase I gene. After transforming into T. reesei QM9414, 43 stable transformants were obtained by PCR amplification and ethylene determination. Southern blot analysis of 14 transformants demonstrated that the efe gene was integrated into chromosomal DNA with copy numbers from 1 to 4. Reverse transcription polymerase chain reaction (RT-PCR analysis of 6 transformants showed that the heterologous gene was transcribed. By using wheat straw as a carbon source, the ethylene production rates of aforementioned 14 transformants were measured. Transformant C30-3 with pgk I promoter had the highest ethylene production (4,012 nl h-1 l-1. This indicates that agricultural wastes could be used to produce ethylene in recombinant filamentous fungus T. reesei.

  15. Morphology and enzyme production of Trichoderma reesei Rut C-30 are affected by the physical and structural characteristics of cellulosic substrates.

    Science.gov (United States)

    Peciulyte, Ausra; Anasontzis, George E; Karlström, Katarina; Larsson, Per Tomas; Olsson, Lisbeth

    2014-11-01

    The industrial production of cellulolytic enzymes is dominated by the filamentous fungus Trichoderma reesei (anamorph of Hypocrea jecorina). In order to develop optimal enzymatic cocktail, it is of importance to understand the natural regulation of the enzyme profile as response to the growth substrate. The influence of the complexity of cellulose on enzyme production by the microorganisms is not understood. In the present study we attempted to understand how different physical and structural properties of cellulose-rich substrates affected the levels and profiles of extracellular enzymes produced by T. reesei. Enzyme production by T. reesei Rut C-30 was studied in submerged cultures on five different cellulose-rich substrates, namely, commercial cellulose Avicel® and industrial-like cellulosic pulp substrates which consist mainly of cellulose, but also contain residual hemicellulose and lignin. In order to evaluate the hydrolysis of the substrates by the fungal enzymes, the spatial polymer distributions were characterised by cross-polarisation magic angle spinning carbon-13 nuclear magnetic resonance (CP/MAS (13)C-NMR) in combination with spectral fitting. Proteins in culture supernatants at early and late stages of enzyme production were labeled by Tandem Mass Tags (TMT) and protein profiles were analysed by liquid chromatography-tandem mass spectrometry. The data have been deposited to the ProteomeXchange with identifier PXD001304. In total 124 proteins were identified and quantified in the culture supernatants, including cellulases, hemicellulases, other glycoside hydrolases, lignin-degrading enzymes, auxiliary activity 9 (AA9) family (formerly GH61), supporting activities of proteins and enzymes acting on cellulose, proteases, intracellular proteins and several hypothetical proteins. Surprisingly, substantial differences in the enzyme profiles were found even though there were minor differences in the chemical composition between the cellulose-rich substrates.

  16. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

  17. Construction of a cellulase hyper-expression system in Trichoderma reesei by promoter and enzyme engineering

    Directory of Open Access Journals (Sweden)

    Zou Gen

    2012-02-01

    Full Text Available Abstract Background Trichoderma reesei is the preferred organism for producing industrial cellulases. However, a more efficient heterologous expression system for enzymes from different organism is needed to further improve its cellulase mixture. The strong cbh1 promoter of T. reesei is frequently used in heterologous expression, however, the carbon catabolite repressor CREI may reduce its strength by binding to the cbh1 promoter at several binding sites. Another crucial point to enhance the production of heterologous enzymes is the stability of recombinant mRNA and the prevention of protein degradation within the endoplasmic reticulum, especially for the bacteria originated enzymes. In this study, the CREI binding sites within the cbh1 promoter were replaced with the binding sites of transcription activator ACEII and the HAP2/3/5 complex to improve the promoter efficiency. To further improve heterologous expression efficiency of bacterial genes within T. reesei, a flexible polyglycine linker and a rigid α-helix linker were tested in the construction of fusion genes between cbh1 from T. reesei and e1, encoding an endoglucanase from Acidothermus cellulolyticus. Results The modified promoter resulted in an increased expression level of the green fluorescent protein reporter by 5.5-fold in inducing culture medium and 7.4-fold in repressing culture medium. The fusion genes of cbh1 and e1 were successfully expressed in T. reesei under the control of promoter pcbh1m2. The higher enzyme activities and thermostability of the fusion protein with rigid linker indicated that the rigid linker might be more suitable for the heterologous expression system in T. reesei. Compared to the parent strain RC30-8, the FPase and CMCase activities of the secreted enzyme mixture from the corresponding transformant R1 with the rigid linker increased by 39% and 30% at 60°C, respectively, and the reduced sugar concentration in the hydrolysate of pretreated corn stover

  18. Influence of the carbon source on production of cellulases, hemicellulases and pectinases by Trichoderma reesei Rut C-30

    DEFF Research Database (Denmark)

    Olsson, Lisbeth; Christensen, T.M.I.E.; Hansen, K.P.;

    2003-01-01

    The growth and enzyme production by Trichoderma reesei Rut C-30 using different lignocellulosic materials as carbon source were investigated. Cellulose, sugar beet pulp and alkaline extracted sugar beet pulp (resulting in partial removal of hemicellulose, lignin and pectin) or mixtures thereof were...

  19. Comparative Secretome Analysis of Aspergillus niger, Trichoderma reesei, and Penicillium oxalicum During Solid-State Fermentation.

    Science.gov (United States)

    Gong, Weili; Zhang, Huaiqiang; Liu, Shijia; Zhang, Lili; Gao, Peiji; Chen, Guanjun; Wang, Lushan

    2015-11-01

    Filamentous fungi such as Aspergillus spp., Trichoderma spp., and Penicillium spp. are frequently used to produce high concentrations of lignocellulosic enzymes. This study examined the discrepancies in the compositions and dynamic changes in the extracellular enzyme systems secreted by Aspergillus niger ATCC1015, Trichoderma reesei QM9414, and Penicillium oxalicum 114-2 cultured on corn stover and wheat bran. The results revealed different types and an abundance of monosaccharides and oligosaccharides were released during incubation, which induced the secretion of diverse glycoside hydrolases. Both the enzyme activities and isozyme numbers of the three fungal strains increased with time. A total of 279, 161, and 183 secretory proteins were detected in A. niger, T. reesei, and P. oxalicum secretomes, respectively. In the A. niger secretomes, more enzymes involved in the degradation of (galacto)mannan, xyloglucan, and the backbone of pectin distributed mostly in dicots were detected. In comparison, although P. oxalicum 114-2 hardly secreted any xyloglucanases, the diversities of enzymes involved in the degradation of xylan and β-(1,3;1,4)-D-glucan commonly found in monocots were higher. The cellulase system of P. oxalicum 114-2 was more balanced. The degradation preference provided a new perspective regarding the recomposition of lignocellulosic enzymes based on substrate types.

  20. Generation of a glucose de-repressed mutant of Trichoderma reesei using disparity mutagenesis.

    Science.gov (United States)

    Iwakuma, Hidekazu; Koyama, Yoshiyuki; Miyachi, Ayako; Nasukawa, Masashi; Matsumoto, Hitoshi; Yano, Shuntaro; Ogihara, Jun; Kasumi, Takafumi

    2016-01-01

    We obtained a novel glucose de-repressed mutant of Trichoderma reesei using disparity mutagenesis. A plasmid containing DNA polymerase δ lacking proofreading activity, and AMAI, an autonomously replicating sequence was introduced into T. reesei ATCC66589. The rate of mutation evaluated with 5-fluoroorotic acid resistance was approximately 30-fold higher than that obtained by UV irradiation. The transformants harboring incompetent DNA polymerase δ were then selected on 2-deoxyglucose agar plates with hygromycin B. The pNP-lactoside hydrolyzing activities of mutants were 2 to 5-fold higher than the parent in liquid medium containing glucose. Notably, the amino acid sequence of cre1, a key gene involved in glucose repression, was identical in the mutant and parent strains, and further, the cre1 expression levels was not abolished in the mutant. Taken together, these results demonstrate that the strains of T. reesei generated by disparity mutagenesis are glucose de-repressed variants that contain mutations in yet-unidentified factors other than cre1.

  1. Quantitative Site-Specific Phosphoproteomics of Trichoderma reesei Signaling Pathways upon Induction of Hydrolytic Enzyme Production.

    Science.gov (United States)

    Nguyen, Elizabeth V; Imanishi, Susumu Y; Haapaniemi, Pekka; Yadav, Avinash; Saloheimo, Markku; Corthals, Garry L; Pakula, Tiina M

    2016-02-01

    The filamentous fungus Trichoderma reesei is used for industrial production of secreted enzymes including carbohydrate active enzymes, such as cellulases and hemicellulases. The production of many of these enzymes by T. reesei is influenced by the carbon source it grows on, where the regulation system controlling hydrolase genes involves various signaling pathways. T. reesei was cultivated in the presence of sorbitol, a carbon source that does not induce the production of cellulases and hemicellulases, and then exposed to either sophorose or spent-grain extract, which are efficient inducers of the enzyme production. Specific changes at phosphorylation sites were investigated in relation to the production of cellulases and hemicellulases using an MS-based framework. Proteome-wide phosphorylation following carbon source exchange was investigated in the early stages of induction: 0, 2, 5, and 10 min. The workflow involved sequential trypsin digestion, TiO2 enrichment, and MS analysis using a Q Exactive mass spectrometer. We report on the identification and quantitation of 1721 phosphorylation sites. Investigation of the data revealed a complex signaling network activated upon induction involving components related to light-mediated cellulase induction, osmoregulation, and carbon sensing. Changes in protein phosphorylation were detected in the glycolytic pathway, suggesting an inhibition of glucose catabolism at 10 min after the addition of sophorose and as early as 2 min after the addition of spent-grain extract. Differential phosphorylation of factors related to carbon storage, intracellular trafficking, cytoskeleton, and cellulase gene regulation were also observed. PMID:26689635

  2. Quantitative Site-Specific Phosphoproteomics of Trichoderma reesei Signaling Pathways upon Induction of Hydrolytic Enzyme Production.

    Science.gov (United States)

    Nguyen, Elizabeth V; Imanishi, Susumu Y; Haapaniemi, Pekka; Yadav, Avinash; Saloheimo, Markku; Corthals, Garry L; Pakula, Tiina M

    2016-02-01

    The filamentous fungus Trichoderma reesei is used for industrial production of secreted enzymes including carbohydrate active enzymes, such as cellulases and hemicellulases. The production of many of these enzymes by T. reesei is influenced by the carbon source it grows on, where the regulation system controlling hydrolase genes involves various signaling pathways. T. reesei was cultivated in the presence of sorbitol, a carbon source that does not induce the production of cellulases and hemicellulases, and then exposed to either sophorose or spent-grain extract, which are efficient inducers of the enzyme production. Specific changes at phosphorylation sites were investigated in relation to the production of cellulases and hemicellulases using an MS-based framework. Proteome-wide phosphorylation following carbon source exchange was investigated in the early stages of induction: 0, 2, 5, and 10 min. The workflow involved sequential trypsin digestion, TiO2 enrichment, and MS analysis using a Q Exactive mass spectrometer. We report on the identification and quantitation of 1721 phosphorylation sites. Investigation of the data revealed a complex signaling network activated upon induction involving components related to light-mediated cellulase induction, osmoregulation, and carbon sensing. Changes in protein phosphorylation were detected in the glycolytic pathway, suggesting an inhibition of glucose catabolism at 10 min after the addition of sophorose and as early as 2 min after the addition of spent-grain extract. Differential phosphorylation of factors related to carbon storage, intracellular trafficking, cytoskeleton, and cellulase gene regulation were also observed.

  3. Comparison between the cellulase systems of Trichoderma harzianum E58 and Trichoderma reesei C30

    Energy Technology Data Exchange (ETDEWEB)

    Saddler, J.N.; Hogan, C.M.; Louis-Seize, G.

    1985-06-01

    Nearly all of the filter paper, endoglucanase and ..beta..-glucosidase activities of T. harzianum E58 were located extracellularly, with low amounts of these activities detected in the cell extracts and relatively little associated with the cell wall. Most of the filter paper and endoglucanase activities of T. reesei C30 were detected extracellularly. The half lives of the different cellulase activities were assayed at various temperatures over a period of time. When the pH of the filtrate was adjusted to 4.8, the cellulase activities were considerably enhanced, with the average half-life at 50/sup 0/C extended to 25 hrs. When various lignocellulosic substrates were hydrolyzed by T. harzianum E58 cellulases approximately 90% of the reducing sugars were present as glucose while 50 - 60% of the reducing sugars were detected as glucose when T. reesei C30 cellulases were used.

  4. Exploring the Synergy between Cellobiose Dehydrogenase from Phanerochaete chrysosporium and Cellulase from Trichoderma reesei.

    Science.gov (United States)

    Wang, Min; Lu, Xuefeng

    2016-01-01

    Recent demands for the production of lignocellulose biofuels boosted research on cellulase. Hydrolysis efficiency and production cost of cellulase are two bottlenecks in "biomass to biofuels" process. The Trichoderma cellulase mixture is one of the most commonly used enzymes for cellulosic hydrolysis. During hydrolytic process cellobiose accumulation causes feedback inhibition against most cellobiohydrolases and endoglucanases. In this study, we demonstrated the synergism effects between cellobiose dehydrogenase (CDH) and cellulase both in vitro and in vivo. The CDH from Phanerochaete chrysosporium was heterologously expressed in Pichia pastoris. Supplementation of the purified CDH in Trichoderma cellulase increased the cellulase activities. Especially β-glucosidase activity was increased by 30-100% varying at different time points. On the other hand, the cdh gene was heterologously expressed in Trichoderma reesei to explore the synergism between CDH and cellulases in vivo. The analyses of gene expression and enzymatic profiles of filter paper activity, carboxymethylcellulase (CMCase) and β-glucosidase show the increased cellulase activity and the enhanced cellulase production in the cdh-expressing strains. The results elucidate a possible mechanism for diminishing the cellobiose inhibition of cellulase by CDH. These findings provide a novel perspective to make more economic enzyme cocktails for commercial application or explore alternative strategies for generating cellulase-producing strains with higher efficiency. PMID:27199949

  5. Disruption of Trichoderma reesei cre2, encoding an ubiquitin C-terminal hydrolase, results in increased cellulase activity

    Directory of Open Access Journals (Sweden)

    Denton Jai A

    2011-11-01

    Full Text Available Abstract Background The filamentous fungus Trichoderma reesei (Hypocrea jecorina is an important source of cellulases for use in the textile and alternative fuel industries. To fully understand the regulation of cellulase production in T. reesei, the role of a gene known to be involved in carbon regulation in Aspergillus nidulans, but unstudied in T. reesei, was investigated. Results The T. reesei orthologue of the A. nidulans creB gene, designated cre2, was identified and shown to be functional through heterologous complementation of a creB mutation in A. nidulans. A T. reesei strain was constructed using gene disruption techniques that contained a disrupted cre2 gene. This strain, JKTR2-6, exhibited phenotypes similar to the A. nidulans creB mutant strain both in carbon catabolite repressing, and in carbon catabolite derepressing conditions. Importantly, the disruption also led to elevated cellulase levels. Conclusions These results demonstrate that cre2 is involved in cellulase expression. Since the disruption of cre2 increases the amount of cellulase activity, without severe morphological affects, targeting creB orthologues for disruption in other industrially useful filamentous fungi, such as Aspergillus oryzae, Trichoderma harzianum or Aspergillus niger may also lead to elevated hydrolytic enzyme activity in these species.

  6. Improved heterologous gene expression in Trichoderma reesei by cellobiohydrolase I gene (cbh1) promoter optimization

    Institute of Scientific and Technical Information of China (English)

    Ti Liu; Tianhong Wang; Xian Li; Xuan Liu

    2008-01-01

    To improve heterologous gene expression in Trichoderma reesei, a set of optimal artificial cellobiohydrolase I gene (cbh1) promoters was obtained. The region from-677 to -724 with three potential glucose repressor binding sites was deleted. Then the region from-620 to-820 of the modified cbh1 promoter, including the CCAAT box and the Ace2 binding site, was repeatedly inserted into the modified cbh1 promoter, obtaining promoters with copy numbers 2, 4,and 6. The results showed that the glucose repression effects were abolished and the expression level of the glucuronidase (gus) reporter gene regulated by these multi-copy promoters was markedly enhanced as the copy number increased simultaneously. The data showed the great promise of using the promoter artificial modification strategy to increase heterologous gene expression in filamentous fungi and provided a set of optional high-expression vectors for gene function investigation and strain modification.

  7. Nutrient control for stationary phase cellulase production in Trichoderma reesei Rut C-30.

    Science.gov (United States)

    Callow, Nicholas V; Ray, Christopher S; Kelbly, Matthew A; Ju, Lu-Kwang

    2016-01-01

    This work describes the use of nutrient limitations with Trichoderma reesei Rut C-30 to obtain a prolonged stationary phase cellulase production. This period of non-growth may allow for dependable cellulase production, extended fermentation periods, and the possibility to use pellet morphology for easy product separation. Phosphorus limitation was successful in halting growth and had a corresponding specific cellulase production of 5±2 FPU/g-h. Combined with the addition of Triton X-100 for fungal pellet formation and low shear conditions, a stationary phase cellulase production period in excess of 300 h was achieved, with a constant enzyme production rate of 7±1 FPU/g-h. While nitrogen limitation was also effective as a growth limiter, it, however, also prevented cellulase production.

  8. Thermoascus aurantiacus CBHI/Cel7A Production in Trichoderma reesei on Alternative Carbon Sources

    Science.gov (United States)

    Benkő, Zsuzsa; Drahos, Eszter; Szengyel, Zsolt; Puranen, Terhi; Vehmaanperä, Jari; Réczey, Kati

    To develop functional enzymes in cellulose hydrolysis at or above 70°C the cellobiohydrolase (CBHI/Cel7A) of Thermoascus aurantiacus was cloned and expressed in Trichoderma reesei Rut-C30 under the strong cbh1 promoter. Cellulase production of the parental strain and the novel strain (RF6026) was examined in submerged fermentation experiments using various carbon sources, which were lactose, Solka Floc 200 cellulose powder, and steam pretreated corn stover. An industrially feasible production medium was used containing only distiller's spent grain, KH2PO4, and (NH4)2SO4. Enzyme production was followed by measurements of protein concentration, total cellulase enzyme activity (filter paper activity), β-glucosidase activity, CBHI activity, and endogenase I (EGI) activity. The Thermoascus CBHI/Cel7A activity was taken as an indication of the heterologous gene expression under the cbh1 promoter.

  9. Molecular simulation evidence for processive motion of Trichoderma reesei Cel7A during cellulose depolymerization

    Science.gov (United States)

    Zhao, Xiongce; Rignall, Tauna R.; McCabe, Clare; Adney, William S.; Himmel, Michael E.

    2008-07-01

    We present free energy calculations for the Trichoderma reesei Cel7A (cellobiohydrolase I) linker peptide from molecular dynamics simulations directed towards understanding the linker role in cellulose hydrolysis. The calculations predict an energy storage mechanism of the linker under stretching/compression that is consistent with processive depolymerization. The linker exhibits two stable states at lengths of 2.5 nm and 5.5 nm during extension/compression, with a free energy difference of 10.5 kcal/mol between the two states separated by an energy barrier. The switching between stable states supports the hypothesis that the linker peptide has the capacity to store energy in a manner similar to a spring.

  10. Is an organic nitrogen source needed for cellulase production by Trichoderma reesei Rut-C30?

    DEFF Research Database (Denmark)

    Rodríguez Gómez, Divanery; Hobley, Timothy John

    2013-01-01

    The effect of organic and inorganic nitrogen sources on Trichoderma reesei Rut-C30 cellulase production was investigated in submerged cultivations. Stirred tank bioreactors and shake flasks, with and without pH control, respectively, were employed. The experimental design involved the addition...... of individual organic nitrogen sources (soy peptone, glutamate, glycine and alanine) within a basal medium containing Avicel (i.e. micro crystalline cellulose) and ammonium sulphate. It was found that in the shake flask experiments, the highest cellulase activities (~0.1 ± 0.02 FPU ml−1) were obtained...... with media containing soy peptone (3–6 g l−1) and glutamate (3.6 g l−1). However, these improvements in the cellulase titers in the presence of the organic nitrogen sources appeared to be related to smaller changes in the pH of the medium. This was confirmed using stirred tank bioreactors with pH control...

  11. Systems analysis of lactose metabolism in Trichoderma reesei identifies a lactose permease that is essential for cellulase induction.

    Directory of Open Access Journals (Sweden)

    Christa Ivanova

    Full Text Available Trichoderma reesei colonizes predecayed wood in nature and metabolizes cellulose and hemicellulose from the plant biomass. The respective enzymes are industrially produced for application in the biofuel and biorefinery industry. However, these enzymes are also induced in the presence of lactose (1,4-0-ß-d-galactopyranosyl-d-glucose, a waste product from cheese manufacture or whey processing industries. In fact, lactose is the only soluble carbon source that induces these enzymes in T. reesei on an industrial level but the reason for this unique phenomenon is not understood. To answer this question, we used systems analysis of the T. reesei transcriptome during utilization of lactose. We found that the respective CAZome encoded all glycosyl hydrolases necessary for cellulose degradation and particularly for the attack of monocotyledon xyloglucan, from which ß-galactosides could be released that may act as the inducers of T. reesei's cellulases and hemicellulases. In addition, lactose also induces a high number of putative transporters of the major facilitator superfamily. Deletion of fourteen of them identified one gene that is essential for lactose utilization and lactose uptake, and for cellulase induction by lactose (but not sophorose in pregrown mycelia of T. reesei. These data shed new light on the mechanism by which T. reesei metabolizes lactose and offers strategies for its improvement. They also illuminate the key role of ß-D-galactosides in habitat specificity of this fungus.

  12. Heterologous expression of codon optimized Trichoderma reesei Cel6A in Pichia pastoris.

    Science.gov (United States)

    Sun, Fubao Fuelbiol; Bai, Renhui; Yang, Huimin; Wang, Fei; He, Jing; Wang, Chundi; Tu, Maobing

    2016-10-01

    The Cel6A deficiency has become one of the limiting factors for cellulose saccharification in biochemical conversion of cellulosic biomass to fuels and chemicals. The work attempted to use codon optimization to enhance Trichoderma reesei Cel6A expression in Pichia pastoris. Two recombinants P. pastoris GS115 containing AOX1 and GAP promotors were successfully constructed, respectively. The optimal temperatures and pHs of the expressed Cel6A from two recombinants were consistent with each other, were also in the extremely similar range to that reported on the native Cel6A from T. reesei. Based on the shake flask fermentation, AOX1 promotor enabled the recombinant to produce 265U/L and 300mg/L of the Cel6A enzyme, and the GAP promotor resulted in 145U/L and 200mg/L. High cell density fed batch (HCDFB) fermentation significantly improved the enzyme titer (1100U/L) and protein yield (2.0g/L) for the recombinant with AOX1 promotor. Results have showed that the AOX1 promotor is more suitable than the GAP for the Cel6A expression in P. pastoris. And the HCDFB cultivation is a favorable way to express the Cel6A highly in the methanol inducible yeast. PMID:27542751

  13. Regulation of cellulase expression, sporulation, and morphogenesis by velvet family proteins in Trichoderma reesei.

    Science.gov (United States)

    Liu, Kuimei; Dong, Yanmei; Wang, Fangzhong; Jiang, Baojie; Wang, Mingyu; Fang, Xu

    2016-01-01

    Homologs of the velvet protein family are encoded by the ve1, vel2, and vel3 genes in Trichoderma reesei. To test their regulatory functions, the velvet protein-coding genes were disrupted, generating Δve1, Δvel2, and Δvel3 strains. The phenotypic features of these strains were examined to identify their functions in morphogenesis, sporulation, and cellulase expression. The three velvet-deficient strains produced more hyphal branches, indicating that velvet family proteins participate in the morphogenesis in T. reesei. Deletion of ve1 and vel3 did not affect biomass accumulation, while deletion of vel2 led to a significantly hampered growth when cellulose was used as the sole carbon source in the medium. The deletion of either ve1 or vel2 led to the sharp decrease of sporulation as well as a global downregulation of cellulase-coding genes. In contrast, although the expression of cellulase-coding genes of the ∆vel3 strain was downregulated in the dark, their expression in light condition was unaffected. Sporulation was hampered in the ∆vel3 strain. These results suggest that Ve1 and Vel2 play major roles, whereas Vel3 plays a minor role in sporulation, morphogenesis, and cellulase expression.

  14. An ethanolamine kinase Eki1 affects radial growth and cell wall integrity in Trichoderma reesei.

    Science.gov (United States)

    He, Ronglin; Guo, Wei; Zhang, Dongyuan

    2015-09-01

    Ethanolamine kinase (ATP:ethanolamine O-phosphotransferase, EC 2.7.1.82) catalyzes the committed step of phosphatidylethanolamine synthesis via the CDP-ethanolamine pathway. The functions of eki genes that encode ethanolamine kinase have been intensively studied in mammalian cells, fruit flies and yeast. However, the role of the eki gene has not yet been characterized in filamentous fungi. In this study, Treki1, an ortholog of Saccharomyces cerevisiae EKI1, was identified and functionally characterized using a target gene deletion strategy in Trichoderma reesei. A Treki deletion mutant was less sensitive to cell wall stressors calcofluor white and Congo red and released fewer protoplasts during cell wall digestion than the parent strain QM9414. Further transcription analysis showed that the expression levels of five genes that encode chitin synthases were drastically increased in the ΔTreki1 mutant. The chitin content was also increased in the null mutant of Treki1 comparing to the parent strain. In addition, the ΔTreki1 mutant exhibited defects in radial growth, conidiation and the accumulation of ethanolamine. The results indicate that Treki1 plays a key role in growth and development and in the maintenance of cell wall integrity in T. reesei.

  15. Trichoderma reesei XYN VI--a novel appendage-dependent eukaryotic glucuronoxylan hydrolase.

    Science.gov (United States)

    Biely, Peter; Puchart, Vladimír; Stringer, Mary Ann; Mørkeberg Krogh, Kristian B R

    2014-09-01

    Expression of a Trichoderma reesei gene coding for a putative GH30 xylanase in Aspergillus oryzae led to isolation and purification of a novel xylanase exhibiting catalytic properties different from those of the previously characterized GH30 xylanase XYN IV of T. reesei. The novel enzyme, named XYN VI, exhibited catalytic properties similar to appendage-dependent GH30 glucuronoxylanases previously recognized only in bacteria. XYN VI showed high specific activity only on xylans or xylooligosaccharides containing 4-O-methyl-D-glucuronic acid or D-glucuronic acid side substituents. The cleavage of the main chain takes place primarily at the second glycosidic linkage from the branch towards the reducing end of the polysaccharides or aldouronic acids. These catalytic properties resemble bacterial GH30 glucuronoxylanases, although the recognition of the uronic acid side chains by XYN VI is apparently based on interaction of the substrate with other amino acids. Moreover, in contrast to bacterial enzymes, XYN VI is also capable of slower but significant cleavage of unsubstituted parts of xylan and acidic xylooligosaccharides. The data point to a great catalytic diversity of xylanases produced by the most extensively studied cellulolytic fungus. PMID:25041335

  16. Influence of sorbitol on protein production and glycosylation and cell wall formation in Trichoderma reesei.

    Science.gov (United States)

    Górka-Nieć, Wioletta; Perlińska-Lenart, Urszula; Zembek, Patrycja; Palamarczyk, Grażyna; Kruszewska, Joanna S

    2010-10-01

    Sorbitol is often used at 1 mol/liter as an osmotic stabilizer for cultivation of fungi with a fragile cell wall phenotype. On the other hand, at this concentration sorbitol causes an osmotic stress in fungal cells resulting in intensive production of intracellular glycerol. The highly increased consumption of glucose for glycerol synthesis may lead to changes in processes requiring carbohydrate residues. This study provides new information on the consequences of osmotic stress to the cell wall composition, protein production and glycosylation, and cell morphology of Trichoderma reesei. We observed that high osmolarity conditions enhanced biomass production and strongly limited synthesis of cell wall glucans and chitin. Moreover, in these conditions the amount of secreted protein decreased nearly ten-fold and expression of cbh1 and cbh2 genes coding for cellobiohydrolase I and cellobiohydrolase II, the main secretory proteins in T. reesei, was inhibited resulting in a lack of the proteins in the cell and cultivation medium. The activity of DPM synthase, enzyme engaged in both N- and O-glycosylation pathways, was reduced two-fold, suggesting an overall inhibition of protein glycosylation. However, the two modes of glycosylation were affected divergently: O-glycosylation of secreted proteins decreased in the early stages of growth while N-glycosylation significantly increased in the stationary phase.

  17. Hydrolytic properties of two cellulases of Trichoderma reesei expressed in yeast.

    Science.gov (United States)

    Bailey, M J; Siika-aho, M; Valkeajärvi, A; Penttilä, M E

    1993-02-01

    Two cellulases of the filamentous fungus Trichoderma reesei, cellobiohydrolase II (CBHII, EC 3.2.1.91) and endoglucanase I (EGI, EC 3.2.1.4), produced in recombinant strains of the yeast Saccharomyces cerevisiae, were tested in the hydrolysis of cellulose, xylan and other polymeric substrates. Both enzymes were active against unsubstituted, insoluble cellulose. CBHII had greater activity than EGI against crystalline cellulose, whereas in the case of amorphous substrate the order was reversed. Evidence for synergism was obtained when mixtures of the two enzymes were used with a constant total protein dosage. The EGI was also active against soluble substituted cellulose derivatives, whereas the activity of CBHII against these substrates was insignificant. Both enzymes were active against barley (1-->3,1-->4)-beta-glucan, but were inactive against (1-->3,1-->6)-beta-glucan (laminarin). An apparent low mannan-degrading activity of EGI against locust-bean (Ceratonia siliqua) gum galactomannan was not confirmed when homopolymeric mannan was used as substrate in a prolonged hydrolysis test. EGI exhibited considerably greater activity against insoluble, unsubstituted hardwood xylan than against amorphous cellulose. Soluble 4-O-methyl-glucuronoxylan was also attacked by EGI, although to a somewhat lesser extent than the unsubstituted xylan. By comparison with two purified xylanases of T. reesei, EGI produced xylo-oligosaccharides with a longer mean chain length when acting on both substituted and unsubstituted xylan substrates. CBHII was inactive against xylan.

  18. Trichoderma reesei FS10-C enhances phytoremediation by Sedum plumbizincicola for Cd-contaminated soils and associated soil microbial activities

    Directory of Open Access Journals (Sweden)

    Ying eTeng

    2015-06-01

    Full Text Available This study aimed to explore the effects of Trichoderma reesei FS10-C on the phytoremediation of Cd-contaminated soil by Sedum plumbizincicola as well as soil fertility. After characterizing the Cd tolerance of T. reesei FS10-C, a pot experiment was carried out to investigate the plant growth and Cd uptake of S. plumbizincicola with the addition of inoculation agents with/without T. reesei FS10-C. The soil samples in pots were analyzed for pH, available phosphorus (P, microbial biomass C, enzyme activities, microbial community functional diversity and Trichoderma colonization ability. The results indicated that FS10-C possessed high Cd resistance up to 300 mg L-1. All inoculation agents enhanced the biomass of plant shoots by 6-53% fresh weight and 16-61% dry weight as well as Cd uptake in plant shoots by 10-53% compared with the control. In addition, soil biomass C, enzyme activities and microbial community evenness were all increased to varying degrees by all inoculation agents, indicating that soil microbial biomass and activities were both enhanced. It was also found that the two inoculation agents accompanied by FS10-C were better compared with the inoculation agents without FS10-C on all accounts, from which it could be concluded that T. reesei FS10-C was effective in improving Cd phytoremediation of S. plumbizincicola and soil fertility. Furthermore, among all the inoculation agents, solid fermentation powder of FS10-C demonstrated the greatest capacity to enhance plant growth, Cd uptake, nutrient release, and microbial biomass and activities, as indicated by its superior ability to colonize Trichoderma. Thus, we could also conclude that solid fermentation powder of FS10-C was a good candidate for use as an inoculation agent for T. reesei FS10-C to improve the phytoremediation of Cd-contaminated soil and soil fertility.

  19. Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei) for Effective Wastewater Treatment and Bio-Oil Production

    OpenAIRE

    Bhanja, Anshuman; Minde, Gauri; Magdum, Sandip; V. Kalyanraman

    2014-01-01

    Biological wastewater treatment typically requires the use of bacteria for degradation of carbonaceous and nitrogenous compounds present in wastewater. The high lipid containing biomass can be used to extract oil and the contents can be termed as bio-oil (or biodiesel or myco-diesel after transesterification). The separate experiments were conducted on actual wastewater samples with 5% v/v inoculum of Mucor circinelloides MTCC1297 and Trichoderma reesei NCIM992 strains. The observed reduction...

  20. Kinetic transcriptome analysis reveals an essentially intact induction system in a cellulase hyper-producer Trichoderma reesei strain

    OpenAIRE

    Poggi-Parodi, Dante; Bidard, Frédérique; Pirayre, Aurélie; Portnoy, Thomas; Blugeon, Corinne; Seiboth, Bernhard; Kubicek, Christian P.; Le Crom, Stéphane; Margeot, Antoine

    2014-01-01

    Background The filamentous fungus Trichoderma reesei is the main industrial cellulolytic enzyme producer. Several strains have been developed in the past using random mutagenesis, and despite impressive performance enhancements, the pressure for low-cost cellulases has stimulated continuous research in the field. In this context, comparative study of the lower and higher producer strains obtained through random mutagenesis using systems biology tools (genome and transcriptome sequencing) can ...

  1. Genetic and metabolic biodiversity of Trichoderma from Colombia and adjacent neotropic regions.

    Science.gov (United States)

    Hoyos-Carvajal, Lilliana; Orduz, Sergio; Bissett, John

    2009-09-01

    The genus Trichoderma has been studied for production of enzymes and other metabolites, as well as for exploitation as effective biological control agents. The biodiversity of Trichoderma has seen relatively limited study over much of the neotropical region. In the current study we assess the biodiversity of 183 isolates from Mexico, Guatemala, Panama, Ecuador, Peru, Brazil and Colombia, using morphological, metabolic and genetic approaches. A comparatively high diversity of species was found, comprising 29 taxa: Trichoderma asperellum (60 isolates), Trichoderma atroviride (3), Trichoderma brevicompactum (5), Trichoderma crassum (3), Trichoderma erinaceum (3), Trichoderma gamsii (2), Trichoderma hamatum (2), Trichoderma harzianum (49), Trichoderma koningiopsis (6), Trichoderma longibrachiatum (3), Trichoderma ovalisporum (1), Trichoderma pubescens (2), Trichoderma rossicum (4), Trichoderma spirale (1), Trichoderma tomentosum (3), Trichoderma virens (8), Trichoderma viridescens (7) and Hypocrea jecorina (3) (anamorph: Trichoderma reesei), along with 11 currently undescribed species. T. asperellum was the prevalent species and was represented by two distinct genotypes with different metabolic profiles and habitat preferences. The second predominant species, T. harzianum, was represented by three distinct genotypes. The addition of 11 currently undescribed species is evidence of the considerable unresolved biodiversity of Trichoderma in neotropical regions. Sequencing of the internal transcribed spacer regions (ITS) of the ribosomal repeat could not differentiate some species, and taken alone gave several misidentifications in part due to the presence of nonorthologous copies of the ITS in some isolates.

  2. Familiar Stranger: Ecological Genomics of the Model Saprotroph and Industrial Enzyme Producer Trichoderma reesei Breaks the Stereotypes.

    Science.gov (United States)

    Druzhinina, I S; Kubicek, C P

    2016-01-01

    The filamentous fungus Trichoderma reesei (Hypocreales, Ascomycota) has properties of an efficient cell factory for protein production that is exploited by the enzyme industry, particularly with respect to cellulase and hemicellulase formation. Under conditions of industrial fermentations it yields more than 100g secreted protein L(-1). Consequently, T. reesei has been intensively studied in the 20th century. Most of these investigations focused on the biochemical characteristics of its cellulases and hemicellulases, on the improvement of their properties by protein engineering, and on enhanced enzyme production by recombinant strategies. However, as the fungus is rare in nature, its ecology remained unknown. The breakthrough in the understanding of the fundamental biology of T. reesei only happened during 2000s-2010s. In this review, we compile the current knowledge on T. reesei ecology, physiology, and genomics to present a holistic view on the natural behavior of the organism. This is not only critical for science-driven further improvement of the biotechnological applications of this fungus, but also renders T. reesei as an attractive model of filamentous fungi with superior saprotrophic abilities. PMID:27261782

  3. A high performance Trichoderma reesei strain that reveals the importance of xylanase III in cellulosic biomass conversion.

    Science.gov (United States)

    Nakazawa, Hikaru; Kawai, Tetsushi; Ida, Noriko; Shida, Yosuke; Shioya, Kouki; Kobayashi, Yoshinori; Okada, Hirofumi; Tani, Shuji; Sumitani, Jun-ichi; Kawaguchi, Takashi; Morikawa, Yasushi; Ogasawara, Wataru

    2016-01-01

    The ability of the Trichoderma reesei X3AB1strain enzyme preparations to convert cellulosic biomass into fermentable sugars is enhanced by the replacement of xyn3 by Aspergillus aculeatus β-glucosidase 1 gene (aabg1), as shown in our previous study. However, subsequent experiments using T. reesei extracts supplemented with the glycoside hydrolase (GH) family 10 xylanase III (XYN III) and GH Family 11 XYN II showed increased conversion of alkaline treated cellulosic biomass, which is rich in xylan, underscoring the importance of XYN III. To attain optimal saccharifying potential in T. reesei, we constructed two new strains, C1AB1 and E1AB1, in which aabg1 was expressed heterologously by means of the cbh1 or egl1 promoters, respectively, so that the endogenous XYN III synthesis remained intact. Due to the presence of wild-type xyn3 in T. reesei E1AB1, enzymes prepared from this strain were 20-30% more effective in the saccharification of alkaline-pretreated rice straw than enzyme extracts from X3AB1, and also outperformed recent commercial cellulase preparations. Our results demonstrate the importance of XYN III in the conversion of alkaline-pretreated cellulosic biomass by T. reesei.

  4. Isotherms for adsorption of cellobiohydrolase I and II from Trichoderma reesei on microcrystalline cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Medve, J.; Tjerneld, F. [Univ. of Lund (Sweden); Stahlberg, J. [Univ. of Uppsala (Sweden)

    1997-04-01

    Adsorption to microcrystalline cellulose (Avicel) of pure cellobiohydrolase I and II (CBH I and CBH II) from Trichoderma reesei has been studied. Adsorption isotherms of the enzymes were measured at 4{degree}C using CBH I and CBH II alone and in reconstituted equimolar mixtures. Several models (Langmuir, Freundlich, Temkin, Jovanovic) were tested to describe the experimental adsorption isotherms. The isotherms did not follow the basic (one site) Langmuir equation that has often been used to describe adsorption isotherms of cellulases; correlation coefficients (R{sup 2}) were only 0.926 and 0.947, for CBH I and II, respectively. The experimental isotherms were best described by a model of Langmuir type with two adsorption sites and by a combined Langmuir-Freundlich model (analogous to the Hill equation); using these models the correlation coefficients were in most cases higher than 0.995. Apparent binding parameters derived from the two sites Langmuir model indicated stronger binding of CBH II compared to CBH I; the distribution coefficients were 20.7 and 3.7 L/g for the two enzymes, respectively. The binding capacity was higher for CBH I than for CBH II. The isotherms when analyzed with the combined model indicated presence of unequal binding sites on cellulose and/or negative cooperativity in the binding of the enzyme molecules. 39 refs., 3 figs., 3 tabs.

  5. Adsorption and synergism of cellobiohydrolase I and II of Trichoderma reesei during hydrolysis of microcrystalline cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Medve, J.; Tjerneld, F. (Univ. of Lund (Sweden). Dept. of Biochemistry); Staahlberg, J. (Univ. of Uppsala (Sweden). Dept. of Molecular Biology)

    1994-11-05

    Hydrolysis of microcrystalline cellulose (Avicel) by cellobiohydrolase I and II (CBH I and II) from Trichoderma reesei has been studied. Adsorption and synergism of the enzymes were investigated. Experiments were performed at different temperatures and enzyme/substrate ratios using CBH I and CBH II alone and in reconstituted equimolar mixtures. Fast protein liquid chromatography (FPLC) analysis was found to be an accurate and reproducible method to follow the enzyme adsorption. A linear correlation was found between the conversion and the amount of adsorbed enzyme when Avicel was hydrolyzed by increasing amounts of CBH I and/or CBH II. CBH I had lower specific activity compared to CBH II although, over a wide concentration range, more CBH I was adsorbed than CBH II. Synergism between the cellobiohydrolases during hydrolysis of the amorphous fraction of Avicel showed a maximum as a function of total enzyme concentration. Synergism measured as a function of bound enzyme showed a continuous increase, which indicates that by decreasing the distance between the two enzymes the synergism is enhanced. The adsorption process for both enzymes was slow. Depending on the enzyme/substrate ratio it took 30--90 min to reach 95% of the equilibrium binding. The amount of bound enzyme decreased with increasing temperature. The two enzymes compete for the adsorption sites but also bind to specific sites. Stronger competition for adsorption sites was shown by CBH I.

  6. The Relation Between Promoter Chromatin Status, Xyr1 and Cellulase Ex-pression in Trichoderma reesei.

    Science.gov (United States)

    Mello-de-Sousa, Thiago M; Rassinger, Alice; Derntl, Christian; Poças-Fonseca, Marcio J; Mach, Robert L; Mach-Aigner, Astrid R

    2016-04-01

    The ascomycete Trichoderma reesei is used for the production of plant cell wall-degrading enzymes in industrial scale. The interplay of the transactivator Xyr1 and the repressor Cre1 mainly regulates the expression of these enzymes. During induc-ing conditions, such as in the presence of sophorose, the transcription of the two major cellulase-encoding genes, cbh1 and cbh2, is activated as well as the expression of xyr1. In the presence of D-glucose carbon catabolite repression mediated by Cre1 takes place and the expression of Xyr1 and the plant cell wall-degrading enzymes is down-regulated. In this study we compare the chromatin status of xyr1, cbh1, and cbh2 promoters in the wild-type strain and the Cre1-deficient strain Rut-C30. Chromatin rearrangement occurs in the xyr1 promoter during induction on sophorose. Chromatin opening and protein-DNA interactions in the xyr1 promoter were detected especially in a region located 0.9 kb upstream the translation start co-don, which bears several putative Cre1-binding sites and a CCAAT-box. Moreover, the xyr1 promoter is overall more acces-sible in a cre1-truncated background, no matter which carbon source is present. This makes the xyr1 regulatory sequence a good target for promoter engineering aiming at the enhancement of cellulase production. PMID:27226770

  7. Computational Investigations of Trichoderma Reesei Cel7A Suggest New Routes for Enzyme Activity Improvements

    Energy Technology Data Exchange (ETDEWEB)

    Beckham, G. T.; Payne, C. M.; Bu, L.; Taylor, C. B.; McCabe, C.; Chu, J. W.; Himmel, M. E.; Crowley, M. F.

    2012-01-01

    The Trichoderma reesei Family 7 cellulase (Cel7A) is a key industrial enzyme in the production of biofuels from lignocellulosic biomass. It is a multi-modular enzyme with a Family 1 carbohydrate-binding module, a flexible O-glycosylated linker, and a large catalytic domain. We have used simulation to elucidate new functions for the 3 sub-domains, which suggests new routes to increase the activity of this central enzyme. These findings include new roles for glycosylation, which we have shown can be used to tune the binding affinity. We have also examined the structures of the catalytically-active complex of Cel7A and its non-processive counterpart, Cel7B, engaged on cellulose, which suggests allosteric mechanisms involved in chain binding when these cellulases are complexed on cellulose. Our computational results also suggest that product inhibition varies significantly between Cel7A and Cel7B, and we offer a molecular-level explanation for this observation. Finally, we discuss simulations of the absolute and relative binding free energy of cellulose ligands and various mutations along the CD tunnel, which will affect processivity and the ability of Cel7A (and related enzymes) to digest cellulose. These results highlight new considerations in protein engineering for processive and non-processive cellulases for production of lignocellulosic biofuels.

  8. Cold adaptation of a mesophilic cellulase, EG III from Trichoderma reesei, by directed evolution

    Institute of Scientific and Technical Information of China (English)

    XIAO; Zhizhuang(肖志壮); WANG; Pan(王攀); QU; Yinbo(曲音波); GAO; Peiji(高培基); WANG; Tianhong(汪天虹)

    2002-01-01

    Cold-active enzymes have received little research attention although they are very useful in industries. Since the structure bases of cold adaptation of enzymes are still unclear, it is also very difficult to obtain cold-adapted enzymes for industrial applications using routine protein engineering methods. In this work, we employed directed evolution method to randomly mutate a mesophilic cellulase, endoglucanase III (EG III) from Trichoderma reesei, and obtained a cold- adapted mutant, designated as w-3. DNA sequence analysis indicates that w-3 is a truncated form of native EG III with a deletion of 25 consecutive amino acids at C-terminus. Further examination of enzymatic kinetics and thermal stability shows that mutant w-3 has a higher Kcat value and becomes more thermolabile than its parent. In addition, activation energies of w-3 and wild type EG III calculated from Arrhenius equation are 13.3 kJ@mol-1 and 26.2 kJ@mol-1, respectively. Therefore, the increased specific activity of w-3 at lower temperatures could result from increased Kcat value and decreased activation energy.

  9. Cellobiohydrolase 1 from Trichoderma reesei degrades cellulose in single cellobiose steps

    Science.gov (United States)

    Brady, Sonia K.; Sreelatha, Sarangapani; Feng, Yinnian; Chundawat, Shishir P. S.; Lang, Matthew J.

    2015-12-01

    Cellobiohydrolase 1 from Trichoderma reesei (TrCel7A) processively hydrolyses cellulose into cellobiose. Although enzymatic techniques have been established as promising tools in biofuel production, a clear understanding of the motor's mechanistic action has yet to be revealed. Here, we develop an optical tweezers-based single-molecule (SM) motility assay for precision tracking of TrCel7A. Direct observation of motility during degradation reveals processive runs and distinct steps on the scale of 1 nm. Our studies suggest TrCel7A is not mechanically limited, can work against 20 pN loads and speeds up when assisted. Temperature-dependent kinetic studies establish the energy requirements for the fundamental stepping cycle, which likely includes energy from glycosidic bonds and other sources. Through SM measurements of isolated TrCel7A domains, we determine that the catalytic domain alone is sufficient for processive motion, providing insight into TrCel7A's molecular motility mechanism.

  10. Application of Statistical Design for the Production of Cellulase by Trichoderma reesei Using Mango Peel.

    Science.gov (United States)

    Saravanan, P; Muthuvelayudham, R; Viruthagiri, T

    2012-01-01

    Optimization of the culture medium for cellulase production using Trichoderma reesei was carried out. The optimization of cellulase production using mango peel as substrate was performed with statistical methodology based on experimental designs. The screening of nine nutrients for their influence on cellulase production is achieved using Plackett-Burman design. Avicel, soybean cake flour, KH(2)PO(4), and CoCl(2)·6H(2)O were selected based on their positive influence on cellulase production. The composition of the selected components was optimized using Response Surface Methodology (RSM). The optimum conditions are as follows: Avicel: 25.30 g/L, Soybean cake flour: 23.53 g/L, KH(2)PO(4): 4.90 g/L, and CoCl(2)·6H(2)O: 0.95 g/L. These conditions are validated experimentally which revealed an enhanced Cellulase activity of 7.8 IU/mL.

  11. Application of Statistical Design for the Production of Cellulase by Trichoderma reesei Using Mango Peel

    Directory of Open Access Journals (Sweden)

    P. Saravanan

    2012-01-01

    Full Text Available Optimization of the culture medium for cellulase production using Trichoderma reesei was carried out. The optimization of cellulase production using mango peel as substrate was performed with statistical methodology based on experimental designs. The screening of nine nutrients for their influence on cellulase production is achieved using Plackett-Burman design. Avicel, soybean cake flour, KH2PO4, and CoCl2·6H2O were selected based on their positive influence on cellulase production. The composition of the selected components was optimized using Response Surface Methodology (RSM. The optimum conditions are as follows: Avicel: 25.30 g/L, Soybean cake flour: 23.53 g/L, KH2PO4: 4.90 g/L, and CoCl2·6H2O: 0.95 g/L. These conditions are validated experimentally which revealed an enhanced Cellulase activity of 7.8 IU/mL.

  12. Antioxidant activity and ACE-inhibitory of Class II hydrophobin from wild strain Trichoderma reesei.

    Science.gov (United States)

    Khalesi, Mohammadreza; Jahanbani, Raheleh; Riveros-Galan, David; Sheikh-Hassani, Vahid; Sheikh-Zeinoddin, Mahmoud; Sahihi, Mehdi; Winterburn, James; Derdelinckx, Guy; Moosavi-Movahedi, Ali Akbar

    2016-10-01

    There are several possible uses of the Class II hydrophobin HFBII in clinical applications. To fully understand and exploit this potential however, the antioxidant activity and ACE-inhibitory potential of this protein need to be better understood and have not been previously reported. In this study, the Class II hydrophobin HFBII was produced by the cultivation of wild type Trichoderma reesei. The crude hydrophobin extract obtained from the fermentation process was purified using reversed-phase liquid chromatography and the identity of the purified HFBII verified by MALDI-TOF (molecular weight: 7.2kDa). Subsequently the antioxidant activities of different concentrations of HFBII (0.01-0.40mg/mL) were determined. The results show that for HFBII concentrations of 0.04mg/mL and upwards the protein significantly reduced the presence of ABTS(+) radicals in the medium, the IC50 value found to be 0.13mg/mL. Computational modeling highlighted the role of the amino acid residues located in the conserved and exposed hydrophobic patch on the surface of the HFBII molecule and the interactions with the aromatic rings of ABTS. The ACE-inhibitory effect of HFBII was found to occur from 0.5mg/mL and upwards, making the combination of HFBII with strong ACE-inhibitors attractive for use in the healthcare industry. PMID:27211298

  13. Improving the activity of Trichoderma reesei cel7B through stabilizing the transition state.

    Science.gov (United States)

    Wang, Yefei; Song, Xiangfei; Zhang, Shujun; Li, Jingwen; Shu, Zhiyu; He, Chunyan; Huang, Qingshan; Yao, Lishan

    2016-06-01

    Trichoderma reesei (Tr.) cellulases, which convert cellulose to reducing sugars, are a promising catalyst used in the lignocellulosic biofuel production. Improving Tr. cellulases activity, though very difficult, is highly desired due to the recalcitrance of lignocellulose. Meanwhile, it is preferable to enhance the cellulase's promiscuity so that substrates other than cellulose can also be hydrolyzed. In this work, an attempt is made to improve the catalytic activity of a major endogluanase Tr. Cel7B against xylan which crosslinks with cellulose in lignocellulose. By using quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations, the transition state of the xylo-oligosaccharide hydrolysis is identified. Then, mutations are introduced and their effect on the transition state stabilization is ranked based on the free energy calculations. Seven top ranked mutants are evaluated experimentally. Three mutants A208Q, A222D, and G230R show a higher activity than the wild-type Tr. Cel7B in the hydrolysis of xylan (by up to 47%) as well as filter paper (by up to 50%). The combination of the single mutants can further improve the enzyme activity. Our work demonstrates that the free energy method is effective in engineering the Tr. Cel7B activity against xylan and cellulose, and thus may also be useful for improving the activity of other Tr. cellulases. Biotechnol. Bioeng. 2016;113: 1171-1177. © 2015 Wiley Periodicals, Inc. PMID:26616246

  14. Purification and some properties of a β-glucanase from a strain, Trichoderma reesei GXC

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    β-glucanase was purified from a solid-state culture of Trichoderma reesei on wheat bran in three steps which comprised ammonium sulfate precipitation, Sephadex G-100 chromatography, and DEAE-Sephadex A-50 chromatography. The molecular mass was determined to be 35.21 kilodaltons by sodium dodecyl sulfate-12.5% polyacrylamide gel electrophoresis. The β-glucanase at low pHs was more stable than that at high pHs, and optimum pH was 5.0. The optimum temperature was 60 ℃, and β-glucanase was relatively stable at below 40 ℃ for 60 min. The Km of the enzyme on β-glucan was 10.86 mg/ml, and the Vmax on β-glucan was 14286 μmol of glucose equivalents per mg of the pure enzyme per min. The β-glucanase activity was significantly inhibited by Fe3+ ions, and was reduced in the presence of Cu2+ ions, Mn2+ ions and Mg2+ ions at 5 mmol/L and 10 mmol/L, respectively. The β-glucanase activity was stimulated by Co2+ ions, Ca2+ ions, Zn2+ ions, and Fe2+ ions at 1 mmol/L and 5 mmol/L, respectively.

  15. Improvement of thermostability and activity of Trichoderma reesei endo-xylanase Xyn III on insoluble substrates.

    Science.gov (United States)

    Matsuzawa, Tomohiko; Kaneko, Satoshi; Yaoi, Katsuro

    2016-09-01

    Trichoderma reesei Xyn III, an endo-β-1,4-xylanase belonging to glycoside hydrolase family 10 (GH10), is vital for the saccharification of xylans in plant biomass. However, its enzymatic thermostability and hydrolytic activity on insoluble substrates are low. To overcome these difficulties, the thermostability of Xyn III was improved using random mutagenesis and directed evolution, and its hydrolytic activity on insoluble substrates was improved by creating a chimeric protein. In the screening of thermostable Xyn III mutants from a random mutagenesis library, we identified two amino acid residues, Gln286 and Asn340, which are important for the thermostability of Xyn III. The Xyn III Gln286Ala/Asn340Tyr mutant showed xylanase activity even after heat treatment at 60 °C for 30 min or 50 °C for 96 h, indicating a dramatic enhancement in thermostability. In addition, we found that the addition of a xylan-binding domain (XBD) to the C-terminal of Xyn III improved its hydrolytic activity on insoluble xylan. PMID:27138202

  16. Two major facilitator superfamily sugar transporters from Trichoderma reesei and their roles in induction of cellulase biosynthesis.

    Science.gov (United States)

    Zhang, Weixin; Kou, Yanbo; Xu, Jintao; Cao, Yanli; Zhao, Guolei; Shao, Jing; Wang, Hai; Wang, Zhixing; Bao, Xiaoming; Chen, Guanjun; Liu, Weifeng

    2013-11-15

    Proper perception of the extracellular insoluble cellulose is key to initiating the rapid synthesis of cellulases by cellulolytic Trichoderma reesei. Uptake of soluble oligosaccharides derived from cellulose hydrolysis represents a potential point of control in the induced cascade. In this study, we identified a major facilitator superfamily sugar transporter Stp1 capable of transporting cellobiose by reconstructing a cellobiose assimilation system in Saccharomyces cerevisiae. The absence of Stp1 in T. reesei resulted in differential cellulolytic response to Avicel versus cellobiose. Transcriptional profiling revealed a different expression profile in the Δstp1 strain from that of wild-type strain in response to Avicel and demonstrated that Stp1 somehow repressed induction of the bulk of major cellulase and hemicellulose genes. Two other putative major facilitator superfamily sugar transporters were, however, up-regulated in the profiling. Deletion of one of them identified Crt1 that was required for growth and enzymatic activity on cellulose or lactose, but was not required for growth or hemicellulase activity on xylan. The essential role of Crt1 in cellulase induction did not seem to rely on its transporting activity because the overall uptake of cellobiose or sophorose by T. reesei was not compromised in the absence of Crt1. Phylogenetic analysis revealed that orthologs of Crt1 exist in the genomes of many filamentous ascomycete fungi capable of degrading cellulose. These data thus shed new light on the mechanism by which T. reesei senses and transmits the cellulose signal and offers potential strategies for strain improvement.

  17. Direct Ethanol Production from Lignocellulosic Sugars and Sugarcane Bagasse by a Recombinant Trichoderma reesei Strain HJ48

    Directory of Open Access Journals (Sweden)

    Jun Huang

    2014-01-01

    Full Text Available Trichoderma reesei can be considered as a candidate for consolidated bioprocessing (CBP microorganism. However, its ethanol yield needs to be improved significantly. Here the ethanol production of T. reesei CICC 40360 was improved by genome shuffling while simultaneously enhancing the ethanol resistance. The initial mutant population was generated by nitrosoguanidine treatment of the spores, and an improved population producing more than fivefold ethanol than wild type was obtained by genome shuffling. The results show that the shuffled strain HJ48 can efficiently convert lignocellulosic sugars to ethanol under aerobic conditions. Furthermore, it was able to produce ethanol directly from sugarcane bagasse, demonstrating that the shuffled strain HJ48 is a suitable microorganism for consolidated bioprocessing.

  18. Effect of temperature on the production of cellulases, xylanases and lytic enzymes by selected Trichoderma reesei mutants

    OpenAIRE

    Piotr Janas; Zdzisław Targoński

    2014-01-01

    The effect of temperature in the rangę of 26-38°C on the production of cellulases, xylanases and lytic enzymes by four mutant strains of Trichoderma reesei was analysed. On the basis of these investigations three thermosensitive strains (M-7. RUT C 30 and VTT-D-78085) which showed reduced excretion of the above mentioned enzymes as well as protein and a thermoresistant mutant (VTT-D-79I24) which grew within a temperature range of 26-34°C were characterized. Higher temperature caused an increa...

  19. A kinetic study of Trichoderma reesei Cel7B catalyzed cellulose hydrolysis.

    Science.gov (United States)

    Song, Xiangfei; Zhang, Shujun; Wang, Yefei; Li, Jingwen; He, Chunyan; Yao, Lishan

    2016-06-01

    One prominent feature of Trichoderma reesei (Tr) endoglucanases catalyzed cellulose hydrolysis is that the reaction slows down quickly after it starts (within minutes). But the mechanism of the slowdown is not well understood. A structural model of Tr- Cel7B catalytic domain bound to cellulose was built computationally and the potentially important binding residues were identified and tested experimentally. The 13 tested mutants show different binding properties in the adsorption to phosphoric acid swollen cellulose and filter paper. Though the partitioning parameter to filter paper is about 10 times smaller than that to phosphoric acid swollen cellulose, a positive correlation is shown for two substrates. The kinetic studies show that the reactions slow down quickly for both substrates. This slowdown is not correlated to the binding constant but anticorrelated to the enzyme initial activity. The amount of reducing sugars released after 24h by Cel7B in phosphoric acid swollen cellulose, Avicel and filter paper cellulose hydrolysis is correlated with the enzyme activity against a soluble substrate p-nitrophenyl lactoside. Six of the 13 tested mutants, including N47A, N52D, S99A, N323D, S324A, and S346A, yield ∼15-35% more reducing sugars than the wild type (WT) Cel7B in phosphoric acid swollen cellulose and filter paper hydrolysis. This study reveals that the slowdown of the reaction is not due to the binding of the enzyme to cellulose. The activity of Tr- Cel7B against the insoluble substrate cellulose is determined by the enzyme's capability in hydrolyzing the soluble substrate. PMID:27178789

  20. Comparison of catalytic properties of multiple β-glucosidases of Trichoderma reesei.

    Science.gov (United States)

    Guo, Boyang; Sato, Nobuaki; Biely, Peter; Amano, Yoshihiko; Nozaki, Kouichi

    2016-06-01

    Ten putative Trichoderma reesei β-glucosidase (BGL) isozymes were heterologously expressed in Escherichia coli and Aspergillus oryzae and purified to homogeneity. Catalytic properties of nine enzymes which showed hydrolytic activity on cellobiose and p-nitrophenyl-β-D-glucopyranoside (pNPG) were investigated. Three BGLs, encoded by the genes cel3A, cel3B, and cel3E, contained a predicted signal peptide, showed higher hydrolytic activity on cello-oligosaccharides than on pNPG, and preferred longer oligosaccharides. Another three putative extracellular BGLs, Cel3B, Cel3F, and Cel3G, and two intracellular enzymes, Cel3C and Cel3D, exhibited preference for pNPG. Intracellular Cel1A showed the highest affinity for cellobiose as a typical cellobiase. Four BGLs, Cel3A, Cel3B, Cel3E, Cel1A, that showed high activity against cello-oligosaccharides were capable of catalyzing transglycosylation reactions from cellobiose, leading to formation of cellotriose and isomeric glucobioses. While Cel3A, Cel3B, and Cel3E synthesized mainly gentiobiose, glycosyl transfer reactions of Cel1A led mainly to sophorose and laminaribiose. Conversion of cellobiose to sophorose by Cel1A reached about 3.6 and 10 % at 1 and 10 % cellobiose concentration, respectively. The formation and persistence of individual cellobiose isomers in incubation mixtures of four BGLs (Cel3A, Cel3B, Cel3E, and Cel1A) with cellobiose correlated well with the k cat values for isomeric glucobioses. Cel1A also showed the lowest sensitivity to inhibition by glucose. Based on all studied catalytic properties, Cel1A appears to be unambiguously the best candidate for site-directed mutations or directed evolution toward improvement of activity, thermostability, and, eventually, efficiency of sophorose synthesis. PMID:26846743

  1. The role of subsite +2 of the Trichoderma reesei beta-mannanase TrMan5A in hydrolysis and transglycosylation

    DEFF Research Database (Denmark)

    Rosengren, Anna; Hägglund, Per; Anderson, Lars Steen;

    2012-01-01

    The N-terminal catalytic module of beta-mannanase TrMan5A from the filamentous fungus Trichoderma reesei is classified into family 5 of glycoside hydrolases. It is further classified in clan A with a (beta/alpha)(8) barrel configuration and has two catalytic glutamates (E169 and E276). It has...

  2. Novel Cellulase Profile of Trichoderma reesei Strains Constructed by cbh1 Gene Replacement with eg3 Gene Expression Cassette

    Institute of Scientific and Technical Information of China (English)

    Tian-Hong WANG; Ti LIU; Zhi-Hong WU; Shi-Li LIU; Yi LU; Yin-Bo QU

    2004-01-01

    To construct strains of the filamentous fungus Trichoderma reesei with low cellobiohydrolases while high endoglucanase activity, the Pcbh1-eg3-Tcbh1 cassette was constructed and the coding sequence of the cellobiohydrolase I (CBHI) gene was replaced with the coding sequence of the eg3 gene by homologous recombination. Disruption of the cbh1 gene was confirmed by PCR, Southern dot blot and Western hybridization analysis in two transforments denoted as L 13 and L29. The filter paper-hydrolyzing activity of strain L29 was 60% of the parent strain Rut C30, and the CMCase activity was increased by 33%. This relatively modest increase suggested that the eg3 cDNA under the control of the cbh1 promoter was not efficiently transcribed as the wild type cbhl gene. However our results confirmed that homologous recombination could be used to construct strains of the filamentous fungus Trichoderma reesei with novel cellulase profile. Such strains are of interest from the basic science perspective and also have potential industrial applications.

  3. Heterologous expression, purification, crystallization and preliminary X-ray analysis of Trichoderma reesei xylanase II and four variants

    International Nuclear Information System (INIS)

    The wild-type protein and four active-site mutants of xylanase II from Trichoderma reesei that catalyzes the hydrolysis of glycosidic bonds in xylan have successfully been crystallized. The crystallization of several structures including ligand-free and protein ligand complexes containing the substrate (xylohexaose) or product (xylotriose) are detailed. Xylanase II from Trichoderma reesei catalyzes the hydrolysis of glycosidic bonds in xylan. Crystallographic studies of this commercially important enzyme have been initiated to investigate its reaction mechanism, substrate binding and dependence on basic pH conditions. The wild-type protein was heterologously expressed in an Escherichia coli host using the defined medium and four active-site amino acids were replaced to abolish its activity (E177Q and E86Q) or to change its pH optimum (N44D and N44H). Cation-exchange and size-exclusion chromatography were used to obtain >90% protein purity. The ligand-free proteins and variant complexes containing substrate (xylohexaose) or product (xylotriose) were crystallized in several different space groups and diffracted to high resolutions (from 1.07 to 1.55 Å)

  4. Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei for Effective Wastewater Treatment and Bio-Oil Production

    Directory of Open Access Journals (Sweden)

    Anshuman Bhanja

    2014-01-01

    Full Text Available Biological wastewater treatment typically requires the use of bacteria for degradation of carbonaceous and nitrogenous compounds present in wastewater. The high lipid containing biomass can be used to extract oil and the contents can be termed as bio-oil (or biodiesel or myco-diesel after transesterification. The separate experiments were conducted on actual wastewater samples with 5% v/v inoculum of Mucor circinelloides MTCC1297 and Trichoderma reesei NCIM992 strains. The observed reductions in chemical oxygen demand (COD were 88.72% and 86.75% in 96 hrs and the observed substrate based biomass yields were 0.21 mg VSS/mg COD and 0.22 mg VSS/mg COD for M. circinelloides reactor and for T. reesei reactor, respectively. The resulted bio-oil production from wastewater treatment by M. circinelloides and T. reesei reactors was 142.2 mg/L and 74.1 mg/L, whereas biomass containing bio-oil contents (%w/w were 22.11% and 9.82%, respectively. In this experiment, the fungal wastewater treatment was also compared with conventional bacterial process with respect to specific growth rate, biomass production, and oil content. This study suggests that wastewater can be used as a potential feedstock for bio-oil production with the use of oleaginous fungal strains and which could be a possible route of waste to energy.

  5. PEA PEEL WASTE: A LIGNOCELLULOSIC WASTE AND ITS UTILITY IN CELLULASE PRODUCTION BY Trichoderma reesei UNDER SOLID STATE CULTIVATION

    Directory of Open Access Journals (Sweden)

    Nitin Verma

    2011-03-01

    Full Text Available A wide variety of waste bioresources are available on our planet for conversion into bioproducts. In the biological systems, microorganisms are used to utilize waste as an energy source for the synthesis of valuable products such as biomass proteins and enzymes. The large quantities of byproducts generated during the processing of plant food involve an economic and environmental problem due to their high volumes and elimination costs. After isolation of the main constituent, there are abundant remains which represent an inexpensive material that has been undervalued until now. Pea peel waste is one of the undervalued, unused sources of energy that can serve as a potential source for cellulase production. Batch experiments have been performed, using pea peel waste as a carbon source for cellulase production under solid state cultivation by Trichoderma reesei. It was observed that 30 oC temperature and pH 5.0 are the most favorable conditions for cellulase production by T. reesei. FPase activity significantly increases by incorporation of whey as well as wheat starch hydrolysate in the basal salt media used in the production study. The present study describes the utility of pea peel waste, whey as well as wheat starch hydrolysate in cellulase production by T. reesei. The utilization of economically cheap, pea peel waste for cellulase production could be a novel, cost effective, and valuable approach in cellulase production as well as in solid waste management.

  6. Identification of the Main Regulator Responsible for Synthesis of the Typical Yellow Pigment Produced by Trichoderma reesei

    Science.gov (United States)

    Derntl, Christian; Rassinger, Alice; Srebotnik, Ewald; Mach, Robert L.

    2016-01-01

    ABSTRACT The industrially used ascomycete Trichoderma reesei secretes a typical yellow pigment during cultivation, while other Trichoderma species do not. A comparative genomic analysis suggested that a putative secondary metabolism cluster, containing two polyketide-synthase encoding genes, is responsible for the yellow pigment synthesis. This cluster is conserved in a set of rather distantly related fungi, including Acremonium chrysogenum and Penicillium chrysogenum. In an attempt to silence the cluster in T. reesei, two genes of the cluster encoding transcription factors were individually deleted. For a complete genetic proof-of-function, the genes were reinserted into the genomes of the respective deletion strains. The deletion of the first transcription factor (termed yellow pigment regulator 1 [Ypr1]) resulted in the full abolishment of the yellow pigment formation and the expression of most genes of this cluster. A comparative high-pressure liquid chromatography (HPLC) analysis of supernatants of the ypr1 deletion and its parent strain suggested the presence of several yellow compounds in T. reesei that are all derived from the same cluster. A subsequent gas chromatography/mass spectrometry analysis strongly indicated the presence of sorbicillin in the major HPLC peak. The presence of the second transcription factor, termed yellow pigment regulator 2 (Ypr2), reduces the yellow pigment formation and the expression of most cluster genes, including the gene encoding the activator Ypr1. IMPORTANCE Trichoderma reesei is used for industry-scale production of carbohydrate-active enzymes. During growth, it secretes a typical yellow pigment. This is not favorable for industrial enzyme production because it makes the downstream process more complicated and thus increases operating costs. In this study, we demonstrate which regulators influence the synthesis of the yellow pigment. Based on these data, we also provide indication as to which genes are under the control

  7. Trichoderma reesei CRE1-mediated Carbon Catabolite Repression in Re-sponse to Sophorose Through RNA Sequencing Analysis.

    Science.gov (United States)

    Antoniêto, Amanda Cristina Campos; de Paula, Renato Graciano; Castro, Lílian Dos Santos; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2016-04-01

    Carbon catabolite repression (CCR) mediated by CRE1 in Trichoderma reesei emerged as a mechanism by which the fungus could adapt to new environments. In the presence of readily available carbon sources such as glucose, the fungus activates this mechanism and inhibits the production of cellulolytic complex enzymes to avoid unnecessary energy expenditure. CCR has been well described for the growth of T. reesei in cellulose and glucose, however, little is known about this process when the carbon source is sophorose, one of the most potent inducers of cellulase production. Thus, we performed high-throughput RNA sequencing to better understand CCR during cellulase formation in the presence of sophorose, by comparing the mutant ∆cre1 with its parental strain, QM9414. Of the 9129 genes present in the genome of T. reesei, 184 were upregulated and 344 downregulated in the mutant strain ∆cre1 compared to QM9414. Genes belonging to the CAZy database, and those encoding transcription factors and transporters are among the gene classes that were repressed by CRE1 in the presence of sophorose; most were possible indirectly regulated by CRE1. We also observed that CRE1 activity is carbon-dependent. A recent study from our group showed that in cellulose, CRE1 repress different groups of genes when compared to sophorose. CCR differences between these carbon sources may be due to the release of cellodextrins in the cellulose polymer, resulting in different targets of CRE1 in both carbon sources. These results contribute to a better understanding of CRE1-mediated CCR in T. reesei when glucose comes from a potent inducer of cellulase production such as sophorose, which could prove useful in improving cellulase production by the biotechnology sector. PMID:27226768

  8. Overexpression of D-Xylose Reductase (xyl1) Gene and Antisense Inhibition of D-Xylulokinase (xyiH) Gene Increase Xylitol Production in Trichoderma reesei

    Science.gov (United States)

    Hong, Yuanyuan; Dashtban, Mehdi; Kepka, Greg; Chen, Sanfeng; Qin, Wensheng

    2014-01-01

    T. reesei is an efficient cellulase producer and biomass degrader. To improve xylitol production in Trichoderma reesei strains by genetic engineering, two approaches were used in this study. First, the presumptive D-xylulokinase gene in T. reesei (xyiH), which has high homology to known fungi D-xylulokinase genes, was silenced by transformation of T. reesei QM9414 strain with an antisense construct to create strain S6-2-2. The expression of the xyiH gene in the transformed strain S6-2-2 decreased at the mRNA level, and D-xylulokinase activity decreased after 48 h of incubation. This led to an increase in xylitol production from undetectable levels in wild-type T. reesei QM9414 to 8.6 mM in S6-2-2. The T. reesei Δxdh is a xylose dehydrogenase knockout strain with increased xylitol production compared to the wild-type T. reesei QM9414 (22.8 mM versus undetectable). The copy number of the xylose reductase gene (xyl1) in T. reesei Δxdh strain was increased by genetic engineering to create a new strain Δ9-5-1. The Δ9-5-1 strain showed a higher xyl1 expression and a higher yield of xylose reductase, and xylitol production was increased from 22.8 mM to 24.8 mM. Two novel strains S6-2-2 and Δ9-5-1 are capable of producing higher yields of xylitol. T. reesei has great potential in the industrial production of xylitol. PMID:25013760

  9. Overexpression of D-Xylose Reductase (xyl1 Gene and Antisense Inhibition of D-Xylulokinase (xyiH Gene Increase Xylitol Production in Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    Yuanyuan Hong

    2014-01-01

    Full Text Available T. reesei is an efficient cellulase producer and biomass degrader. To improve xylitol production in Trichoderma reesei strains by genetic engineering, two approaches were used in this study. First, the presumptive D-xylulokinase gene in T. reesei (xyiH, which has high homology to known fungi D-xylulokinase genes, was silenced by transformation of T. reesei QM9414 strain with an antisense construct to create strain S6-2-2. The expression of the xyiH gene in the transformed strain S6-2-2 decreased at the mRNA level, and D-xylulokinase activity decreased after 48 h of incubation. This led to an increase in xylitol production from undetectable levels in wild-type T. reesei QM9414 to 8.6 mM in S6-2-2. The T. reesei Δxdh is a xylose dehydrogenase knockout strain with increased xylitol production compared to the wild-type T. reesei QM9414 (22.8 mM versus undetectable. The copy number of the xylose reductase gene (xyl1 in T. reesei Δxdh strain was increased by genetic engineering to create a new strain Δ9-5-1. The Δ9-5-1 strain showed a higher xyl1 expression and a higher yield of xylose reductase, and xylitol production was increased from 22.8 mM to 24.8 mM. Two novel strains S6-2-2 and Δ9-5-1 are capable of producing higher yields of xylitol. T. reesei has great potential in the industrial production of xylitol.

  10. The phosducin-like protein PhLP1 impacts regulation of glycoside hydrolases and light response in Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    Tisch Doris

    2011-12-01

    Full Text Available Abstract Background In the biotechnological workhorse Trichoderma reesei (Hypocrea jecorina transcription of cellulase genes as well as efficiency of the secreted cellulase mixture are modulated by light. Components of the heterotrimeric G-protein pathway interact with light-dependent signals, rendering this pathway a key regulator of cellulase gene expression. Results As regulators of heterotrimeric G-protein signaling, class I phosducin-like proteins, are assumed to act as co-chaperones for G-protein beta-gamma folding and exert their function in response to light in higher eukaryotes. Our results revealed light responsive transcription of the T. reesei class I phosducin-like protein gene phlp1 and indicate a light dependent function of PhLP1 also in fungi. We showed the functions of PhLP1, GNB1 and GNG1 in the same pathway, with one major output being the regulation of transcription of glycoside hydrolase genes including cellulase genes in T. reesei. We found no direct correlation between the growth rate and global regulation of glycoside hydrolases, which suggests that regulation of growth does not occur only at the level of substrate degradation efficiency. Additionally, PhLP1, GNB1 and GNG1 are all important for proper regulation of light responsiveness during long term exposure. In their absence, the amount of light regulated genes increased from 2.7% in wild type to 14% in Δphlp1. Besides from the regulation of degradative enzymes, PhLP1 was also found to impact on the transcription of genes involved in sexual development, which was in accordance with decreased efficiency of fruiting body formation in Δphlp1. The lack of GNB1 drastically diminished ascospore discharge in T. reesei. Conclusions The heterotrimeric G-protein pathway is crucial for the interconnection of nutrient signaling and light response of T. reesei, with the class I phosducin-like protein PhLP1, GNB1 and GNG1 acting as important nodes, which influence light

  11. Direct Conversion of Pretreated Straw Cellulose into Citric Acid by Co-cultures of Yarrowia lipolytica SWJ-1b and Immobilized Trichoderma reesei Mycelium

    OpenAIRE

    Liu, Xiaoyan; Lv, Jinshun; Zhang, Tong; Deng, Yuanfang

    2014-01-01

    The immobilized cellulase-producing mycelium of Trichoderma reesei was found to produce 2.9 U/ml of cellulase activity within 144 h while 2.1 U/ml of cellulase activity was produced within 120 h by the free mycelium of the same strain. When the immobilized mycelium of T. reesei was co-cultivated with the free cells of Yarrowia lipolytica SWJ-1b in flask, Y. lipolytica SWJ-1b could yield 10.7 g/l of citric acid and 3.9 g/l of isocitric acid from 40.0 g/l pretreated straw within 240 h. Under th...

  12. Transcriptional monitoring of steady state and effects of anaerobic phases in chemostat cultures of the filamentous fungus Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    Penttilä Merja

    2006-10-01

    Full Text Available Abstract Background Chemostat cultures are commonly used in production of cellular material for systems-wide biological studies. We have used the novel TRAC (transcript analysis with aid of affinity capture method to study expression stability of approximately 30 process relevant marker genes in chemostat cultures of the filamentous fungus Trichoderma reesei and its transformant expressing laccase from Melanocarpus albomyces. Transcriptional responses caused by transient oxygen deprivations and production of foreign protein were also studied in T. reesei by TRAC. Results In cultures with good steady states, the expression of the marker genes varied less than 20% on average between sequential samples for at least 5 or 6 residence times. However, in a number of T. reesei cultures continuous flow did not result in a good steady state. Perturbations to the steady state were always evident at the transcriptional level, even when they were not measurable as changes in biomass or product concentrations. Both unintentional and intentional perturbations of the steady state demonstrated that a number of genes involved in growth, protein production and secretion are sensitive markers for culture disturbances. Exposure to anaerobic conditions caused strong responses at the level of gene expression, but surprisingly the cultures could regain their previous steady state quickly, even after 3 h O2 depletion. The main effect of producing M. albomyces laccase was down-regulation of the native cellulases compared with the host strain. Conclusion This study demonstrates the usefulness of transcriptional analysis by TRAC in ensuring the quality of chemostat cultures prior to costly and laborious genome-wide analysis. In addition TRAC was shown to be an efficient tool in studying gene expression dynamics in transient conditions.

  13. Enzyme production by filamentous fungi: analysis of the secretome of Trichoderma reesei grown on unconventional carbon source

    Directory of Open Access Journals (Sweden)

    Kieselbach Thomas

    2011-08-01

    Full Text Available Abstract Background Spent hydrolysates from bioethanolic fermentation processes based on agricultural residues have potential as an abundant and inexpensive source of pentose sugars and acids that could serve as nutrients for industrial enzyme-producing microorganisms, especially filamentous fungi. However, the enzyme mixtures produced in such media are poorly defined. In this study, the secretome of Trichoderma reesei Rut C-30 grown either on a spent hydrolysate model medium (SHMM or on a lactose-based standard medium (LBSM was explored using proteomics. Results Our results show that both the SHMM and LBSM serve as excellent growth media for T. reesei Rut C-30. In total, 52 protein spots on 2-D gels were identified by using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS and electrospray ionization liquid chromatography tandem mass spectrometry (ESI-LC MS/MS. As expected, a considerable number of the identified proteins were related to the degradation of lignocellulosic biomass. The enzyme production profiles in the two media were similar, but β-glucosidase and β-galactosidase were only produced in LBSM. The main cellobiohydrolases (Cel7A/Cel6A and endoglucanases (Cel7B/Cel5A were identified in both media and the cellobiohydrolases, i.e. Cel7A and Cel6A, were the most abundant cellulolytic enzymes. Moreover, both media can also serve as a potent inducer of xylanolytic enzymes. Several key enzymes involved in sugar assimilation and regulation of cellulase formation were identified, and were found to be differentially expressed in the two growth media. Conclusions This study not only provides a catalogue of the prevalent proteins secreted by T. reesei in the two media, but the results also suggest that production of hydrolytic enzymes using unconventional carbon sources, such as components in spent hydrolysates, deserves further attention in the future.

  14. Trpac1, a pH response transcription regulator, is involved in cellulase gene expression in Trichoderma reesei.

    Science.gov (United States)

    He, Ronglin; Ma, Lijuan; Li, Chen; Jia, Wendi; Li, Demao; Zhang, Dongyuan; Chen, Shulin

    2014-12-01

    Fungi grow over a relatively wide pH range and adapt to extracellular pH through a genetic regulatory system mediated by a key component PacC, which is a pH transcription regulator. The cellulase production of the filamentous fungi Trichoderma reesei is sensitive to ambient pH. To investigate the connection between cellulase expression regulation and ambient pH, an ortholog of Aspergillus nidulans pacC, Trpac1, was identified and functionally characterized using a target gene deletion strategy. Deleting Trpac1 dramatically increased the cellulase production and the transcription levels of the major cellulase genes at neutral pH, which suggested Trpac1 is involved in the regulation of cellulase production. It was further observed that the expression levels of transcription factors xyr1 and ace2 also increased in the ΔTrpac1 mutant at neutral pH. In addition, the ΔTrpac1 mutant exhibited conidiation defects under neutral and alkaline pH. These results implied that Trpac1 in involved in growth and development process and cellulase gene expression in T. reesei.

  15. Proteome scale census of major facilitator superfamily transporters in Trichoderma reesei using protein sequence and structure based classification enhanced ranking.

    Science.gov (United States)

    Chaudhary, Nitika; Kumari, Indu; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

    2016-07-01

    Trichoderma spp. have been acknowledged as potent bio-control agents against microbial pathogens and also as plant growth promoters. Various secondary metabolites are attributed for these beneficial activities. Major facilitator superfamily (MFS) includes the large proportion of efflux-pumps which are linked with membrane transport of these secondary metabolites. We have carried out a proteome-wide identification of MFS transporters using protein sequence and structure based hierarchical method in Trichoderma reesei. 448 proteins out of 9115 were detected to carry transmembrane helices. MFS specific intragenic gene duplication and its context with transport function have been presented. Finally, using homology based techniques, domains and motifs of MFS families have been identified and utilized to classify them. From query dataset of 448 transmembrane proteins, 148 proteins are identified as potential MFS transporters. Sugar porter, drug: H(+) antiporter-1, monocarboxylate porter and anion: cation symporter emerged as major MFS families with 51, 35, 17 and 11 members respectively. Representative protein tertiary structures of these families are homology modeled for structure-function analysis. This study may help to understand the molecular basis of secretion and transport of agriculturally valuable secondary metabolites produced by these bio-control fungal agents which may be exploited in future for enhancing its biotechnological applications in eco-friendly sustainable development.

  16. Proteome scale census of major facilitator superfamily transporters in Trichoderma reesei using protein sequence and structure based classification enhanced ranking.

    Science.gov (United States)

    Chaudhary, Nitika; Kumari, Indu; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

    2016-07-01

    Trichoderma spp. have been acknowledged as potent bio-control agents against microbial pathogens and also as plant growth promoters. Various secondary metabolites are attributed for these beneficial activities. Major facilitator superfamily (MFS) includes the large proportion of efflux-pumps which are linked with membrane transport of these secondary metabolites. We have carried out a proteome-wide identification of MFS transporters using protein sequence and structure based hierarchical method in Trichoderma reesei. 448 proteins out of 9115 were detected to carry transmembrane helices. MFS specific intragenic gene duplication and its context with transport function have been presented. Finally, using homology based techniques, domains and motifs of MFS families have been identified and utilized to classify them. From query dataset of 448 transmembrane proteins, 148 proteins are identified as potential MFS transporters. Sugar porter, drug: H(+) antiporter-1, monocarboxylate porter and anion: cation symporter emerged as major MFS families with 51, 35, 17 and 11 members respectively. Representative protein tertiary structures of these families are homology modeled for structure-function analysis. This study may help to understand the molecular basis of secretion and transport of agriculturally valuable secondary metabolites produced by these bio-control fungal agents which may be exploited in future for enhancing its biotechnological applications in eco-friendly sustainable development. PMID:27041239

  17. The Cellulases Endoglucanase I and Cellobiohydrolase II of Trichoderma reesei Act Synergistically To Solubilize Native Cotton Cellulose but Not To Decrease Its Molecular Size

    OpenAIRE

    Kleman-Leyer, K. M.; Siika-Aho, M.; Teeri, T. T.; Kirk, T K

    1996-01-01

    Degradation of cotton cellulose by Trichoderma reesei endoglucanase I (EGI) and cellobiohydrolase II (CBHII) was investigated by analyzing the insoluble cellulose fragments remaining after enzymatic hydrolysis. Changes in the molecular-size distribution of cellulose after attack by EGI, alone and in combination with CBHII, were determined by size exclusion chromatography of the tricarbanilate derivatives. Cotton cellulose incubated with EGI exhibited a single major peak, which with time shift...

  18. Continuous Degradation of Chitosan in a Convoluted Fibrous Bed Bioreactor with Immobilized Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    WUMianbin; XIALiming; 等

    2002-01-01

    Continuous hydrolysis of chitosan was performed in a convoluted fibrous bed bioreactor (CFBB) with immobilized T. reesei. At dilution rate of 0.4d-1 and substrate concentration of 2% (mass vs. volume), the average degree of polymerization of hydrolysate can be kept at 1.25-1.35, which can be easily regulated by changing dilution rate or inlet chitosan concentration.

  19. KINETIKA FERMENTASI SELULOSA MURNI OLEH Trichoderma reesi QM 9414 MENJADI GLUKOSA DAN PENERAPANNYA PADA JERAMI PADI BEBAS LIGNIN [Kinetics of Pure Cellulose Fermentation by Trichoderma Reesei QM 9414 to Glucose and Its Application of on Lignin Free Rice Straw

    Directory of Open Access Journals (Sweden)

    M Iyan Sofyan

    2004-12-01

    Full Text Available The objectives of this research were: 1 to determine aeration rate and substrate concentration of pure cellulose to produce maximum glucose by Trichoderma reesei QM 9414 at 30 oC, and agitation 150 rpm; 2 to study the kinetics of pure cellulose fermentation by Trichoderma reesei QM 9414 to glucose and its implication upon fermentation of the lignin free rice straw. The experiment was arranged in factorial randomized complete design in three times replication. Treatments consisted of three levels of aeration (1,00 vvm; 1,5 vvm; 2,0 vvm and three levels of substrate concentration (0,75 ; 1,00 ; 1,25 % w/v. The results showed that at the exponential phase the average specific growth of Trichoderma reesei QM 9414 was 0,05374 hour-1, the maximum glucose product concentration of pure cellulose was 0.1644 gL-1,and the oxygen transfer was 0,0328 mg L-1 hour-1. According to t-test, the kinetics of pure cellulose fermentation model just the same as the lignin free rice straw fermentation.The enzymes produced by Trichoderma reesei QM 9414 in pure cellulose fermentation media followed the Michaelis-Menten model. The enzyme kinetic parameters were the maximum growth rate was 37x10-3 hour-1 and Michaelis-Menten constant was ½ maximum μ =17,5x10-3 hour-1. The volumetric oxygen transfer (KLa using rice straw was 0,0337 mg.hour-1. The value of KLa could be used for conversion from bioreactor at laboratory scale to commercial scale design.

  20. Optimization of a synthetic mixture composed of major Trichoderma reesei enzymes for the hydrolysis of steam-exploded wheat straw

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    Billard Hélène

    2012-02-01

    Full Text Available Abstract Background An efficient hydrolysis of lignocellulosic substrates to soluble sugars for biofuel production necessitates the interplay and synergistic interaction of multiple enzymes. An optimized enzyme mixture is crucial for reduced cost of the enzymatic hydrolysis step in a bioethanol production process and its composition will depend on the substrate and type of pretreatment used. In the present study, an experimental design was used to determine the optimal composition of a Trichoderma reesei enzyme mixture, comprising the main cellulase and hemicellulase activities, for the hydrolysis of steam-exploded wheat straw. Methods Six enzymes, CBH1 (Cel7a, CBH2 (Cel6a, EG1 (Cel7b, EG2 (Cel5a, as well as the xyloglucanase Cel74a and the xylanase XYN1 (Xyl11a were purified from a T. reesei culture under lactose/xylose-induced conditions. Sugar release was followed in milliliter-scale hydrolysis assays for 48 hours and the influence of the mixture on initial conversion rates and final yields is assessed. Results The developed model could show that both responses were strongly correlated. Model predictions suggest that optimal hydrolysis yields can be obtained over a wide range of CBH1 to CBH2 ratios, but necessitates a high proportion of EG1 (13% to 25% which cannot be replaced by EG2. Whereas 5% to 10% of the latter enzyme and a xylanase content above 6% are required for highest yields, these enzymes are predicted to be less important in the initial stage of hydrolysis. Conclusions The developed model could reliably predict hydrolysis yields of enzyme mixtures in the studied domain and highlighted the importance of the respective enzyme components in both the initial and the final hydrolysis phase of steam-exploded wheat straw.

  1. Improvement and scale-down of a Trichoderma reesei shake flask protocol to microtiter plates enables high-throughput screening.

    Science.gov (United States)

    Giese, Heiner; Kruithof, Paulien; Meier, Kristina; Sieben, Michaela; Antonov, Elena; Hommes, Ronald W J; Büchs, Jochen

    2014-12-01

    Nowadays, high-throughput screening is essential for determining the best microbial strains and fermentation conditions. Although microtiter plates allow higher throughput in screening than shake flasks, they do not guarantee sufficient oxygen supply if operated at unsuitable conditions. This is especially the case in viscous fermentations, potentially leading to poor liquid movement and surface growth. Therefore, in this study, two aims were pursued. First, an industrial Trichoderma reesei shake flask protocol is improved with respect to oxygen supply and production. Second, this improved shake flask protocol is scaled down into microtiter plate under consideration of similar oxygen supply. For this purpose, the respiration activity monitoring system (RAMOS) was applied. An approach based on a sulfite system was introduced to ensure equal maximum oxygen transfer capacities (OTRmax) in microtiter plates and shake flasks. OTRmax-values of 250 mL shake flasks and 24-well microtiter plates were determined in a wide range of operating conditions. These sulfite datasets were used to identify operating conditions leading to the same oxygen supply for T. reesei in shake flasks and 24-well microtiter plates. For 24-well microtiter plates, the shake flask OTRmax of 20 mmol/L/h of an industrial protocol was obtained under the following optimal operating conditions: 1 mL filling volume per well, 200 rpm shaking frequency and 50 mm shaking diameter. With these conditions almost identical oxygen transfer rates and product concentrations were measured in both scales. The proposed approach is a fast and accurate means to scale-down established screening procedures into microtiter plates to achieve high-throughput.

  2. COMPARISON OF THE EXPRESSION IN Saccharomyces cerevisiae OF ENDOGLUCANASE II FROM Trichoderma reesei AND ENDOGLUCANASE I FROM Aspergillus aculeatus

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    Weina Zhang,

    2012-07-01

    Full Text Available Two distinct expression cassettes were synthesized by overlapping PCR for expressing the endoglucanase I gene (egl1 from Aspergillus aculeatus and the endoglucanase II gene (egl2 from Trichoderma reesei in a Saccharomyces cerevisiae host. One contained the anchored sequence from the S. cerevisiae cwp2 gene, while the other did not. The low and high copy number plasmids YCplac33 and YEplac195 were used. The enzymatic activities and viscosity changes in the YP-CMC medium varied between the eight recombinant yeast strains produced, and the greatest values were obtained with the YE-TrEII’ strain, which had an activity of 347.7 U/g dry cell weight (DCW and viscosity at 12 h of 4.7% of the initial control value, respectively; YE-TrEII’ was YEplac195-based and contained T. reesei egl2 and no Cwp2 sequence. Strains YC-AaEI and YC-TrEII showed the lowest enzyme activitiy (80.5 and 30.4 U/g DCW, respectively and viscosity changes at 12 h (20.5 and 26.2% of the initial control viscosity, respectively, which were YCplac33-based and contained the Cwp2 sequence. The results showed that gene copy number was the most significant factor to influence the expression of endoglucanases in S. cerevisiae, and the existence of Cwp2 sequence led to decreased enzymatic level and viscosity-reducing performance, while it was shown not to realize efficient surface display of these two endoglucanases.Keywords

  3. Recombinant Expression of Trichoderma reesei Cel61A in Pichia pastoris: Optimizing Yield and N-terminal Processing.

    Science.gov (United States)

    Tanghe, Magali; Danneels, Barbara; Camattari, Andrea; Glieder, Anton; Vandenberghe, Isabel; Devreese, Bart; Stals, Ingeborg; Desmet, Tom

    2015-12-01

    The auxiliary activity family 9 (AA9, formerly GH61) harbors a recently discovered group of oxidative enzymes that boost cellulose degradation. Indeed, these lytic polysaccharide monooxygenases (LPMOs) are able to disrupt the crystalline structure of cellulose, thereby facilitating the work of hydrolytic enzymes involved in biomass degradation. Since these enzymes require an N-terminal histidine residue for activity, their recombinant production as secreted protein is not straightforward. We here report the expression optimization of Trichoderma reesei Cel61A (TrCel61A) in the host Pichia pastoris. The use of the native TrCel61A secretion signal instead of the alpha-mating factor from Saccharomyces cerevisiae was found to be crucial, not only to obtain high protein yields (>400 mg/L during fermentation) but also to enable the correct processing of the N-terminus. Furthermore, the LPMO activity of the enzyme is demonstrated here for the first time, based on its degradation profile of a cellulosic substrate.

  4. Effect of Temperature on Xylanase II from Trichoderma reesei QM 9414: A Calorimetric, Catalytic, and Conformational Study

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    Gloria López

    2014-01-01

    Full Text Available The secondary structure of xylanase II from Trichoderma reesei is lost in an apparent irreversible cooperative process as temperature is increased with a midpoint transition of 58.8 ± 0.1°C. The shift of the spectral centre of mass above 50°C is also apparently cooperative with midpoint transition of 56.3 ± 0.2°C, but the existence of two isofluorescent points in the fluorescence emission spectra suggests a non-two-state process. Further corroboration comes from differential scanning calorimetry experiments. At protein concentrations ≤0.56 mg·mL−1 the calorimetric transition is reversible and the data were fitted to a non-two-state model and deconvoluted into six transitions, whereas at concentrations greater than 0.56 mg·mL−1 the calorimetric transition is irreversible with an exothermic contribution to the thermogram. The apparent Tm increased linearly with the scan rate according to first order inactivation kinetics. The effect of additives on the calorimetric transition of xylanase is dependent on their nature. The addition of sorbitol transforms reversible transitions into irreversible transitions while stabilizing the protein as the apparent Tm increases linearly with sorbitol concentration. d-Glucono-1,5-lactone, a noncompetitive inhibitor in xylanase kinetics, and soluble xylan change irreversible processes into reversible processes at high protein concentration.

  5. Isolation of Trichoderma reesei pyrG Negative Mutant by UV Mutagenesis and Its Application in Transformation

    Institute of Scientific and Technical Information of China (English)

    LONG Hao; WANG Tian-hong; ZHANG Ying-kuan

    2008-01-01

    Two uridine auxotrophic mutants of Trichoderma reesei were isolated by resistance to 5-fluoroorotic acid after UV mutagenesis.One mutant,called M23,was complemented with the Aspergillus niger pyrG gene carded by plasmid pAB4-1.A mutated pyrG gene of M23 was cloned and DNA sequencing analysis indicated that a cytosine was inserted into the 934-939 oligo dC position of the pyrG coding region,resulted in a frameshift mutation.Transformation efficiency was approximately 200-300 transformants per microgram of DNA with plasmid pAB4-1.Stable transformants were obtained by monosporic culture and showed to be prototroph after successive propagation.Vitreoscilla hemoglobin expression plasmid pUCVHb was cotransformed with plasmid pAB4-1 and attained a transformation efficiency of 71.8% or of 26.1% with pAN7-1.Southern blot analysis of the transformants demonstrated that plasmid pUCVHb was integrated into the chromosomal DNA.The experimental results demonstrated that the pyrG-based system was more efficient and timesaving than the conventional hygromycin B resistance-based transformation system.

  6. High-level expression and characterization of a thermostable xylanase mutant from Trichoderma reesei in Pichia pastoris.

    Science.gov (United States)

    Li, Yang-yuan; Zhong, Kai-xin; Hu, Ai-hong; Liu, Dan-ni; Chen, Li-zhi; Xu, Shu-de

    2015-04-01

    A gene encoding xylanase 2 mutant from Trichoderma reesei (T2C/T28C, named mxyn2) was cloned into the Pichia pastoris X33 strain using the vector pPICZαA. Recombinant Mxyn2p was functionally expressed in P. pastoris X33 and secreted into the supernatant. Real time qPCR demonstrated that an increase in gene copy number correlated with higher levels of expression. Supernatant from methanol induced cells was concentrated by ultrafiltration with a 10kDa cut off membrane, and purified with ion exchange chromatography using SP Sepharose Fast Flow chromatography. Recombinant Mxyn2p protein had the highest activity at 75°C, while recombinant protein encoded by the "wild type" xylanase gene xyn2, also expressed in Pichia, was 20°C lower. The Mxyn2p enzyme retained more than 70% of its activity after incubation at 80°C for 10min. The effects of the optimal pH and temperature for higher expression levels in P. pastoris were also determined, 6.0 and 22°C, respectively. The maximum xylanase activity of Mxyn2p was 13,000nkat/mg (9.88g/l) in fed-batch cultivation after 168h induction with methanol in a 50l bioreactor. PMID:25434687

  7. Enabling Low Cost Biopharmaceuticals: A Systematic Approach to Delete Proteases from a Well-Known Protein Production Host Trichoderma reesei

    Science.gov (United States)

    Landowski, Christopher P.; Huuskonen, Anne; Wahl, Ramon; Westerholm-Parvinen, Ann; Kanerva, Anne; Hänninen, Anna-Liisa; Salovuori, Noora; Penttilä, Merja; Natunen, Jari; Ostermeier, Christian; Helk, Bernhard; Saarinen, Juhani; Saloheimo, Markku

    2015-01-01

    The filamentous fungus Trichoderma reesei has tremendous capability to secrete proteins. Therefore, it would be an excellent host for producing high levels of therapeutic proteins at low cost. Developing a filamentous fungus to produce sensitive therapeutic proteins requires that protease secretion is drastically reduced. We have identified 13 major secreted proteases that are related to degradation of therapeutic antibodies, interferon alpha 2b, and insulin like growth factor. The major proteases observed were aspartic, glutamic, subtilisin-like, and trypsin-like proteases. The seven most problematic proteases were sequentially removed from a strain to develop it for producing therapeutic proteins. After this the protease activity in the supernatant was dramatically reduced down to 4% of the original level based upon a casein substrate. When antibody was incubated in the six protease deletion strain supernatant, the heavy chain remained fully intact and no degradation products were observed. Interferon alpha 2b and insulin like growth factor were less stable in the same supernatant, but full length proteins remained when incubated overnight, in contrast to the original strain. As additional benefits, the multiple protease deletions have led to faster strain growth and higher levels of total protein in the culture supernatant. PMID:26309247

  8. pH effects on the structural dynamics of cutinase from Trichoderma reesei: insights from molecular dynamics simulations.

    Science.gov (United States)

    Duan, Mei Lin; Liu, Lin; Du, Juan; Yao, Xiao Jun

    2015-11-01

    Cutinases are utilized in a variety of industries for the hydrolysis of a broad range of substrates, such as cutin, polyesters, soluble esters, insoluble short- and long-chain triglycerides. The novel cutinase from Trichoderma reesei (Tr) attracted much attention due to its two rare characteristics distinct from the classical cutinases: it possesses a lid covering its active site and its optimal activity at acidic pH. However, the structural basis for pH preference and the function of lid is still not well understood. In this work, total of six initial systems were set up either under acidic or basic pH conditions (closed-apo, open-apo and open-holo). Then, molecular dynamics (MD) simulations were performed to make a better understanding of structural dynamics of Tr cutinase under different pH conditions for the first time. The results mainly suggest that it is easier to open for the lid under an acidic pH condition. In addition, the binding of long-chain triglyceride is more stable at lower pH than higher pH. These findings elucidate that how pH influences Tr cutinase at the atomistic level. The structural and dynamic details would be useful for rational enzyme design for acidic cutinase. PMID:26387959

  9. Utilization of recombinant Trichoderma reesei expressing Aspergillus aculeatus β-glucosidase I (JN11) for a more economical production of ethanol from lignocellulosic biomass.

    Science.gov (United States)

    Treebupachatsakul, Treesukon; Shioya, Koki; Nakazawa, Hikaru; Kawaguchi, Takashi; Morikawa, Yasushi; Shida, Yosuke; Ogasawara, Wataru; Okada, Hirofumi

    2015-12-01

    The capacity of Trichoderma reesei cellulase to degrade lignocellulosic biomass has been enhanced by the construction of a recombinant T. reesei strain expressing Aspergillus aculeatus β-glucosidase I. We have confirmed highly efficient ethanol production from converge-milled Japanese cedar by recombinant T. reesei expressing A. aculeatus β-glucosidase I (JN11). We investigated the ethanol productivity of JN11 and compared it with the cocktail enzyme T. reesei PC-3-7 with reinforced cellobiase activity by the commercial Novozyme 188. Results showed that the ethanol production efficiency under enzymatic hydrolysis of JN11 was comparable to the cocktail enzyme both on simultaneous saccharification and fermentation (SSF) or separate hydrolysis and fermentation (SHF) processes. Moreover, the cocktail enzyme required more protein loading for attaining similar levels of ethanol conversion as JN11. We propose that JN11 is an intrinsically economical enzyme that can eliminate the supplementation of BGL for PC-3-7, thereby reducing the cost of industrial ethanol production from lignocellulosic biomass.

  10. Overexpression of erg20 gene encoding farnesyl pyrophosphate synthase has contrasting effects on activity of enzymes of the dolichyl and sterol branches of mevalonate pathway in Trichoderma reesei.

    Science.gov (United States)

    Piłsyk, Sebastian; Perlińska-Lenart, Urszula; Górka-Nieć, Wioletta; Graczyk, Sebastian; Antosiewicz, Beata; Zembek, Patrycja; Palamarczyk, Grażyna; Kruszewska, Joanna S

    2014-07-10

    The mevalonate pathway is the most diverse metabolic route resulting in the biosynthesis of at least 30,000 isoprenoid compounds, many of which, such as sterols or dolichols, are indispensable for living cells. In the filamentous fungus Trichoderma of major biotechnological interest isoprenoid metabolites are also involved in the biocontrol processes giving the mevalonate pathway an additional significance. On the other hand, little is known about genes coding for enzymes of the mevalonate pathway in Trichoderma. Here, we present cloning and functional analysis of the erg20 gene from Trichoderma reesei coding for farnesyl pyrophosphate (FPP) synthase (EC 2.5.1.10), an enzyme located at the branching point of the mevalonate pathway. Expression of the gene in a thermosensitive erg20-2 mutant of Saccharomyces cerevisiae impaired in the FPP synthase activity suppressed the thermosensitive phenotype. The same gene overexpressed in T. reesei significantly enhanced the FPP synthase activity and also stimulated the activity of cis-prenyltransferase, an enzyme of the dolichyl branch of the mevalonate pathway. Unexpectedly, the activity of squalene synthase from the other, sterol branch, was significantly decreased without, however, affecting ergosterol level.

  11. A mitogen-activated protein kinase Tmk3 participates in high osmolarity resistance, cell wall integrity maintenance and cellulase production regulation in Trichoderma reesei.

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    Mingyu Wang

    Full Text Available The mitogen-activated protein kinase (MAPK pathways are important signal transduction pathways conserved in essentially all eukaryotes, but haven't been subjected to functional studies in the most important cellulase-producing filamentous fungus Trichoderma reesei. Previous reports suggested the presence of three MAPKs in T. reesei: Tmk1, Tmk2, and Tmk3. By exploring the phenotypic features of T. reesei Δtmk3, we first showed elevated NaCl sensitivity and repressed transcription of genes involved in glycerol/trehalose biosynthesis under higher osmolarity, suggesting Tmk3 participates in high osmolarity resistance via derepression of genes involved in osmotic stabilizer biosynthesis. We also showed significant downregulation of genes encoding chitin synthases and a β-1,3-glucan synthase, decreased chitin content, 'budded' hyphal appearance typical to cell wall defective strains, and increased sensitivity to calcofluor white/Congo red in the tmk3 deficient strain, suggesting Tmk3 is involved in cell wall integrity maintenance in T. reesei. We further observed the decrease of cellulase transcription and production in T. reesei Δtmk3 during submerged cultivation, as well as the presence of MAPK phosphorylation sites on known transcription factors involved in cellulase regulation, suggesting Tmk3 is also involved in the regulation of cellulase production. Finally, the expression of cell wall integrity related genes, the expression of cellulase coding genes, cellulase production and biomass accumulation were compared between T. reesei Δtmk3 grown in solid state media and submerged media, showing a strong restoration effect in solid state media from defects resulted from tmk3 deletion. These results showed novel physiological processes that fungal Hog1-type MAPKs are involved in, and present the first experimental investigation of MAPK signaling pathways in T. reesei. Our observations on the restoration effect during solid state cultivation suggest

  12. Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

  13. Efficient cellulase production by Trichoderma reesei in continuous cultivation on lactose medium with a computer-controlled feeding strategy.

    Science.gov (United States)

    Bailey, M J; Tähtiharju, J

    2003-08-01

    A low-foaming hydrophobin II deletant of the Trichoderma reesei strain Rut-C30 was used for production of cellulases by continuous cultivation on lactose medium in a laboratory fermenter. The control paradigm of the addition of new medium to the continuous process was based on the growth dynamics of the fungus. A decrease in the rate of base addition to the cultivation for pH-minimum control was used as an indicator of imminent exhaustion of carbon source for growth and enzyme induction. When the amount of base added per 5 min computation cycle decreased below a given value, new medium was added to the fermenter. When base addition for pH control thereafter increased above the criterion value, due to increased growth, the medium feed was discontinued or decreased. The medium feeding protocol employed was successful in locking the fungus in the stage of imminent, but not actual, exhaustion of carbon source. According to the results of a batch cultivation of the same strain on the same medium, this is the phase of maximal enzyme productivity. The medium addition protocol used in this work resulted in a very stable continuous process, in which cellulase productivity was maintained for several hundred hours at the maximum level observed in a batch cultivation for only about 10 h. Despite a major technical disturbance after about 420 h, the process was restored to stability. When the cultivation was terminated after 650 h, the level of enzyme production was still maximal, with no signs of instability of the process.

  14. 13C-metabolic flux ratio and novel carbon path analyses confirmed that Trichoderma reesei uses primarily the respirative pathway also on the preferred carbon source glucose

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    Saloheimo Markku

    2009-10-01

    Full Text Available Abstract Background The filamentous fungus Trichoderma reesei is an important host organism for industrial enzyme production. It is adapted to nutrient poor environments where it is capable of producing large amounts of hydrolytic enzymes. In its natural environment T. reesei is expected to benefit from high energy yield from utilization of respirative metabolic pathway. However, T. reesei lacks metabolic pathway reconstructions and the utilization of the respirative pathway has not been investigated on the level of in vivo fluxes. Results The biosynthetic pathways of amino acids in T. reesei supported by genome-level evidence were reconstructed with computational carbon path analysis. The pathway reconstructions were a prerequisite for analysis of in vivo fluxes. The distribution of in vivo fluxes in both wild type strain and cre1, a key regulator of carbon catabolite repression, deletion strain were quantitatively studied by performing 13C-labeling on both repressive carbon source glucose and non-repressive carbon source sorbitol. In addition, the 13C-labeling on sorbitol was performed both in the presence and absence of sophorose that induces the expression of cellulase genes. Carbon path analyses and the 13C-labeling patterns of proteinogenic amino acids indicated high similarity between biosynthetic pathways of amino acids in T. reesei and yeast Saccharomyces cerevisiae. In contrast to S. cerevisiae, however, mitochondrial rather than cytosolic biosynthesis of Asp was observed under all studied conditions. The relative anaplerotic flux to the TCA cycle was low and thus characteristic to respiratory metabolism in both strains and independent of the carbon source. Only minor differences were observed in the flux distributions of the wild type and cre1 deletion strain. Furthermore, the induction of the hydrolytic gene expression did not show altered flux distributions and did not affect the relative amino acid requirements or relative anabolic

  15. Hydrodynamic and kinetic study of cellulase production by Trichoderma reesei with pellet morphology.

    Science.gov (United States)

    Yu, Liang; Chao, Yapeng; Wensel, Pierre; Chen, Shulin

    2012-07-01

    Numerical simulations and experimental validation were performed to understand the effects of hydrodynamics on pellet formation and cellulase production by filamentous T. reesei. The constructed model combined a steady-state multiple reference frame (MRF) approach describing mechanical mixing, oxygen mass transfer, and non-Newtonian flow field with a transient sliding mesh approach and kinetics of oxygen consumption, pellet formation, and enzyme production. The model was experimentally validated at various agitation speeds in a two-impeller Rushton turbine fermentor. Results from simulation and experimentation showed that higher agitation speeds led to increases in the pellet diameter and the proportion of pelletized (vs. filamentous) forms of the biomass. It also led to increase in dissolved oxygen mass transfer rate in shear-thinning fluid and cellulase productivity. The extent of these increases varied considerably among agitation speeds. Pellet formation and morphology were presumably affected within a viscosity-dependent shear-rate range. Cellulase activity and cell viability were shown to be sensitive to impeller shear. A maximum cellulase activity of 3.5 IU/mL was obtained at 400 rpm, representing a twofold increase over that at 100 rpm. PMID:22252572

  16. Characterization and Strain Improvement of a Hypercellulytic Variant, Trichoderma reesei SN1, by Genetic Engineering for Optimized Cellulase Production in Biomass Conversion Improvement.

    Science.gov (United States)

    Qian, Yuanchao; Zhong, Lixia; Hou, Yunhua; Qu, Yinbo; Zhong, Yaohua

    2016-01-01

    The filamentous fungus Trichoderma reesei is a widely used strain for cellulolytic enzyme production. A hypercellulolytic T. reesei variant SN1 was identified in this study and found to be different from the well-known cellulase producers QM9414 and RUT-C30. The cellulose-degrading enzymes of T. reesei SN1 show higher endoglucanase (EG) activity but lower β-glucosidase (BGL) activity than those of the others. A uracil auxotroph strain, SP4, was constructed by pyr4 deletion in SN1 to improve transformation efficiency. The BGL1-encoding gene bgl1 under the control of a modified cbh1 promoter was overexpressed in SP4. A transformant, SPB2, with four additional copies of bgl1 exhibited a 17.1-fold increase in BGL activity and a 30.0% increase in filter paper activity. Saccharification of corncob residues with crude enzyme showed that the glucose yield of SPB2 is 65.0% higher than that of SP4. These results reveal the feasibility of strain improvement through the development of an efficient genetic transformation platform to construct a balanced cellulase system for biomass conversion. PMID:27621727

  17. Study of a High-Yield Cellulase System Created by Heavy-Ion Irradiation-Induced Mutagenesis of Aspergillus niger and Mixed Fermentation with Trichoderma reesei.

    Directory of Open Access Journals (Sweden)

    Shu-Yang Wang

    Full Text Available The aim of this study was to evaluate and validate the efficiency of 12C6+ irradiation of Aspergillus niger (A. niger or mutagenesis via mixed Trichoderma viride (T. viride culturing as well as a liquid cultivation method for cellulase production via mixed Trichoderma reesei (T. reesei and A. niger culture fermentation. The first mutagenesis approach was employed to optimize yield from a cellulase-producing strain via heavy-ion mutagenesis and high-throughput screening, and the second was to effectively achieve enzymatic hydrolysis of cellulase from a mixed culture of mutant T. viride and A. niger. We found that 12C6+-ion irradiation induced changes in cellulase biosynthesis in A. niger but had no effect on the time course of the synthesis. It is notable that the exoglucanases (CBH activities of A. niger strains H11-1 and H differed (6.71 U/mL vs. 6.01 U/mL and were significantly higher than that of A. niger mutant H3-1. Compared with strain H, the filter paper assay (FPA, endoglucanase (EG and β-glucosidase (BGL activities of mutant strain H11-1 were increased by 250.26%, 30.26% and 34.91%, respectively. A mixed culture system was successfully optimized, and the best ratio of T. reesei to A. niger was 5:1 for 96 h with simultaneous inoculation. The BGL activity of the mixed culture increased after 72 h. At 96 h, the FPA and BGL activities of the mixed culture were 689.00 and 797.15 U/mL, respectively, significantly higher than those of monocultures, which were 408.70 and 646.98 U/mL for T. reesei and 447.29 and 658.89 U/mL for A. niger, respectively. The EG activity of the mixed culture was 2342.81 U/mL, a value that was significantly higher than that of monocultures at 2206.57 U/mL for T. reesei and 1727.62 U/mL for A. niger. In summary, cellulose production and hydrolysis yields were significantly enhanced by the proposed combination scheme.

  18. Characterization, optimization, and scale-up of cellulases production by trichoderma reesei cbs 836.91 in solid-state fermentation using agro-industrial products.

    Science.gov (United States)

    Ortiz, Gastón E; Guitart, María E; Cavalitto, Sebastián F; Albertó, Edgardo O; Fernández-Lahore, Marcelo; Blasco, Martín

    2015-11-01

    The application of cellulases in saccharification processes is restricted by its production cost. Consequently, new fungal strains able to elaborate higher cellulases titers and with special activity profiles are required to make the process economical. The aim of this investigation was to find a promising wild-type Trichoderma strain for cellulases production. The Trichoderma reesei strain 938 (CBS 836.91) was selected among twenty strains on the basis of cellulase-agar-plate screening. Evaluation of the selected strain on six solid substrates indicated the highest activities to be obtained from wheat bran. Statistical analyses of the experimental design indicated a significant effect of pH and moisture on the generation of endoglucanase (EGA) and filter-paper (FPA) activity. Furthermore, a central-composite design-based optimization revealed that pH values between 6.4 and 6.6 and moisture from 74 to 94% were optimal for cellulases production. Under these conditions, 8-10 IU gds(-1) of FPA and 15.6-17.8 IU gds(-1) of EGA were obtained. In addition, cultivation in a rotating-drum reactor under optimal conditions gave 8.2 IU gds(-1) FPA and 13.5 IU gds(-1) EGA. Biochemical characterization of T. reesei 938 cellulases indicated a substantially higher resistance to 4 mM Fe(+2) and a slightly greater tolerance to alkaline pH in comparison to Celluclast(®). These results suggest that T. reesei 938 could be a promising candidate for improved cellulases production through direct-evolution strategies.

  19. Characterization, optimization, and scale-up of cellulases production by trichoderma reesei cbs 836.91 in solid-state fermentation using agro-industrial products.

    Science.gov (United States)

    Ortiz, Gastón E; Guitart, María E; Cavalitto, Sebastián F; Albertó, Edgardo O; Fernández-Lahore, Marcelo; Blasco, Martín

    2015-11-01

    The application of cellulases in saccharification processes is restricted by its production cost. Consequently, new fungal strains able to elaborate higher cellulases titers and with special activity profiles are required to make the process economical. The aim of this investigation was to find a promising wild-type Trichoderma strain for cellulases production. The Trichoderma reesei strain 938 (CBS 836.91) was selected among twenty strains on the basis of cellulase-agar-plate screening. Evaluation of the selected strain on six solid substrates indicated the highest activities to be obtained from wheat bran. Statistical analyses of the experimental design indicated a significant effect of pH and moisture on the generation of endoglucanase (EGA) and filter-paper (FPA) activity. Furthermore, a central-composite design-based optimization revealed that pH values between 6.4 and 6.6 and moisture from 74 to 94% were optimal for cellulases production. Under these conditions, 8-10 IU gds(-1) of FPA and 15.6-17.8 IU gds(-1) of EGA were obtained. In addition, cultivation in a rotating-drum reactor under optimal conditions gave 8.2 IU gds(-1) FPA and 13.5 IU gds(-1) EGA. Biochemical characterization of T. reesei 938 cellulases indicated a substantially higher resistance to 4 mM Fe(+2) and a slightly greater tolerance to alkaline pH in comparison to Celluclast(®). These results suggest that T. reesei 938 could be a promising candidate for improved cellulases production through direct-evolution strategies. PMID:26256022

  20. Secretome analysis of Trichoderma reesei and Aspergillus niger cultivated by submerged and sequential fermentation processes: Enzyme production for sugarcane bagasse hydrolysis.

    Science.gov (United States)

    Florencio, Camila; Cunha, Fernanda M; Badino, Alberto C; Farinas, Cristiane S; Ximenes, Eduardo; Ladisch, Michael R

    2016-08-01

    Cellulases and hemicellulases from Trichoderma reesei and Aspergillus niger have been shown to be powerful enzymes for biomass conversion to sugars, but the production costs are still relatively high for commercial application. The choice of an effective microbial cultivation process employed for enzyme production is important, since it may affect titers and the profile of protein secretion. We used proteomic analysis to characterize the secretome of T. reesei and A. niger cultivated in submerged and sequential fermentation processes. The information gained was key to understand differences in hydrolysis of steam exploded sugarcane bagasse for enzyme cocktails obtained from two different cultivation processes. The sequential process for cultivating A. niger gave xylanase and β-glucosidase activities 3- and 8-fold higher, respectively, than corresponding activities from the submerged process. A greater protein diversity of critical cellulolytic and hemicellulolytic enzymes were also observed through secretome analyses. These results helped to explain the 3-fold higher yield for hydrolysis of non-washed pretreated bagasse when combined T. reesei and A. niger enzyme extracts from sequential fermentation were used in place of enzymes obtained from submerged fermentation. An enzyme loading of 0.7 FPU cellulase activity/g glucan was surprisingly effective when compared to the 5-15 times more enzyme loadings commonly reported for other cellulose hydrolysis studies. Analyses showed that more than 80% consisted of proteins other than cellulases whose role is important to the hydrolysis of a lignocellulose substrate. Our work combined proteomic analyses and enzymology studies to show that sequential and submerged cultivation methods differently influence both titers and secretion profile of key enzymes required for the hydrolysis of sugarcane bagasse. The higher diversity of feruloyl esterases, xylanases and other auxiliary hemicellulolytic enzymes observed in the enzyme

  1. Expression of Trichoderma reesei β-mannanase in tobacco chloroplasts and its utilization in lignocellulosic woody biomass hydrolysis.

    Directory of Open Access Journals (Sweden)

    Pankaj Agrawal

    Full Text Available Lignocellulosic ethanol offers a promising alternative to conventional fossil fuels. One among the major limitations in the lignocellulosic biomass hydrolysis is unavailability of efficient and environmentally biomass degrading technologies. Plant-based production of these enzymes on large scale offers a cost-effective solution. Cellulases, hemicellulases including mannanases and other accessory enzymes are required for conversion of lignocellulosic biomass into fermentable sugars. β-mannanase catalyzes endo-hydrolysis of the mannan backbone, a major constituent of woody biomass. In this study, the man1 gene encoding β-mannanase was isolated from Trichoderma reesei and expressed via the chloroplast genome. PCR and Southern hybridization analysis confirmed site-specific transgene integration into the tobacco chloroplast genomes and homoplasmy. Transplastomic plants were fertile and set viable seeds. Germination of seeds in the selection medium showed inheritance of transgenes into the progeny without any Mendelian segregation. Expression of endo-β-mannanase for the first time in plants facilitated its characterization for use in enhanced lignocellulosic biomass hydrolysis. Gel diffusion assay for endo-β-mannanase showed the zone of clearance confirming functionality of chloroplast-derived mannanase. Endo-β-mannanase expression levels reached up to 25 units per gram of leaf (fresh weight. Chloroplast-derived mannanase had higher temperature stability (40 °C to 70 °C and wider pH optima (pH 3.0 to 7.0 than E.coli enzyme extracts. Plant crude extracts showed 6-7 fold higher enzyme activity than E.coli extracts due to the formation of disulfide bonds in chloroplasts, thereby facilitating their direct utilization in enzyme cocktails without any purification. Chloroplast-derived mannanase when added to the enzyme cocktail containing a combination of different plant-derived enzymes yielded 20% more glucose equivalents from pinewood than the

  2. Malting of barley with combinations of Lactobacillus plantarum, Aspergillus niger, Trichoderma reesei, Rhizopus oligosporus and Geotrichum candidum to enhance malt quality.

    Science.gov (United States)

    Hattingh, M; Alexander, A; Meijering, I; van Reenen, C A; Dicks, L M T

    2014-03-01

    Good quality malt is characterised by the presence of high levels of fermentable sugars, amino acids and vitamins. To reach the starch-rich endosperm of the kernel, β-glucan- and arabinoxylan-rich cell walls have to be degraded. β-Glucanase is synthesized in vast quantities by the aleurone layer and scutellum during germination. Secretion of hydrolytic enzymes is often stimulated by addition of the plant hormone gibberellic acid (GA3) during germination. We have shown an enhanced β-glucanase and α-amylase activity in malt when germinating barley was inoculated with a combination of Lactobacillus plantarum B.S1.6 and spores of Aspergillus niger MH1, Rhizopus oligosporus MH2 or Trichoderma reesei MH3, and L. plantarum B.S1.6 combined with cell-free culture supernatants from each of these fungi. Highest malt β-glucanase activity (414 Units/kg malt) was recorded with a combination of L. plantarum B.S1.6 and spores of A. niger MH1. Highest α-amylase activities were recorded with a combination of L. plantarum B.S1.6 and spores of R. oligosporus MH2 (373 Ceralpha Units/g malt). Highest FAN levels were recorded when L. plantarum was inoculated in combination with spores of either R. oligosporus MH2 or T. reesei MH3 (259 and 260 ppm, respectively). This is the first study showing that cell-free culture supernatants of Aspergillus, Rhizopus and Trichoderma have a stimulating effect on β-glucanase and α-amylase production during malting. A combination of L. plantarum B.S1.6, and spores of A. niger MH1 and R. oligosporus MH2 may be used as starter cultures to enhance malt quality.

  3. Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, Lindsey N.; Culley, David E.; Hofstad, Beth A.; Chauvigne-Hines, Lacie M.; Zink, Erika M.; Purvine, Samuel O.; Smith, Richard D.; Callister, Stephen J.; Magnuson, Jon M.; Wright, Aaron T.

    2013-12-01

    Development of alternative, non-petroleum based sources of bioenergy that can be applied in the short-term find great promise in the use of highly abundant and renewable lignocellulosic plant biomass.1 This material obtained from different feedstocks, such as forest litter or agricultural residues, can yield liquid fuels and other chemical products through biorefinery processes.2 Biofuels are obtained from lignocellulosic materials by chemical pretreatment of the biomass, followed by enzymatic decomposition of cellulosic and hemicellulosic compounds into soluble sugars that are converted to desired chemical products via microbial metabolism and fermentation.3, 4 To release soluble sugars from polymeric cellulose multiple enzymes are required, including endoglucanase, exoglucanase, and β-glucosidase.5, 6 However, the enzymatic hydrolysis of cellulose into soluble sugars remains a significant limiting factor to the efficient and economically viable utilization of lignocellulosic biomass for transport fuels.7, 8 The primary industrial source of cellulose and hemicellulases is the mesophilic soft-rot fungus Trichoderma reesei,9 having widespread applications in food, feed, textile, pulp, and paper industries.10 The genome encodes 200 glycoside hydrolases, including 10 cellulolytic and 16 hemicellulolytic enzymes.11 The hypercellulolytic catabolite derepressed strain RUT-C30 was obtained through a three-step UV and chemical mutagenesis of the original T. reesei strain QM6a,12, 13 in which strains M7 and NG14 were intermediate, having higher cellulolytic activity than the parent strain but less activity and higher catabolite repression than RUT-C30.14 Numerous methods have been employed to optimize the secreted enzyme cocktail of T. reesei including cultivation conditions, operational parameters, and mutagenesis.3 However, creating an optimal and economical enzyme mixture for production-scale biofuels synthesis may take thousands of experiments to identify.

  4. Genome sequencing of the Trichoderma reesei QM9136 mutant identifies a truncation of the transcriptional regulator XYR1 as the cause for its cellulase-negative phenotype

    Energy Technology Data Exchange (ETDEWEB)

    Lichius, Alexander; Bidard, Frederique; Buchholz, Franziska; Le Crom, Stphane; Martin, Joel X.; Schackwitz, Wendy; Austerlitz, Tina; Grigoriev, Igor V.; Baker, Scott E.; Margeot, Antoine; Seiboth, Bernhard; Kubicek, Christian P.

    2015-12-01

    Background: Trichoderma reesei is the main industrial source of cellulases and hemicellulases required for the hydrolysis of biomass to simple sugars, which can then be used in the production of biofuels and biorefineries. The highly productive strains in use today were generated by classical mutagenesis. As byproducts of this procedure, mutants were generated that turned out to be unable to produce cellulases. In order to identify the mutations responsible for this inability, we sequenced the genome of one of these strains, QM9136, and compared it to that of its progenitor T. reesei QM6a. Results: In QM9136, we detected a surprisingly low number of mutagenic events in the promoter and coding regions of genes, i.e. only eight indels and six single nucleotide variants. One of these indels led to a frame-shift in the Zn2Cys6 transcription factor XYR1, the general regulator of cellulase and xylanase expression, and resulted in its C-terminal truncation by 140 amino acids. Retransformation of strain QM9136 with the wild-type xyr1 allele fully recovered the ability to produce cellulases, and is thus the reason for the cellulase-negative phenotype. Introduction of an engineered xyr1 allele containing the truncating point mutation into the moderate producer T. reesei QM9414 rendered this strain also cellulase-negative. The correspondingly truncated XYR1 protein was still able to enter the nucleus, but failed to be expressed over the basal constitutive level. Conclusion: The missing 140 C-terminal amino acids of XYR1 are therefore responsible for its previously observed auto-regulation which is essential for cellulases to be expressed. Our data present a working example of the use of genome sequencing leading to a functional explanation of the QM9136 cellulase-negative phenotype.

  5. On-site enzymes produced from Trichoderma reesei RUT-C30 and Aspergillus saccharolyticus for hydrolysis of wet exploded corn stover and loblolly pine.

    Science.gov (United States)

    Rana, Vandana; Eckard, Anahita D; Teller, Philip; Ahring, Birgitte K

    2014-02-01

    Cellulase production by two filamentous fungi Trichoderma reesei RUT-C30 and novel fungal strain, Aspergillus saccharolyticus on pretreated corn stover was investigated. Cellulase production was followed by the hydrolysis of two feedstocks, wet-exploded corn stover (WECS) and wet-exploded loblolly pine (WELP) by on-site produced enzyme cocktails containing cellulase from T. reesei RUT-C30 and β-glucosidase from A. saccharolyticus. The sugar yields using the on-site enzyme cocktails were compared with commercial enzymes preparations, Celluclast 1.5L and Novozym 188 at two substrate concentrations, 5% and 10% (w/w) and enzyme loading at 5 and 15 FPU/g glucan for WECS and WELP. The highest sugar yields were obtained at 5% (w/w) substrate concentration and 15 FPU/g glucan for both feedstocks. Glucose yields of 81% and 88% were obtained from on-site and commercial enzymes, respectively using WECS as feed stock. The sugar yields were 55% and 58% for WELP samples hydrolyzed with on-site and commercial enzymes, respectively.

  6. Saccharification of pumpkin residues by coculturing of Trichoderma reesei RUT-C30 and Phanerochaete chrysosporium Burdsall with delayed inoculation timing.

    Science.gov (United States)

    Yang, Rui; Meng, Demei; Hu, Xiaosong; Ni, Yuanying; Li, Quanhong

    2013-09-25

    Trichoderma reesei and Phanerochaete chrysosporium with different lignocellulose-degrading enzyme systems have received much attention due to their ability to biodegrade lignocellulosic biomass. However, the synergistic effect of the two fungi on lignocellulose degradation is unknown. Herein, a cocultivation of T. reesei RUT-C30 and P. chrysosporium Burdsall for biodegradation of lignocellulosic pumpkin residues (PRS) was developed to produce soluble saccharide. Results indicated that a cocultivation of the two fungi with P. chrysosporium Burdsall inoculation delayed for 1.5 days produced the highest saccharide yield of 53.08% (w/w), and only 20.83% (w/w) of PRS were left after one batch of fermentation. In addition, this strategy increased the activities of secreted cellulases (endoglucanase, cellobiohydrolase, and β-glucosidase) and ligninases (lignin peroxidase and manganese peroxidase), which correlated to the increased saccharide yield. Besides, the resulting monosaccharides including glucose (1.23 mg/mL), xylose (0.13 mg/mL), arabinose (0.46 mg/mL), and fructose (0.21 mg/mL) from cocultures exhibited much higher yields than those from monoculture, which provides basal information for further fermentation research. This bioconversion of PRS into soluble sugars by cocultured fungal species provides a low cost method based on lignocellulose for potential biofuels or other bioproduct production. PMID:24020787

  7. Identifying beneficial qualities of Trichoderma parareesei for plants.

    Science.gov (United States)

    Rubio, M Belén; Quijada, Narciso M; Pérez, Esclaudys; Domínguez, Sara; Monte, Enrique; Hermosa, Rosa

    2014-03-01

    Trichoderma parareesei and Trichoderma reesei (teleomorph Hypocrea jecorina) produce cellulases and xylanases of industrial interest. Here, the anamorphic strain T6 (formerly T. reesei) has been identified as T. parareesei, showing biocontrol potential against fungal and oomycete phytopathogens and enhanced hyphal growth in the presence of tomato exudates or plant cell wall polymers in in vitro assays. A Trichoderma microarray was used to examine the transcriptomic changes in T6 at 20 h of interaction with tomato plants. Out of a total 34,138 Trichoderma probe sets deposited on the microarray, 250 showed a significant change of at least 2-fold in expression in the presence of tomato plants, with most of them being downregulated. T. parareesei T6 exerted beneficial effects on tomato plants in terms of seedling lateral root development, and in adult plants it improved defense against Botrytis cinerea and growth promotion under salt stress. Time course expression patterns (0 to 6 days) observed for defense-related genes suggest that T6 was able to prime defense responses in the tomato plants against biotic and abiotic stresses. Such responses undulated, with a maximum upregulation of the jasmonic acid (JA)/ethylene (ET)-related LOX1 and EIN2 genes and the salt tolerance SOS1 gene at 24 h and that of the salicylic acid (SA)-related PR-1 gene at 48 h after T6 inoculation. Our study demonstrates that the T. parareesei T6-tomato interaction is beneficial to both partners.

  8. Paracelsin; characterization by NMR spectroscopy and circular dichroism, and hemolytic properties of a peptaibol antibiotic from the cellulolytically active mold Trichoderma reesei. Part B.

    Science.gov (United States)

    Brückner, H; Graf, H; Bokel, M

    1984-11-15

    Paracelsin, a hemolytic and membrane active polypeptide antibiotic of the peptaibol class which is excreted by the mold Trichoderma reesei, was obtained by a simplified and rapid isolation procedure utilizing hydrophobic adsorber resins. Investigation by 13C nuclear magnetic resonance spectroscopy and circular dichroism revealed considerable helical portions in solution, and the very recently accomplished sequence determination of paracelsin allows the discussion of the results with regard to the closely related analogues, alamethicin and suzukacillin. A selective cleavage of the peptide was achieved by careful treatment with various acids, and a buffer of pH 8.25 and of high ionic strength made possible the quantitative determination of the C-terminal phenylalaninol released by means of ion-exchange chromatography. The significance of the production of paracelsin and related mycotoxins of the peptaibol class, exhibiting various kinds of biological activity, is discussed with respect to the extensive effort being made towards biotechnological applications of species, strains and cellulolytically highly active mutants of the fungus Trichoderma. PMID:6500005

  9. 里氏木霉Trichoderma reesei产纤维素酶的发酵培养基碳氮源优化%Culture medium optimization for producing cellulase by Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    祖彩霞; 李志敏; 叶勤

    2011-01-01

    The influence of carbon and nitrogen sources on cellulase production was investigated. Single factor method and response surface methodology were used to optimize the medium for cellulase production by Trichoderma reesei. Firstly, it showed that soybean powder, corn cob and corn steep liquor had positive effects on cellulase production in one factor at a time. Then, the optimal concentrations of soybean powder (32. 21 g/L) , corn cob (42. 29 g/L) and corn steep liquor (4.45 g/L) were determined by central compostion design and response surface analysis. Under these optimizations conditions, the cellulase production reached (10. 65 ±0. 50) U/mL on the flask cultivation for 7 d.%研究C、N源对里氏木霉(Trichoderma reesei)生产纤维素酶的影响,采用单因素实验方法和中心复合方法对发酵培养基进行优化.单因素实验表明:黄豆饼粉、玉米芯、玉米浆对纤维素酶的影响显著.通过响应面优化,得到最优培养基C、N源的组成:黄豆饼粉32.21g/L,玉米芯42.29g/L,玉米浆4.45g/L.优化条件下,摇瓶发酵7 d的比酶活达到(10.65±0.50)U/mL.

  10. Optimization of Cellulose Production by Trichoderma reesei%里氏木霉纤维素酶生产工艺的优化

    Institute of Scientific and Technical Information of China (English)

    王涫; 王明钰; 穆子铭; 侯少莉; 张杰; 伏小平; 方诩

    2012-01-01

    提高纤维素酶生产效率,降低纤维素酶生产成本是纤维素乙醇生产技术的关键之一。而碳源、氮源和无机盐等产酶培养基成分以及接种时间、产酶温度、培养初始pH等产酶条件是纤维素酶生产过程中的关键因素。为充分利用里氏木霉生产纤维素酶,研究了纤维素酶高产菌株里氏木酶FST-1产酶培养基和产酶条件对纤维素酶产酶的影响。结果表明:麸皮、蛋白胨和磷酸二氢钾的含量对于纤维素酶的生产影响较大,并且确定了最优产酶培养基为4号培养基。通过对不同产酶条件的研究,确定最佳接种时间为24h、最佳产酶温度为32℃、最佳初始pH为5.5,优化后的生产工艺可以将滤纸酶活力和蛋白含量提高3倍。%Reducing cellulose production cost through improving cellulose titers during the fermentation is a key strategy for developing economically competitive bioethanol production from lignocellulosic biomass.Medium composition and inoculate time,temperature,initial pH and other factors were key factors to cellulose production.For the purpose to fully used Trichoderma reesei to produce cellulose,the influence of medium composition and growth condition of the cellulose-producing strain Trichoderma reesei FST-1on its enzyme production was investigated.The result showed that the contents of wheat bran,peptone and KH 2 PO 4 had the significant effect on cellulose titer,the No.4medium was the best medium.Through the study on different enzyme production condition,it found that the optimum inoculation time was 24hours,the optimum temperature was 32℃and the optimum initial pH was 5.5.Cellulose titer and soluble protein content were increased 3times with optimized culture condition.

  11. Trichoderma genes

    Science.gov (United States)

    Foreman, Pamela; Goedegebuur, Frits; Van Solingen, Pieter; Ward, Michael

    2012-06-19

    Described herein are novel gene sequences isolated from Trichoderma reesei. Two genes encoding proteins comprising a cellulose binding domain, one encoding an arabionfuranosidase and one encoding an acetylxylanesterase are described. The sequences, CIP1 and CIP2, contain a cellulose binding domain. These proteins are especially useful in the textile and detergent industry and in pulp and paper industry.

  12. Impact of alg3 gene deletion on growth, development, pigment production, protein secretion, and functions of recombinant Trichoderma reesei cellobiohydrolases in Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Dai, Ziyu; Aryal, Uma K.; Shukla, Anil; Qian, Wei-Jun; Smith, Richard D.; Magnuson, Jon K.; Adney, William S.; Beckham, Gregg T.; Brunecky, Roman; Himmel, Michael E.; Decker, Stephen R.; Ju, Xiaohui; Zhang, Xiao; Baker, Scott E.

    2013-12-01

    ALG3 is a Family 58 glycosyltransferase enzyme involved in early N-linked glycan synthesis. Here, we investigated the effect of the alg3 gene disruption on growth, development, metabolism, and protein secretion in Aspergillus niger. The alg3 gene deletion resulted in a significant reduction of growth on complete (CM) and potato dextrose agar (PDA) media and a substantial reduction of spore production on CM. It also delayed spore germination in the liquid cultures of both CM and PDA media, but led to a significant accumulation of red pigment on both CM and liquid modified minimal medium (MM) supplemented with yeast extract. The relative abundance of 55 proteins of the total 190 proteins identified in the secretome was significantly different as a result of alg3 gene deletion. Comparison of a Trichoderma reesei cellobiohydrolase (Cel7A) heterologously expressed in A. niger parental and Δalg3 strains showed that the recombinant Cel7A expressed in the mutant background was smaller in size than that from the parental strains. This study suggests that ALG3 is critical for growth and development, pigment production, and protein secretion in A. niger. Functional analysis of recombinant Cel7A with aberrant glycosylation demonstrates the feasibility of this alternative approach to evaluate the role of N-linked glycosylation in glycoprotein secretion and function.

  13. Increases thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase by fusion of cellulose binding domain derived from Trichoderma reesei

    Energy Technology Data Exchange (ETDEWEB)

    Thongekkaew, Jantaporn, E-mail: jantaporn_25@yahoo.com [Department of Biological Science, Faculty of Science, Ubon-Ratchathani University, Warinchumrab, Ubon-Ratchathani 34190 (Thailand); Ikeda, Hiroko; Iefuji, Haruyuki [Application Research Division, National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima 739-0046 (Japan)

    2012-03-30

    Highlights: Black-Right-Pointing-Pointer The CSLP and fusion enzyme were successfully expressed in the Pichia pastoris. Black-Right-Pointing-Pointer The fusion enzyme was stable at 80 Degree-Sign C for 120-min. Black-Right-Pointing-Pointer The fusion enzyme was responsible for cellulose-binding capacity. Black-Right-Pointing-Pointer The fusion enzyme has an attractive applicant for enzyme immobilization. -- Abstract: To improve the thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase (CSLP), the cellulose-binding domain originates from Trichoderma reesei cellobiohydrolase I was engineered into C-terminal region of the CSLP (CSLP-CBD). The CSLP and CSLP-CBD were successfully expressed in the Pichia pastoris using the strong methanol inducible alcohol oxidase 1 (AOX1) promoter and the secretion signal sequence from Saccharomyces cerevisiae ({alpha} factor). The recombinant CSLP and CSLP-CBD were secreted into culture medium and estimated by SDS-PAGE to be 22 and 27 kDa, respectively. The fusion enzyme was stable at 80 Degree-Sign C and retained more than 80% of its activity after 120-min incubation at this temperature. Our results also found that the fusion of fungal exoglucanase cellulose-binding domain to CSLP is responsible for cellulose-binding capacity. This attribute should make it an attractive applicant for enzyme immobilization.

  14. COMPARED ANALYSIS OF CATALASE AND PEROXIDASE ACTIVITY IN CELLULOLYTIC FUNGUS TRICHODERMA REESEI GROWN ON MEDIUM WITH DIFFERENT CONCENTRATIONS OF GRINDED WHEAT AND BARLEY STRAWS

    Directory of Open Access Journals (Sweden)

    Mihaela Cristica

    2010-09-01

    Full Text Available The purpose of this study was to assess the evolution of catalase and peroxidase activity in Trichoderma reesei grown on medium containing grinded wheat and barley straws. Carbon source of cultivation medium - glucose was replaced by various concentrations of grinded wheat and barley straws, finally resulting three experimental variants as follows: V1 = 20 g/l, V2 = 30 g/l, V3 = 40 g/l. ĂŽn addition to these variants a control sample was added in which composition remainded unchanged. The catalase activity was determined by spectrophotometric Sinha method (Artenie et al., 2008 while peroxidase activity was assesed using the o-dianisidine method (Cojocaru, 2009. Enzymatic determinations were carried out at 7 and 14 days from inoculation, in both fungus mycelium and culture liquid. The enzymatic assay showed significant differences between determinations intervals and work variants. Enzyme activity is influenced by the age of fungus and by the different nature of the substrate used.

  15. Heterologously expressed Aspergillus aculeatus β-glucosidase in Saccharomyces cerevisiae is a cost-effective alternative to commercial supplementation of β-glucosidase in industrial ethanol production using Trichoderma reesei cellulases.

    Science.gov (United States)

    Treebupachatsakul, Treesukon; Nakazawa, Hikaru; Shinbo, Hideaki; Fujikawa, Hiroki; Nagaiwa, Asami; Ochiai, Nobuhiro; Kawaguchi, Takashi; Nikaido, Mitsuru; Totani, Kazuhide; Shioya, Koki; Shida, Yosuke; Morikawa, Yasushi; Ogasawara, Wataru; Okada, Hirofumi

    2016-01-01

    Trichoderma reesei is a filamentous organism that secretes enzymes capable of degrading cellulose to cellobiose. The culture supernatant of T. reesei, however, lacks sufficient activity to convert cellobiose to glucose using β-glucosidase (BGL1). In this study, we identified a BGL (Cel3B) from T. reesei (TrCel3B) and compared it with the active β-glucosidases from Aspergillus aculeatus (AaBGL1). AaBGL1 showed higher stability and conversion of sugars to ethanol compared to TrCel3B, and therefore we chose to express this recombinant protein for use in fermentation processes. We expressed the recombinant protein in the yeast Saccharomyces cerevisiae, combined it with the superb T. reesei cellulase machinery and used the combination in a simultaneous saccharification and fermentation (SSF) process, with the hope that the recombinant would supplement the BGL activity. As the sugars were processed, the yeast immediately converted them to ethanol, thereby eliminating the problem posed by end product inhibition. Recombinant AaBGL1 activity was compared with Novozyme 188, a commercially available supplement for BGL activity. Our results show that the recombinant protein is as effective as the commercial supplement and can process sugars with equal efficiency. Expression of AaBGL1 in S. cerevisiae increased ethanol production effectively. Thus, heterologous expression of AaBGL1 in S. cerevisiae is a cost-effective and efficient process for the bioconversion of ethanol from lignocellulosic biomass.

  16. Overproduction of cellulase by Trichoderma reesei RUT C30 through batch-feeding of synthesized low-cost sugar mixture.

    Science.gov (United States)

    Li, Yonghao; Liu, Chenguang; Bai, Fengwu; Zhao, Xinqing

    2016-09-01

    Cellulase is a prerequisite for the bioconversion of lignocellulosic biomass, but its high cost presents the biggest challenge. In this article, low-cost mixture was produced from glucose through the transglycosylation reaction catalyzed by β-glucosidase for cellulase overproduction by Trichodema reesei RUT C30. As a result, cellulase titer of 90.3FPU/mL, which was more than 10 folds of that achieved with lactose as inducer, was achieved at 144h. Meanwhile, cellulase productivity was drastically increased to 627.1FPU/L/h, at least 3-5 folds higher than previously reported by the fungal species. The crude enzyme was further tested by hydrolyzing NaOH-pretreated corn stover with 15% solid loading, and 96.6g/L glucose was released with 92.6% sugar yield at 96h and 44.8g/L ethanol was obtained. PMID:27268435

  17. Impact of alg3 gene deletion on growth, development, pigment production, protein secretion, and functions of recombinant Trichoderma reesei cellobiohydrolases in Aspergillus niger.

    Science.gov (United States)

    Dai, Ziyu; Aryal, Uma K; Shukla, Anil; Qian, Wei-Jun; Smith, Richard D; Magnuson, Jon K; Adney, William S; Beckham, Gregg T; Brunecky, Roman; Himmel, Michael E; Decker, Stephen R; Ju, Xiaohui; Zhang, Xiao; Baker, Scott E

    2013-12-01

    Dolichyl-P-Man:Man(5)GlcNAc(2)-PP-dolichyl α-1,3-mannosyltransferase (also known as "asparagine-linked glycosylation 3", or ALG3) is involved in early N-linked glycan synthesis and thus is essential for formation of N-linked protein glycosylation. In this study, we examined the effects of alg3 gene deletion (alg3Δ) on growth, development, pigment production, protein secretion and recombinant Trichoderma reesei cellobiohydrolase (rCel7A) expressed in Aspergillus niger. The alg3Δ delayed spore germination in liquid cultures of complete medium (CM), potato dextrose (PD), minimal medium (MM) and CM with addition of cAMP (CM+cAMP), and resulted in significant reduction of hyphal growth on CM, potato dextrose agar (PDA), and CM+cAMP and spore production on CM. The alg3Δ also led to a significant accumulation of red pigment on both liquid and solid CM cultures. The relative abundances of 54 of the total 215 proteins identified in the secretome were significantly altered as a result of alg3Δ, 63% of which were secreted at higher levels in alg3Δ strain than the parent. The rCel7A expressed in the alg3Δ mutant was smaller in size than that expressed in both wild-type and parental strains, but still larger than T. reesei Cel7A. The circular dichroism (CD)-melt scans indicated that change in glycosylation of rCel7A does not appear to impact the secondary structure or folding. Enzyme assays of Cel7A and rCel7A on nanocrystalline cellulose and bleached kraft pulp demonstrated that the rCel7As have improved activities on hydrolyzing the nanocrystalline cellulose. Overall, the results suggest that alg3 is critical for growth, sporulation, pigment production, and protein secretion in A. niger, and demonstrate the feasibility of this alternative approach to evaluate the roles of N-linked glycosylation in glycoprotein secretion and function. PMID:24076077

  18. 海洋真菌Trichoderma reesei的次级代谢产物的分离与鉴定%Secondary metabolism product of fungus Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    孙奕; 吕阿丽; 田黎; 魏岚; 裴月湖

    2007-01-01

    目的 系统研究海洋真菌Trlchoderma reesei的次级代谢产物.方法 利用硅胶柱色谱、高效液相色谱等手段进行分离,根据理化性质及波谱数据对所得结构进行了鉴定.结果 从海洋真菌Trichoderma reesei发酵液的乙酸乙酯提取物中共分离得到6个成分,分别鉴定为cyclonerodiol(1)、8,9-dihydro-dihydroxymegastigrrmtrienone(2)、harzialactone A(3)、3,6-二苄基哌嗪-2,5-二酮(4)、3-异丁基-8-羟基吡咯并哌嗪-2,5-二酮(5)、3-苄基-8-羟基吡咯并哌嗪-2,5-二酮(6).结论 所分得的化合物均为从海洋真菌Trichoderma reesei的次级代谢产物中首次分离得到.

  19. The Trichoderma reesei Cry1 protein is a member of the cryptochrome/photolyase family with 6-4 photoproduct repair activity.

    Directory of Open Access Journals (Sweden)

    Jesús Guzmán-Moreno

    Full Text Available DNA-photolyases use UV-visible light to repair DNA damage caused by UV radiation. The two major types of DNA damage are cyclobutane pyrimidine dimers (CPD and 6-4 photoproducts (6-4PP, which are repaired under illumination by CPD and 6-4 photolyases, respectively. Cryptochromes are proteins related to DNA photolyases with strongly reduced or lost DNA repair activity, and have been shown to function as blue-light photoreceptors and to play important roles in circadian rhythms in plants and animals. Both photolyases and cryptochromes belong to the cryptochrome/photolyase family, and are widely distributed in all organisms. Here we describe the characterization of cry1, a member of the cryptochrome/photolyase protein family of the filamentous fungus Trichoderma reesei. We determined that cry1 transcript accumulates when the fungus is exposed to light, and that such accumulation depends on the photoreceptor Blr1 and is modulated by Envoy. Conidia of cry1 mutants show decreased photorepair capacity of DNA damage caused by UV light. In contrast, strains over-expressing Cry1 show increased repair, as compared to the parental strain even in the dark. These observations suggest that Cry1 may be stimulating other systems involved in DNA repair, such as the nucleotide excision repair system. We show that Cry1, heterologously expressed and purified from E. coli, is capable of binding to undamaged and 6-4PP damaged DNA. Photorepair assays in vitro clearly show that Cry1 repairs 6-4PP, but not CPD and Dewar DNA lesions.

  20. The Widescreen Anamorphic Lens

    NARCIS (Netherlands)

    Willemsen, Steven; Fossati, Giovanna; van den Oever, Annie

    2016-01-01

    This article examines the transformative impact of anamorphic widescreen technology on film aesthetics, in two parts: first, I discuss how the affordances of early anamorphic widescreen technology (primarily CinemaScope) played a key role in standardizing widescreen formats in the Hollywood film ind

  1. Process technological effects of deletion and amplification of hydrophobins I and II in transformants of Trichoderma reesei.

    Science.gov (United States)

    Bailey, M J; Askolin, S; Hörhammer, N; Tenkanen, M; Linder, M; Penttilä, M; Nakari-Setälä, T

    2002-05-01

    Transformants of the Trichoderma reeseistrains QM9414 and Rut-C30 were constructed in which the genes for the two major hydrophobin proteins, hydrophobins I (HFBI) and II (HFBII), were deleted or amplified by molecular biological techniques. Growth parameters and foam production of the transformant strains were compared with the corresponding properties of the parent strains by cultivation in laboratory bioreactors under conditions of catabolite repression (glucose medium) or induction of cellulolytic enzymes and other secondary metabolites (cellulose and lactose media). All the transformed strains exhibited vegetative growth properties similar to those of their parent. The Delta hfb2 (but not the Delta hfb1) transformant showed reduced tendency to foam, whereas both strains overproducing hydrophobins foamed extensively, particularly in the case of HFBII. Enzyme production on cellulose medium was unaltered in the Delta hfb2 transformant VTT D-99676, but both the Delta hfb2 and HFBII-overproducing transformants exhibited somewhat decreased enzyme production properties on lactose medium. Production of HFBI by the multi-copy transformant VTT D-98692 was almost 3-fold that of the parent strain QM9414. Overproduction of HFBII by the transformant VTT D-99745, obtained by transformation with three additional copies of the hfb2 gene under the cbh1 promoter, was over 5-fold compared to production by the parent strain Rut-C30. The Delta hfb2transformant VTT D-99676 produced a greatly increased number of spores on lactose medium compared with the parent strain, whereas the HFBII-overproducing transformant VTT D-99745 produced fewer spores.

  2. Optimization of covalent immobilization of Trichoderma reesei cellulase onto modified ReliZyme HA403 and Sepabeads EC-EP supports for cellulose hydrolysis, in buffer and ionic liquids/buffer media.

    Science.gov (United States)

    Bilgin, Ramazan; Yalcin, M Serkan; Yildirim, Deniz

    2016-08-01

    The covalent immobilization of Trichoderma reesei cellulase onto modified ReliZyme HA403 and Sepabeads EC-EP supports were carried out. The optimal immobilization conditions were determined using response surface methodology. The hydrolysis of cellulose using the free and immobilized cellulase preparations in ionic liquids (IL) using cosolvents was investigated. The hydrolytic activities in buffer medium containing 25% (v/v) of 1-butyl-3-methylimidazolium hexafluorophosphate were around 2.6-, 1.6-, and 5.5-fold higher than the activities in buffer medium. The retained initial activities were 32% and 57%, respectively for cellulase preparations immobilized onto Sepabeads EC-EP support and onto modified ReliZyme HA403 support after 5 reuses. PMID:25811997

  3. Engineering cellulase mixtures by varying the mole fraction of Thermomonospora fusca E[sub 5] and E[sub 3], Trichoderma reesei CBHI, and Caldocellum saccharolyticum [beta]-glucosidase

    Energy Technology Data Exchange (ETDEWEB)

    Walker, L.P.; Belair, C.D.; Wilson, D.B.; Irwin, D.C. (Cornell Univ., Ithaca, NY (United States))

    1993-11-05

    In this study, different mole fractions of pure Thermomonospora fusca E[sub 5] and E[sub 3], plus Trichoderma reesei CBHI were studied for reducing sugar production at 2 h, degree of synergism, and cellulose binding. In addition, the effects of introducing the Caldocellum saccharolyticum [Beta]-glucosidase into this cellulase system were investigated. The cellulases used were purified to homogeneity. Avicel PH 102 was the substrate. Reactions were run at 50 C for 2 h using total cellulase concentrations of 8.3 or 12.2 [mu]M. A bimixture of T. fusca E[sub 3] and T. reesei CBHI was very effective in hydrolyzing microcrystalline cellulose. The addition of endoglucanase E[sub 5] to the mixture only increased conversion to 9.8%. However, when both E[sub 5] and [Beta]-glucosidase were added, conversion increased to 14%. It was also observed that increasing total cellulase concentration beyond 8.3 [mu]M did little to increase percent conversion of cellulose into glucose. The results of the binding studies indicate no competition for binding sites between the endo- and exocellulases.

  4. Simultaneous Cellulase Production, Saccharification and Detoxification Using Dilute Acid Hydrolysate of S. spontaneum with Trichoderma reesei NCIM 992 and Aspergillus niger.

    Science.gov (United States)

    Sateesh, Lanka; Rodhe, Adivikatla Vimala; Naseeruddin, Shaik; Yadav, Kothagauni Srilekha; Prasad, Yenumulagerard; Rao, Linga Venkateswar

    2012-06-01

    Bioethanol production from lignocellulosic materials has several limitations. One aspect is the high production cost of cellulases used for saccharification of substrate and inhibition of fermenting yeast due to inhibitors released in acid hydrolysis. In the present work we have made an attempt to achieve simultaneous cellulases production, saccharification and detoxification using dilute acid hydrolysate of Saccharum spontaneum with and without addition of nutrients, supplemented with acid hydrolyzed biomass prior to inoculation in one set and after 3 days of inoculation in another set. Organisms used were T. reesei NCIM 992, and Aspergillus niger isolated in our laboratory. Cellulase yield obtained was 0.8 IU/ml on fourth day with T. reesei. Sugars were found to increase from fourth to fifth day, when hydrolysate was supplemented with nutrients and acid hydrolyzed biomass followed by inoculation with T. reesei. Phenolics were also found to decrease by 67%. PMID:23729891

  5. Cyclic anamorphic cosmology

    CERN Document Server

    Ijjas, Anna

    2016-01-01

    Cyclic models of the universe have the advantage of avoiding initial conditions problems related to postulating any sort of beginning in time. To date, the only known viable examples of cyclic models have been ekpyrotic. In this paper, we show that the recently proposed anamorphic scenario can also be made cyclic. The key to the cyclic completion is a classically stable, non-singular bounce. Remarkably, even though the bounce construction was originally developed to connect a period of contraction with a period of expansion both described by Einstein gravity, we show here that it can naturally be modified to connect an ordinary contracting phase described by Einstein gravity with a phase of anamorphic smoothing. The paper will present the basic principles and steps in constructing cyclic anamorphic models.

  6. The Effect of Microwave-NaOH Pretreatment and Hydrolysis Enzyme Using Trichoderma reesei-Aspergillus niger on Rice Straw Bioethanol Production

    Directory of Open Access Journals (Sweden)

    Bambang Dwi Argo

    2016-01-01

    Full Text Available The process of bioethanol production from rice straw consists of two steps: (1 conversion of cellulose into simple sugars which is conducted by using microwave-NaOH pretreatment and straw hydrolysis using mold catalyst T.reesei and A. niger; and (2 fermentation of simple sugars into ethanol. In a microwave-NaOH pretreatment process has been obtained the best value content of cellulose in straw size of 100 mesh and a long exposure of 40 minutes for 72.70+1:10%. Crude cellulase enzyme activity of T.reesei isolation, A.niger and it mixtures were optimum at temperature of 50°C. The addition of crude enzyme from A.niger and T.reesei on a comparison of 1: 2 (v/v was able to increase the yield of the rice straw cellulose hydrolysis which is produces sugar at 12.89 mg/ml (1.29% w/v or 0.15% (w/v when converted into ethanol. The glucose yield from rice straw was 25.47% with 3% ethanol.

  7. 里氏木霉产纤维素酶的诱导和合成机理研究进展%Research progress of the mechanism of induction and synthesis of cellulase by Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    李辉; 王义强; 陈介南; 张伟涛

    2011-01-01

    Cellulase is a kind of inducible enzyme.The inducer is required for cellulase expression.Cellulose, sophorose, lactose, cellobiose and sorbitol are common inducers.Trichoderma reesei mainly expresses three cellulases, endoglucanase, exoglucanase and β-1,4-glucosidase, in which the expression level of exoglucanase Ⅰ is highest, followed by endoglucanase and β-1,4-glucosidase.The synthesis and expression of cellulase is affected by active elements and inhibitory factors.In details, the combination of inducer and active element induces the synthesis and expression of cellulose.When the decomposition products of cellulase accumulate to a certain level, the intracellular inhibitory factor will combine with promoter to prevent the expression of cellulase.%纤维素酶是一类诱导酶,诱导剂对纤维素酶的表达是必需的.常用的诱导剂如纤维素、槐糖、乳糖、纤维二糖、山梨糖等.里氏木霉主要表达3种纤维素酶:内切葡聚糖酶、外切葡聚糖酶和β-1,4-葡萄糖苷酶,其中以外切葡聚糖苷酶Ⅰ的合成量最多,其次是内切葡聚糖苷酶和β-1,4-葡萄糖苷酶.纤维素酶的合成和表达是激活原件和抑制因子共同作用的结果:诱导物质和激活元件结合激活细胞内纤维素酶的合成和表达;当纤维素酶的分解产物达到一定水平时,细胞内的抑制因子和启动子结合,阻止纤维素酶的表达.

  8. Analysis of Cellulase Production withTrichoderma reeseiRut C30 Induced by Corn Cob%玉米芯诱导里氏木霉Rut C30产纤维素酶的分析

    Institute of Scientific and Technical Information of China (English)

    马立娟; 蔡瑞; 崔有志; 赫荣琳; 陈树林; 肖冬光

    2015-01-01

    分别以不同木质纤维素类为碳源(质量浓度均为10,g/L)诱导里氏木霉Rut C30产酶,通过测定诱导过程的产酶曲线,结果表明:以玉米芯为诱导碳源时,各酶组分的产酶水平最高,诱导120,h 后纤维素酶滤纸酶活、β–葡萄糖苷酶和木聚糖酶酶活分别为1.53,FPU/mL、0.61,IU/mL 和72.86,IU/mL.采用高效液相色谱(HPLC)法分析了玉米芯水解过程中糖的组成及变化,可检测到的单糖包括葡萄糖、木糖、阿拉伯糖、甘露糖和半乳糖以及纤维二糖等寡糖.诱导产酶实验表明单糖中只有半乳糖可诱导纤维素酶的合成.%Trichoderma reesei Rut C30 was induced by different lignocellulosic inducers with the same concentration of 10,g/L to produce enzymes,and the enzyme-producing curves were determined. Results indicated that the production of cel-lulase,β-glucosidase and xylanase were the highest when corn cob was used as the inducing carbon source,and the produc-tion was up to 1.53,FPU/mL,0.61 IU/mL and 72.86 IU/mL respectively after inducing 120 hours. The sugar composition and variation during the enzymatic hydrolysis of corn cob were analyzed with high performance liquid chromatography(HPLC). Monosaccharide detected included glucose,xylose,arabinose,mannose and galactose. The results of induction experiments indicated that only galactose among monosaccharides could induce cellulase.

  9. 木霉 -1,3-1,4-葡聚糖酶性质及其cDNA片段克隆%Study on the Properties of  -1,3-1,4-glucanase;Cloning and Sequence Analysis of cDNA Fragment from Trichoderma reesei

    Institute of Scientific and Technical Information of China (English)

    孙建义; 李卫芬; 许梓荣; 廖玉华

    2001-01-01

    本研究探讨了里氏木霉GXC的 -1,3-1,4-葡聚糖酶特性,克隆和分析了酶基因片段。结果表明,粗酶液经硫酸铵沉淀、Sephadex G-25、Sephadex G-100和DEAE-Sephadex A-50 柱层析得到纯 -1,3-1,4-葡聚糖酶;经12.5%SDS-PAGE凝胶电泳表明,该酶的分子量为35.21 kD;酶最适反应pH5.0,最适反应温度为60℃;Michaelis-Menten 动力学分析表明, -1,3-1,4-葡聚糖酶的Km 和 Vmax 分别为10.86 mg/mL和 14 286 mol/(min mg)。通过RT- PCR方法扩增并克隆了 -1,3-1,4-葡聚糖酶cDNA片段,测序表明,该片段长度为280 bp;同源性分析显示,该cDNA片段与水解淀粉芽孢杆菌、厌氧真菌 Orpinomyces strain PC-2、枯草芽孢杆菌中的 -1,3-1,4-葡聚糖酶的基因片段有较高的同源性,分别为90%,79%,91%。%-1,3-1,4-glucanase was purified from a solid-state culture of Trichoderma reesei GXC on wheat bran in three steps which comprisedammonium sulfate precipitation,Sephadex G-100 chromatography,and DEAE-Sephadex A-50 chromatography.The molecular mass was determined to be 35.21 kilodaltons by 12.5%sodium dodecyl sulfate polyacrylamide gel electrophoresis.The optimal temperature and pH for purified -1,3-1,4-glucanase reaction were 50℃ and 6.0,respectively.The Km of the enzyme on  -glucan was 10.86 mg/mL,and the Vmax on  -glucan was 14 286  molof glucose equivalents per mg of the pure enzyme per min.A partial  -1,3-1,4-glucanase cDNA Fragment from Trichoderma reesei GXC was amplified and cloned by Reverse Transcription-PCR strategy. Sequence analysis showed that the cDNA Fragment was 280 bp,it had 90%,79%,91% identity with the sequences of  -1,3-1,4-glucanase from Bacillus amyloliquefaciens,Orpinomyces Strain PC-2 and Bacillus subtilis,respectively.

  10. Biotransformation of Trichoderma spp. and their tolerance to aromatic amines, a major class of pollutants.

    Science.gov (United States)

    Cocaign, Angélique; Bui, Linh-Chi; Silar, Philippe; Chan Ho Tong, Laetitia; Busi, Florent; Lamouri, Aazdine; Mougin, Christian; Rodrigues-Lima, Fernando; Dupret, Jean-Marie; Dairou, Julien

    2013-08-01

    Trichoderma spp. are cosmopolitan soil fungi that are highly resistant to many toxic compounds. Here, we show that Trichoderma virens and T. reesei are tolerant to aromatic amines (AA), a major class of pollutants including the highly toxic pesticide residue 3,4-dichloroaniline (3,4-DCA). In a previous study, we provided proof-of-concept remediation experiments in which another soil fungus, Podospora anserina, detoxifies 3,4-DCA through its arylamine N-acetyltransferase (NAT), a xenobiotic-metabolizing enzyme that enables acetyl coenzyme A-dependent detoxification of AA. To assess whether the N-acetylation pathway enables AA tolerance in Trichoderma spp., we cloned and characterized NATs from T. virens and T. reesei. We characterized recombinant enzymes by determining their catalytic efficiencies toward several toxic AA. Through a complementary approach, we also demonstrate that both Trichoderma species efficiently metabolize 3,4-DCA. Finally, we provide evidence that NAT-independent transformation is solely (in T. virens) or mainly (in T. reesei) responsible for the observed removal of 3,4-DCA. We conclude that T. virens and, to a lesser extent, T. reesei likely utilize another, unidentified, metabolic pathway for the detoxification of AA aside from acetylation. This is the first molecular and functional characterization of AA biotransformation in Trichoderma spp. Given the potential of Trichoderma for cleanup of contaminated soils, these results reveal new possibilities in the fungal remediation of AA-contaminated soil.

  11. Overexpression and characterization of a laccase gene from Pleurotus ostreatus in Trichoderma reesei%糙皮侧耳(Pleurotus ostreatus)漆酶POXA1在里氏木霉中的高效表达及酶学性质

    Institute of Scientific and Technical Information of China (English)

    董欣睿; 秦丽娜; 陶勇; 黄建忠; 董志扬

    2012-01-01

    [Objective] Expression, purification and characterization of a laccase gene from Pleurotus ostreatus in Trichoderma reesei. [Methods] The strong promoter and terminator of cellobiohydrolase I (cbhl) gene from T. reesei were amplified by PCR and inserted into pBluescriptIISK(+) to form vector pSKCST. The laccase gene from Pleurotus ostreatus was de novo synthesized according to T. reesei condon bias and cloned into the vector pLacdt resulting in the expression vector pSKLDT. The linearized pSKLDT was introduced into T. reesei strain Tu6 by protoplast-mediated transformation. The screened laccase expression transformants were grown in shake flasks on minimal medium and the recombinant laccase was purified and characterized. [Results] Transformants were isolated in selective screening medium plate and identified by PCR. The enzyme activity of laccase in transformant LC-7 was 237.134 U/mL which was 28. 6 — fold higher than that in P. ostreatus. The specific activity of the purified enzyme was 9852 IU/mg. Enzymatic assay revealed that the optimum temperature for its activity was 501 and pH was 3.0. The optimum substrate was ABTS and the Km and Vmax for ABTS were 7. 58 × 10-2 mmol/L and 9. 752 × 10-3 mmol/L/min. Metal ions like Cu2+ , Zn2+ , Fe3+ , Mn2+ , Ba2+ , Mg2+ and Fe2+ had different inhibitory effect on purified laccase. [Conclusions] Under the regulation of cbhl promoter and cbhl signal peptide, heterologous laccase was successfully overexpressed in T. reesei.%[目的]将优化后的糙皮侧耳(Pleurotus ostreatus)漆酶基因poxl,在里氏木霉中进行高效表达并对重组表达的漆酶进行酶学性质测定.[方法]:根据里氏木霉的密码子偏好性,对漆酶POXA1密码子进行优化并合成.以质粒pBluscriptⅡSK(+)为骨架,利用里氏木霉纤维二糖水解酶基因(cbhl)的启动子和终止子序列构建里氏木霉外源蛋白表达载体pSKLDT.PEG介导的原生质体转化方法转化里氏木霉菌株Tu6,筛选获得漆酶表

  12. The Terpenoid Biosynthesis Toolkit of Trichoderma.

    Science.gov (United States)

    Bansal, Ravindra; Mukherjee, Prasun Kumar

    2016-04-01

    The widely used biotechnologically important fungi belonging to the genus Trichoderma are rich sources of secondary metabolites. Even though the genomes of several Trichoderma spp. have been published, and data are available on the genes involved in biosynthesis of non-ribosomal peptide synthetases and polyketide synthases, no genome-wide data are available for the terpenoid biosynthesis machinery in these organisms. In the present study, we have identified the genes involved in terpene biosynthesis in the genomes of three Trichoderma spp., viz., T. virens, T. atroviride and T. reesei. While the genes involved in the condensation steps are highly conserved across the three species, these fungi differed in the number and organization of terpene cyclases. T. virens genome harbours eleven terpene cyclases, while T. atroviride harbours seven, and T. reeseisix in their genomes; seven, three and two being part of putative secondary metabolism related gene clusters. PMID:27396184

  13. The Terpenoid Biosynthesis Toolkit of Trichoderma.

    Science.gov (United States)

    Bansal, Ravindra; Mukherjee, Prasun Kumar

    2016-04-01

    The widely used biotechnologically important fungi belonging to the genus Trichoderma are rich sources of secondary metabolites. Even though the genomes of several Trichoderma spp. have been published, and data are available on the genes involved in biosynthesis of non-ribosomal peptide synthetases and polyketide synthases, no genome-wide data are available for the terpenoid biosynthesis machinery in these organisms. In the present study, we have identified the genes involved in terpene biosynthesis in the genomes of three Trichoderma spp., viz., T. virens, T. atroviride and T. reesei. While the genes involved in the condensation steps are highly conserved across the three species, these fungi differed in the number and organization of terpene cyclases. T. virens genome harbours eleven terpene cyclases, while T. atroviride harbours seven, and T. reeseisix in their genomes; seven, three and two being part of putative secondary metabolism related gene clusters.

  14. Structural insights into the inhibition of cellobiohydrolase Cel7A by xylo‐oligosaccharides

    DEFF Research Database (Denmark)

    Momeni, Majid Haddad; Ubhayasekera, Wimal; Sandgren, Mats;

    2015-01-01

    The filamentous fungus Hypocrea jecorina (anamorph of Trichoderma reesei) is the predominant source of enzymes for industrial saccharification of lignocellulose biomass. The major enzyme, cellobiohydrolase Cel7A, constitutes nearly half of the total protein in the secretome. The performance...

  15. New genera in the Calosphaeriales: Togniniella and its anamorph Phaeocrella, and Calosphaeriophora as anamorph of Calosphaeria

    NARCIS (Netherlands)

    Réblová, M.; Mostert, L.; Gams, W.; Crous, P.W.

    2004-01-01

    During a survey of perithecial ascomycetes in New Zealand, two collections of a Togninia-like fungus were made on decayed wood. In culture, colonies produced a Phaeoacremonium-like anamorph. In order to reveal the phylogenetic relationships of the unknown fungus and its affinity to Togninia and othe

  16. [The isolation and characterization of beta-glucosidase gene and beta-glucosidase of Trichoderma viride]: Progress report

    International Nuclear Information System (INIS)

    Our project was to isolate and characterize the enzyme β-glucosidase and to clone and characterize the β-glucosidase gene; our goal is to clone and characterize each of the cellulase genes from Trichoderma. The induction of the Trichoderma reesei cellulase complex by cellulose and by the soluble inducer, sophorose, has been demonstrated. Although the induction of the cellulase complex has previously been well documented, the induction of β-glucosidase had been questioned. 49 refs., 6 figs., 2 tabs

  17. 77 FR 35331 - Trichoderma reesei; Proposed Significant New Use Rule

    Science.gov (United States)

    2012-06-13

    ... Register issue of April 11, 1997 (62 FR 17910) (FRL- 5577-2). C. Applicability of General Provisions... the Federal Register issue of April 24, 1990 (55 FR 17376), EPA has decided that the intent of TSCA..., entitled ``Regulatory Planning and Review'' (58 FR 51735, October 4, 1993). B. Paperwork Reduction...

  18. Structural insights into the β-xylosidase from Trichoderma reesei

    International Nuclear Information System (INIS)

    Xylan is a major structural polysaccharide in plant cells, and is the second most abundant polysaccharide in nature, accounting for approximately one-third of all renewable organic carbon on earth. Xylan together with cellulose (1,4-β-glucan) and lignin (a complex polyphenolic compound) make up the major polymeric constituents of plant cell walls, recently, there was a significant industrial interest in Xylan and its hydrolytic enzymatic complex, as a supplement in animal feed, for the manufacture of bread, food and drinks, textiles, bleaching of cellulose pulp, ethanol and xylitol production. (author)

  19. Structural insights into the {beta}-xylosidase from Trichoderma reesei

    Energy Technology Data Exchange (ETDEWEB)

    Rojas, Adriana L.; Fischer, Hannes; Polikarpov, Igor [Sao Paulo Univ. (USP), Sao Carlos, SP (Brazil). Inst. de Fisica; Eneiskaya, Elena V.; Kulminskaya, Anna A.; Shabalin, Konstantin A.; Neustroev, Kirill N.; Golubev, Alexander M. [Petersburg Nuclear Physics Inst., Moskow (Russian Federation); Craievich, Aldo Felix [Sao Paulo Univ. (USP), SP (Brazil). Inst. de Fisica

    2005-07-01

    Xylan is a major structural polysaccharide in plant cells, and is the second most abundant polysaccharide in nature, accounting for approximately one-third of all renewable organic carbon on earth. Xylan together with cellulose (1,4-{beta}-glucan) and lignin (a complex polyphenolic compound) make up the major polymeric constituents of plant cell walls, recently, there was a significant industrial interest in Xylan and its hydrolytic enzymatic complex, as a supplement in animal feed, for the manufacture of bread, food and drinks, textiles, bleaching of cellulose pulp, ethanol and xylitol production. (author)

  20. Redisposition of phoma-like anamorphs in Pleosporales

    NARCIS (Netherlands)

    de Gruyter, J; Woudenberg, J H C; Aveskamp, M M; Verkley, G J M; Groenewald, J Z; Crous, P W

    2013-01-01

    UNLABELLED: The anamorphic genus Phoma was subdivided into nine sections based on morphological characters, and included teleomorphs in Didymella, Leptosphaeria, Pleospora and Mycosphaerella, suggesting the polyphyly of the genus. Recent molecular, phylogenetic studies led to the conclusion that Pho

  1. Biodegradation of polycyclic aromatic hydrocarbons by Trichoderma species: a mini review.

    Science.gov (United States)

    Zafra, German; Cortés-Espinosa, Diana V

    2015-12-01

    Fungi belonging to Trichoderma genus are ascomycetes found in soils worldwide. Trichoderma has been studied in relation to diverse biotechnological applications and are known as successful colonizers of their common habitats. Members of this genus have been well described as effective biocontrol organisms through the production of secondary metabolites with potential applications as new antibiotics. Even though members of Trichoderma are commonly used for the commercial production of lytic enzymes, as a biological control agent, and also in the food industry, their use in xenobiotic biodegradation is limited. Trichoderma stands out as a genus with a great range of substrate utilization, a high production of antimicrobial compounds, and its ability for environmental opportunism. In this review, we focused on the recent advances in the research of Trichoderma species as potent and efficient aromatic hydrocarbon-degrading organisms, as well as aimed to provide insight into its potential role in the bioremediation of soils contaminated with heavy hydrocarbons. Several Trichoderma species are associated with the ability to metabolize a variety of both high and low molecular weight polycyclic aromatic hydrocarbons (PAHs) such as naphthalene, phenanthrene, chrysene, pyrene, and benzo[a]pyrene. PAH-degrading species include Trichoderma hamatum, Trichoderma harzianum, Trichoderma reesei, Trichoderma koningii, Trichoderma viride, Trichoderma virens, and Trichoderma asperellum using alternate enzyme systems commonly seen in other organisms, such as multicooper laccases, peroxidases, and ring-cleavage dioxygenases. Within these species, T. asperellum stands out as a versatile organism with remarkable degrading abilities, high tolerance, and a remarkable potential to be used as a remediation agent in polluted soils.

  2. Biodegradation of polycyclic aromatic hydrocarbons by Trichoderma species: a mini review.

    Science.gov (United States)

    Zafra, German; Cortés-Espinosa, Diana V

    2015-12-01

    Fungi belonging to Trichoderma genus are ascomycetes found in soils worldwide. Trichoderma has been studied in relation to diverse biotechnological applications and are known as successful colonizers of their common habitats. Members of this genus have been well described as effective biocontrol organisms through the production of secondary metabolites with potential applications as new antibiotics. Even though members of Trichoderma are commonly used for the commercial production of lytic enzymes, as a biological control agent, and also in the food industry, their use in xenobiotic biodegradation is limited. Trichoderma stands out as a genus with a great range of substrate utilization, a high production of antimicrobial compounds, and its ability for environmental opportunism. In this review, we focused on the recent advances in the research of Trichoderma species as potent and efficient aromatic hydrocarbon-degrading organisms, as well as aimed to provide insight into its potential role in the bioremediation of soils contaminated with heavy hydrocarbons. Several Trichoderma species are associated with the ability to metabolize a variety of both high and low molecular weight polycyclic aromatic hydrocarbons (PAHs) such as naphthalene, phenanthrene, chrysene, pyrene, and benzo[a]pyrene. PAH-degrading species include Trichoderma hamatum, Trichoderma harzianum, Trichoderma reesei, Trichoderma koningii, Trichoderma viride, Trichoderma virens, and Trichoderma asperellum using alternate enzyme systems commonly seen in other organisms, such as multicooper laccases, peroxidases, and ring-cleavage dioxygenases. Within these species, T. asperellum stands out as a versatile organism with remarkable degrading abilities, high tolerance, and a remarkable potential to be used as a remediation agent in polluted soils. PMID:26498812

  3. Validation of a novel sequential cultivation method for the production of enzymatic cocktails from Trichoderma strains.

    Science.gov (United States)

    Florencio, C; Cunha, F M; Badino, A C; Farinas, C S

    2015-02-01

    The development of new cost-effective bioprocesses for the production of cellulolytic enzymes is needed in order to ensure that the conversion of biomass becomes economically viable. The aim of this study was to determine whether a novel sequential solid-state and submerged fermentation method (SF) could be validated for different strains of the Trichoderma genus. Cultivation of the Trichoderma reesei Rut-C30 reference strain under SF using sugarcane bagasse as substrate was shown to be favorable for endoglucanase (EGase) production, resulting in up to 4.2-fold improvement compared with conventional submerged fermentation. Characterization of the enzymes in terms of the optimum pH and temperature for EGase activity and comparison of the hydrolysis profiles obtained using a synthetic substrate did not reveal any qualitative differences among the different cultivation conditions investigated. However, the thermostability of the EGase was influenced by the type of carbon source and cultivation system. All three strains of Trichoderma tested (T. reesei Rut-C30, Trichoderma harzianum, and Trichoderma sp INPA 666) achieved higher enzymatic productivity when cultivated under SF, hence validating the proposed SF method for use with different Trichoderma strains. The results suggest that this bioprocess configuration is a very promising development for the cellulosic biofuels industry.

  4. Novel traits of Trichoderma predicted through the analysis of its secretome.

    Science.gov (United States)

    Druzhinina, Irina S; Shelest, Ekaterina; Kubicek, Christian P

    2012-12-01

    Mycotrophic species of Trichoderma are among the most common fungi isolated from free soil, dead wood and as parasites on sporocarps of other fungi (mycoparasites). In addition, they undergo various other biotrophic associations ranging from rhizosphere colonization and endophytism up to facultative pathogenesis on such animals as roundworms and humans. Together with occurrence on a variety of less common substrata (marine invertebrates, artificial materials, indoor habitats), these lifestyles illustrate a wealthy opportunistic potential of the fungus. One tropical species, Trichoderma reesei, has become a prominent producer of cellulases and hemicellulases, whereas several other species are applied in agriculture for the biological control of phytopathogenic fungi. The sequencing of the complete genomes of the three species (T. reesei, T. virens, and T. atroviride) has led to a deepened understanding of Trichoderma lifestyle and its molecular physiology. In this review, we present the in silico predicted secretome of Trichoderma, and - in addition to the unique features of carbohydrate active enzymes - demonstrate the importance of such protein families as proteases, oxidative enzymes, and small cysteine-rich proteins, all of that received little attention in Trichoderma genetics so far. We also discuss the link between Trichoderma secretome and biology of the fungus.

  5. Selection of Trichoderma mutants with enhanced cellulase production and resistant to catabolite repression

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Megyeri L; Kovacs K; Zacchi G

    2004-01-01

    @@ Due to high cost and relatively low efficiency of cellulase enzymes used for the saccharification of pretreated lignocelluloses, the improvement of cellulase secreting microorganisms is of vital importance. Trichoderma reesei QM 6a, an excellent source of cellulase was selected in the late 1960's at Natick Laboratories by its performance on pure cellulose (Solka Floc, Avicel) . QM 6a is the wild parent strain of best existing hypercellulolytic mutants such as Rut C30, VTT-D-80133,L27, CL-847 and others. Utilization of cheaper carbon sources (e. g. , pretreated wood or straw) both in enzyme production and in hydrolysis necessitates to investigate fungal species other than T. reesei.

  6. Microjet printing of anamorphic microlens arrays

    Science.gov (United States)

    Cox, Weldon R.; Chen, Ting; Ussery, Daryl W.; Hayes, Donald J.; Hoenigman, R. F.; MacFarlane, Duncan L.; Rabinovich, Emmanuil M.

    1996-03-01

    The microjet printing method of micro-optical element fabrication is being used to make arrays of high-performance hemi-elliptical and hemi-cylindrical microlenses for potential use in applications such as collimation of edge-emitting diode laser array beams. The printing method enables both the fabrication of very fast (e.g., f/0.75) microlenses and the potential for reducing costs and increasing flexibility in micro-optics manufacture. The process for fabricating anamorphic microlenses, including those of square or rectangular shape, involves the dispensing and placing of precisely sized microdroplets of optical material onto optical substrates, and then controlling their coalescence and solidification. By varying the number, diameter and spacing of adjacent microdroplets of optical materials deposited at elevated temperatures onto heated substrate, both the dimensional aspect ratios and the ratio of `fast'- to-`slow' focal lengths of a printed hemi-elliptical microlens may be varied over a very wide range. Arrays of hemi-elliptical and hemi-cylindrical microlenses on the order of 100 - 300 micrometers in width and 150 micrometers to 20 mm long, with focal length ratios (fast/slow) from 1 (circular) to 0 (cylindrical), have been printed. A model for predicting printed hemi-elliptical microlens focal lengths from printed lenslet geometry is illustrated, along with an interferometric method of detecting lenslet defects and aberrations.

  7. Cordyceps bassiana and production of stromata in vitro showing Beauveria anamorph in Korea

    Science.gov (United States)

    A Cordyceps species was found with Beauveria anamorph state on larval insect cadavers on Obong Mountsin in Gangwon Pronvince, Republic of Korea. Cultures from discharged ascospores formed an anamorph identifiable as Beauveria bassiana. This teleomorph-anamorph connection was also confirmed by the in...

  8. Comparative analysis of microsatellites in five different antagonistic Trichoderma species for diversity assessment.

    Science.gov (United States)

    Rai, Shalini; Kashyap, Prem Lal; Kumar, Sudheer; Srivastava, Alok Kumar; Ramteke, Pramod W

    2016-01-01

    Microsatellites provide an ideal molecular markers system to screen, characterize and evaluate genetic diversity of several fungal species. Currently, there is very limited information on the genetic diversity of antagonistic Trichoderma species as determined using a range of molecular markers. In this study, expressed and whole genome sequences available in public database were used to investigate the occurrence, relative abundance and relative density of SSRs in five different antagonistic Trichoderma species: Trichoderma atroviride, T. harzianum, T. reesei, T. virens and T. asperellum. Fifteen SSRs loci were used to evaluate genetic diversity of twenty isolates of Trichoderma spp. from different geographical regions of India. Results indicated that relative abundance and relative density of SSRs were higher in T. asperellum followed by T. reesei and T. atroviride. Tri-nucleotide repeats (80.2%) were invariably the most abundant in all species. The abundance and relative density of SSRs were not influenced by the genome sizes and GC content. Out of eighteen primer sets, only 15 primer pairs showed successful amplification in all the test species. A total of 24 alleles were detected and five loci were highly informative with polymorphism information content values greater than 0.40, these markers provide useful information on genetic diversity and population genetic structure, which, in turn, can exploit for establishing conservation strategy for antagonistic Trichoderma isolates.

  9. Comparative analysis of microsatellites in five different antagonistic Trichoderma species for diversity assessment.

    Science.gov (United States)

    Rai, Shalini; Kashyap, Prem Lal; Kumar, Sudheer; Srivastava, Alok Kumar; Ramteke, Pramod W

    2016-01-01

    Microsatellites provide an ideal molecular markers system to screen, characterize and evaluate genetic diversity of several fungal species. Currently, there is very limited information on the genetic diversity of antagonistic Trichoderma species as determined using a range of molecular markers. In this study, expressed and whole genome sequences available in public database were used to investigate the occurrence, relative abundance and relative density of SSRs in five different antagonistic Trichoderma species: Trichoderma atroviride, T. harzianum, T. reesei, T. virens and T. asperellum. Fifteen SSRs loci were used to evaluate genetic diversity of twenty isolates of Trichoderma spp. from different geographical regions of India. Results indicated that relative abundance and relative density of SSRs were higher in T. asperellum followed by T. reesei and T. atroviride. Tri-nucleotide repeats (80.2%) were invariably the most abundant in all species. The abundance and relative density of SSRs were not influenced by the genome sizes and GC content. Out of eighteen primer sets, only 15 primer pairs showed successful amplification in all the test species. A total of 24 alleles were detected and five loci were highly informative with polymorphism information content values greater than 0.40, these markers provide useful information on genetic diversity and population genetic structure, which, in turn, can exploit for establishing conservation strategy for antagonistic Trichoderma isolates. PMID:26712623

  10. 77 FR 4903 - Trichoderma

    Science.gov (United States)

    2012-02-01

    ... Trichoderma virens strain GL-21) (40 CFR 180.1100)--see the Federal Register of September 20, 1995 (60 FR 48657) (FRL-4974-1) and October 5, 1995 (60 FR 52248) (FRL-4974-1). 2. Trichoderma harzianum Rifai.... Trichoderma harzianum strain T-39 (40 CFR 180.1201)--see the Federal Register of June 22, 2000 (65 FR...

  11. Comparative genomics provide insights into evolution of trichoderma nutrition style.

    Science.gov (United States)

    Xie, Bin-Bin; Qin, Qi-Long; Shi, Mei; Chen, Lei-Lei; Shu, Yan-Li; Luo, Yan; Wang, Xiao-Wei; Rong, Jin-Cheng; Gong, Zhi-Ting; Li, Dan; Sun, Cai-Yun; Liu, Gui-Ming; Dong, Xiao-Wei; Pang, Xiu-Hua; Huang, Feng; Liu, Weifeng; Chen, Xiu-Lan; Zhou, Bai-Cheng; Zhang, Yu-Zhong; Song, Xiao-Yan

    2014-02-01

    Saprotrophy on plant biomass is a recently developed nutrition strategy for Trichoderma. However, the physiology and evolution of this new nutrition strategy is still elusive. We report the deep sequencing and analysis of the genome of Trichoderma longibrachiatum, an efficient cellulase producer. The 31.7-Mb genome, smallest among the sequenced Trichoderma species, encodes fewer nutrition-related genes than saprotrophic T. reesei (Tr), including glycoside hydrolases and nonribosomal peptide synthetase-polyketide synthase. Homology and phylogenetic analyses suggest that a large number of nutrition-related genes, including GH18 chitinases, β-1,3/1,6-glucanases, cellulolytic enzymes, and hemicellulolytic enzymes, were lost in the common ancestor of T. longibrachiatum (Tl) and Tr. dN/dS (ω) calculation indicates that all the nutrition-related genes analyzed are under purifying selection. Cellulolytic enzymes, the key enzymes for saprotrophy on plant biomass, are under stronger purifying selection pressure in Tl and Tr than in mycoparasitic species, suggesting that development of the nutrition strategy of saprotrophy on plant biomass has increased the selection pressure. In addition, aspartic proteases, serine proteases, and metalloproteases are subject to stronger purifying selection pressure in Tl and Tr, suggesting that these enzymes may also play important roles in the nutrition. This study provides insights into the physiology and evolution of the nutrition strategy of Trichoderma.

  12. Anamorphic integral field spectrometer for diffuse ultraviolet astronomy.

    Science.gov (United States)

    Cook, Timothy

    2013-12-20

    We present the design of a novel anamorphic integral field spectrometer for diffuse ultraviolet astronomy. The system is designed to be able to measure emissions from faint diffuse astrophysical sources across a large field with good spectral resolving power. The design uses a standard focal plane image slicer and an anamorphic relay mirror and spectrometer to achieve an exceptional area, field of view, resolving power (AΩR) product, the key figure of merit for a spectroscopic system intended to study line emission from extended or diffuse sources. We present a typical design with R∼1000 and an effective etendue (AΩ) of 1.5×10(-4)cm2 sr.

  13. Trichoderma species from China

    Institute of Scientific and Technical Information of China (English)

    ZHANG Chu-long; XU Tong

    2004-01-01

    @@ Seventeen species of Trichoderma, isolated from soil or tree bark from China are identified based on morphological and physiological characters, and from their phylogenetic position inferred from parsimony analyses of nucleotide sequences of the internal transcribed spacer regions of the rDNA cluster (ITS1 and 2) and partial sequences of translation elongation factor 1-alpha (tef1) . There were T. citrinoviride, T. longibrachiatum, T. sinensis in section Longibrachiatum, T. atroviride, T.koningii, T. viride, T. asperellum, T. hamatum, T. erinaceum in section Trichoderma, T.harzianum (H.lixii) , T. inhamatum, T. velutinum , T. cerinum , T. strictipile , T. spirale ,T. virens, H. nigrovirens (Trichoderma sp.) in section Pachybasium. Among them four species:T. asperellum , T. velutinum , T. cerinum , T. spirale were reported firstly in China. In addition, two suspected new taxa (Trichoderma spp.) in Trichoderma section were proposed:Trichoderma sp. 1 (ZAUT261, 4, 4A, 15A, 2C), Trichoderma sp. 2 (2B, 5, 7A, 7B, 9A).Trichoderma sp. 1 was similar to T. hamatum , but the temperature optimum for mycelial growth was lower than that of T. hamatum and the species tended to form hemisphaerical pustule with Telatively larger conidia (average length 4.6 μm × 2.8 μm). Trichoderma sp. 2 was distinguished morphologically from related species T. strigosum, T. pubescens, T. erinaceum, T. hamatum and Trichoderma sp. 1 in pustules on CMD without fertile or sterile conidiophore elongation and distinctive phialide shape, the conidiophore branches similar to T. koningii, but the conidia similar to T. viride, subglobose, conspicuously tuberculate.

  14. Induction of mutation in Trichoderma viride for conversion of natural cellulose into glucose

    Energy Technology Data Exchange (ETDEWEB)

    Tahoun, M.K.; Khalil, A.I.; Helmi, S.; Khairy, A.H. [Univ. of Alexandria Research Centre, Alexandria (Egypt)

    1991-12-31

    The production of cellulolytic enzymes from fungi has been extensively studied. Several mutants of Trichoderma reesei were selected. Most of the studies were carried out on T. reesei, T. viride, T. harzianum, Penicillium funiculosum, Altemaria alternata. Aspergillus phoenicis, A. ustus, A. tamarii, A. japonicus, and A. niger. T. koningii is one of the most active producers of the so-called C, factor, which is indispensable for the rapid and extensive attack on crystal-line cellulose. However, Trichodenna is known to excrete only small amounts of {beta}-glucosidase. Therefore, Trichoderma is supplemented with {beta}-glucosidase from Aspergillus to increase the saccharification rate of cellulose to glucose as the main sugar. Induction of mutations in Trichodenna spp. rather than T. viride as a tool for the enhancement of {beta}-glucosidase activity was reported. Unfortunately, T. reesei is a poor producer of {beta}-glucosidase. On the other hand, T. harzianum M{sub 5}, a mutant that was induced by gamma radiation, produced high yields, not only of Avicelase and carboxy methyl cellulose, but also of {beta}-glucosidase, than its respective wild type.

  15. Fatal cutaneous mycosis in tentacled snakes caused by the chrysosporium anamorph of nannizziposis vriesii

    DEFF Research Database (Denmark)

    Bertelsen, Mads Frost; Crawshaw, Graham J.; Sigler, Lynne;

    2005-01-01

    The fungus Chrysosporium anamorph of Nannizziopsis vriesii was identified as the caurse of fatal, multifocal, heterophilic dermatitis in for freshwater aquatic captive-bred tentacled snakes......The fungus Chrysosporium anamorph of Nannizziopsis vriesii was identified as the caurse of fatal, multifocal, heterophilic dermatitis in for freshwater aquatic captive-bred tentacled snakes...

  16. Species concepts and biodiversity in Trichoderma and Hypocrea: from aggregate species to species clusters?

    Institute of Scientific and Technical Information of China (English)

    DRUZHININA Irina; KUBICEK Christian P.

    2005-01-01

    Trichoderma/Hypocrea is a genus of soil-borne or wood-decaying fungi containing members important to mankind as producers of industrial enzymes and biocontrol agents against plant pathogens, but also as opportunistic pathogens of immunocompromised humans. Species identification, while essential in view of the controversial properties of taxa ofthis genus, has been problematic by traditional methods. Here we will present a critical survey of the various identification methods in use. In addition,we will present an update on the taxonomy and phylogeny of the 88 taxa (which occur as 14 holomorphs, 49 teleomorphs and 25 anamorphs in nature) of Trichoderma/Hypocrea that have been confirmed by a combination of morphological, physiological and genetic approaches.

  17. The significance of cellulolytic enzymes produced by Trichoderma in opportunistic lifestyle of this fungus.

    Science.gov (United States)

    Strakowska, Judyta; Błaszczyk, Lidia; Chełkowski, Jerzy

    2014-07-01

    The degradation of native cellulose to glucose monomers is a complex process, which requires the synergistic action of the extracellular enzymes produced by cellulolytic microorganisms. Among fungi, the enzymatic systems that can degrade native cellulose have been extensively studied for species belonging to the genera of Trichoderma. The majority of the cellulolytic enzymes described so far have been examples of Trichoderma reesei, extremely specialized in the efficient degradation of plant cell wall cellulose. Other Trichoderma species, such as T. harzianum, T. koningii, T. longibrachiatum, and T. viride, known for their capacity to produce cellulolytic enzymes, have been isolated from various ecological niches, where they have proved successful in various heterotrophic interactions. As saprotrophs, these species are considered to make a contribution to the degradation of lignocellulosic plant material. Their cellulolytic potential is also used in interactions with plants, especially in plant root colonization. However, the role of cellulolytic enzymes in species forming endophytic associations with plants or in those existing in the substratum for mushroom cultivation remains unknown. The present review discusses the current state of knowledge about cellulolytic enzymes production by Trichoderma species and the encoding genes, as well as the involvement of these proteins in the lifestyle of Trichoderma.

  18. Snuffelen aan Trichoderma

    NARCIS (Netherlands)

    Baars, J.J.P.

    2011-01-01

    Problemen met groene schimmel in champignon-compost blijven zich voor doen, ondanks alle genomen maatregelen. In een onderzoek van Plant Research International wordt nu getracht, via geurprofielen Trichoderma aggressivum al tijdens het doorgroeien aan te tonen.

  19. 木霉菌属的定义及其属下分类%Delineation of the genus Trichoderma and its sub-genus division

    Institute of Scientific and Technical Information of China (English)

    杨合同; 唐文华; 徐砚珂; 王加宁; 姚碗生

    2002-01-01

    本文总结了木霉菌属的界定以及木霉菌属的属下分类.Bissett等人把木霉菌属(Trichoderma)分为5个组,共31个种.Hypocreanum组包括一个种,即T.lactea;Longibrachiatum组包括4个种,即T.longibrachiatum, T. citroviride, T. pseudokoingii 和 T. parceramosum; Saturnisporum 组包括2个种, 即T. saturnisporum 和 T. ghaneuse ; Pach ybasium 组包括20个种, 即T. crassum , T. croceum , T. f asciculatum , T. fertile, T. flavo f uscum , Trichoderma anamorph of Hypocrea gelatinosa, T. hamatum, T. harzianum, T. longipilis, T. minutisporum, T. oblongisporum, T. polysporum, T. pubescens, Trichoderma anamorph of Hypocrea semiorbis, T. spirale,T. strictipilis, T. striggosum, T. tomentosum 和 T. virens; Trichoderma 组包括4个种, 即T. viride,T.aureoviride,T.koningii和T.atroviride.以上是木霉菌属目前最完整的分类体系.

  20. Cellulolytic and xylanolytic potential of high β-glucosidase-producing Trichoderma from decaying biomass.

    Science.gov (United States)

    Okeke, Benedict C

    2014-10-01

    Availability, cost, and efficiency of microbial enzymes for lignocellulose bioconversion are central to sustainable biomass ethanol technology. Fungi enriched from decaying biomass and surface soil mixture displayed an array of strong cellulolytic and xylanolytic activities. Strains SG2 and SG4 produced a promising array of cellulolytic and xylanolytic enzymes including β-glucosidase, usually low in cultures of Trichoderma species. Nucleotide sequence analysis of internal transcribed spacer 2 (ITS2) region of rRNA gene revealed that strains SG2 and SG4 are closely related to Trichoderma inhamatum, Trichoderma piluliferum, and Trichoderma aureoviride. Trichoderma sp. SG2 crude culture supernatant correspondingly displayed as much as 9.84 ± 1.12, 48.02 ± 2.53, and 30.10 ± 1.11 units mL(-1) of cellulase, xylanase, and β-glucosidase in 30 min assay. Ten times dilution of culture supernatant of strain SG2 revealed that total activities were about 5.34, 8.45, and 2.05 orders of magnitude higher than observed in crude culture filtrate for cellulase, xylanase, and β-glucosidase, respectively, indicating that more enzymes are present to contact with substrates in biomass saccharification. In parallel experiments, Trichoderma species SG2 and SG4 produced more β-glucosidase than the industrial strain Trichoderma reesei RUT-C30. Results indicate that strains SG2 and SG4 have potential for low cost in-house production of primary lignocellulose-hydrolyzing enzymes for production of biomass saccharides and biofuel in the field.

  1. Novel Endophytic Trichoderma spp. Isolated from Healthy Coffea arabica Roots are Capable of Controlling Coffee Tracheomycosis

    Directory of Open Access Journals (Sweden)

    Temesgen Belayneh Mulaw

    2013-10-01

    Full Text Available One of the biggest threats to coffee growers in East Africa are emerging vascular wilt diseases (tracheomycosis caused by Fusarium spp. Many Trichoderma species are known to be natural antagonists of these pathogens and are widely used in biological control of fungal plant diseases. More recently, several Trichoderma spp., which exhibited high antifungal activity have been isolated as endophytes. Consequently, we have investigated the presence and the antagonistic activity of endophytic Trichoderma isolated from roots of healthy coffee plants (Coffea arabica from the major coffee growing regions of Ethiopia. Our results showed that community of Trichoderma spp. in roots of C. arabica contains fungi from coffee rhizosphere, as well as putatively obligate endophytic fungi. The putatively “true” endophytic species, until now, isolated only from coffee plant ecosystems in Ethiopia and recently described as T. flagellatum and novel T. sp. C.P.K. 1812 were able to antagonize Fusarium spp., which cause coffee tracheomycosis. Moreover, we found that strains of these species are also highly antagonistic against other phytopathogenic fungi, such as Alternaria alternata, Botryotinia fuckeliana (anamorph: Botrytis cinerea, and Sclerotinia sclerotiorum.

  2. Trichoderma volatiles effecting Arabidopsis

    DEFF Research Database (Denmark)

    Ramadan, Metwaly; Gigolashvili, Tamara; Grosskinsky, Dominik Kilian;

    2015-01-01

    Trichoderma species are present in many ecosystems and some strains have the ability to reduce the severity of plant diseases by activating various defense pathways via specific biologically active signaling molecules. Hence we investigated the effects of low molecular weight volatile compounds...... of Trichoderma asperellum IsmT5 on Arabidopsis thaliana. During co-cultivation of T. asperellum IsmT5 without physical contact to A. thaliana we observed smaller but vital and robust plants. The exposed plants exhibit increased trichome numbers, accumulation of defense-related compounds such as H2O2, anthocyanin......, camalexin, and increased expression of defense-related genes. We conclude that A. thaliana perceives the Trichoderma volatiles as stress compounds and subsequently initiates multilayered adaptations including activation of signaling cascades to withstand this environmental influence. The prominent headspace...

  3. Crystallization and Preliminary X-ray Diffraction Analysis of the Glucuronoyl Esterase Catalytic Domain from Hypocrea jecorina

    Science.gov (United States)

    The catalytic domain of the glucuronoyl esterase from Hypocrea jecorina (anamorph Trichoderma reesei) was over-expressed, purified, and crystallized by sitting-drop vapor-diffusion method using 1.4 M sodium/potassium phosphate pH 6.9. Crystals had space group P212121 and X-ray diffraction data were...

  4. Trichoderma inoculation augments grain amino acids and mineral nutrients by modulating arsenic speciation and accumulation in chickpea (Cicer arietinum L.).

    Science.gov (United States)

    Tripathi, Pratibha; Singh, Poonam C; Mishra, Aradhana; Tripathi, Rudra D; Nautiyal, Chandra S

    2015-07-01

    Trichoderma reesei is an industrially important fungi which also imparts stress tolerance and plant growth promotion in various crops. Arsenic (As) contamination of field soils is one of the challenging problems in agriculture, posing potential threats for both human health and the environment. Plants in association with microbes are a liable method to improve metal tolerance and enhance crop productivity. Chickpea (Cicer arietinum L.), is an important grain legume providing cheap source of protein in semi-arid regions including As affected areas. In this study we report the role of T. reesei NBRI 0716 (NBRI 0716) in supporting chickpea growth and improving soil quality in As simulated conditions. NBRI 0716 modulated the As speciation and its availability to improve grain yield and quality (amino acids and mineral content) in chickpea (C. arietinum L.) plants grown in As spiked soil (100 mg As kg(-1) soil). Arsenic accumulation and speciation results indicate that arsenate [As(V)] was the dominant species in chickpea seeds and rhizosphere soil. The Trichoderma reduced total grain inorganic As (Asi) by 66% and enhanced dimethylarsonic acid (DMA) and monomethylarsinic acid (MMA) content of seed and rhizosphere soil. The results indicate a probable role of NBRI 0716 in As methylation as the possible mechanism for maneuvering As stress in chickpea. Analysis of functional diversity using carbon source utilization (Biolog) showed significant difference in diversity and evenness indices among the soil microbial rhizosphere communities. Microbial diversity loss caused by As were prevented in the presence of Trichoderma NBRI 0716.

  5. Optimization of solid fermentation of cellulase from Trichoderma koningii

    Institute of Scientific and Technical Information of China (English)

    LI Pei-jun; JING De-bing; ZHOU Qi-xing; ZHANG Chun-gui

    2004-01-01

    To exploit peashrub resources in Ordos as fodders, it is very crucial to realize industrial production of cheap cellulase of high activity by optimizing culture technology, especially culture substrate. In this study, a new prescription experiment based on uniform design ideal was invented and successfully applied in the solid fermentation of Trichoderma koningii F244, which being performed with two different temperature degrees. The activities of FPA, cotton lyase, CMCase and β-glucosidase were assayed and then mathematical models of enzymatic activities, which were figured out by Unconstraint Mathematical Programming, were developed by Multivariate Regression Program of SPSS10.0. Enzymatic activities of optimized substrate prescriptions corresponding to mathematical models were forecasted to determine an ideal substrate prescription. It is revealed that in solid fermentation, Tween80 has negative effect on cellulase production. Furthermore, the ideal prescription for cellulase complex production by Trichoderma koningii F244 was straw powder 16.9%,wheat bran 26.5%, (NH4)2SO4 9.5% and water 47.1%, whose corresponding cellulase activity was expected to be at the same high level with that of Trichoderma reesei Q9414 on its own recommended substrate. Especially, goats mainly fed on peashrub tissues mixed with cellulase complex of this prescription and culture technology, got an incremental ratio of 0.3 kg/d, which brought a very promising feeding prospect for local peashrub resource. By populization of this cellulase complex, it can integrate living standard, economic construction of local residents into vegetational restoration tightly and thus this paper will be very meaningful to be use for reference for western China like Ordos to realize its sustainable development of economy, society and environment.

  6. Genome Sequencing and Comparative Analysis of the Biocontrol Agent Trichoderma harzianum sensu stricto TR274

    Energy Technology Data Exchange (ETDEWEB)

    Steindorff, Andrei S.; Noronha, Elilane F.; Ulhoa, Cirano J.; Kuo, Alan; Salamov, Asaf A.; Haridas, Sajeet; Riley, Robert W.; Druzhinina, Irina S.; Kubicek, Christian P.; Grigoriev, Igor V.

    2015-03-17

    Biological control is a complex process which requires many mechanisms and a high diversity of biochemical pathways. The species of Trichoderma harzianum are well known for their biocontrol activity against many plant pathogens. To gain new insights into the biocontrol mechanism used by T. harzianum, we sequenced the isolate TR274 genome using Illumina. The assembly was performed using AllPaths-LG with a maximum coverage of 100x. The assembly resulted in 2282 contigs with a N50 of 37033bp. The genome size generated was 40.8 Mb and the GC content was 47.7%, similar to other Trichoderma genomes. Using the JGI Annotation Pipeline we predicted 13,932 genes with a high transcriptome support. CEGMA tests suggested 100% genome completeness and 97.9% of RNA-SEQ reads were mapped to the genome. The phylogenetic comparison using orthologous proteins with all Trichoderma genomes sequenced at JGI, corroborates the Trichoderma (T. asperellum and T. atroviride), Longibrachiatum (T. reesei and T. longibrachiatum) and Pachibasium (T. harzianum and T. virens) section division described previously. The comparison between two Trichoderma harzianum species suggests a high genome similarity but some strain-specific expansions. Analyses of the secondary metabolites, CAZymes, transporters, proteases, transcription factors were performed. The Pachybasium section expanded virtually all categories analyzed compared with the other sections, specially Longibrachiatum section, that shows a clear contraction. These results suggests that these proteins families have an important role in their respective phenotypes. Future analysis will improve the understanding of this complex genus and give some insights about its lifestyle and the interactions with the environment.

  7. The Genomes of Three Uneven Siblings: Footprints of the Lifestyles of Three Trichoderma Species.

    Science.gov (United States)

    Schmoll, Monika; Dattenböck, Christoph; Carreras-Villaseñor, Nohemí; Mendoza-Mendoza, Artemio; Tisch, Doris; Alemán, Mario Ivan; Baker, Scott E; Brown, Christopher; Cervantes-Badillo, Mayte Guadalupe; Cetz-Chel, José; Cristobal-Mondragon, Gema Rosa; Delaye, Luis; Esquivel-Naranjo, Edgardo Ulises; Frischmann, Alexa; Gallardo-Negrete, Jose de Jesus; García-Esquivel, Monica; Gomez-Rodriguez, Elida Yazmin; Greenwood, David R; Hernández-Oñate, Miguel; Kruszewska, Joanna S; Lawry, Robert; Mora-Montes, Hector M; Muñoz-Centeno, Tania; Nieto-Jacobo, Maria Fernanda; Nogueira Lopez, Guillermo; Olmedo-Monfil, Vianey; Osorio-Concepcion, Macario; Piłsyk, Sebastian; Pomraning, Kyle R; Rodriguez-Iglesias, Aroa; Rosales-Saavedra, Maria Teresa; Sánchez-Arreguín, J Alejandro; Seidl-Seiboth, Verena; Stewart, Alison; Uresti-Rivera, Edith Elena; Wang, Chih-Li; Wang, Ting-Fang; Zeilinger, Susanne; Casas-Flores, Sergio; Herrera-Estrella, Alfredo

    2016-03-01

    The genus Trichoderma contains fungi with high relevance for humans, with applications in enzyme production for plant cell wall degradation and use in biocontrol. Here, we provide a broad, comprehensive overview of the genomic content of these species for "hot topic" research aspects, including CAZymes, transport, transcription factors, and development, along with a detailed analysis and annotation of less-studied topics, such as signal transduction, genome integrity, chromatin, photobiology, or lipid, sulfur, and nitrogen metabolism in T. reesei, T. atroviride, and T. virens, and we open up new perspectives to those topics discussed previously. In total, we covered more than 2,000 of the predicted 9,000 to 11,000 genes of each Trichoderma species discussed, which is >20% of the respective gene content. Additionally, we considered available transcriptome data for the annotated genes. Highlights of our analyses include overall carbohydrate cleavage preferences due to the different genomic contents and regulation of the respective genes. We found light regulation of many sulfur metabolic genes. Additionally, a new Golgi 1,2-mannosidase likely involved in N-linked glycosylation was detected, as were indications for the ability of Trichoderma spp. to generate hybrid galactose-containing N-linked glycans. The genomic inventory of effector proteins revealed numerous compounds unique to Trichoderma, and these warrant further investigation. We found interesting expansions in the Trichoderma genus in several signaling pathways, such as G-protein-coupled receptors, RAS GTPases, and casein kinases. A particularly interesting feature absolutely unique to T. atroviride is the duplication of the alternative sulfur amino acid synthesis pathway.

  8. The Genomes of Three Uneven Siblings: Footprints of the Lifestyles of Three Trichoderma Species.

    Science.gov (United States)

    Schmoll, Monika; Dattenböck, Christoph; Carreras-Villaseñor, Nohemí; Mendoza-Mendoza, Artemio; Tisch, Doris; Alemán, Mario Ivan; Baker, Scott E; Brown, Christopher; Cervantes-Badillo, Mayte Guadalupe; Cetz-Chel, José; Cristobal-Mondragon, Gema Rosa; Delaye, Luis; Esquivel-Naranjo, Edgardo Ulises; Frischmann, Alexa; Gallardo-Negrete, Jose de Jesus; García-Esquivel, Monica; Gomez-Rodriguez, Elida Yazmin; Greenwood, David R; Hernández-Oñate, Miguel; Kruszewska, Joanna S; Lawry, Robert; Mora-Montes, Hector M; Muñoz-Centeno, Tania; Nieto-Jacobo, Maria Fernanda; Nogueira Lopez, Guillermo; Olmedo-Monfil, Vianey; Osorio-Concepcion, Macario; Piłsyk, Sebastian; Pomraning, Kyle R; Rodriguez-Iglesias, Aroa; Rosales-Saavedra, Maria Teresa; Sánchez-Arreguín, J Alejandro; Seidl-Seiboth, Verena; Stewart, Alison; Uresti-Rivera, Edith Elena; Wang, Chih-Li; Wang, Ting-Fang; Zeilinger, Susanne; Casas-Flores, Sergio; Herrera-Estrella, Alfredo

    2016-03-01

    The genus Trichoderma contains fungi with high relevance for humans, with applications in enzyme production for plant cell wall degradation and use in biocontrol. Here, we provide a broad, comprehensive overview of the genomic content of these species for "hot topic" research aspects, including CAZymes, transport, transcription factors, and development, along with a detailed analysis and annotation of less-studied topics, such as signal transduction, genome integrity, chromatin, photobiology, or lipid, sulfur, and nitrogen metabolism in T. reesei, T. atroviride, and T. virens, and we open up new perspectives to those topics discussed previously. In total, we covered more than 2,000 of the predicted 9,000 to 11,000 genes of each Trichoderma species discussed, which is >20% of the respective gene content. Additionally, we considered available transcriptome data for the annotated genes. Highlights of our analyses include overall carbohydrate cleavage preferences due to the different genomic contents and regulation of the respective genes. We found light regulation of many sulfur metabolic genes. Additionally, a new Golgi 1,2-mannosidase likely involved in N-linked glycosylation was detected, as were indications for the ability of Trichoderma spp. to generate hybrid galactose-containing N-linked glycans. The genomic inventory of effector proteins revealed numerous compounds unique to Trichoderma, and these warrant further investigation. We found interesting expansions in the Trichoderma genus in several signaling pathways, such as G-protein-coupled receptors, RAS GTPases, and casein kinases. A particularly interesting feature absolutely unique to T. atroviride is the duplication of the alternative sulfur amino acid synthesis pathway. PMID:26864432

  9. A Comprehensive Characterization of Simple Sequence Repeats in the Sequenced Trichoderma Genomes Provides Valuable Resources for Marker Development.

    Science.gov (United States)

    Mahfooz, Sahil; Singh, Satyendra P; Rakh, Ramraje; Bhattacharya, Arpita; Mishra, Nishtha; Singh, Poonam C; Chauhan, Puneet S; Nautiyal, Chandra S; Mishra, Aradhana

    2016-01-01

    Members of genus Trichoderma are known worldwide for mycoparasitism. To gain a better insight into the organization and evolution of their genomes, we used an in silico approach to compare the occurrence, relative abundance and density of SSRs in Trichoderma atroviride, T. harzianum, T. reesei, and T. virens. Our analysis revealed that in all the four genome sequences studied, the occurrence, relative abundance, and density of microsatellites varied and was not influenced by genome sizes. The relative abundance and density of SSRs positively correlated with the G + C content of their genomes. The maximum frequency of SSRs was observed in the smallest genome of T. reesei whereas it was least in second smallest genome of T. atroviride. Among different classes of repeats, the tri-nucleotide repeats were abundant in all the genomes and accounts for ∼38%, whereas hexa-nuceotide repeats were the least (∼10.2%). Further evaluation of the conservation of motifs in the transcript sequences shows a 49.5% conservation among all the motifs. In order to study polymorphism in Trichoderma isolates, 12 polymorphic SSR markers were developed. Of the 12 markers, 6 markers are from T. atroviride and remaining 6 belong to T. harzianum. SSR markers were found to be more polymorphic from T. atroviride with an average polymorphism information content value of 0.745 in comparison with T. harzianum (0.615). Twelve polymorphic markers obtained in this study clearly demonstrate the utility of newly developed SSR markers in establishing genetic relationships among different isolates of Trichoderma. PMID:27199911

  10. Nucleosome transactions on the Hypocrea jecorina (Trichoderma reesei) cellulase promoter cbh2 associated with cellulase induction.

    Science.gov (United States)

    Zeilinger, S; Schmoll, M; Pail, M; Mach, R L; Kubicek, C P

    2003-10-01

    The 5' regulatory region of the cbh2 gene of Hypocrea jecorina contains the cbh2 activating element (CAE) which is essential for induction of cbh2 gene expression by sophorose and cellulose. The CAE consists of two motifs, a CCAAT box on the template strand and a GTAATA box on the coding strand, which cooperate during induction. Northern analyses of cbh2 gene expression has revealed an absolute dependence on induction, but no direct effect of Cre1-mediated carbon catabolite repression. Investigation of the chromatin structure in the wild-type strain showed that, under repressing conditions, there is a nucleosome free region (nfr) around the CAE, which is flanked by strictly positioned nucleosomes. Induction results in a loss of positioning of nucleosomes -1 and -2 downstream of the CAE, thus making the TATA box accessible. Simultaneous mutation of both motifs of the CAE, or of the CCAAT-box alone, also leads to shifting of nucleosome -1, which normally covers the TATA-box under repressing conditions, whereas mutation of the GTAATA element results in a narrowing of the nfr, indicating that the proteins that bind to both motifs in the CAE interact with chromatin, although in different ways. A cellulase-negative mutant strain, which has previously been shown to be altered in protein binding to the CAE, still displayed the induction-specific changes in nucleosome structure, indicating that none of the proteins that directly interact with CAE are affected, and that nucleosome rearrangement and induction of cbh2 expression are uncoupled. Interestingly, the carbon catabolite repressor Cre1 is essential for strict nucleosome positioning in the 5' regulatory sequences of cbh2 under all of the conditions tested, and induction can occur in a promoter that lacks positioned nucleosomes. These data suggest that Cre1, the Hap2/3/5 complex and the GTAATA-binding protein are all involved in nucleosome assembly on the cbh2 promoter, and that the latter two respond to inducing conditions by repositioning nucleosome -1. PMID:12905071

  11. The properties of catalytically-inactivated Trichoderma reesei cellobiohydrolase I: Role of the cellulose binding domain

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J.; Donner, T.R.; Affholter, K.A. [Oak Ridge National Lab., TN (United States)

    1993-12-31

    Cellobiohydrolase I (CBH I) was purified from a crude cellulase by preparative isoelectric focusing. Treatment of CBH I with 1-ethyl-3-3(3-dimethylaminopropyl)-carbodiimide (EDC) resulted in its catalytic inactivation but did not abolish its ability to be absorbed to microcrystalline cellulose (Avicel). CBH I thus modified possessed a pI of between 8.5 and 9.3 and decreased tryptophan fluorescence compared to native CBH I. A comparison of the effect of native and modified CBH I on the morphology of crystalline cotton cellulose fibers was made using scanning electron microscopy.

  12. Trichoderma reesei CE16 acetyl esterase and its role in enzymatic degradation of acetylated hemicellulose

    DEFF Research Database (Denmark)

    Biely, Peter; Cziszarava, Maria; Agger, Jane W.;

    2014-01-01

    Results The combined action of GH10 xylanase and acetylxylan esterases (AcXEs) leads to formation of neutral and acidic xylooligosaccharides with a few resistant acetyl groups mainly at their non-reducing ends. We show here that these acetyl groups serve as targets for TrCE16 AcE. The most promin...

  13. IONIC LIQUID SALT-INDUCED INACTIVATION AND UNFOLDING OF CELLULASE FROM TRICHODERMA REESEI. (R828257)

    Science.gov (United States)

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  14. Construction of a cellulase hyper-expression system in Trichoderma reesei by promoter and enzyme engineering

    NARCIS (Netherlands)

    Zou, G.; Shi, S.; Jiang, Y.; van den Brink, J.; de Vries, R.P.; Chen, L.; Zhang, J.; Ma, L.; Wang, C.; Zhou, Z.

    2012-01-01

    Background A bacterial strain previously isolated from pyrite mine drainage and named BAS-10 was tentatively identified as Klebsiella oxytoca. Unlikely other enterobacteria, BAS-10 is able to grow on Fe(III)-citrate as sole carbon and energy source, yielding acetic acid and CO2 coupled with Fe(III)

  15. A study of Cyathicula strobilina and its Chalara anamorph in vitro

    OpenAIRE

    Gams, Walter; Philippi, Susanne

    1992-01-01

    The connection of Cyathicula strobilina (Fr.: Fr.) Korf & Dixon with the Chalara strobilina Sacc. anamorph is confirmed by means of ascospore isolations. The in vitro morphology of the latter is described in detail and the heterogeneity of the genus Chalara is pointed out.

  16. The production of Multiple Small Peptaibol Families by Single 14-Module Peptide Synthetases in Trichoderma/Hypocrea

    Energy Technology Data Exchange (ETDEWEB)

    Degenkolb, Thomas; Aghchehb, Razieh Karimi; Dieckmann, Ralf; Neuhof, Torsten; Baker, Scott E.; Druzhinina, Irina S.; Kubicek, Christian P.; Brückner, Hans; von Dohren, Hans

    2012-03-01

    The most common peptaibibiotic structures are 11-residue peptaibols found widely distributed in the genus Trichoderma/Hypocrea. Frequently associated are 14-residue peptaibols sharing partial sequence identity. Genome sequencing projects of 3 Trichoderma strains of the major clades reveal the presence of up to 3 types of nonribosomal peptide synthetases with 7, 14, or 18-20 amino acid adding modules. We here provide evidence that the 14-module NRPS type found in T. virens, T. reesei (teleomorph Hypocrea jecorina) and T. atroviride produces both 11- and 14- residue peptaibols based on the disruption of the respective NRPS gene of T. reesei, and bioinformatic analysis of their amino acid activating domains and modules. The structures of these peptides may be predicted from the gene structures and have been confirmed by analysis of families of 11- and 14-residue peptaibols from the strain 618, termed hypojecorins A (23 sequences determined, 4 new) and B (3 new sequences), and the recently established trichovirins A from T. virens. The distribution of 11- and 14-residue products is strain-specific and depends on growth conditions as well. Possible mechanisms of module skipping are discussed.

  17. Copper tolerance of Trichoderma species

    Directory of Open Access Journals (Sweden)

    Jovičić-Petrović Jelena

    2014-01-01

    Full Text Available Some Trichoderma strains can persist in ecosystems with high concentrations of heavy metals. The aim of this research was to examine the variability of Trichoderma strains isolated from different ecosystems, based on their morphological properties and restriction analysis of ITS fragments. The fungal growth was tested on potato dextrose agar, amended with Cu(II concentrations ranging from 0.25 to 10 mmol/l, in order to identify copper-resistant strains. The results indicate that some isolated strains of Trichoderma sp. show tolerance to higher copper concentrations. Further research to examine the ability of copper bioaccumulation by tolerant Trichoderma strains is needed. [Projekat Ministarstva nauke Republike Srbije, br. TR 31080 i br. III 43010

  18. Filter Paper Degrading Ability of a Trichoderma Strain With Multinucleate Conidia

    Science.gov (United States)

    Toyama, Hideo; Yano, Makiko; Hotta, Takeshi; Toyama, Nobuo

    The multinucleate conidia were produced from the green mature conidia of Trichoderma reesei Rut C-30 strain by colchicine treatment. The strain with higher Filter paper degrading ability was selected among those conidia using a double layer selection medium. The selected strain, JS-2 was able to collapse the filter paper within 15 min but the original strain took 25 min to collapse it completely. Moreover, the amount of reducing sugar in the L-type glass tube of the strain, JS-2, was greater than that of the original strain. The Avicel, CMC-Na, and Salicin hydrolyzing activity of the strain, JS-2, increased 2.1 times, 1.2 times, and 3.6 times higher than that of the original strain.

  19. Enzymes of Trichoderma and other origin in biopreparation of cotton

    Institute of Scientific and Technical Information of China (English)

    Anita Losonczi; George Szakacs; Emilia Csiszar; Outi Kareela

    2004-01-01

    @@ Preparation and bleaching are among the most energy and chemical intensive steps of the traditional cotton finishing. About 75 % of the organic pollutants arising from textile finishing are derived from the preparation of cotton goods. Biopreparation may be a valuable and environmentally friendly alternative to harsh alkaline chemicals for preparing of cotton. Different enzymes i. e. cellulases,pectinases, lipases and proteases have been tested for biopreparation. Many papers have been published how enzymes effect on the properties of cotton fabrics in respect to their effectiveness to replace alkaline scouring and how enzymes modify the various cell wall components of cotton fibers. However,very few data have been reported on bleachability and dyeing behavior of the bioscoured fabrics. In this work an acidic cellulase (Celluclast 1.5 L, Trichoderma reesei origin), an acidic pectinasehemicellulase (Viscozyme 120 L , Aspergillus aculeatus ) and a cellulase-free xylanase (Pulpzyme HC,Bacillus sp. ) enzymes were used for bioscouring of greige cotton fabric. Selected properties of the pretreated samples, which are important for effective dyeing, are characterized in detail. Effect of hydrogen peroxide bleaching, applied subsequent to the biopretreatment on the bleachability and dyeing properties of the samples is also elucidated. A reactive dye was used at four different concentrations to compare the dyeing behavior of biopretreated as well as biopretreated and subsequently bleached cotton fabrics with that of the alkaline scoured as well as alkaline scoured and bleached fabrics, respectively. Color evenness and uniformity, and wash fastness properties of the dyed samples are also determined[1].

  20. Nonminimally coupled inflation with initial conditions from a preinflation anamorphic contracting era

    Science.gov (United States)

    McDonald, John

    2016-08-01

    Inflation due to a nonminimally coupled scalar field, as first proposed by Salopek, Bardeen and Bond (SBB), is in good agreement with the observed value of the spectral index and constraints on the tensor-to-scalar ratio. Here we explore the possibility that SBB inflation represents the late stage of a Universe which emerges from an early contracting era. We present a model in which the Universe smoothly transitions from an anamorphic contracting era to late-time SBB inflation without encountering a singular bounce. This corresponds to a continuous expansion in the Einstein frame throughout. We show that the anamorphic contracting era is able to provide the smooth superhorizon initial conditions necessary for subsequent SBB inflation to occur. The model predicts corrections to the nonminimal coupling, kinetic term and potential of SBB inflation which can observably increase the spectral index relative to its SBB prediction.

  1. Non-Minimally Coupled Inflation with a Pre-Inflation Anamorphic Contracting Era

    CERN Document Server

    McDonald, John

    2015-01-01

    Inflation due to a non-minimally coupled scalar field, as first proposed by Salopek, Bardeen and Bond (SBB), is in good agreement with the observed value of spectral index and constraints on the tensor-to-scalar ratio. Here we explore the possibility that SBB inflation represents the late stage of a Universe which emerges from an early contracting era. We present a model in which the Universe smoothly transitions from an anamorphic contracting era to late-time SBB inflation without encountering a singular bounce. This corresponds to a continuous expansion in the Einstein frame throughout. We show that the anamorphic contracting era is able to provide the smooth superhorizon initial conditions necessary for subsequent SBB inflation to occur. The model predicts corrections to the non-minimal coupling, kinetic term and potential of SBB inflation which can observably increase the observed spectral index relative to its SBB prediction.

  2. A comprehensive characterization of simple sequence repeats in the sequenced Trichoderma genomes provides valuable resources for marker development

    Directory of Open Access Journals (Sweden)

    Sahil eMahfooz

    2016-04-01

    Full Text Available Members of genus Trichoderma are known worldwide for mycoparasitism. To gain a better insight into the organization and evolution of their genomes, we used an in-silico approach to compare the occurrence, relative abundance and density of SSRs in T.atroviride, T. harzianum, T. reesei, and T. virens. Our analysis revealed that in all the four genome sequences studied, the occurrence, relative abundance, and density of microsatellites varied and was not influenced by genome sizes. The relative abundance and density of SSRs positively correlated with the G+C content of their genomes. The maximum frequency of SSRs was observed in the smallest genome of T. reesei whereas it was least in second smallest genome of T. atroviride. Among different classes of repeats, the tri-nucleotide repeats were abundant in all the genomes and accounts for ~38%, whereas hexa-nuceotide repeats were the least (~10.2%. Further evaluation of the conservation of motifs in the transcript sequences shows a 49.5% conservation among all the motifs. In order to study polymorphism in Trichoderma isolates, 12 polymorphic SSR markers were developed. Of the 12 markers, 6 markers are from T. atroviride and remaining 6 belong to T. harzianum. SSR markers were found to be more polymorphic from T. atroviride with an average polymorphism information content value of 0.745 in comparison with T. harzianum (0.615. Twelve polymorphic markers obtained in this study clearly demonstrate the utility of newly developed SSR markers in establishing genetic relationships among different isolates of Trichoderma.

  3. Disentangling the Trichoderma viridescens complex

    NARCIS (Netherlands)

    Jaklitsch, W.M.; Samuels, G.J.; Ismaiel, A.; Voglmayr, H.

    2013-01-01

    Trichoderma viridescens is recognised as a species complex. Multigene analyses based on the translation elongation factor 1-alpha encoding gene (tef1), a part of the rpb2 gene, encoding the second largest RNA polymerase subunit and the larger subunit of ATP citrate lyase (acl1) reveals 13 phylogenet

  4. Rapid Isolation of the Trichoderma Strain with Higher Degrading Ability of a Filter Paper and Superior Proliferation Characteristics Using Avicel Plates and the Double-Layer Selection Medium

    Science.gov (United States)

    Toyama, Hideo; Nakano, Megumi; Satake, Yuuki; Toyama, Nobuo

    The cost of cellulase is still a problem for bioethanol production. As the cellulase of Trichoderma reesei is applicable for producing ethanol from cellulosic materials, the cellulase productivity of this fungus should be increased. Therefore, we attempted to develop a system to isolate the strain with higher degrading ability of a filter paper and superior proliferation characteristics among the conidia treated with the mitotic arrester, colchicine. When green mature conidia of T. reesei RUT C-30 were swollen, autopolyploidized, and incubated in the double-layer selection medium containing Avicel, colonies appeared on the surface earlier than the original strain. When such colonies and the original colony were incubated on the Avicel plates, strain B5, one of the colonies derived from the colchicinetreated conidia, showed superior proliferation characteristics. Moreover, when strain B5 and the original strain were compared in the filter paper degrading ability and the cellulose hydrolyzing activity, strain B5 was also superior to the original strain. It was suspected that superior proliferation characteristics of strain B5 reflects higher filter paper degrading ability. Thus, we concluded that the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics can be isolated using Avicel plates and the double-layer selection medium.

  5. Rapid isolation of the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics using avicel plates and the double-layer selection medium.

    Science.gov (United States)

    Toyama, Hideo; Nakano, Megumi; Satake, Yuuki; Toyama, Nobuo

    2008-03-01

    The cost of cellulase is still a problem for bioethanol production. As the cellulase of Trichoderma reesei is applicable for producing ethanol from cellulosic materials, the cellulase productivity of this fungus should be increased. Therefore, we attempted to develop a system to isolate the strain with higher degrading ability of a filter paper and superior proliferation characteristics among the conidia treated with the mitotic arrester, colchicine. When green mature conidia of T. reesei RUT C-30 were swollen, autopolyploidized, and incubated in the double-layer selection medium containing Avicel, colonies appeared on the surface earlier than the original strain. When such colonies and the original colony were incubated on the Avicel plates, strain B5, one of the colonies derived from the colchicine-treated conidia, showed superior proliferation characteristics. Moreover, when strain B5 and the original strain were compared in the filter paper degrading ability and the cellulose hydrolyzing activity, strain B5 was also superior to the original strain. It was suspected that superior proliferation characteristics of strain B5 reflects higher filter paper degrading ability. Thus, we concluded that the Trichoderma strain with higher degrading ability of a filter paper and superior proliferation characteristics can be isolated using Avicel plates and the double-layer selection medium.

  6. Factors that contribute to the mycoparasitism stimulus in Trichoderma atroviride strain P1

    Institute of Scientific and Technical Information of China (English)

    Woo S L; Lorito M; Formisano E; Fogliano V; Cosenza C; Mauro A; Turrà D; Soriente I; Ferraioli S; Scala F

    2004-01-01

    @@ Trichoderma atroviride strain P1 has been used extensively to study the mycoparasitic mechanisms in the interaction between plant pathogenic host and beneficial antagonistic fungi. Mutants of P1 containing the green fluorescent protein (gfp) or glucose oxidase (gox) reporter systems and different inducible promoters (from the exochitinase nag1 gene, or the endochitinase ech42 gene of P1) were used to determine the factors that activate the biocontrol gene expression cascade in the antagonist. The following compounds were tested singly and in various combinations: purified Trichoderma P1 enzymes (endochitinase, exochitinase, chitobiosidase,glucanase); antagonist culture filtrates (T. atroviride P1 wild-type and relative knock-out mutants, T.harzianum, T. reesei); pathogen culture filtrates (Botrytis, Pythium, Rhizoctonia); purified fungal cell walls (CWs) from Trichoderma , Botrytis, Pythium, Rhizoctonia; colloidal crab shell chitin; and plant extracts from cucumber leaves, stems or roots. Strong induction of mycoparasitism was found with the various digestion products produced by treating fungal CWs and colloidal chitin with purified enzymes or fungal culture filtrates. Filtrates from chitinase knock-out mutants, as well as CWs from Oomycetes fungi, were less active in producing the stimulus for mycoparasitism. The host CW digestion products were separated by molecular weight (MW) to determine which compounds were able to activate Trichoderma. Micromolecules of MW less than 3 kDa were found to trigger mycoparasitism gene expression before physical contact with the host pathogen. These compounds stimulated mycelial growth and spore germination of the antagonist. Purification of these host-derived compounds was conducted by HPLC and in vivo assay. The obtained inducers were able to stimulate both the production of endochitinase and exochitinase enzymes, even under repressing conditions in the presence of glucose. Inducers stimulated the biocontrol effect of P1 in

  7. The anamorphic genus Monotosporella (Ascomycota) from Eocene amber and from modern Agathis resin.

    Science.gov (United States)

    Sadowski, Eva-Maria; Beimforde, Christina; Gube, Matthias; Rikkinen, Jouko; Singh, Hukam; Seyfullah, Leyla J; Heinrichs, Jochen; Nascimbene, Paul C; Reitner, Joachim; Schmidt, Alexander R

    2012-10-01

    The anamorphic fungal genus Monotosporella (Ascomycota, Sordariomycetes) has been reco-vered from a piece of Early Eocene Indian amber, as well as from the surface of extant resin flows in New Caledonia. The fossil fungus was obtained from the Tarkeshwar Lignite Mine of Gujarat State, western India, and was part of the biota of an early tropical angiosperm rainforest. The amber inclusion represents the second fossil record of Sordariomycetes, as well as the first fossil of its particular order (either Savoryellales or Chaetosphaeriales). The fossil fungus is distinguished from extant representatives by possessing both short conidiophores and small two-septate pyriform conidia, and is described as Monotosporella doerfeltii sp. nov. Inside the amber, the anamorph is attached to its substrate, which is likely the degraded thallus of a cladoniform lichen. The extant New Caledonian species is assigned to Monotosporella setosa. It was found growing on semi-solidified resin flows of Agathis ovata (Araucariaceae), and is the first record of Monotosporella from modern resin substrates. PMID:23063189

  8. New advances in the science and use of Trichoderma spp.

    Institute of Scientific and Technical Information of China (English)

    Gary Harman

    2004-01-01

    @@ Trichoderna spp. have been known for their strong abilities to produce extracellular enzymes,especially cellulases and glucanases by T. reesei, and also for their abilities to control plant pathogens.Most notably, a number of lines of evidence indicate that the agriculturally relevant strains are synergistic plant symbionts. As plant symbionts they possess the following characteristics: (a) they infect plant roots but cause the plant to restrict their growth to outer layers of the plant cortex (an exception is the recent discoveries of plant endophytic strains of Trichoderma and Gliocladium), (b)they produce bioactive plant molecules in the zone of interaction that (c) induce localized systemic resistance to plant diseases and (d) systemic plant resistance to diseases that in some respects is similar to that induced by rhizobacteria, (e) they induce changes in plant proteomes, (f) increase plant growth and yield at least in part by enhancing root growth, and (g) they increase plant nutrient uptake. The increase in plant growth and yields is strongly interactive with plant genotype, at least in maize. There also are very significant increases in the knowledge of events that occur in mycoparasitism that in many respects have features in common with their interactions with plants.They are highly resistant to toxicants and have recently been shown to degrade cyanide and take up,and then degrade, metallocyanides. These discoveries allow new uses for the organisms in managed plant systems, including agriculture. For example, the induced systemic resistance system allows control of fungi, bacteria, Oomycetes and even one virus at sites temporally and spatially distant from the site of application. The abilities of the fungi to resist/degrade toxicants and increase plant growth indicate they will highly useful components in plant-microbe or microbe-only strategies for remediation of soil and water pollution. Finally, there are new uses of their enzyme systems for

  9. Trichoderma species collected from Iran

    Institute of Scientific and Technical Information of China (English)

    Doostmorad Zafari

    2004-01-01

    @@ In order to identify Trichoderma species isolated from Iran, Trichoderma selective media and malt extract agar (MEA) were used to isolate Trichoderma species from the soil samples. All the cultures were purified on 2% water agar by hyphal tip method prior to morphological examination.Morphological observations were carried out on the cultures grown on 2% MEA and oat meal agar at 20℃ under ambient laboratory conditions. Macroscopic features of colony and microscopic features of conidiophore, phialid and conidium including position of phialids on conidiophore and shape and size of phialids and conidia were studied and recorded 3-5 days after inoculation. Out of 36 tested isolates, using morphological features and molecular data obtained from ITS1, ITS2 and 5.8S regions fourteen species were identified as follow: T. atroviride, T. ghanense, T. spirale, T. erinaceum, T. citrinoviride, T. saturnisporum,T. longibrachiatum , T. hamatum , T. harzianum, T. inhamatum , T. tomentosum , T.virens, T. asperellum, T. koningii. Among the species T. harzianum and T. virens isolates were the most frequent species. In addition of the mentioned species two Tichoderma sp. were collected from walnut rhizospher that they are not fit to any described species so far. Although one of them are T. brevicumpactum introduced informally.

  10. Multiple gene genealogies and phenotypic characters differentiate several novel species of Mycosphaerella and related anamorphs on banana

    NARCIS (Netherlands)

    Arzanlou, M.; Groenewald, J.Z.; Fullerton, R.A.; Abeln, E.C.A.; Carlier, J.; Zapater, M.-F.; Buddenhagen, I.W.; Viljoen, A.; Crous, P.W.

    2008-01-01

    Three species of Mycosphaerella, namely M. eumusae, M. fijiensis, and M. musicola are involved in the Sigatoka disease complex of bananas. Besides these three primary pathogens, several additional species of Mycosphaerella or their anamorphs have been described from Musa. However, very little is kno

  11. Biodiversity and distribution of Hypocrea/Trichoderma species in New Zealand

    Institute of Scientific and Technical Information of China (English)

    Sarah L Dodd; Alison Stewart

    2004-01-01

    @@ With increased imports of foreign microbes either as commercial biocontrol products or for the purposes of research, there is potentially an increased threat to indigenous beneficial microflora. In the present study, indigenous species of the fungal genus Hypocrea/Trichoderma are being used as a model system to determine the impact of foreign microbes on the native microflora of New Zealand. In order to protect such microflora, one has to first be aware of what is currently present and what sites, if any,are most vulnerable. A preliminary survey for the presence and diversity of species of Hypocrea/Trichoderma is currently underway in New Zealand and samples are being assessed from forest soils,agricultural soils, orchards, garden soils, sclerotia of various plant pathogens and pasture land. To date 238 isolates have been identified using both morphological characters and DNA sequence data from the ITS regions of the ribosomal gene cluster (ITS1 & ITS2) and, in some instances, sequence of the elongation factor gene (EF1-α) . Isolates were found to represent 16 known species plus three species as yet undescribed. In forest soils T. harzianum /T. inhamatum (31%) and T. viride (29%)followed by T. fertile (13%), were clearly the most abundant species and the remaining five species found in forests ( T. atroviride, T. koningii, T. aureoviride, H. cf. flavovirens anamorph and one unknown) each accounting for <8% of the total. Dominance by the species T. harzianum/inhamatum is consistent with studies done in South-East Asia, a mid-European primeval floodplainforest and Moscow. In contrast, when isolations were conducted with a bias for biocontrol capabilities it was found that the species T. atroviride (29%), T. koningii (17%), T. harzianum (15%)and T. viride (12%) dominated respectively. This survey is currently ongoing in New Zealand.Future studies will monitor indigenous species and strains following inoculation of specific microbes to assess the impact of the

  12. Anamorphic Spectrum Transform and its Application to Time-Bandwidth Compression

    CERN Document Server

    Asghari, Mohammad H

    2013-01-01

    A mathematical transform for compressing the time-bandwidth product of analog signals is introduced. As one of its applications, this physics-based signal grooming allows a conventional digitizer to sample and digitize the analog signal with variable resolution. The net result is that frequency components that were beyond the digitizer bandwidth can now be captured and, at the same time, the total digital data volume is reduced. This analog compression is loss-less and is made possible because the proposed transformation, performed prior to sampling, causes more samples to be allocated to higher frequencies, where they are needed, and less to lower frequencies, where they are redundant. Our method is inspired by operation of Fovea centralis in human eye and by anamorphic transformation in visual arts. The proposed transform can also be performed in digital domain as a data compression algorithm to alleviate the storage and transmission bottlenecks of big data.

  13. Implications of Planck2015 for inflationary, ekpyrotic and anamorphic bouncing cosmologies

    CERN Document Server

    Ijjas, Anna

    2015-01-01

    The results from Planck2015, when combined with earlier observations from WMAP, ACT, SPT and other experiments, were the first observations to disfavor the "classic" inflationary paradigm. To satisfy the observational constraints, inflationary theorists have been forced to consider plateau-like inflaton potentials that introduce more parameters and more fine-tuning, problematic initial conditions, multiverse-unpredictability issues, and a new 'unlikeliness problem.' Some propose turning instead to a "postmodern" inflationary paradigm in which the cosmological properties in our observable universe are only locally valid and set randomly, with completely different properties (and perhaps even different physical laws) existing in most regions outside our horizon. By contrast, the new results are consistent with the simplest versions of ekpyrotic cyclic models in which the universe is smoothed and flattened during a period of slow contraction followed by a bounce, and another promising bouncing theory, anamorphic...

  14. Trichoderma: the genomics of opportunistic success

    Energy Technology Data Exchange (ETDEWEB)

    Druzhinina, Irina S.; Seiboth, Verena Seidl; Estrella, Alfredo Herrera; Horwitz, Benjamin A.; Kenerley, Charles M.; Monte, Enrique; Mukherjee, Prasun K.; Zeilinger, Susanne; Grigoriev, Igor V.; Kubicek, Christian P.

    2011-01-01

    Trichoderma is a genus of common filamentous fungi that display a remarkable range of lifestyles and interactions with other fungi, animals and plants. Because of their ability to antagonize plant-pathogenic fungi and to stimulate plant growth and defence responses, some Trichoderma strains are used for biological control of plant diseases. In this Review, we discuss recent advances in molecular ecology and genomics which indicate that the interactions of Trichoderma spp. with animals and plants may have evolved as a result of saprotrophy on fungal biomass (mycotrophy) and various forms of parasitism on other fungi (mycoparasitism), combined with broad environmental opportunism.

  15.   The fungus Trichoderma spp. on vegetables

    DEFF Research Database (Denmark)

    Winding, Anne

      The fungus Trichoderma spp. on vegetables   Anne Winding, National Environmental Research Institute, Roskilde, AU   Trichoderma spp. is a naturally occurring fungus in soil and T. harzianum is an active ingredient in microbial pest control agents (MPCA) active against root pathogenic fungi....... The MPCA is administered to the plants by watering. The fungus establishes in the root zone and exerts its beneficial effect by general increase of resistance against pathogenic fungi. The natural occurrence of Trichoderma spp. and the fate and survival of the introduced T. harzianum on vegetables...... are important for risk assessment of MPCA and are the objectives of this project. Trichoderma spp. on tomatoes and cucumbers grown in greenhouses and on broccoli and celery leaf grown outdoors were quantified during a growth season. A MPCA with T. harzianum was applied in a greenhouse growing tomatoes...

  16. Isolation and Mass Production of Trichoderma.

    Science.gov (United States)

    Mendoza-Mendoza, Artemio; Clouston, Annabel; Li, Jin-Hua; Nieto-Jacobo, Maria Fernanda; Cummings, Nicholas; Steyaert, Johanna; Hill, Robert

    2016-01-01

    Members of the genus Trichoderma comprise the majority of commercial fungal biocontrol agents of plant diseases. As such, there is a wealth of information available on the analysis of their biocontrol potential and the mechanisms behind their superior abilities. This chapter aims to summarize the most common methods utilized within a Trichoderma biocontrol program for the isolation, identification, and mass propagation of individual strains. PMID:27565488

  17. A monograph of the entomopathogenic genera Hypocrella, Moelleriella, and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their aschersonia-like anamorphs in the Neotropics

    Science.gov (United States)

    Chaverri, P.; Liu, M.; Hodge, K.T.

    2008-01-01

    The present taxonomic revision deals with Neotropical species of three entomopathogenic genera that were once included in Hypocrella s. l.: Hypocrella s. str. (anamorph Aschersonia), Moelleriella (anamorph aschersonia-like), and Samuelsia gen. nov (anamorph aschersonia-like). Species of Hypocrella, Moelleriella, and Samuelsia are pathogens of scale insects (Coccidae and Lecaniidae, Homoptera) and whiteflies (Aleyrodidae, Homoptera) and are common in tropical regions. Phylogenetic analyses of DNA sequences from nuclear ribosomal large subunit (28S), translation elongation factor 1-α (TEF 1-α), and RNA polymerase II subunit 1 (RPB1) and analyses of multiple morphological characters demonstrate that the three segregated genera can be distinguished by the disarticulation of the ascospores and shape and size of conidia. Moelleriella has filiform multi-septate ascospores that disarticulate at the septa within the ascus and aschersonia-like anamorphs with fusoid conidia. Hypocrella s. str. has filiform to long-fusiform ascospores that do not disarticulate and Aschersonia s. str. anamorphs with fusoid conidia. The new genus proposed here, Samuelsia, has filiform to long-fusiform ascospores that do not disarticulate and aschersonia-like anamorphs with small allantoid conidia. In addition, the present study presents and discusses the evolution of species, morphology, and ecology in Hypocrella, Moelleriella, and Samuelsia based on multigene phylogenetic analyses. PMID:18490956

  18. PEA PEEL WASTE: A LIGNOCELLULOSIC WASTE AND ITS UTILITY IN CELLULASE PRODUCTION BY Trichoderma reesei UNDER SOLID STATE CULTIVATION

    OpenAIRE

    Nitin Verma; Mukesh C. Bansal; Vivek Kumar

    2011-01-01

    A wide variety of waste bioresources are available on our planet for conversion into bioproducts. In the biological systems, microorganisms are used to utilize waste as an energy source for the synthesis of valuable products such as biomass proteins and enzymes. The large quantities of byproducts generated during the processing of plant food involve an economic and environmental problem due to their high volumes and elimination costs. After isolation of the main constituent, there are abundan...

  19. Taxonomy and phylogeny of the ascomycetous yeast genus Zygoascus, with proposal of Zygoascus meyerae sp. nov. and related anamorphic varieties.

    Science.gov (United States)

    Smith, Maudy Th; Robert, V; Poot, G A; Epping, Wendy; de Cock, A W A M

    2005-05-01

    Physiological characters, mating compatibility, PCR-RAPD fingerprints, mol% G + C content, DNA-DNA relatedness, and large-subunit and internal transcribed spacer rRNA gene sequences of strains assigned to the genus Zygoascus were re-examined. On the basis of those data, and after phylogenetic analyses, an emendation of Zygoascus hellenicus (type material is a cross of CBS 6736(T) x CBS 5839(T)) is proposed, comprising two novel anamorphic varieties, Candida steatolytica var. steatolytica (CBS 6736(T)) and C. steatolytica var. inositophila (CBS 5839(T)). A novel teleomorphic species, Zygoascus meyerae sp. nov. (type material is a cross of CBS 4099(T) x CBS 7521(T)) is described, together with two novel anamorphic varieties corresponding to it, Candida hellenica var. hellenica (CBS 4099(T)) and C. hellenica var. acidophila (CBS 7115(T)).

  20. 打造出色的光学性能 Schneider Cine-Digitar Anamorphic 1.33x

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    Cine-Digitar Anamorphic 1.33x是Schneider推出的高品质变形镜头,采用全肖特(SCHOTT)玻璃镜片,具有最高的MIF值,控制四角变形达到最低值;减损的亮度控制在极限的-2%内,忠实再现接近100%的画面本质。

  1. Identification of Anamorph Yeast of Tremella aurantialba and Optimization of Medium Composition for Production of Exopolysaccharides

    Institute of Scientific and Technical Information of China (English)

    刘春卉; 杨秀伟; 俞建国; 马维新; 瞿伟菁; 包艳洁; 武文斌; 庄秀园

    2012-01-01

    A yeast-like fungus strain B1 isolated from wild fungus Tremella aurantialba was identified and initially characterized.Two phylogenetic trees were generated based on the sequences of large subunit ribosomal RNA gene D1/D2 regions and internal transcribed spacer (ITS) regions of related fungi,respectively.The analysis of D1/D2 regions and ITS sequences showed that fungus B1 was clustered together with T.aurantialba,T.aurantia and T.microspore in the phylogenetic trees.Both the morphological characteristic and phylogenetic analysis established that fungus B1 was one of the anamorph strains of T.aurantialba and belongs to Tremella genus.A fermentation medium for exopolysaccharides (EPS)production by T.aurantialba B1.Plackett-Burmen design was used to evaluate the effects of different components in the culture medium.Glucose and yeast extract have significant influence on the EPS production.The concentrations of two factors were optimized subsequently using central composite design and response surface analysis.The results showed that 49.2 g/L glucose and 10.4 g/L yeast extract could lead to the maximum production of EPS (4.99 g/L).The optimized medium led to a 1.5-fold enhancement of the production of EPS by T.aurantialba B1,as compared with that without optimization.

  2. Uji Antagonisme Jamur Trichoderma koningii dan Trichoderma harzianum Terhadap Penyakit Bidang Sadap Tanaman Karet Mouldy rot (Ceratocystis fimbriata) di Laboratorium

    OpenAIRE

    Marbun, Harif Nepen

    2016-01-01

    The objective of the research wasto test antagonism ability of Trichoderma koningii andTrichoderma harzianumto control Ceratocystis fimbriatain laboratory. The research was conducted at Laboratory of Research Agency of Rubber, Sungei Putih from April to December 2015. It was done by using Completely Randomized Design (CRD) non factorial with three treatments and eight replications. The results showed Trichoderma koningii andTrichoderma harzianumpotential as biological agents to control Cerato...

  3. Gliocladium and Trichoderma in agricultural soil

    Institute of Scientific and Technical Information of China (English)

    LIANG Chen; LI Bao-du; LU Guo-zhong

    2004-01-01

    @@ Gliocladium and Trichoderma are common fungi in agricultural soil. Several species of them were isolated and identified, great diversity was displayed in different agricultural soils of different crops,agricultural climate zones, different seasons, depths, different treated soybean cyst nematode soil,healthy and diseased crop soil. Among five crops soil samples, wheat and corn soil were found to possess the largest number of Gliocladium and Trichoderma separately. Gliocladium and Trichoderma of three major crops showed consistent changing patterns with seasonal variation. Corn soil displayed distinct vertical distribution of Trichoderna. There is a different distribution of the two fungi in diseased and healthy plant soil. Among the various isolated methods, diluted plate method is the best for isolating Gliocladium, and Trichoderma could be found in plant residue method and be tolerant to steam for two minutes. In the soybean cyst nematode soil mycobiota, the frequency of Gliocladium is higher than that of the others fungi, and Trichoderma may have the role of bioremediation in herbicide treated soil. Similarly, Gliocladium occurred frequently in different climate zones.

  4. Screening of Trichoderma strains tolerant to benzimidazole

    Institute of Scientific and Technical Information of China (English)

    LIU Kai-qi; XIANG Mei-mei; LIU Ren; ZENG Yong-san; ZHOU Hong-zi; YU Jin-feng; JIANG Xin-yin; ZHANG Yue-li

    2004-01-01

    @@ The screening of isolates and the assay of biocontrol mechanisms of Trichoderma were studied systematically in laboratory and greenhouse in vivo. The proteins tolerant to benzimidazole in Trichoderma strains were purified, and their physical and chemical properties were detected. Compared their biological activities in vitro and vivo in greenhouse, nine biocontrol strains (including Ty- 10-2, LTR-2, Tj-5-1, Tj-5-4, Ty- 11-1, Tj-11-3, Ty- 11-3, Tj-3-3-2, Tj-3-3-4) were screened. These biocontrol strains had faster rates of growth and higher inhibition to gray mould (Bortrytis cinerea),and the inhibition was stable. The effects of controlling gray mould in greenhouse with the screened Trichoderma strains were 70 % and 50 % in vivo.

  5. [Improvement of Trichoderma strains for biocontrol].

    Science.gov (United States)

    Benítez, T; Rey, M; Delgado-Jarana, J; Rincón, A M; Limón, M C

    2000-03-01

    The use of the fungal genus Trichoderma to control fungal plant diseases is a promising alternative to the use of chemical compounds. The aim of this work has been to obtain Trichoderma strains with improved capacity as biological control agents. To do so, the hydrolytic capacity on fungal cell walls of strains of the fungus Trichoderma harzianum has been increased. On one hand, transformation experiments with genes which coded for chitinases and glucanases have been carried out in T. harzianumstra ins. On the other hand, the medium composition has also been modified in order to eliminate proteolytic degradation of some of the overproduced enzymes. Finally, hybrid chitinolytic enzymes with substrate-binding domains have been produced as an alternative to obtain improved biocontrol strains. The transformant strains, when compared with the wild type, showed improved antifungal capacity against the phytopathogenic fungus Rhizoctonia solani, in in vitro experiments. PMID:15762779

  6. Use of a new Trichoderma harzianum strain isolated from the Amazon rainforest with pretreated sugar cane bagasse for on-site cellulase production.

    Science.gov (United States)

    Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; da Silva, Mateus Ribeiro; Azzoni, Sindelia Freitas; Pradella, José Geraldo da Cruz

    2012-03-01

    The on-site production of cellulases is an important strategy for the development of sustainable second-generation ethanol production processes. This study concerns the use of a specific cellulolytic enzyme complex for hydrolysis of pretreated sugar cane bagasse. Glycosyl hydrolases (FPase, xylanase, and β-glucosidase) were produced using a new strain of Trichoderma harzianum, isolated from the Amazon rainforest and cultivated under different conditions. The influence of the carbon source was first investigated using shake-flask cultures. Selected carbon sources were then further studied under different pH conditions using a stirred tank bioreactor. Enzymatic activities up to 121 FPU/g, 8000 IU/g, and 1730 IU/g of delignified steam-exploded bagasse+sucrose were achieved for cellulase, xylanase and β-glucosidase, respectively. This enzymatic complex was used to hydrolyze pretreated sugar cane bagasse. A comparative evaluation, using an enzymatic extract from Trichoderma reesei RUTC30, indicated similar performance of the T. harzianum enzyme complex, being a potential candidate for on-site production of enzymes.

  7. Applications of Trichoderma formulations in crop protection

    Institute of Scientific and Technical Information of China (English)

    Monte E; Rodríguez A; Rey M; Axpilicueta A; Gómez M I; de la Vina G; Grondona I; Llobell A

    2004-01-01

    @@ The choice of active Trichoderma strains is important in designing effective and safe biocontrol applications. Many species of Trichoderma have multiple strategies for fungal antagonism and indirect effects on plant health, such as growth promotion, systemic resistance induction and fertility improvements. Some strains are powerful antibiotic producers, and their suitability for use in biocontrol systems must be carefully assessed. However, many other active strains have no antibiotic capacity, and these are likely to be more useful in food production systems since they have not adverse effects on important groups of beneficial soil organisms. We have assessed the performance of selected naturally occurring Trichoderma strains (singly and in combination) and developed TUSAL , a mixture of Trichoderma harzianum and T. viride that has demonstrated to be effective against major pathogens in sugar beet and horticulture. TUSAL , has been bulked up and tested under field conditions, showing positive effects on precocity and root development, and increasing the crop production in field trials carried out in different pathosystems. The environmental impact of TUSAL strains on beneficial organisms in the environment were assessed before release, and molecular detection methods were developed to monitor the presence and performance of strains in the field.

  8. Cyclonerol derivatives from Trichoderma longibrachiatum YM311505.

    Science.gov (United States)

    Xuan, Qi-Cun; Huang, Rong; Chen, You-Wei; Miao, Cui-Ping; Ma, Kai-Xia; Wang, Tang; Wu, Shao-Hua

    2014-03-01

    A new sesquiterpene, 10,11-dihydrocyclonerotriol (1), together with two known compounds, catenioblin C (2) and sohirnone A (3), were isolated from Trichoderma longibrachiatum YM311505, an endophytic fungus of Azadirachta indica. Their structures were characterized and identified by spectral analysis. Compounds 1-3 exhibited antifungal activities against Pyricularia oryzae and Candida albicans. PMID:24689203

  9. Novel understanding of Trichoderma interaction mechanisms

    Institute of Scientific and Technical Information of China (English)

    Matteo Lorito

    2004-01-01

    @@ Trichoderma- based biofungicides are a reality in commercial agriculture, with more than 50formulations registered worldwide as biopesticides or biofertilizers. Several research strategies have been applied to identify the main genes and compounds involved in the complex, three-way interactions between fungal antagonists, plants and microbial pathogens. Proteome and genome analyses have greatly enhanced our ability to conduct targeted and genome-based functional studies. We have obtained reproducible 2-D maps of the entire fungal proteome in various conditions of interaction,which permitted the isolation of many proteins related to specific functions. Many differential proteins from several biocontrol strains of Trichoderma spp. during the in vivo interaction with different plants and/or several phytopathogenic fungi have been isolated and analyzed by MALDI-TOF.Relevant genes have been cloned and specifically inactivated, to demonstrate their function in biocontrol and induction of disease resistance. GFP-based reporter systems with interaction-inducible promoters allowed the characterization of regulatory sequences activated by the presence of the pathogen or the plant. From extensive cDNA and EST libraries of genes expressed during Trichoderma-pathogen-plant interactions, we are identified and determined the role of a variety of novel genes and gene-products, including ABC transporters specifically induced during antagonism with other microbes; enzymes and other proteins that produce or act as novel elicitors of Induced Resistance in plant and promote root growth and crop yield; proteins possibly responsible of a gene-forgene avirulent interaction between Trichoderma and plants; mycoparasitism-related inducers released from fungal pathogens and that activate biocontrol in Trichoderma; fungal promoters specifically induced during mycoparasitism and plant colonization; plant proteins and a novel phytoalexin induced by the presence of the fungal antagonist; etc

  10. Enhancement of Rice Seed Germination and Vigour by Trichoderma spp.

    OpenAIRE

    Febri Doni; I. Anizan; C.M.Z. Che Radziah; Ahmad Hilmi Salman; Muhammad Hidayat Rodzihan; Wan Mohtar Wan Yusoff

    2014-01-01

    The present study was undertaken to examine the effectiveness of Trichoderma spp. to enhance rice germination and vigour. An in vitro experiment was carried out to assess the effect of seven isolates of Trichoderma spp. in enhancing rice germination and vigour. The results showed that all isolates of Trichoderma spp. significantly increased rice seedling growth, germination rate, vigour index and speed of germination with sp., SL2 showing the greatest increase in all the four parameters. Tric...

  11. Uji Pengaruh Beberapa Herbisida Terhadap Trichoderma sp Secara In Vitro

    OpenAIRE

    Majid, Muhammad

    2016-01-01

    Muhammad Majid: In Vitro Test on the Effect of Trichoderma sp Through Application of some Herbicides. Under Supervision ofHasanuddin and Mukhtar Iskandar Pinem. Herbicide is most commonly used pesticide, yet contained multiple negative effect on environment, especially towards soil microorganisms. This research’s objective is to study effect of herbicide with different concentration towards Trichoderma sp. Trichoderma sp has been inoculated in PDA which have been induced with six different...

  12. Transpiration rates of rice plants treated with Trichoderma spp.

    Science.gov (United States)

    Doni, Febri; Anizan, I.; Che Radziah C. M., Z.; Yusoff, Wan Mohtar Wan

    2014-09-01

    Trichoderma spp. are considered as successful plant growth promoting fungi and have positive role in habitat engineering. In this study, the potential for Trichoderma spp. to regulate transpiration process in rice plant was assessed experimentally under greenhouse condition using a completely randomized design. The study revealed that Trichoderma spp. have potential to enhance growth of rice plant through transpirational processes. The results of the study add to the advancement of the understanding as to the role of Trichoderma spp. in improving rice physiological process.

  13. Antioxident activity of the mangrove endophytic fungus (Trichoderma sp.)

    Institute of Scientific and Technical Information of China (English)

    Saravanakumar Kandasamy; Kathiresan Kandasamy

    2014-01-01

    Objective: To test antioxidant property of the endophytic Trichoderma species isolated from the leaves of 12 mangroves of Andaman Nicobar Islands. Methods: Eight strains of Trichoderma species were found predominant and their crude extracts were assessed for antioxidant activity by using seven assays.Results:EMFCAS8 and other strains also showed considerable activity. Total antioxidant activity varied with the strains and it was maximum in Trichoderma Conclusions: This work concluded that mangroves are rich in endophytic Trichoderma species with potential for antioxidant activity.

  14. Cryptococcus randhawai sp nov., a novel anamorphic basidiomycetous yeast isolated from tree trunk hollow of Ficus religiosa (peepal tree) from New Delhi, India

    NARCIS (Netherlands)

    Khan, Z.U.; Ahmad, S.; Hagen, F.; Fell, J.W.; Kowshik, T.; Chandy, R.; Boekhout, T.

    2010-01-01

    A novel anamorphic Cryptococcus species is described, which was isolated in New Delhi (India) from decaying wood of a tree trunk hollow of Ficus religiosa. On the basis of sequence analysis of the D1/D2 domains of the 26S rRNA gene and the internally transcribed spacer (ITS)-1 and ITS-2 region seque

  15. Seed-borne Botryosphaeria spp. from native Prunus and Podocarpus trees in Ethiopia, with a description of the anamorph Diplodia rosulata sp. nov.

    Science.gov (United States)

    Gure, Abdella; Slippers, Bernard; Stenlid, Jan

    2005-09-01

    Botryosphaeria spp. from seeds of the native afromontane forest tree species, Podocarpus falcatus and Prunus africana, in Ethiopia have been identified. This is achieved by combining anamorph morphological characters and ITS sequence data. From a relatively small sample, four Botryosphaeria spp. were encountered. Two of the species from P. falcatus and the one from P. africana have not been previously described. The two species from P. falcatus were represented by only one isolate each and are not named here. The morphology of their Diplodia and Dothiorella anamorphs is, however, characterised. The anamorph of the other Botryosphaeria sp. from P. africana is described here as Diplodia rosulata sp. nov. Furthermore, B. parva is identified from P. falcatus based on ITS phylogeny. This species is also an important pathogen of various commercially important tree species in Ethiopia and elsewhere. This study highlights the ability of Botryosphaeria spp. to infect seeds and the possibility that they might be distributed in this way. The study also contributes to recent attempts to stabilize the taxonomy of Botryosphaeria anamorphs, especially regarding Diplodia, which is currently in taxonomic disarray.

  16. Effects of different inoculum densities of Trichoderma harzianum and Trichoderma viride against Meloidogyne javanica on tomato.

    Science.gov (United States)

    Al-Hazmi, Ahmad Saad; TariqJaveed, Muhammad

    2016-03-01

    A greenhouse experiment was conducted to evaluate the effects of different inoculum densities of two Saudi isolates of Trichoderma harzianum and Trichoderma viride against Meloidogyne javanica on tomato. Four densities (10(4), 10(6), 10(8) and 10(10) spores/g of soil) of each fungus were used. The results indicate that all four inoculum densities of the two Trichoderma species suppressed the nematode reproduction and root galling; and increased the growth of tomato plants, compared to controls. Efficacy of both fungi increased as their inoculum densities increased. Generally, efficacy of T. harzianum was better than that of T. viride, especially at the highest used density (10(10) spore/g soil) which resulted in the best control. PMID:26981012

  17. Diversity of Trichoderma in greenhouse soil

    Institute of Scientific and Technical Information of China (English)

    ZHAO Zhi-hui; SUN Xiao-dong; YANG Rui-xiu; YANG Hong; LU Guo-zhong

    2004-01-01

    @@ The protected agricultural production has become one of the fast growing and widespread cultivation technology in the north parts of China. Continuous cultivation of single crop or less rotation of crops usually resulted in the large amount of accumulation of soilborne pathogens and serious crop diseases in the greenhouse. After a few years of investigation of soilborne fungi in the north parts of China, nearly one hundred species of mictosporic fungi have been identified by the authors. Among these fungi 11species of Trichoderma have been morphologically identified, namely T. atroviride, T.aureoviride, T. citrinoviride , T. fertile, T. harzianum , T. inhamatum , T.longibrachiatum, T. parceramosum, T. reeseii, T. virens and T. viride. Trichoderm is found to be a frequently occurring genus of fungi in greenhouse soil. As an important component of effective beneficial antibiotic mycoparasites in soil Trichoderma plays an important part to regulate the balance of beneficial and harmful soilborne microorganisms.

  18. Cytostatic effect of L-lysine-. cap alpha. -oxidase from Trichoderma harzianum Rifai and Trichoderma viride

    Energy Technology Data Exchange (ETDEWEB)

    Khaduev, S.Kh.; Zhukova, O.S.; Dobrynin, Ya.V.; Soda, K.; Berezov, T.T.

    1987-09-01

    Comparative data is given on the effect of the new antitumor enzyme LO, obtained from a Soviet strain of Trichoderma harzianum Rifai and from Trichoderma viride from Japan, on DNA and RNA synthesis in human ovarian carcinoma cells in culture and also the results of the action of LO from Tr. harzianum Rifai on protein synthesis. Specific precursors were added to the samples 1 hour before the end of incubation time: /sup 3/H-thymidine, the precursor for DNA synthesis, /sup 3/H-uridine for RNA synthesis, and /sup 3/H-leucine for protein synthesis.

  19. Deep fungal dermatitis caused by the Chrysosporium anamorph of Nannizziopsis vriesii in captive coastal bearded dragons (Pogona barbata).

    Science.gov (United States)

    Johnson, R S P; Sangster, C R; Sigler, L; Hambleton, S; Paré, J A

    2011-12-01

    Deep fungal dermatitis caused by the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) was diagnosed in a group of coastal bearded dragons (Pogona barbata). The outbreak extended over a 6-month period, with four of six lizards from the same zoological outdoor enclosure succumbing to infection. A fifth case of dermatomycosis was identified in a pet lizard originally sourced from the wild. Diagnosis of infection with the CANV was based on similar clinical signs and histopathology in all animals and confirmed by culture and sequencing of the fungus from one animal. This is the first report of the CANV causing disease in a terrestrial reptile species in Australia and the first in the coastal bearded dragon. PMID:22103953

  20. Anthraquinone dyes decolorization capacity of anamorphic Bjerkandera adusta CCBAS 930 strain and its HRP-like negative mutants.

    Science.gov (United States)

    Korniłłowicz-Kowalska, Teresa; Rybczyńska, Kamila

    2014-06-01

    Cultures of the anamorphic fungus Bjerkandera adusta CCBAS 930 decolorizing, in stationary cultures, 0.01 % solutions of carminic acid and Poly R-478, were characterised by a strong increase in the activity of the horseradish peroxidase (HRP-like) and manganese-dependent peroxidase (MnP) at a low activity of lignin peroxidase. Genotypically modified mutants of B. adusta CCBAS 930: 930-5 and 930-14, with total or partial loss of decolorization capabilities relative to anthraquinonic dyes, showed inhibition of the activity of HRP-like peroxidase and MnP. Whereas, compared to the parental strain, in the mutant cultures there was an increase in the activity of lignin peroxidase and laccase. The paper presents a discussion of the role of the studied enzymatic activities in the process of decolorization of anthraquinonic dyes by the strain B. adusta CCBAS 930. PMID:24415463

  1. Trichoderma (Hypocrea) species with green ascospores from China

    NARCIS (Netherlands)

    Zhu, Z.X.; Zhuang, W.Y.

    2015-01-01

    Stromata of Trichoderma species having green ascospores were collected in various regions of China. Based on morphology of the sexual and asexual morph, culture characteristics, and sequence analyses of rpb2 and tef1 genes, 17 species with green ascospores were identified. Among them, Trichoderma ro

  2. Trichoderma genes in plants for stress tolerance- status and prospects.

    Science.gov (United States)

    Nicolás, Carlos; Hermosa, Rosa; Rubio, Belén; Mukherjee, Prasun K; Monte, Enrique

    2014-11-01

    Many filamentous fungi from the genus Trichoderma are well known for their anti-microbial properties. Certain genes from Trichoderma spp. have been identified and transferred to plants for improving biotic and abiotic stress tolerance, as well for applications in bioremediation. Several Trichoderma genomes have been sequenced and many are in the pipeline, facilitating high throughput gene analysis and increasing the availability of candidate transgenes. This, coupled with improved plant transformation systems, is expected to usher in a new era in plant biotechnology where several genes from these antagonistic fungi can be transferred into plants to achieve enhanced stress tolerance, bioremediation activity, herbicide tolerance, and reduction of phytotoxins. In this review, we illustrate the major achievements made by transforming plants with Trichoderma genes as well as their possible mode of action. Moreover, examples of efficient application of genetically modified plants as biofactories to produce active Trichoderma enzymes are indicated.

  3. PERBAIKAN PERTUMBUHAN DAN HASIL STEVIA (Stevia rebaudiana BERTONI M) MELALUI APLIKASI Trichoderma sp.

    OpenAIRE

    Haryuni -

    2013-01-01

    AbstrakTujuan penelitian ini adalah menguji perbaikan pertumbuhan dan hasil stevia (Stevia rebaudiana Bertoni M) melalui penggunaan  Trichoderma sp.  Perbanyakan  Trichoderma sp. dilakukan di laboratorium Balai Proteksi Perkebunan di Salatiga Jawa Tengah. Penelitian dirancang menggunakan rancangan factorial dengan dua faktor. Faktor pertama adalah inokulasi Trichoderma sp (To = tanpa  Trichoderma sp. & T1 = menggunakan Trichoderma sp. 100 g.  Faktor kedua adalah variasi aplikasi perlakuan...

  4. Antibiosis functions during interactions of Trichoderma afroharzianum and Trichoderma gamsii with plant pathogenic Rhizoctonia and Pythium.

    Science.gov (United States)

    Zhang, Xinjian; Harvey, Paul R; Stummer, Belinda E; Warren, Rosemary A; Zhang, Guangzhi; Guo, Kai; Li, Jishun; Yang, Hetong

    2015-09-01

    Trichoderma afroharzianum is one of the best characterized Trichoderma species, and strains have been utilized as plant disease suppressive inoculants. In contrast, Trichoderma gamsii has only recently been described, and there is limited knowledge of its disease suppressive efficacies. Comparative studies of changes in gene expression during interactions of these species with their target plant pathogens will provide fundamental information on pathogen antibiosis functions. In the present study, we used complementary DNA amplified fragment length polymorphism (cDNA-AFLP) analysis to investigate changes in transcript profiling of T. afroharzianum strain LTR-2 and T. gamsii strain Tk7a during in vitro interactions with plant pathogenic Rhizoctonia solani and Pythium irregulare. Considerable differences were resolved in the overall expression profiles of strains LTR-2 and Tk7a when challenged with either plant pathogen. In strain LTR-2, previously reported mycoparasitism-related genes such as chitinase, polyketide synthase, and non-ribosomal peptide synthetase were found to be differentially expressed. This was not so for strain Tk7a, with the only previously reported antibiosis-associated genes being small secreted cysteine-rich proteins. Although only one differentially expressed gene was common to both strains LTR-2 and Tk7a, numerous genes reportedly associated with pathogen antibiosis processes were differentially expressed in both strains, including degradative enzymes and membrane transport proteins. A number of novel potential antibiosis-related transcripts were found from strains LTR-2 and Tk7a and remain to be identified. The expression kinetics of 20 Trichoderma (10 from strain LTR-2, 10 from strain Tk7a) transcript-derived fragments (TDFs) were quantified by quantitative reverse transcription PCR (RT-qPCR) at pre- and post-mycelia contact stages of Trichoderma-prey interactions, thereby confirming differential gene expression. Collectively, this research

  5. Purifying Selection and Birth-and-Death Evolution in the Class II Hydrophobin Gene Families of the Ascomycete Trichoderma/Hypocrea

    Energy Technology Data Exchange (ETDEWEB)

    kubicek, Christian P.; Baker, Scott E.; Gamauf, Christian; Kenerley, Chuck; Druzhinina, Irina S.

    2008-01-10

    Hydrophobins are proteins containing eight conserved cysteine residues that occur uniquely in mycelial fungi, where their main function is to confer hydrophobicity to fungal surfaces in contact with air and during attachment of hyphae to hydrophobic surfaces of hosts, symbiotic partners or of themselves resulting in morphogenetic signals. Based on their hydropathy patterns and their solubility characteristics, they are classified in class I and class II hydrophobins, the latter being found only in ascomycetes. Here we have investigated the mechanisms driving the evolution of the class II hydrophobins in nine species of the mycoparasitic ascomycetous genus Trichoderma/Hypocrea, using three fully sequenced genomes (H. jecorina=T. reesei, H. atroviridis=T. atroviride; H. virens=T. virens) and a total of 14.000 ESTs of six others (T. asperellum, H. lixii=T. harzianum, T. aggressivum var. europeae, T. longibrachiatum, T. cf. viride). The former three contained six, ten and nine members, which is the highest number found in any other ascomycete so far. They all showed the conserved four beta-strands/one helix structure, which is stabilized by four disulfide bonds. In addition, a small number of these HFBs contained an extended N-terminus rich in either praline and aspartate, or glycine-asparagine. Phylogenetic analysis reveals a mosaic of terminal clades contain duplicated genes and shows only three reasonably supported clades. Calculation of the ratio of differences in synonymous vs. non-synonymous nucleotide substitutions provides evidence for strong purifying selection (KS/Ka >> 1). A genome database search for class II HFBs from other ascomycetes retrieved a much smaller number of hydrophobins (2-4) from each species, and most of them were from Pyrenomycetes. A combined phylogeny of these sequences with those of Trichoderma showed that the Trichoderma HFBs mostly formed their own clades, whereas those of other pyrenomycetes occured in shared clades. Our study shows

  6. Genetic regulation of conidiation in Trichoderma hamatun

    Institute of Scientific and Technical Information of China (English)

    Johanna Steyaert; Travis Glare; Alison Stewart; Margaret Carpenter; Hayley Ridgway

    2004-01-01

    @@ Achieving a balance between vegetative growth and spore production is essential for successful biocontrol by fungi. Low sporulation rates in the field can result in poor establishment and survival,whereas failure of conidia to recognise hosts can lead to persistence without efficacy. Commercial biocontrol products involve bulk preparations of conidia, however considerable variability in conidiation rates exists between biocontrol agents, which can restrict choice of strain for production. The majority of studies on Trichoderma conidiation have focused on the species T. viride and T. atroviride.These species form conidia in response to blue and near-UV light and/or nutrient deprivation and conidiation proceeds in a highly co-ordinated fashion, however relatively little is known on the genetic basis of Trichoderrma conidiation. In addition, whilst photoconidiation appears to be a general response detailed studies in other Trichoderma species are absent. In this study, conidiation in the lesser known biocontrol species T. hamatum is being investigated using a combined morphological and molecular approach. In contrast to T. atroviride, conidiation in response to blue-light was weaker and variable and suggested that additional triggers may be required for the T. hamatum photoresponse. A series of comparative photoconidiation assays are currently being undertaken investigating the effect of inoculum type and abiotic factors on timing and intensity of the response.Results will be discussed in relation to the current knowledge on conidial morphogenesis in Trichoderma. In addition to these morphological assays, a selection of genes implicated in sporulation and the blue-light responses are currently being isolated and characterised from T. hamatum. Two genes, phr1 and cmp1 , which were isolated previously from T. atroviride will be used as early and late markers of gene expression during the photoresponse in T. hamatum in order to define time points for harvesting

  7. Vision and development in Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

    Casas S; Cortés C; Ríos M; Rosales T; Bibbins M; Olmedo V; Herrera-Estrella A

    2004-01-01

    @@ Phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma is used as a photomorphogenetic model due to its ability to conidiate upon exposure to light. In total darkness, T. atroviride grows indefinitely as a mycelium provided that nutrients are not limiting. However, nutrient deprivation and light trigger the conidiation process. A pulse of blue light given to a radially growing colony induces synchronous sporulation. A ring of conidiophores bearing green conidia is produced at what had been the colony perimeter at the time of the light pulse. All known responses to blue light in N. crassa are initiated by a couple of transcription factors encoded by the white-collar genes (wc -1 and wc-2). WC-1 and WC-2 bind to the promoters of light regulated genes to rapidly activate transcription in response to light. In T. atroviride the photolyase encoding gene phr1 undergoes fast transcriptional activation in response to light. The presence of putative WCC binding boxes in the promoter of phr1 , suggested that light responses in Trichoderma could be under the control of white-collar homologues. We cloned two genes and demonstrated by gene replacement that both are essential for photoconidiation and photolyase gene expression. Therefore, they were named blue-light regulator one and two (blr1 and blr2 ). The BLR1 protein has all the characteristics of a blue-light photoreceptor. The generation of subtractive cDNA libraries allowed us to identify novel, BLR independent, light responses including the regulation of gene expression by blue-light. In addition, we recently initiated a Trichoderma ESTs sequencing project. Until now, we have sequenced above 3000 ESTs, from which we have obtained approximately 1800 unigenes. This unigene set was printed in microarrays and used to search for light induced genes. Twenty five clearly induced and around thirty repressed genes have been

  8. Exopolysaccharide from Trichoderma pseudokoningii induces macrophage activation.

    Science.gov (United States)

    Wang, Guodong; Zhu, Lei; Yu, Bo; Chen, Ke; Liu, Bo; Liu, Jun; Qin, Guozheng; Liu, Chunyan; Liu, Huixia; Chen, Kaoshan

    2016-09-20

    In this study, we evaluated the immunomodulatory activity of an exopolysaccharide (EPS) derived from Trichoderma pseudokoningii and investigated the molecular mechanism of EPS-mediated activation of macrophages. Results revealed that EPS could significantly induce the production of nitric oxide (NO), tumor necrosis factor (TNF)-α and interleukin (IL)-1β and enhance phagocytic activity in RAW 264.7 cells. Immunofluorescence staining indicated that EPS promoted the nuclear translocation of nuclear factor (NF)-κB p65 subunit. Western blot analysis showed that EPS increased the expression of inducible nitric oxide synthase (iNOS) protein, the degradation of IκB-α and the phosphorylation of mitogen-activated protein kinases (MAPKs). Furthermore, pretreatment of RAW 264.7 cells with specific inhibitors of NF-κB and MAPKs significantly attenuated EPS-induced TNF-α and IL-1β production. EPS also induced the inhibition of cytokine secretion by special antibodies against Toll-like receptor-4 (TLR4) and Dectin-1. These data suggest that EPS from Trichoderma pseudokoningii activates RAW 264.7 cells through NF-κB and MAPKs signaling pathways via TLR4 and Dectin-1. PMID:27261736

  9. Growth Inhibition of Colletotrichum gloeosporioides by Trichoderma harzianum, Trichoderma koningii, Bacillus subtilis and Pseudomonas fluorescens

    OpenAIRE

    Febrilia Nur ‘Aini; Sri Sukamto; Dwi Wahyuni; Risma Galuh Suhesti; Qurrotun Ayunin

    2015-01-01

    Colletotrichum  gloeosporioides is  a  disease  which  can  cause  significant yield  loss  of  cocoa.  The  objective  of  this  research  is  to  investigate  the  abilityof  antagonist  microbes,  Trichoderma  harzianum,  Trichoderma  koningii,  Bacillus subtilis  and Pseudomonas  fluorescens  in  controlling  gloeosporioides  biologically  in  laboratorium  condition.  The  experiment  was  carried  out  in  Crop  Protection  Laboratory,  Indonesian  Coffee  and  Cocoa  Research  Institut...

  10. Cytological Characterization of Anamorphic Fungus Lecanicillium pui and Its Relationship with Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes).

    Science.gov (United States)

    Lei, Wei; Zhang, Guren; Wu, Guangguo; Liu, Xin

    2016-01-01

    Ophiocordyceps sinensis (syn. Cordyceps sinensis), one of the most valuable medicinal mushrooms, has great economic importance on the Tibetan Plateau. We isolated an anamorphic fungus Lecanicillium pui from natural O. sinensis specimens and found that the optimal temperature for its culture on potato dextrose agar media was 25°C. Cell ultrastructure in L. pui hyphae and spores was characterized by transmission electron microscopy, and it was observed that some primary organelles showed the typical fungal features. Five chemical elements were determined in this fungus and niobium was discovered for the first time even with trace amounts. A species-specific method, nested polymerase chain reaction, was established to investigate the colonization of this fungus. Thus, the extensive distribution of L. pui on O. sinensis, in the shape of hyphae or mycelia, suggested that it may have subtle and chronic effects on the growth of the O. sinensis teleomorphic stage. These findings provide a potential reference, in the view of microbial ecology, for the study on the occurrence and mechanism of development of O. sinensis. PMID:27279447

  11. Analysis of expressed sequence tags from the anamorphic basidiomycetous yeast, Pseudozyma antarctica, which produces glycolipid biosurfactants, mannosylerythritol lipids.

    Science.gov (United States)

    Morita, Tomotake; Konishi, Masaaki; Fukuoka, Tokuma; Imura, Tomohiro; Kitamoto, Dai

    2006-07-15

    Pseudozyma antarctica T-34 secretes a large amount of biosurfactants (BS), mannosylerythritol lipids (MEL), from different carbon sources such as hydrocarbons and vegetable oils. The detailed biosynthetic pathway of MEL remained unknown due to lack of genetic information on the anamorphic basidiomycetous yeasts, including the genus Pseudozyma. Here, in order to obtain genetic information on P. antarctica T-34, we constructed a cDNA library from yeast cells producing MEL from soybean oil and identified the genes expressed through the creation of an expressed sequence tags (EST) library. We generated 398 ESTs, assembled into 146 contiguous sequences. Based upon a BLAST search similarity cut-off of E

  12. Growth Inhibition of Colletotrichum gloeosporioides by Trichoderma harzianum, Trichoderma koningii, Bacillus subtilis and Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Febrilia Nur ‘Aini

    2015-11-01

    Full Text Available Colletotrichum  gloeosporioides is  a  disease  which  can  cause  significant yield  loss  of  cocoa.  The  objective  of  this  research  is  to  investigate  the  abilityof  antagonist  microbes,  Trichoderma  harzianum,  Trichoderma  koningii,  Bacillus subtilis  and Pseudomonas  fluorescens  in  controlling  gloeosporioides  biologically  in  laboratorium  condition.  The  experiment  was  carried  out  in  Crop  Protection  Laboratory,  Indonesian  Coffee  and  Cocoa  Research  Institute.  Results of  this  research  showed  that  antagonist  fungi,  T.  harzianum,  T.  koningii,  had  a stronger  ability  in  inhibiting  growth  of  C.  gloeosporioides about  83%  compared  to  the  ability  of  antagonist  bacteria,  B.  subtilis  and P.  fluorescens,  only about  49%. Key words: Growth  inhibition,  Colletotrichum  gloeosporioides,  Trichoderma  harzianum, Trichoderma koningii,  Bacillus subtilis, Pseudomonas fluorescens.

  13. Trichoharzianol, a new antifungal from Trichoderma harzianum F031.

    Science.gov (United States)

    Jeerapong, Chotika; Phupong, Worrapong; Bangrak, Phuwadol; Intana, Warin; Tuchinda, Patoomratana

    2015-04-15

    A new decalin derivative, trichoharzianol (1), together with three known compounds, eujavanicol A (2), 5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone (3), and 4,6-dihydroxy-5-methylphthalide (4), were isolated from Trichoderma harzianum F031. For the first time, compounds 2-4 were reported from the Trichoderma species. Their structures were characterized by spectroscopic methods. Trichoharzianol (1) showed the highest antifungal activity against Colletotrichum gloeosporioides, with a minimum inhibitory concentration (MIC) of 128 μg/mL.

  14. Trichoderma spp. decrease Fusarium root rot in common bean

    Directory of Open Access Journals (Sweden)

    Hudson Teixeira

    2012-12-01

    Full Text Available The effectiveness of six Trichoderma-based commercial products (TCP in controlling Fusarium root rot (FRR in common bean was assessed under field conditions. Three TCP, used for seed treatment or applied in the furrow, increased seedling emergence as much as the fungicide fludioxonil. FRR incidence was not affected, but all TCP and fludioxonil reduced the disease severity, compared to control. Application of Trichoderma-based products was as effective as that of fludioxonil in FRR management.

  15. Enhancement of Rice Seed Germination and Vigour by Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Febri Doni

    2014-05-01

    Full Text Available The present study was undertaken to examine the effectiveness of Trichoderma spp. to enhance rice germination and vigour. An in vitro experiment was carried out to assess the effect of seven isolates of Trichoderma spp. in enhancing rice germination and vigour. The results showed that all isolates of Trichoderma spp. significantly increased rice seedling growth, germination rate, vigour index and speed of germination with sp., SL2 showing the greatest increase in all the four parameters. Trichoderma sp., SL2 treated rice seeds attained values of 4.48 and 6.00 cm, 0.0084 and 0.0048 g and 1016.56 and 44.75 seeds/day for seedling shoot length seedling root length, shoot weight, root weight, vigour index and speed of germination respectively. We may conclude that Trichoderma spp. is able to enhance seed germination and vigour. The results of the study adds to the further understanding of the role of beneficial fungi in improving rice resistance to stress, yield and quality through seed invigoration. Trichoderma

  16. Impacto de herbicidas em isolados de Trichoderma spp. Impact of herbicides on strains of Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    M.R. Reis

    2013-06-01

    Full Text Available O uso de microrganismos é uma alternativa para o controle de doenças em plantas. Todavia, é prudente verificar a interação desse com os demais métodos de controle empregados em determinada cultura. Dessa forma, objetivou-se avaliar a fungitoxicidade dos herbicidas sobre o crescimento e desenvolvimento dos isolados de Trichoderma spp. Utilizou-se o delineamento inteiramente casualizado, em esquema fatorial 6 x 6 x 4, com quatro repetições. O fator A correspondeu aos herbicidas pendimethalin, clomazone, carfentrazone-ethyl, oxadiazon, thiobencarb + propanil e byspiribac-sodium; o fator B, às doses dos herbicidas - 0, 25, 50, 75, 100 e 200% da dose recomendada; e o fator C, aos isolados de Trichoderma spp. AJAM 18, CE 66, TRI 01 e TRI 02. O ensaio foi realizado em condições in vitro; avaliaram-se o crescimento micelial radial (CMR e a esporulação dos isolados após aplicação dos herbicidas. Observaram-se diferenças de sensibilidade dos isolados para o mesmo produto testado. O oxadiazon reduziu o CMR dos isolados AJAM 18 e TRI 01 em 66 e 35%, respectivamente. No entanto, reduziu apenas 16% do CMR do isolado TRI 02 e não alterou o CMR do isolado CE 66 mesmo em 200% da dose recomendada. Verificaram-se diferentes efeitos dos produtos em cada isolado. A mistura comercial de thiobencarb+propanil foi altamente tóxica aos isolados de Trichoderma spp., com reduções em torno de 85% no CMR e no número de esporos. Por outro lado, o byspiribac-sodium pouco afetou os isolados, apresentando reduções inferiores a 10% no CMR e na esporulação. O carfentrazone-ethyl e byspiribac-sodium demonstraram ser compatíveis com os isolados de Trichoderma spp. estudados.The use of microorganisms is an alternative for the control of plant diseases. However, one should verify its interaction with other methods of control used for a particular crop. The objective of this work was to evaluate the effect of herbicide fungitoxicity on the growth and

  17. Biocontrol of Rhizoctonia solani with Trichoderma Spp.

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ From over 800 fungal strains of Trichoderma Spp. , 6 strains were found to greatly inhibit the growing of Rhizocotonia solani, the pathogen of rice sheath blight in dual culture. Among them, strain T3 was the best antagonist,which reduced the growing of the pathogen by 52.54% (Table 1). In field, both the pesticide Jinggangmycin and the mixture of T1 T6 could reduce the severity of rice sheath blight(Table 2), which resulted in the increases of seed setting rate and 1000 grain weight. Because the effect of the antagonists on the control of the pathogen could be partially realized in the watery environment, studies on the biocontrol mechanism of the fungi should be strengthened to help the establishment of a best way of antagonist utilization.

  18. Xylanase production by Trichoderma harzianum E58

    Energy Technology Data Exchange (ETDEWEB)

    Senior, D.J.; Mayers, P.R.; Saddler, J.N. (Fortintek Canada Corp., Ottawa, ON (Canada). Dept. of Biotechnology and Chemistry)

    1989-12-01

    Growth of Trichoderma harzianum E58 on hemicellulose-rich media, both in batch and fermentor cultures, resulted in independent profiles of the production of xylanase and endoglucanase enzymes. Dramatic differences in the ratio of xylanase to endoglucanase activities were observed among cultures grown on cellulose-rich Solka Floc and xylan. These results indicated that the induction of xylanases and cellulases was likely to be under separate regulatory control. The specific activity and amount of xylanases produced were found to be dependent on the concentration of xylan in the growth media. Growth on oat spelts xylan or the hemicellulose-rich, watersoluble fraction from steam-treated aspenwood (SEA-WS) greatly enhanced the production of xylanases and xylosidase in the culture filtrates. Constitutive levels of xylanase and endoglucanase enzymes were detected during growth of the fungus on glucose. (orig.).

  19. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Directory of Open Access Journals (Sweden)

    Suchada Chantrapromma

    2014-04-01

    Full Text Available The title compound, C20H28O3, known as `trichodermaerin' [systematic name: (4E-4,9,15,16,16-pentamethyl-6-oxatetracyclo[10.3.1.01,10.05,9]hexadec-4-ene-7,13-dione], is a diterpene lactone which was isolated from Trichoderma asperellum. The structure has a tetracycic 6–5–7–5 ring system, with the cyclohexanone ring adopting a twisted half-chair conformation and the cyclopentane ring adopting a half-chair conformation, whereas the cycloheptene and tetrahydrofurananone rings are in chair and envelope (with the methyl-substituted C atom as the flap conformations, respectively. The three-dimensional architecture is stabilized by C—H...O interactions.

  20. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Science.gov (United States)

    Chantrapromma, Suchada; Jeerapong, Chotika; Phupong, Worrapong; Quah, Ching Kheng; Fun, Hoong-Kun

    2014-01-01

    The title compound, C20H28O3, known as ‘trichodermaerin’ [systematic name: (4E)-4,9,15,16,16-penta­methyl-6-oxa­tetra­cyclo­[10.3.1.01,10.05,9]hexa­dec-4-ene-7,13-dione], is a diterpene lactone which was isolated from Trichoderma asperellum. The structure has a tetra­cycic 6–5–7–5 ring system, with the cyclo­hexa­none ring adopting a twisted half-chair conformation and the cyclo­pentane ring adopting a half-chair conformation, whereas the cyclo­heptene and tetra­hydro­furan­anone rings are in chair and envelope (with the methyl-substituted C atom as the flap) conformations, respectively. The three-dimensional architecture is stabilized by C—H⋯O inter­actions. PMID:24826124

  1. Effect of infesting soil with Trichoderma harzianum and amendment with coffee pulp on survival of Armillaria

    NARCIS (Netherlands)

    Otieno, W.; Jeger, M.J.; Termorshuizen, A.J.

    2003-01-01

    Six isolates of Trichoderma were screened for antagonism to Armillaria in tea stem sections buried in the soil. The inability of Armillaria to invade Trichoderma-colonized stem sections and the reduction of its viability in the plant materials following invasion of these by Trichoderma were used as

  2. Advances in biocontrol mechanism and application of Trichoderma spp. for plant diseases

    Institute of Scientific and Technical Information of China (English)

    HUANG Caihong; YANG Qian

    2007-01-01

    Trichoderma spp. is a filamentous soil fungus known as an effective biocontrol agent of a range of important airborne and soilborne pathogens, it has universal distribution and economic importance. This article reviewed the researches on biocontrol mechanism for plant diseases and application of Trichoderma spp., especially Trichoderma harzianum in recent years.

  3. Cellulose hydrolysis by Trichoderma reesei cellulases: studies on adsorption, sugar production and synergism of cellobiohydrolase I,II and endoglucanase II

    Energy Technology Data Exchange (ETDEWEB)

    Medve, J.

    1997-02-01

    Three major cellulases have been purified by ion-exchange chromatography in an FPLC system. Microcrystalline cellulose (Avicel) was hydrolyzed by the single enzymes and by equimolar mixtures of CBH I-CBH II and CBH I-EG II. Enzyme adsorption was followed indirectly by selectively quantifying the enzymes in the supernatant by ion-exchange chromatography in an FPLC system. The (synergistic) production of small, soluble sugars (glucose, cellobiose and cellotriose) by the enzymes was followed by HPLC. 76 refs

  4. A Native Threonine Coordinates Ordered Water to Tune Light-Oxygen-Voltage (LOV) Domain Photocycle Kinetics and Osmotic Stress Signaling in Trichoderma reesei ENVOY.

    Science.gov (United States)

    Lokhandwala, Jameela; Silverman Y de la Vega, Rafael I; Hopkins, Hilary C; Britton, Collin W; Rodriguez-Iglesias, Aroa; Bogomolni, Roberto; Schmoll, Monika; Zoltowski, Brian D

    2016-07-01

    Light-oxygen-voltage (LOV) domain-containing proteins function as small light-activated modules capable of imparting blue light control of biological processes. Their small modular nature has made them model proteins for allosteric signal transduction and optogenetic devices. Despite intense research, key aspects of their signal transduction mechanisms and photochemistry remain poorly understood. In particular, ordered water has been identified as a possible key mediator of photocycle kinetics, despite the lack of ordered water in the LOV active site. Herein, we use recent crystal structures of a fungal LOV protein ENVOY to interrogate the role of Thr(101) in recruiting water to the flavin active site where it can function as an intrinsic base to accelerate photocycle kinetics. Kinetic and molecular dynamic simulations confirm a role in solvent recruitment to the active site and identify structural changes that correlate with solvent recruitment. In vivo analysis of T101I indicates a direct role of the Thr(101) position in mediating adaptation to osmotic stress, thereby verifying biological relevance of ordered water in LOV signaling. The combined studies identify position 101 as a mediator of both allostery and photocycle catalysis that can impact organism physiology. PMID:27226624

  5. The Hypocrea jecorina (syn. Trichoderma reesei) lxr1 gene encodes a D-mannitol dehydrogenase and is not involved in L-arabinose catabolism

    NARCIS (Netherlands)

    Metz, Benjamin; de Vries, Ronald P; Polak, Stefan; Seidl, Verena; Seiboth, Bernhard

    2009-01-01

    The Hypocrea jecorina LXR1 was described as the first fungal L-xylulose reductase responsible for NADPH dependent reduction of L-xylulose to xylitol in L-arabinose catabolism. Phylogenetic analysis now reveals that LXR1 forms a clade with fungal D-mannitol 2-dehydrogenases. Lxr1 and the orthologous

  6. Three new species of Trichoderma with hyaline ascospores from China.

    Science.gov (United States)

    Zhu, Z X; Zhuang, W Y

    2015-01-01

    Collections of Trichoderma having hyaline ascospores from different areas of China were examined. Using combined analyses of morphological data, culture characters and phylogenetic information based on rDNA sequences of partial nuc translation elongation factor 1-α encoding gene (TEF1-α) and the gene encoding the second largest nuc RNA polymerase subunit (RPB2), three new species, Trichoderma applanatum, T. oligosporum and T. sinoluteum, were discovered and are described. Trichoderma applanatum produces continuous flat to pulvinate, white to cream stromata with dense orange or pale brown ostioles, and simple acremonium-like to verticillium-like conidiophores, belongs to the Hypocreanum clade and is closely related to T. decipiens. Trichoderma oligosporum forms reddish brown stromata with a downy surface, hyaline conidia and gliocladium-like conidiophores, and is closely related to but distinct from T. crystalligenum in the Psychrophila clade. Trichoderma sinoluteum, as a member of the Polysporum clade, is characterized by pale yellow stromata, white pustulate conidiomata, pachybasium-like conidiophores, and hyaline conidia. Differences between the new species and their close relatives are discussed.

  7. Trichoderma (Hypocrea) species with green ascospores from China.

    Science.gov (United States)

    Zhu, Z X; Zhuang, W Y

    2015-06-01

    Stromata of Trichoderma species having green ascospores were collected in various regions of China. Based on morphology of the sexual and asexual morph, culture characteristics, and sequence analyses of rpb2 and tef1 genes, 17 species with green ascospores were identified. Among them, Trichoderma rosulatum, T. rufobrunneum and T. stipitatum are described as new species, and seven other species are reported for the first time from China. Trichoderma rosulatum produces small bright yellow or pale greenish stromata with dense dark green ostioles and gliocladium-like conidiophores, shows a close relationship to T. thelephoricola, and belongs to the Chlorospora clade. Trichoderma rufobrunneum, which typically forms reddish brown stromata, is recognised as a member of the Harzianum clade. Trichoderma stipitatum is characterised by turbinate, pale yellow to nearly orange stromata and verticillium-like conidiophores; it forms a distinct, independent lineage with strong bootstrap support in the phylogenetic trees. The distinctions between the new species and their close relatives are discussed, and their phylogenetic positions are explored. PMID:26240449

  8. Two new Chinese record of the genus Trichoderma: Trichoderma pleuroticola and T.pleurotum%木霉属中国新纪录种Trichoderma pleuroticola和T.pleurotum

    Institute of Scientific and Technical Information of China (English)

    张广志; 杨合同; 张新建; 李纪顺; 陈凯; 黄玉杰

    2013-01-01

    [Objective] Identification of two Trichoderma isolates were isolated from the soil in vegetable greenhouses and the pileus of Asafoetida mushroom. [Methods] By combination of morphological charaters and application of internal transcribed spacer (ITS). [Results] Two Trichoderma isolates were identified as Trichoderma pleuroticola S.H.Yu & Park sp. nov. and Trichoderma pleurotum S.H.Yu & Park. The morphological charaters of T. pleuroticola is similar with T. harzianum, but its conidiospore is obviously more than T. harzianum, secretes dark brown pigment, and forms yellow crystal on PDA medium. The typical characteristics of Trichoderma pleurotum is that its conidiophores are mostly solitary and more or less prostrate, branches scattered, arising separately and bearing crowded whorls of appressed phialides at the apex rsembling the conidiophore in Gliocladium. [Conclusion] Two Trichoderma isolates are T. pleuroticola and T. pleurotum respectively, which are two new record species in China.%[目的]对蔬菜大棚土壤中和阿魏菇腐烂的菌盖上分离的两株木霉菌进行分类鉴定.[方法]结合形态学分类特征和ITS序列分析的方法进行鉴定.[结果]从蔬菜大棚的土壤中和阿魏菇腐烂的菌盖上分离的两株木霉菌分别为Trichoderma pleuroticola和T.pleurotum.T.pleuroticola的形态特征与T.harzianum相似,但其分生孢子显著大于T.harzianum的分生孢子,且在PDA上产生黑褐色的色素以及黄色的结晶物.T.pleurotum 典型特征是分生孢子梗单生,有时匍匐,分枝散生,初级分枝和分生孢子梗顶端聚生,类似粘帚霉.[结论]分离的两株木霉分别是T.pleuroticola和T.pleurotum,为木霉菌中国新纪录种.

  9. Trichoderma songyi sp. nov., a new species associated with the pine mushroom (Tricholoma matsutake).

    Science.gov (United States)

    Park, Myung Soo; Oh, Seung-Yoon; Cho, Hae Jin; Fong, Jonathan J; Cheon, Woo-Jae; Lim, Young Woon

    2014-10-01

    A new species, Trichoderma songyi, was found to be associated with the pine mushroom (Tricholoma matsutake) in Korea. This species was isolated from three different substrates: Tricholoma matsutake basidiomata, as well as roots of Pinus densiflora and soil in the fairy ring. Based on its molecular and phenotypic characteristics, we demonstrate that Trichoderma songyi is unique and distinguishable from closely related species. We performed phylogenetic analyses based on two molecular markers, the genes for both translation elongation factor 1-alpha and the second largest subunit of RNA polymerase II. Phylogenetic analyses showed that Trichoderma songyi is closely related to Trichoderma koningii aggregate and Trichoderma caerulescens. Morphologically, Trichoderma songyi can be distinguished from these closely related taxa by its growth rates, colony morphology on PDA in darkness, and coconut-like odour. Due to the economic importance of the pine mushroom, the relationship between Trichoderma songyi and Tricholoma matsutake should be studied further.

  10. Trichoderma species occurring on wood with decay symptoms in mountain forests in Central Europe: genetic and enzymatic characterization.

    Science.gov (United States)

    Błaszczyk, Lidia; Strakowska, Judyta; Chełkowski, Jerzy; Gąbka-Buszek, Agnieszka; Kaczmarek, Joanna

    2016-08-01

    The aim of this study was to explore the species diversity of Trichoderma obtained from samples of wood collected in the forests of the Gorce Mountains (location A), Karkonosze Mountains (location B) and Tatra Mountains (location C) in Central Europe and to examine the cellulolytic and xylanolytic activity of these species as an expression of their probable role in wood decay processes. The present study has led to the identification of the following species and species complex: Trichoderma atroviride P. Karst., Trichoderma citrinoviride Bissett, Trichoderma cremeum P. Chaverri & Samuels, Trichoderma gamsii Samuels & Druzhin., Trichoderma harzianum complex, Trichoderma koningii Oudem., Trichoderma koningiopsis Samuels, C. Suárez & H.C. Evans, Trichoderma longibrachiatum Rifai, Trichoderma longipile Bissett, Trichoderma sp. (Hypocrea parapilulifera B.S. Lu, Druzhin. & Samuels), Trichoderma viride Schumach. and Trichoderma viridescens complex. Among them, T. viride was observed as the most abundant species (53 % of all isolates) in all the investigated locations. The Shannon's biodiversity index (H), evenness (E), and the Simpson's biodiversity index (D) calculations for each location showed that the highest species diversity and evenness were recorded for location A-Gorce Mountains (H' = 1.71, E = 0.82, D = 0.79). The preliminary screening of 119 Trichoderma strains for cellulolytic and xylanolytic activity showed the real potential of all Trichoderma species originating from wood with decay symptoms to produce cellulases and xylanases-the key enzymes in plant cell wall degradation. PMID:26586561

  11. Biological control of banana black Sigatoka disease with Trichoderma

    Directory of Open Access Journals (Sweden)

    Poholl Adan Sagratzki Cavero

    2015-06-01

    Full Text Available Black Sigatoka disease caused by Mycosphaerella fijiensis is the most severe banana disease worldwide. The pathogen is in an invasive phase in Brazil and is already present in most States of the country. The potential of 29 isolates of Trichoderma spp. was studied for the control of black Sigatoka disease under field conditions. Four isolates were able to significantly reduce disease severity and were further tested in a second field experiment. Isolate 2.047 showed the best results in both field experiments and was selected for fungicide sensitivity tests and mass production. This isolate was identified as Trichoderma atroviride by sequencing fragments of the ITS region of the rDNA and tef-1α of the RNA polymerase. Trichoderma atroviride was as effective as the fungicide Azoxystrobin, which is recommended for controlling black Sigatoka. This biocontrol agent has potential to control the disease and may be scaled-up for field applications on rice-based solid fermentation

  12. Study on Biological Control Of Rhizoctonia solani via Trichoderma

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Strain T02-25 was selected from approximately 30 rhizosphere isolates of Trichoderma species isolated from roots of crops. Its biological activity against Rhizoctonia solani was determined for the control efficacy to pepper seedling blight caused by R. solani in the field. The assay methods were treating R. solani sclerotia by Trichoderma conidial suspension (106cfu ml-1) and scattering Thichoderma rice bran over the pepper root medium. The results showed that T02-25 was active against R. solani in both ways, and its control efficacy was 82.7% and 78.0%, respectively. In addition to comparison of the efficacy of the two application methods, the relationship of different factors in the control efficacy of Trichoderma against R. solani was discussed.

  13. Mutagenesis of Trichoderma Viride by Ultraviolet and Plasma

    Institute of Scientific and Technical Information of China (English)

    姚日生; 李曼曼; 邓胜松; 胡华佳; 王淮; 李凤和

    2012-01-01

    Considering the importance of a microbial strain capable of increased cellulase production, a mutant strain UP4 of Trichoderma viride was developed by ultraviolet (UV) and plasma mutation. The mutant produced a 21.0 IU/mL FPase which was 98.1% higher than that of the parent strain Trichoderma viride ZY-1. In addition, the effect of ultraviolet and plasma mutagenesis was not merely simple superimposition of single ultraviolet mutation and single plasma mutation. Meanwhile, there appeared a capsule around some of the spores after the ultraviolet and plasma treatment, namely, the spore surface of the strain became fuzzy after ultraviolet or ultraviolet and plasma mutagenesis.

  14. Mutagenesis of Trichoderma Viride by Ultraviolet and Plasma

    Science.gov (United States)

    Yao, Risheng; Li, Manman; Deng, Shengsong; Hu, Huajia; Wang, Huai; Li, Fenghe

    2012-04-01

    Considering the importance of a microbial strain capable of increased cellulase production, a mutant strain UP4 of Trichoderma viride was developed by ultraviolet (UV) and plasma mutation. The mutant produced a 21.0 IU/mL FPase which was 98.1% higher than that of the parent strain Trichoderma viride ZY-1. In addition, the effect of ultraviolet and plasma mutagenesis was not merely simple superimposition of single ultraviolet mutation and single plasma mutation. Meanwhile, there appeared a capsule around some of the spores after the ultraviolet and plasma treatment, namely, the spore surface of the strain became fuzzy after ultraviolet or ultraviolet and plasma mutagenesis.

  15. Trichoharzianol, a new antifungal from Trichoderma harzianum F031.

    Science.gov (United States)

    Jeerapong, Chotika; Phupong, Worrapong; Bangrak, Phuwadol; Intana, Warin; Tuchinda, Patoomratana

    2015-04-15

    A new decalin derivative, trichoharzianol (1), together with three known compounds, eujavanicol A (2), 5-hydroxy-3-hydroxymethyl-2-methyl-7-methoxychromone (3), and 4,6-dihydroxy-5-methylphthalide (4), were isolated from Trichoderma harzianum F031. For the first time, compounds 2-4 were reported from the Trichoderma species. Their structures were characterized by spectroscopic methods. Trichoharzianol (1) showed the highest antifungal activity against Colletotrichum gloeosporioides, with a minimum inhibitory concentration (MIC) of 128 μg/mL. PMID:25817439

  16. Detection of Fusarium spp. and Trichoderma spp. and antagonism of Trichoderma sp. in soybean under no-tillage

    OpenAIRE

    Paola Mendes Milanesi; Elena Blume; Marlove Fátima Brião Muniz; Lia Rejane Silveira Reiniger; Zaida Inês Antoniolli; Emanuele Junges; Manoeli Lupatini

    2013-01-01

    This study aimed i) to quantify the occurrence of Fusarium spp. and Trichoderma spp. in rhizospheric soil, with and without symptoms of Sudden Death Syndrome (SDS) in eight soybean genotypes; ii) morphologically identify isolates of Fusarium spp. from roots with SDS; iii) evaluate the antagonism between Trichoderma spp. and Fusarium spp. isolates from rhizospheric soil and roots from with and without SDS, respectively; and iv) characterize through the ITS1-5.8S-ITS2 region of rDNA the isolate...

  17. Trichosporon wieringae sp.nov., an anamorphic basidiomycetous yeast from soil, and assimilation of some phenolic compounds, polysaccharides and other non-conventional carbon sources by saprophytic Trichosporon species

    NARCIS (Netherlands)

    Middelhoven, W.J.

    2004-01-01

    A morphological and physiological description of an anamorphic basidiomycetous yeast species isolated from soil, named Trichosporon wieringae, is presented. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA and of the ITS re

  18. Identification of Trichoderma strains by image analysis of HPLC chromatograms

    DEFF Research Database (Denmark)

    Thrane, Ulf; Poulsen, S.B.; Nirenberg, H.I.;

    2001-01-01

    Forty-four Trichoderma strains from water-damaged building materials or indoor dust were classified with chromatographic image analysis on full chromatographic matrices obtained by high performance liquid chromatography with UV detection of culture extracts. The classes were compared with morphol...

  19. Functional genomic approach to the study of biodiversitywithin Trichoderma

    Institute of Scientific and Technical Information of China (English)

    Monte E; Hermosa M R; González F J; Rey M; Cardoza R E; Gutiérrez S; Delgado Jarana J; Llobell A

    2004-01-01

    @@ Trichoderma is a fungal genus of great and demonstrable biotechnological value, but its genome is poorly surveyed compared with other model microorganisms. Due to their ubiquity and rapid substrate colonization, Trichoderma species have been widely used as biocontrol organisms for agriculture, and their enzyme systems are widely used in industry. Therefore, there is a clear interest to explore beyond the phenotype to exploit the underlying genetic systems using functional genomics tools. The great diversity of species within the Trichoderma genus, the absence of optimized systems for its exploration, and the great variety of genes expressed under a wide range of ambient conditions are the main challenges to consider when starting a comprehensive functional genomics study. An initial project started by three Spanish groups has been extended into the project TRICHOEST, funded by the EU (FP5, QLRT-2001-02032) to target the transcriptome analysis of selected Trichoderma strains with biocontrol potential, in conditions related to antagonism, nutrient stress and plant interactions. Once specific conditions were defined, cDNA libraries were produced and used for EST sequencing. Nine strains from seven Trichoderma species have been considered in this study and an important amount of gene sequence data has been generated, analyzed and used to compare the gene expression in different strains.In parallel to sequencing, genomic expression studies were carried out by means of macro-arrays to identify genes expressed in specific conditions. In silico analysis of DNA sequencing data together with macro-array expression results have lead to a selection based on the potential use of the gene sequences.The selected clone sequences were completed and cloned in appropriate vectors to initiate functional analysis by means of expression studies in homologous and heterologous systems.

  20. Phylogeny of the clinically relevant species of the emerging fungus Trichoderma and their antifungal susceptibilities.

    Science.gov (United States)

    Sandoval-Denis, Marcelo; Sutton, Deanna A; Cano-Lira, José F; Gené, Josepa; Fothergill, Annette W; Wiederhold, Nathan P; Guarro, Josep

    2014-06-01

    A set of 73 isolates of the emerging fungus Trichoderma isolated from human and animal clinical specimens were characterized morphologically and molecularly using a multilocus sequence analysis that included the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and fragments of the translation elongation factor 1 alpha (Tef1), endochitinase CHI18-5 (Chi18-5), and actin 1 (Act1) genes. The most frequent species was Trichoderma longibrachiatum (26%), followed by Trichoderma citrinoviride (18%), the Hypocrea lixii/Trichoderma harzianum species complex (15%), the newly described species Trichoderma bissettii (12%), and Trichoderma orientale (11%). The most common anatomical sites of isolation in human clinical specimens were the respiratory tract (40%), followed by deep tissue (30%) and superficial tissues (26%), while all the animal-associated isolates were obtained from superficial tissue samples. Susceptibilities of the isolates to eight antifungal drugs in vitro showed mostly high MICs, except for voriconazole and the echinocandins.

  1. Construction of engineering Trichoderma strains and their characteristics against tomato gray mold

    Institute of Scientific and Technical Information of China (English)

    LIU Xian; ZHUANG Jing-hua; GAO Zeng-gui; YANG Chang-cheng; CHEN Jie

    2004-01-01

    @@ The transformed Trichoderma strains Ttrm31, Ttrm34 and Ttrm55 were obtained from Trichoderma wild strain T21 mutated by REMI technique for more effective biocontrol of tomato gray mold (Botrytis cinerea) with Trichoderma agent. Those transformants appeared much better in biocontrol activity in vitro or in vivo against tomato gray mold were better than that of wild strain T21. The main results were as follow:

  2. CHARACTERIZATION OF SOIL TRICHODERMA ISOLATES FOR POTENTIAL BIOCONTROL OF PLANT PATHOGENS

    OpenAIRE

    S. Matei; Gabi-Mirela Matei; Petruta Cornea; Gabriela Popa

    2011-01-01

    Various fungal strains belonging to genus Trichoderma act as biological control agents for soil born plant pathogens. Two new strains of Trichoderma harzianum (T.h.) and Trichoderma viride (T.v.) were isolated from forest soils in Ilfov county and their morphological aspects, enzymatic and antagonistic activity were examined. Current chemical fungicides had constantly, in time, less influence on pathogens due to their diversity, adaptability and increasing resistance.The paper present the mor...

  3. Isolation and expression of two polyketide synthase genes from Trichoderma harzianum 88 during mycoparasitism

    OpenAIRE

    Lin Yao; Chong Tan; Jinzhu Song; Qian Yang; Lijie Yu; Xinling Li

    2016-01-01

    Abstract Metabolites of mycoparasitic fungal species such as Trichoderma harzianum 88 have important biological roles. In this study, two new ketoacyl synthase (KS) fragments were isolated from cultured Trichoderma harzianum 88 mycelia using degenerate primers and analysed using a phylogenetic tree. The gene fragments were determined to be present as single copies in Trichoderma harzianum 88 through southern blot analysis using digoxigenin-labelled KS gene fragments as probes. The complete se...

  4. PERBAIKAN PERTUMBUHAN DAN HASIL STEVIA (Stevia rebaudiana BERTONI M MELALUI APLIKASI Trichoderma sp.

    Directory of Open Access Journals (Sweden)

    Haryuni -

    2013-09-01

    Full Text Available AbstrakTujuan penelitian ini adalah menguji perbaikan pertumbuhan dan hasil stevia (Stevia rebaudiana Bertoni M melalui penggunaan  Trichoderma sp.  Perbanyakan  Trichoderma sp. dilakukan di laboratorium Balai Proteksi Perkebunan di Salatiga Jawa Tengah. Penelitian dirancang menggunakan rancangan factorial dengan dua faktor. Faktor pertama adalah inokulasi Trichoderma sp (To = tanpa  Trichoderma sp. & T1 = menggunakan Trichoderma sp. 100 g.  Faktor kedua adalah variasi aplikasi perlakuan yaitu: 1. S0 =  tanpa perlakuan, 2. S1 = 10 hari sebelum tanam, 3. S3 = tanama dan  4. S3 = 10  hari setelah tanam. Tiap perlakuan diulang tiga ulangan, tiap ulangan terdiri dari 16 tanaman. Hasil penelitian menunjukkan  bahwa inokulasi Trichoderma sp. dan aplikasi perlakuan  S3 = 10 hari setelah tanam dapat meningkatkan pertumbuhan dan hasil dari  tanaman tevia. AbstractThe object of this research was examine to repair of growth and yield of stevia (Stevia rebaudiana Bertoni M by Trichoderma sp. application.  Reproduction of Trichoderma sp. performed in the laboratory center of Protection plantation Central of Java at Salatiga.The experiment was arranged in a completely randomized completely factorial design, consisted of two factors. The first factor was Trichoderma sp inoculation which were To = without Trichoderma sp. and T1 = Trichoderma sp. 100 g. The second factor was variation  of plant application treatment, which were 1. S0 = without treatment, 2. S1 = 10 days before of planting , 3. S3 = planting  and 4. S3 = 10 days after planting. Each treatment was repeated three times and each replicate consisted of 16 plants. The result showed that that the inoculation of Trichoderma sp.  and application of treatmen 4 is S3= 10 days after planting increases plant growth and yields of stevia.

  5. Neosartorya fischeri var fischeri (Wehmer) Malloch and Cain 1972 (anamorph: Aspergillus fischerianus Samson and Gams 1985) as a cause of mycotic keratitis.

    Science.gov (United States)

    Coriglione, G; Stella, G; Gafa, L; Spata, G; Oliveri, S; Padhye, A A; Ajello, L

    1990-12-01

    The first case of mycotic keratitis caused by Neosartorya fischeri var. fischeri the teleomorph of Aspergillus fischerianus, is described. The patient, a 62-year-old man, had no history of trauma to his infected left eye. The infection progressed despite treatment with ketoconazole and the eye had to be eviscerated. Histological studies revealed the presence of hyaline, septate mycelium in the eye tissue. Cultures gave rise to a thermotolerant mould that developed both its asexual and sexual forms. The isolate was identified on the basis of the morphologic features of its anamorphic and teleomorphic states. In the literature only seven other species of Aspergillus have been unequivocally reported as causing mycotic keratitis. PMID:2091938

  6. Isolated sinusitis sphenoidalis caused by Trichoderma longibrachiatum in an immunocompetent patient with headache.

    Science.gov (United States)

    Molnár-Gábor, Etelka; Dóczi, Ilona; Hatvani, Lóránt; Vágvölgyi, Csaba; Kredics, László

    2013-08-01

    We present a case of isolated sinusitis sphenoidalis caused by Trichoderma longibrachiatum, an emerging causal agent of fungal infections with an often fatal outcome. A Trichoderma strain was isolated from secretion obtained from the sinus sphenoidalis of a rhinosinusitis patient and identified by sequence analysis of two loci as Trichoderma longibrachiatum from the Longibrachiatum Clade of the genus Trichoderma. T. longibrachiatum can trigger a fatal pathomechanism in immunodeficient patients, but only rarely causes disease in healthy people. The case presented is unique because the patient was not immunocompromised.

  7. Formulation of Trichoderma sp. SL2 inoculants using different carriers for soil treatment in rice seedling growth

    OpenAIRE

    Doni, Febri; Isahak, Anizan; Che Mohd Zain, Che Radziah; Mohd Ariffin, Salwati; Wan Mohamad, Wan Nur’ashiqin; Wan Yusoff, Wan Mohtar

    2014-01-01

    Background Trichoderma sp. SL2 has been previously reported to enhance rice germination, vigour, growth and physiological characteristics. The use of Potato Dextrose Agar as carrier of Trichoderma sp. SL2 inoculant is not practical for field application due to its short shelf life and high cost. This study focuses on the use of corn and sugarcane bagasse as potential carriers for Trichoderma sp. SL2 inoculants. Findings A completely randomized design was applied for this study. Trichoderma sp...

  8. DNA barcoding survey of Trichoderma diversity in soil and litter of the Colombian lowland Amazonian rainforest reveals Trichoderma strigosellum sp. nov. and other species.

    Science.gov (United States)

    López-Quintero, Carlos A; Atanasova, Lea; Franco-Molano, A Esperanza; Gams, Walter; Komon-Zelazowska, Monika; Theelen, Bart; Müller, Wally H; Boekhout, Teun; Druzhinina, Irina

    2013-11-01

    The diversity of Trichoderma (Hypocreales, Ascomycota) colonizing leaf litter as well as the rhizosphere of Garcinia macrophylla (Clusiaceae) was investigated in primary and secondary rain forests in Colombian Amazonia. DNA barcoding of 107 strains based on the internal transcribed spacers 1 and 2 (ITS1 and 2) of the ribosomal RNA gene cluster and the partial sequence of the translation elongation factor 1 alpha (tef1) gene revealed that the diversity of Trichoderma was dominated (71 %) by three common cosmopolitan species, namely Trichoderma harzianum sensu lato (41 %), Trichoderma spirale (17 %) and Trichoderma koningiopsis (13 %). Four ITS 1 and 2 phylotypes (13 strains) could not be identified with certainty. Multigene phylogenetic analysis and phenotype profiling of four strains with an ITS1 and 2 phylotype similar to Trichoderma strigosum revealed a new sister species of the latter that is described here as Trichoderma strigosellum sp. nov. Sequence similarity searches revealed that this species also occurs in soils of Malaysia and Cameroon, suggesting a pantropical distribution.

  9. Fungal genus Hypocrea/Trichoderma: from barcodes to biodiversity

    Institute of Scientific and Technical Information of China (English)

    Christian P. KUBICEK; Monika KOMON-ZELAZOWSKA; Irina S. DRUZHININA

    2008-01-01

    Hypocrea/Trichoderma is a genus of soil-borne or wood-decaying fungi containing members important to mankind as producers of industrial enzymes and biocontrol agents against plant pathogens, but also as opportunistic pathogens of immuno-compromised humans and animals, while others can cause damage to cultivated mushroom. With the recent advent of a reliable, BarCode-aided identification system for all known taxa of Trichoderma and Hypocrea, it became now possible to study some of the biological fundamentals of the diversity in this fungal genus in more detail. In this article, we will therefore review recent progress in (1) the understanding of the geographic distribution of individual taxa; (2) mechanisms of speciation leading to development of mushroom diseases and facultative human mycoses; and (3) the possible correlation of specific traits of secondary metabolism and molecular phylogeny.

  10. 40 CFR 180.1293 - Trichoderma gamsii strain ICC 080; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma gamsii strain ICC 080... RESIDUES IN FOOD Exemptions From Tolerances § 180.1293 Trichoderma gamsii strain ICC 080; exemption from the requirement of a tolerance. Trichoderma gamsii strain ICC 080 is exempted from the requirement...

  11. 40 CFR 180.1294 - Trichoderma asperellum strain ICC 012; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma asperellum strain ICC 012... RESIDUES IN FOOD Exemptions From Tolerances § 180.1294 Trichoderma asperellum strain ICC 012; exemption from the requirement of a tolerance. Trichoderma asperellum strain ICC 012 is exempted from...

  12. 40 CFR 180.1201 - Trichoderma harzianum strain T-39; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma harzianum strain T-39... RESIDUES IN FOOD Exemptions From Tolerances § 180.1201 Trichoderma harzianum strain T-39; exemption from the requirement of a tolerance. Trichoderma harzianum strain T-39 is exempt from the requirement of...

  13. In Vitro Antifungal Activity of Strains of Trichoderma harzianum

    OpenAIRE

    KÜÇÜK, Çiğdem; Merih KIVANÇ

    2004-01-01

    Interactions between Trichoderma harzianum strains and some soilborne plant pathogens (Gaeumannomyces graminis var. tritici, Fusarium culmorum and F. moniliforme) were studied on PDA medium. All T. harzianum strains tested produced a metabolite that inhibited growth of plant pathogenic fungi on PDA medium. When grown in liquid cultures containing laminarin, chitin or fungal cell walls as sole carbon sources, 2 strains of T. harzianum produced 1,3-b-glucanase and chitinase in the medium. Highe...

  14. Control of lettuce bottom rot by isolates of Trichoderma spp

    Directory of Open Access Journals (Sweden)

    Zayame Vegette Pinto

    2014-06-01

    Full Text Available Bottom rot, caused by Rhizoctonia solani AG 1-IB, is an important disease affecting lettuce in Brazil, where its biological control with Trichoderma was not developed yet. The present study was carried out with the aim of selecting Trichoderma isolates to be used in the control of lettuce bottom rot. Forty-six Trichoderma isolates, obtained with baits containing mycelia of the pathogen, were evaluated in experiments carried out in vitro and in vivo in a greenhouse in two steps. In the laboratory, the isolates were evaluated for their capabilities of parasitizing and producing toxic metabolic substances that could inhibit the pathogen mycelial growth. In the first step of the in vivo experiments, the number and the dry weight of lettuce seedlings of the cultivar White Boston were evaluated. In the second step, 12 isolates that were efficient in the first step and showed rapid growth and abundant sporulation in the laboratory were tested for their capability of controlling bottom rot in two repeated experiments, and had their species identified. The majority of the isolates of Trichoderma spp. (76% showed high capacity for parasitism and 50% of them produced toxic metabolites capable of inhibiting 60-100% of R. solani AG1-IB mycelial growth. Twenty-four isolates increased the number and 23 isolates increased the dry weight of lettuce seedlings inoculated with the pathogen in the first step of the in vivo experiments.In both experiments of the second step, two isolates of T. virens, IBLF 04 and IBLF 50, reduced the severity of bottom rot and increased the number and the dry weight of lettuce seedlings inoculated with R. solani AG1-IB. These isolates had shown a high capacity for parasitism and production of toxic metabolic substances, indicating that the in vitro and in vivo steps employed in the present study were efficient in selecting antagonists to be used for the control of lettuce bottom rot.

  15. In vitro sensitivity of antagonistic Trichoderma atroviride to herbicides

    OpenAIRE

    Patricia Helena Santoro; Silvia Akimi Cavaguchi; Talita Moretto Alexandre; Janaina Zorzetti; Pedro Manuel Oliveira Janeiro Neves

    2014-01-01

    Trichoderma atroviride was tested in vitro for its sensitivity to different herbicides. The dosages tested were recommended dosage (RD), half dosage (½RD), and double dosage (2RD). Germination, colony-forming units (CFU), radial growth, and spore production were evaluated. Carfentrazone-ethyl and sulfentrazone inhibited the germination at RD and 2RD. A reduction in the CFU was observed for glufosinate-ammonium, atrazine, carfentrazone-ethyl, diuron + paraquat dichloride, imazapyr, oxyfluorfen...

  16. ANTAGONISMO IN VITRO DE Trichoderma spp. A Verticillium dahliae KLEB

    Directory of Open Access Journals (Sweden)

    MARTINS-CORDER M.P.

    1998-01-01

    Full Text Available O presente trabalho visou testar e selecionar isolados selvagens de Trichoderma spp. quanto à capacidade antagônica ao fungo fitopatogênico Verticillium dahliae. Os ensaios in vitro consistiram de testes para avaliar a capacidade hiperparasítica e de antibiose através da produção de metabólitos voláteis e não voláteis. Pela técnica de culturas pareadas, 47 isolados de diversas espécies de Trichoderma foram avaliados e, pelos resultados obtidos, 20 foram selecionados. Através do teste de antibiose, selecionaram-se 7 isolados: T15P e Tal-1 (T. viride, TW6 e CNP311A (T. koningii, CNP17 e TCII (T. harzianum e Tal-10 (T. aureoviride os quais inibiram completamente o crescimento micelial de V. dahliae através da produção de metabólitos. Observações microscópicas demonstraram interações de hifas entre Trichoderma sp. e V. dahliae, tais como: enrolamento, crescimento paralelo de ambos, formação de ganchos.

  17. Identification of novel Trichoderma hamatum genes expressed during mycoparasitism

    Institute of Scientific and Technical Information of China (English)

    Margaret Carpenter; Alison Stewart; Hayley Ridgway

    2004-01-01

    @@ Trichoderma species are currently used as biocontrol agents for crop diseases caused by a number of fungal plant pathogens. However, their biocontrol performance in the field can be unreliable and it is likely that more consistent performance could be achieved through knowledge and manipulation of the genes involved. For example, induction of the genes could be optimised for variable environmental and physiological conditions, superior strains could be selected more effectively and novel strains could be created. One method by which Trichoderma species accomplish biocontrol is mycoparasitism. Several genes involved in the mycoparasitic interaction have previously been characterised, however these consist predominantly of those that encode enzymes that degrade fungal cell walls. In the current study subtractive hybridisation was used to target genes expressed when Trichoderma hamatum and the plant pathogen Sclerotinia sclerotiorum were cultured together, subtracting genes expressed when each are grown individually. This experimental design has the potential to yield T.hamatum genes involved in mycoparasitism of S. sclerotiorum, and S. sclerotiorum genes upregulated in host defence. The cDNA fragments yielded by the subtraction were characterised with respect to expression, sequence and species of origin. A number of novel T. hamatum genes which were up-regulated during mycoparasitism were identified.

  18. Ecological plasticity of Trichoderma fungi in leached chernozem

    Science.gov (United States)

    Svistova, I. D.; Senchakova, T. Yu.

    2010-03-01

    The autecological properties of Trichoderma fungi ecotypes isolated from the leached chernozem of the forest-steppe zone of the European part of Russia have been studied. We were the first who carried out the complex study of the synecological relations of micromycetes of such kinds in a system including the soil, microbial community, and plants, i.e., their relations with soil saprotrophic fungi, bacteria, actinomycetes, plants, and pathogenic fungi. It was shown that the ecological plasticity of the Trichoderma genus in the soil of this zone is determined by its growth rate, the optimum pH and temperature, the biosynthesis of extracellular hydrolytic enzymes, the biological action of mycotoxins, and the ability for parasitism. The efficiency of the introduction of Trichoderma species typical and atypical for the leached chernozem into this soil and their influence on the structure of the microbial community were evaluated. The T. pseudokoningii ecotype, which produces cellulolytic enzymes, is very promising for industrial biotechnology, and the T. harzianum ecotype can be used in soil biotechnology for the biocontrol of chernozem. The addition of a commercial trichodermin preparation into the chernozem damages the structure of its microbial community.

  19. Methods for the Evaluation of the Bioactivity and Biocontrol Potential of Species of Trichoderma.

    Science.gov (United States)

    Steyaert, Johanna; Hicks, Emily; Kandula, Janaki; Kandula, Diwakar; Alizadeh, Hossein; Braithwaite, Mark; Yardley, Jessica; Mendoza-Mendoza, Artemio

    2016-01-01

    Members of the genus Trichoderma comprise the majority of commercial fungal biocontrol agents of plant diseases. As such, there is a wealth of information available on the analysis of their biocontrol potential and the mechanisms behind their superior abilities. This chapter aims to summarize the most common methods utilized within a Trichoderma biocontrol program for assessing the biological properties of individual strains. PMID:27565489

  20. Trichoderma species fungemia after high-dose chemotherapy and autologous stem cell transplantation: a case report.

    Science.gov (United States)

    Festuccia, M; Giaccone, L; Gay, F; Brunello, L; Maffini, E; Ferrando, F; Talamo, E; Boccadoro, M; Serra, R; Barbui, A; Bruno, B

    2014-08-01

    We present a case of Trichoderma fungemia with pulmonary involvement in a multiple myeloma patient, who was severely immunocompromised and heavily treated with high-dose melphalan, and underwent autologous hematopoietic cell transplantation. This is the first report, to our knowledge, of proven Trichoderma fungemia, defined by published criteria, successfully treated with voriconazole.

  1. Combined inoculation of Pseudomonas fluorescens and Trichoderma harzianum for enhancing plant growth of vanilla (Vanilla planifolia).

    Science.gov (United States)

    Sandheep, A R; Asok, A K; Jisha, M S

    2013-06-15

    This study was conducted to evaluate the plant growth promoting efficiency of combined inoculation of rhizobacteria on Vanilla plants. Based on the in vitro performance of indigenous Trichoderma spp. and Pseudomonas spp., four effective antagonists were selected and screened under greenhouse experiment for their growth enhancement potential. The maximum percentage of growth enhancement were observed in the combination of Trichoderma harzianum with Pseudomonas fluorescens treatment followed by Pseudomonas fluorescens, Trichoderma harzianum, Pseudomonas putida and Trichoderma virens, respectively in decreasing order. Combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens registered the maximum length of vine (82.88 cm), highest number of leaves (26.67/plant), recorded the highest fresh weight of shoots (61.54 g plant(-1)), fresh weight of roots (4.46 g plant(-1)) and dry weight of shoot (4.56 g plant(-1)) where as the highest dry weight of roots (2.0806 g plant(-1)) were achieved with treatments of Pseudomonas fluorescens. Among the inoculated strains, combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens recorded the maximum nitrogen uptake (61.28 mg plant(-1)) followed by the combined inoculation of Trichoderma harzianum (std) and Pseudomonas fluorescens (std) (55.03 mg plant(-1)) and the highest phosphorus uptake (38.80 mg plant(-1)) was recorded in dual inoculation of Trichoderma harzianum and Pseudomonas fluorescens.

  2. 21 CFR 184.1250 - Cellulase enzyme preparation derived from Trichoderma longibrachiatum.

    Science.gov (United States)

    2010-04-01

    ... incorporated by reference in accordance with 5 U.S.C. 552(a) and 1 CFR part 51. Copies are available from the... Trichoderma longibrachiatum. 184.1250 Section 184.1250 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT....1250 Cellulase enzyme preparation derived from Trichoderma longibrachiatum. (a) Cellulase...

  3. Impact of Trichoderma spp. on Soybean Seed Germination and Potential Antagonistic Effect on Sclerotinia sclerotiorum

    OpenAIRE

    Sonja Tančić; Jelica Skrobonja; Mirjana Lalošević; Radivoje Jevtić; Miloš Vidić

    2013-01-01

    Trichoderma species have been registered as species with important plant growth promoting potential and antagonistic effect against various phytopathogens. Trichoderma isolates originating from different soil types from the Vojvodina region (Serbia) were screened using dual culture test for their antagonistic effect against the pathogen Sclerotinia sclerotiorum. All tested isolates had high radial growth inhibition (RGI) factors of the pathogen and high col...

  4. Trichoderma Biocontrol: Signal Transduction Pathways Involved in Host Sensing and Mycoparasitism

    Directory of Open Access Journals (Sweden)

    Susanne Zeilinger

    2007-01-01

    Full Text Available Fungi of the genus Trichoderma are used as biocontrol agents against several plant pathogenic fungi like Rhizoctonia spp., Pythium spp., Botrytis cinerea and Fusarium spp. which cause both soil-borne and leaf- or flower-borne diseases of agricultural plants. Plant disease control by Trichoderma is based on complex interactions between Trichoderma, the plant pathogen and the plant. Until now, two main components of biocontrol have been identified: direct activity of Trichoderma against the plant pathogen by mycoparasitism and induced systemic resistance in plants. As the mycoparasitic interaction is host-specific and not merely a contact response, it is likely that signals from the host fungus are recognised by Trichoderma and provoke transcription of mycoparasitism-related genes.In the last few years examination of signalling pathways underlying Trichoderma biocontrol started and it was shown that heterotrimeric G-proteins and mitogen-activated protein (MAP kinases affected biocontrol-relevant processes such as the production of hydrolytic enzymes and antifungal metabolites and the formation of infection structures. MAPK signalling was also found to be involved in induction of plant systemic resistance in Trichoderma virens and in the hyperosmotic stress response in Trichoderma harzianum. Analyses of the function of components of the cAMP pathway during Trichoderma biocontrol revealed that mycoparasitism-associated coiling and chitinase production as well as secondary metabolism are affected by the internal cAMP level; in addition, a cross talk between regulation of light responses and the cAMP signalling pathway was found in Trichoderma atroviride.

  5. Cryptococcus randhawai sp. nov., a novel anamorphic basidiomycetous yeast isolated from tree trunk hollow of Ficus religiosa (peepal tree) from New Delhi, India.

    Science.gov (United States)

    Khan, Zia U; Ahmad, Suhail; Hagen, Ferry; Fell, Jack W; Kowshik, Tusharantak; Chandy, Rachel; Boekhout, Teun

    2010-03-01

    A novel anamorphic Cryptococcus species is described, which was isolated in New Delhi (India) from decaying wood of a tree trunk hollow of Ficus religiosa. On the basis of sequence analysis of the D1/D2 domains of the 26S rRNA gene and the internally transcribed spacer (ITS)-1 and ITS-2 region sequences, the isolate belonged to the Cryptococcus albidus cluster (Filobasidiales, Tremellomycetes) and was closely related to Cryptococcus saitoi, Cryptococcus cerealis and Cryptococcus friedmannii with 98% sequence identity. Phenotypically, the species differed from C. saitoi with respect to growth temperature (up to 37degrees C), presence of a thin capsule, ability to grow in the absence of vitamins, and inability to assimilate citrate and ethylamine. With respect to C. friedmannii, it differed in growth temperature, ability to assimilate lactose, raffinose, L: -rhamnose, myo-inositol, and inability to utilize citrate. Furthermore, our isolate also differed from C. cerealis in growth temperature, presence of capsule and inability to assimilate L: -sorbose. In view of the above phenotypic differences and unique rDNA sequences, we consider that our isolate represents a new species of Cryptococcus, and therefore, a new species, Cryptococcus randhawai is proposed for this taxon. The type strain J11/2002 has been deposited in the culture collection of the Centraalbureau voor Schimmelcultures (CBS10160) and CABI Biosciences (IMI 393306). PMID:20091225

  6. An isolate of Arthroderma benhamiae with Trichophyton mentagrophytes var. erinacei anamorph isolated from a four-toed hedgehog (Atelerix albiventris) in Japan.

    Science.gov (United States)

    Takahashi, Yoko; Haritani, Kuniko; Sano, Ayako; Takizawa, Kayoko; Fukushima, Kazutaka; Miyaji, Makoto; Nishimura, Kazuko

    2002-01-01

    A female four-toed hedgehog probably imported from Africa and kept as a pet by a family suffered from depilation and mite (Caparinia tripilis) infection. Depilated quills were inoculated on a commercially available medium and an isolate of the dermatophytes was obtained. A giant colony after 14 days incubation on yeast extract Sabourauds agar had a central umbo with white granular surface and a yellow pigment ring in the reverse. The hedgehog isolate produced numerous elongated microconidia singly attached along the sides of hyphae. Macroconidia were somewhat irregular in shape and size and 2-6 septa. Abundant intermediate sized spores between micro- and macro conidia and few spirals were observed. Hair perforation and urease activity tests were positive. Maximum growth temperature was 40 C. In the mating tests using the tester strains of both African and Americano-European races of Arthroderma benhamiae, the strain produced numerous gymnothecia only when paired with the African race mating type minus(-). In addition, 591 bases of the internal transcribed spacer region of the ribosomal RNA gene including the 5.8S region (ITS1-5.8S-ITS2) were sequenced and corresponded to those of T. mentagrophytes var. erinacei (DDBJ/EMBL/GenBank accession numbers Z97996 and Z97997) by more than 99.7%. Therefore, our case is the first isolation of A. benhamiae with T. mentagrophytes var. erinacei anamorph in Japan. PMID:12402026

  7. Unraveling the efficient applications of secondary metabolites of various Trichoderma spp.

    Science.gov (United States)

    Keswani, Chetan; Mishra, Sandhya; Sarma, Birinchi Kumar; Singh, Surya Pratap; Singh, Harikesh Bahadur

    2014-01-01

    Recent shift in trends of agricultural practices from application of synthetic fertilizers and pesticides to organic farming has brought into focus the use of microorganisms that carryout analogous function. Trichoderma spp. is one of the most popular genera of fungi commercially available as a plant growth promoting fungus (PGPF) and biological control agent. Exploitation of the diverse nature of secondary metabolites produced by different species of Trichoderma augments their extensive utility in agriculture and related industries. As a result, Trichoderma has achieved significant success as a powerful biocontrol agent at global level. The endorsement of Trichoderma spp. by scientific community is based on the understanding of its mechanisms of action against a large set of fungal, bacterial and in certain cases viral infections. However, it is still an agnostic view that there could be any single major mode of operation, although it is argued that all mechanisms operate simultaneously in a synchronized fashion. The central idea behind this review article is to emphasize the potentiality of applications of target specific secondary metabolites of Trichoderma for controlling phytopathogens as a substitute of commercially available whole organism formulations. With the aim to this point, we have compiled an inclusive list of secondary metabolites produced by different species of Trichoderma and their applications in diverse areas with the major emphasis on agriculture. Outlining the importance and diverse activities of secondary metabolites of Trichoderma besides its relevance to agriculture would generate greater understanding of their other important and beneficial applications apart from target specific biopesticides.

  8. The potentiality of Trichoderma harzianum in alleviation the adverse effects of salinity in faba bean plants.

    Science.gov (United States)

    Abd El-Baki, G K; Mostafa, Doaa

    2014-12-01

    The interaction between sodium chloride and Trichoderma harzianum (T24) on growth parameters, ion contents, MDA content, proline, soluble proteins as well as SDS page protein profile were studied in Vicia faba Giza 429. A sharp reduction was found in fresh and dry mass of shoots and roots with increasing salinity. Trichoderma treatments promoted the growth criteria as compared with corresponding salinized plants. The water content and leaf area exhibited a marked decrease with increasing salinity. Trichoderma treatments induced a progressive increase in both parameters. Both proline and MDA contents were increased progressively as the salinity rose in the soil. Trichoderma treatments considerably retarded the accumulation of both parameters in shoots and roots. Both Na+ and K+ concentration increased in both organs by enhancing salinity levels. The treatment with Trichoderma harzianum enhanced the accumulation of both ions. Exposure of plants to different concentrations of salinity, or others treated with Trichoderma harzianum produced marked changes in their protein pattern. Three types of alterations were observed: the synthesis of certain proteins declined significantly, specific synthesis of certain other proteins were markedly observed and synthesis of a set specific protein was induced de novo in plant treated with Trichoderma harzianum.

  9. Trichoderma strains- Silybum marianum hairy root cultures interactions

    Directory of Open Access Journals (Sweden)

    T. Hasanloo

    2015-03-01

    Full Text Available Background and objectives: Silymarin is a unique flavonoid complex with documented hepatoprotective properties. Silybum  marianum hairy root culture as a source for producing silymarin has been an important strategy for study the cell signaling pathway. In the present investigation Trichoderma strains- Silybum marianum hairy root cultures interactions have been studied. Methods: The effects of two Trichoderma Strains (KHB and G46-7 (0, 0.5, 1, 2 and 4 mg/ 50 mL culture in 6 different exposure times (0, 24, 48, 72, 96 and 120 h have been investigated on flavonolignans production. The flavonolignans were analyzed by High Performance Liquid Chromatography method. Cell signaling pathway was evaluated by determination of H2O2 content, peroxidase and ascorbate peroxidase activities. Results:The elicitation effects of two Trichoderma Strains (KHB and G46-7 were examined on flavonolignans accumulation and the activation of cell defense system in S. marianum hairy root cultures. The results indicated that the highest silymarin accumulation (0.45 and 0.33 mg/g DW was obtained in media elicited with 0.5 mg/50 mL cultures of T. harzianum Strains (KHB and G46-3, respectively after 120 h. Feeding time experiments indicated that a significant higher content of silymarin production was achieved after 120 and 72 h in media treated with 0.5 mg/50 mL cultures of KHB and G46-3, respectively. Our results showed that S. marianum treated by KHB strain, increased taxifolin, silychristin, isosilybin and silydianin productions significantly. The H2O2 content in the control hairy root cultures remained lower than the treated cultures. There was significant enhancement in both peroxidase and ascorbate peroxidase activities in treated hairy roots reaching a peak after 72 h. Conclusion: These findings suggested that some Trichoderma strains are positive elicitors for promoting silymarin accumulation in S. marianum hairy root cultures. The results also suggested the presence

  10. Proteomic mapping of secreted proteins of Trichoderma spp.

    Institute of Scientific and Technical Information of China (English)

    Li S; Bramley P M; Smith J; Cannon P F

    2004-01-01

    @@ A series of highly taxonomically diverse Trichoderma strains were investigated using proteomic approaches, to investigate the utility of protein profiles as taxonomic markers and to identify proteins of potential economic importance. Initial studies have focused on a comparison of single strains of T.aureoviride, T. saturnisporum, T. polysporum, T. longbrachiatum and T. spirale, along with two strains of T. harzianum. All seven strains were grown in synthetic medium supplemented with 2 % (w/v) glycerol, to maximize the diversity of extracellular protein production. Samples of secreted protein were separated by 2D gel electrophoresis and will be characterized by MALDI-TOF peptide fingerprinting.

  11. Three tubulin genes of Trichoderma harzianum: alpha, beta and gamma

    Directory of Open Access Journals (Sweden)

    Min Li

    2010-08-01

    Full Text Available Three tubulin genes of Trichoderma harzianum were cloned followed genomic walking procedure. The tubulins showed high degree of amino acid homology with other fungal tubulins and were homologous with each other with 32 to 38% amino acid identity. Three measures for the degree of codon usage bias indicated the presence of bias in all the sequences, suggesting high expression levels in all the genes. Protein structures were modeled to provide the basis for understanding the tubulin's properties and its interactions with microtubule-associated proteins. Potential motifs were also postulated.

  12. Genetic improvement of Trichoderma ability to induce systemic resistance

    Institute of Scientific and Technical Information of China (English)

    Ciliento R; Mach R L; Lorito M; Woo S L; Di Benedetto P; Ruocco M; Scala F; Soriente I; Ferraioli S; Brunner K; Zeilinger S

    2004-01-01

    @@ The beneficial applications of Trichoderma spp. in agriculture include not only the control of plant pathogens, but also the improvement of plant growth, micronutrient availability, and plant tolerance to abiotic stress. In addition, it has been suggested that these fungi are able to increase plant disease resistance by activating induced systemic resistance (ISR) . The mode of action of these beneficial fungi in the Trichoderma -plant-pathogen interaction are many, complex and not completely understood. Numerous lytic enzymes have been characterized, the encoding genes (ech42 gluc78,nag1 from T. atroviride strain P1) cloned, and their role in biocontrol demonstrated. The corresponding biocontrol-related inducible promoters have been used in a reporter system based on the Aspergillus niger glucose oxidase gene (goxA) to monitor biocontrol activity. Glucose oxidase catalyzes the oxygen-dependent oxidation of D-glucose to D-glucono-1, 5-lactone and hydrogen peroxide; this latter compound is known to have an antifungal effect and activate the plant defence cascade, thus increasing resistance to pathogen attack. T. atroviride P1 transformants with various promoters gox were tested as seed coating treatments on bean seeds planted in soil infested with a soilborne fungal pathogen. Successively, the emergent leaves were inoculated with a foliar pathogen to determine the effect of the GOX transformants on biocontrol and resistance to pathogen attack.Inoculations with the P1-GOX transformants not only reduced disease symptoms caused by a soil pathogen, but also the lesions of various foliar pathogens applied far from the Trichoderma colonization, thus activating ISR. A similar approach is being use to genetically improve T.harzianum T22, a rhizosphere competent and commercially marketed strain not transformed yet, by using four different gox gene constructs under the control of constitutive and inducible promoters.Plasmids have been introduced in Trichoderma by

  13. Competition in artifical plant growth media by Trichoderma spp

    DEFF Research Database (Denmark)

    Sarocco, Sabrina; Lübeck, Mette; Vannacci, Giovanni

    of the reason why more biocontrol agents are reaching the market place. A comparative evaluation of life strategies of both the pathogen and its antagonists is required to predict the fate of a biopesticide in agricultural systems.The objectives of this work have been: 1) to screen a collection of Trichoderma......, but even after R. solani was able to re-colonize haulm fragments, the positive effects on damping off were still evident, demonstrating a “residual” effect of antagonists. On the whole, competition for the food base explains a large part of the recorded effects....

  14. Trichoderma koningiopsis:A New Chinese Record of the Genus Trichoderma%木霉属中国新记录种Trichoderma koningiopsis记述

    Institute of Scientific and Technical Information of China (English)

    李广记; 陈捷; 刘铜; 刘力行

    2010-01-01

    在华东地区木霉菌资源调查中,利用内转录间隔区(ITS)序列分析和形态学鉴定方法,对从土壤中分离到的木霉菌进行鉴定,发现一个中国新纪录种,即拟康宁木霉.Trichoderma koningiopsis/Hypocrea koningiopsis Samuels,C.Suarez & H.C.Evans sp.nov..该种典型的形态特征是在PDA以及CMD(玉米粉琼脂)上有瓶梗层出现象,而在MA(麦芽提取物)培养基上没有此特征.

  15. Cell wall degrading enzymes in Trichoderma asperellum grown on wheat bran

    DEFF Research Database (Denmark)

    Bech, Lasse; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    Trichoderma asperellum is a filamentous fungus that is able to produce and secrete a wide range of extracellular hydrolytic enzymes used for plant cell wall degradation. The Trichoderma genus has attracted considerable attention from the biorefinery industry due to the production of cell wall...... Pattern Recognition enabling an efficient enzyme discovery. This was furthermore used to re-annotate CAZYmes present in five publically available Trichoderma species, hereby elucidating differences in CAZYmes on a functional level in contrary to glycoside hydrolase family level. This comparison supports...

  16. Isolation, Purification, and Structural Identification of an Antifungal Compound from a Trichoderma Strain.

    Science.gov (United States)

    Li, Chong-Wei; Song, Rui-Qing; Yang, Li-Bin; Deng, Xun

    2015-08-01

    Trichoderma strain T-33 has been demonstrated to have inhibitory effect on the fungus species Cytospora chrysosperma. Here, an active antifungal compound was obtained from Trichoderma strain T-33 extract via combined separation technologies, including organic solvent extraction, liquid chromatography, and thin-layer chromatography. The purified compound was further characterized by advanced analytical technologies to elucidate its chemical structure. Results indicated that the active antifungal compound in Trichoderma strain T-33 extract is 2,5- cyclohexadiene-1,4-dione-2,6-bis (1,1-dimethylethyl).

  17. Antagnism of three strains of Trichoderma spp.against mycelial growth of Rhizoctonia salani

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jing-ze; TU Yan-la

    2004-01-01

    @@ Three strains of Trichoderma spp. TVll2, TX003, TY009 obtained from previous experiments could inhibit the sclerotial formation of two strains of Rhizoctonia salani AG1 (-1A) isolated from the rice paddies in Hanzhou of China. However, it is unclear if there are the antagonism and mycoparasitism of the Trichoderma strains tested against the mycelial growth of R. solani . The objective of this research was to evaluate the ability of the Trichoderma strains to inhibit the mycelial growth of R. solani in vitro .

  18. INFLUENCE OF TRICHODERMA SPP AGAINST ALTERNARIA TENUISSIMA INCITING LEAF SPOT OF RUMEX ACETOSA L

    Directory of Open Access Journals (Sweden)

    M. G. Ambuse, V. S. Chatage and U. N. Bhale

    2012-06-01

    Full Text Available Leaf spot of Sorrel (Rumex acetosa L. is caused by Alternaria tenuissima. This paper describes the efficacy of Trichoderma spp against sensitive and resistant isolates of A. tenuissima by dual culture method under invitro conditions. Trichoderma viride T. harzianum, T. virens, T. koningii and T. pseudokoningii species were used for antagonistic study. Results indicate that all Trichoderma species showed antagonistic activity. But among them, T.viride, T.koningii and T.pseudokoningii showed 80% antagonistic activity than others in case of sensitive isolate of test fungus. Resistant isolate of pathogen was restricting the antagonism in some extent.

  19. Parasitism of Rhizoctonia solani by strains of Trichoderma spp. Parasitismo de Rhizoctonia solani por linhagens de Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Itamar Soares de Melo

    2000-03-01

    Full Text Available Rhizoctonia solani causes serious diseases in a wide range of plant species. The fungus Trichoderma has been shown to be particularly effective in the control of the pathogen. Thus, this research was carried out to screen fourteen Trichoderma strains against R. solani in vitro. All strains tested inhibited the growth of R. solani. Three T. koningii strains produced toxic metabolites with strong activity against R. solani, inhibiting the mycelial growth by 79%. T. harzianum, Th-9 reduced the viability of sclerotia of R. solani by 81.8% and T. koningii, TK-5 reduced by 53%. Electron microscopic observations revealed that all T. harzianum strains interacted with R. solani. Th-9 grew toward and coiled around the host cells, penetrating and destroying the hyphae. Penetration of host cells was apparently accomplished by mechanical activity.Rhizoctonia solani é um dos mais destrutivos patógenos de plantas cultivadas. Métodos alternativos de controle têm sido empregados com sucesso, particularmente, utilizando-se o fungo Trichoderma. Este trabalho visou, portanto, selecionar linhagens efetivas desse micoparasita contra o patógeno. Onze linhagens de T. harzianum e três de T. koningii foram testadas in vitro com relação ao parasitismo de hifas e de escleródios e produção de metabólitos tóxicos. Todas as linhagens de Trichoderma spp. inibiram o crescimento miceliano de R. solani e as três linhagens de T. koningii produziram potentes antibióticos, que inibiram mais de 79% o crescimento do patógeno. Uma linhagem de T. harzianum, Th-9, reduziu a viabilidade dos escleródios em 81,8% e uma de T. koningii em 53%. Microscopia eletrônica de varredura revelou que todas as linhagens de T. harzianum parasitaram R. solani enquanto nenhuma linhagem de T. koningii interagiu com R. solani, possivelmente, devido à forte inibição causada pelos metabólitos que impediu o contato entre os dois fungos. T. harzianum, Th-9, cresceu ao redor, penetrou e

  20. POTENCIAL FARMACOINDUSTRIAL DE Trichoderma harzianum PARA FINS FARMACOTERAPÊUTICOS

    Directory of Open Access Journals (Sweden)

    Luís Fernando Albarello Gellen

    2014-12-01

    Full Text Available Linhagens de Trichoderma estão bem difundidas nos processos de controle de fitopatógenos, além disto, para promoção do desenvolvimento e crescimento das culturas onde são inoculados, estes benefícios dão-se pela gama de processos desempenhados por este organismo, os processos são classificados em parasitismo, antibiose e competição, além de secretarem produtos enzimáticos com ações degradantes, compostos voláteis e antimicrobianos. Por meio de testes de produção enzimática, confronto em placa, metabólitos voláteis e sensibilização do agente, os isolados de Trichoderma harzianum mostraram-se como um potente produtor de substâncias antimicrobianas e antifúngicas perante Sthaphylococcus aureus, Streptococcus pyogenes, E. coli, Pseudomonas aeruginosa, E. faecalis e Rhodotorula sp., Candida albicans, Candida parapsilosis e Candida lusitaniae. Palavras-chave: antimicrobianos, antifúngicas, metabólitos, sensibilização. DOI: http://dx.doi.org/10.18561/2179-5746/biotaamazonia.v4n4p91-96

  1. Quality of Agricultural by Products Fermented by Trichoderma harzianum

    Directory of Open Access Journals (Sweden)

    Nurhayati Nurhayati

    2014-10-01

    Full Text Available (Kualitas limbah produk pertanian yang difermentasi dengan trichoderma harzianum ABSTRAK. Telah dilakukan penelitian untuk mengetahui pengaruh fermentasi menggunakan Trichoderma harzianum terhadap kualitas limbah pertanian sehingga dapat digunakan sebagai bahan pakan non konvensional pada ternak unggas. Sekam padi, kulit pisang dan kulit nanas digunakan dalam penelitian ini dan difermentasi dengan T. harzianum selama 7 hari. Penelitian dirancang menggunakan Rancangan Acak Lengkap pola faktorial dengan 2 faktor yaitu 3 jenis limbah pertanian (sekam padi, kulit pisang dan kulit nanas dan 4 level T. harzianum ( 0, 4, 8 dan 12%. Setiap unit perlakuan diulang 4 kali. Parameter yang diamati yaitu kandungan bahan kering, protein kasar dan serat kasar. Data yang dihimpun dianalisis menggunakan analisis ragam (ANOVA dan Uji Jarak Berganda Duncan. Hasil penelitian memperlihatkan bahwa terdapat perbedaan yang nyata (P<0.05 kualitas kulit nanas, kulit pisang dan sekam padi setelah difermentasi menggunakan T. harzianum. Kandungan protein kasar kulit nanas dan sekam padi lebih tinggi dibandingkan kulit pisang sedangkan kandungan serat kasarnya lebih rendah. Semakin meningkat level penggunaan T. harzianum pada fermentasi kulit nanas dan sekam padi maka semakin meningkat kandungan protein kasar dan semakin menurun kandungan serat kasarnya. Berbeda dengan kulit pisang, yang kualitasnya tidak nyata dipengaruhi oleh meningkatnya level T. harzianum. Disimpulkan bahwa T. harzianum dapat digunakan untuk memfermentasi limbah pertanian yang kaya kandungan serat untuk meningkatkan kandungan protein dan menurunkan serat kasarnya. Kulit nanas dan sekam padi memiliki kualitas lebih tinggi setelah difermentasi menggunakan 12 % T. harzianum. Kulit pisang tidak disarankan untuk difermentasi menggunakan T. harzianum.

  2. Trichoderma spp.对樟子松枯梢病病原菌的影响%Effect of Trichoderma spp.on the Pathogen of Sphaeropsis Blight of Pinus sylvestris var.mongolica

    Institute of Scientific and Technical Information of China (English)

    宋瑞清; 周秀华

    2004-01-01

    通过Trichoderma属3个菌株与樟子松枯梢病菌(Sphaeropsis sapinea)对峙培养试验的结果表明:Trichoderma viride 1, Trichoderma viride 2及Trichoderma harzianum对樟子松枯梢病病原菌都有抑制效果,其中Trichoderma viride 2对病原菌的相对抑制效果最好,且其相对抑制效果随着时间的增加而增长,在40h达到最高,为73.55.Trichoderma viride 1和Trichoderma harzianum对病原菌的相对抑制效果在16h达到最高,分别为4.86及2.59.

  3. Trichoderma chlorosporum,a new record of endophytic fungi from Dendrobium nobile in China%中国木霉属内生真菌一个新记录种Trichoderma chlorosporum

    Institute of Scientific and Technical Information of China (English)

    袁志林; 陈益存; 章初龙; 林福呈; 陈连庆

    2008-01-01

    @@ 1 INTRODUCTION Typically,Trichoderma spp.are free-living fungi most frequently isolated from soils and have been extensively studied due to their remarkable biocontrol and plant-growth promoting capacity.There is increasing evidence that sometimes they display flexible lifestyle and penetrate epidermis of roots and act as opportunistic,avirulent plant symbiont(Harman et al.2004).Recently,a Trichoderma species living as endophyte,Trichoderma taxi C.L.Zhang et al.(2007),has been recognized.

  4. Carbendazim resistance and calculation effective concentration of carbendazim for Trichoderma harzianum

    Institute of Scientific and Technical Information of China (English)

    Elham Siassi; YANG Qian

    2005-01-01

    There is a method for investigating the transformation of resistance gene of carbendazim into Trichoderma harzianum. In order to introduce the resistance to benzimidazole fungicide into bio-control microorganism Trichoderma harzianum was transformed with the resistance gene. In this study, we investigate resistance level and calculate EC 50 ( effective concentration of carbendazim that can survive 50% of Trichoderma harzianum in that concentration) and stability of the resistance for the transformant isolate of Trichoderma harzianum.Results show the transformants can growth on the medium containing more than 1 000 μg/ml carbendazim and the resistance is stabled after 10 times transfer on non-selective medium and have EC 50 average about, 1 200μg/ml.

  5. Comparative molecular evolution of Trichoderma chitinases in response to mycoparasitic interactions

    DEFF Research Database (Denmark)

    Ihrmark, Katarina; Asmail, Nashwan; Ubhayasekera, Wimal;

    2010-01-01

    Certain species of the fungal genus Trichoderma are potent mycoparasites and are used for biological control of fungal diseases on agricultural crops. In Trichoderma, whole-genome sequencing reveal between 20 and 36 different genes encoding chitinases, hydrolytic enzymes that are involved...... in the mycoparasitic attack. Sequences of Trichoderma chitinase genes chi18-5, chi18-13, chi18-15 and chi18-17, which all exhibit specific expression during mycoparasitism-related conditions, were determined from up to 13 different taxa and studied with regard to their evolutionary patterns. Two of them, chi18......-usage and contains five codons that evolve under positive selection and three groups of co-evolving sites. Regions of high amino acid variability are preferentially localized to substrate- or product side of the catalytic clefts. Differences in amino acid diversity/conservation patterns between different Trichoderma...

  6. Studies on the control of Fusarium oxysporum f.sp.cubense with Trichoderma

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yue-li; LIU Kai-qi; XIANG Mei-mei; LIU Ren

    2004-01-01

    @@ One hundred and fifty one isolates of Trichoderma were collected mainly from more than 40 soil samples and other materials in Guangdong Province (including Chigang, Zhanjiang, Wuchuan,Panyu, Zhaoqing, Dongguan, Humen, Qingyuan, Guanzhou) and the soil samples were also from different plant rhizosphere (including rice, different fruits and different vegetables). It was shown that 39 isolates of Trichoderma grew faster than other isolates using growth velocity experiments.The 39 isolates could effectively inhibit Fusarium oxysporum f. sp. cubense (E. F. Sm) Sny. &Hans.by dual cultural experiments. The inhibited activity included the antifungal activities of its metabolite,mycoparasitic activities and the lytic enzymes by dual culture, electronic microcopy and enzyme assay.At present, studies on the taxonomy of the 151 isolates of Trichoderma are carried out in our experiments, some Trichoderma species aggregates will be identified according to the taxonomy system revised by Rifai and Bissett.

  7. Production of chitinases with Trichoderma harzianun isolates using solid substrate fermentation

    Institute of Scientific and Technical Information of China (English)

    Viviana Nagy

    2004-01-01

    @@ Over forty Trichoderma harzianum isolates have been screened in solid substrate fermentation (SSF)for chitinase production. Strains were isolated from Asian soil and tree bark samples. Identification was performed in Canada and Austria by classical and molecular taxonomical methods.

  8. Exploitation of Trichoderma species on the growth of Pythium Aphanidermatum in Chilli.

    Science.gov (United States)

    Muthukumar, A; Eswaran, A; Sanjeevkumas, K

    2011-10-01

    Damping-off of chilli caused by Pythium aphanidermatum is a major nursery disease in vegetables. In vitro experiments evaluated the effect of eight isolates of Trichoderma species (from chilli rhizosphere) were tested against P. aphanidermatum. All the Trichoderma species had varied antagonistic effects against the pathogen. Among them, TVC3 recorded maximum growth inhibition of P. aphanidermatum and produced more amounts of volatile and non-volatile metabolites. The culture filtrate of the Trichoderma isolate TVC3 recorded complete inhibition on the mycelial growth of pathogen at 15% concentration. Moreover, chilli seeds treated with culture filtrate of the isolate TVC3 recorded maximum germination percentage, shoot length, root length and vigour index of chilli. The study identified the Trichoderma isolate (TVC3) performed well in inhibiting the mycelial growth of pathogen as well as increased the plant growth in chilli. PMID:24031794

  9. Trichoderma spp. NA EMERGÊNCIA E CRESCIMENTO DE MUDAS DE CAMBARÁ (Gochnatia polymorpha (Less. Cabrera1

    Directory of Open Access Journals (Sweden)

    Daniele Franco Martins Machado

    2015-02-01

    Full Text Available Neste trabalho, o objetivo foi avaliar o efeito de isolados de Trichoderma spp. na emergência de plântulas e no crescimento de mudas de cambará (Gochnatia polymorpha. Utilizou-se, em casa de vegetação, substrato esterilizado e não esterilizado, sendo avaliados os efeitos de quatro isolados de trichoderma: TSM1 e TSM2 de Trichoderma viride, 2B2 e 2B22 de Trichoderma harzianum mais um mix preparado com a mistura desses quatro isolados, além de dois produtos comerciais à base de trichoderma. A análise dos dados permitiu concluir que os isolados de trichoderma testados não interferem na emergência das plântulas, mas os isolados 2B2 e 2B22 de T. harzianum apresentam potenciais como promotores de crescimento de mudas de cambará.

  10. Molecular Characterization of Reptile Pathogens Currently Known as Members of the Chrysosporium Anamorph of Nannizziopsis vriesii Complex and Relationship with Some Human-Associated Isolates

    Science.gov (United States)

    Hambleton, Sarah; Paré, Jean A.

    2013-01-01

    In recent years, the Chrysosporium anamorph of Nannizziopsis vriesii (CANV), Chrysosporium guarroi, Chrysosporium ophiodiicola, and Chrysosporium species have been reported as the causes of dermal or deep lesions in reptiles. These infections are contagious and often fatal and affect both captive and wild animals. Forty-nine CANV isolates from reptiles and six isolates from human sources were compared with N. vriesii based on their cultural characteristics and DNA sequence data. Analyses of the sequences of the internal transcribed spacer and small subunit of the nuclear ribosomal gene revealed that the reptile pathogens and human isolates belong in well-supported clades corresponding to three lineages that are distinct from all other taxa within the family Onygenaceae of the order Onygenales. One lineage represents the genus Nannizziopsis and comprises N. vriesii, N. guarroi, and six additional species encompassing isolates from chameleons and geckos, crocodiles, agamid and iguanid lizards, and humans. Two other lineages comprise the genus Ophidiomyces, with the species Ophidiomyces ophiodiicola occurring only in snakes, and Paranannizziopsis gen. nov., with three new species infecting squamates and tuataras. The newly described species are Nannizziopsis dermatitidis, Nannizziopsis crocodili, Nannizziopsis barbata, Nannizziopsis infrequens, Nannizziopsis hominis, Nannizziopsis obscura, Paranannizziopsis australasiensis, Paranannizziopsis californiensis, and Paranannizziopsis crustacea. Chrysosporium longisporum has been reclassified as Paranannizziopsis longispora. N. guarroi causes yellow fungus disease, a common infection in bearded dragons and green iguanas, and O. ophiodiicola is an emerging pathogen of captive and wild snakes. Human-associated species were not recovered from reptiles, and reptile-associated species were recovered only from reptiles, thereby mitigating concerns related to zoonosis. PMID:23926168

  11. Efficiency of treatments for controlling Trichoderma spp during spawning in cultivation of lignicolous mushrooms.

    Science.gov (United States)

    Colavolpe, María Belén; Mejía, Santiago Jaramillo; Albertó, Edgardo

    2014-01-01

    Trichoderma spp is the cause of the green mold disease in mushroom cultivation production. Many disinfection treatments are commonly applied to lignocellulose substrates to prevent contamination. Mushroom growers are usually worried about the contaminations that may occur after these treatments during handling or spawning. The aim of this paper is to estimate the growth of the green mold Trichoderma sp on lignocellulose substrates after different disinfection treatments to know which of them is more effective to avoid contamination during spawning phase. Three different treatments were assayed: sterilization (121 °C), immersion in hot water (60 and 80 °C), and immersion in alkalinized water. Wheat straw, wheat seeds and Eucalyptus or Populus sawdust were used separately as substrates. After the disinfection treatments, bagged substrates were sprayed with 3 mL of suspension of conidia of Trichoderma sp (10(5) conidia/mL) and then separately spawned with Pleurotus ostreatus or Gymnopilus pampeanus. The growth of Trichoderma sp was evaluated based on a qualitative scale. Trichoderma sp could not grow on non-sterilized substrates. Immersions in hot water treatments and immersion in alkalinized water were also unfavorable treatments for its growth. Co- cultivation with mushrooms favored Trichoderma sp growth. Mushroom cultivation disinfection treatments of lignocellulose substrates influence on the growth of Trichoderma sp when contaminations occur during spawning phase. The immersion in hot water at 60 °C for 30 min or in alkalinized water for 36 h, are treatments which better reduced the contaminations with Trichoderma sp during spawning phase for the cultivation of lignicolous species.

  12. Trichoderma spp.capable of growing at low temperatures with biocontrol potential

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Tavaszi A

    2004-01-01

    @@ Though there are successful commercial biocontrol products with Trichoderma spp. In many countries including US, Israel and Europe, their usefulness is limited in cold environments such as mid-and northern part of Europe, US and Canada, especially in the late fall, winter and early spring period.Trichoderma isolates capable of growing at low temperatures (5-10 ℃) and showing good antagonistic properties against plant pathogenic fungi may have therefore both scientific and commercial value.

  13. Biological control of rice brown spot with native isolates of three Trichoderma species

    OpenAIRE

    Elham Khalili; Mehdi Sadravi; Shahram Naeimi; Vahid Khosravi

    2012-01-01

    Brown spot caused by Bipolaris oryzae is an important rice disease in Southern coast of Caspian Sea, the major rice growing region in Iran. A total of 45 Trichoderma isolates were obtained from rice paddy fields in Golestan and Mazandaran provinces which belonged to Trichoderma harzianum, T. virens and T. atroviride species. Initially, they were screened against B. oryzae by antagonism tests including dual culture, volatile and nonvolatile metabolites and hyperparasitism. Results showed that ...

  14. Enhanced biocontrol activity of Trichoderma through inactivation of a mitogen-activated protein kinase

    OpenAIRE

    Mendoza-Mendoza, Artemio; Pozo, María J.; Grzegorski, Darlene; Martínez, Pedro; García, Juan M.; Olmedo-Monfil, Vianey; Cortés, Carlos; Kenerley, Charles; Herrera-Estrella, Alfredo

    2003-01-01

    The production of lytic enzymes in Trichoderma is considered determinant in its parasitic response against fungal species. A mitogen-activated protein kinase encoding gene, tvk1, from Trichoderma virens was cloned, and its role during the mycoparasitism, conidiation, and biocontrol was examined in tvk1 null mutants. These mutants showed a clear increase in the level of the expression of mycoparasitism-related genes under simulated mycoparasitism and during direct confrontation with the plant ...

  15. Uji Efektifitas Jamur Antagonis Trichoderma sp. Dan Gliocladium sp. Untuk Mengendalikan Penyakit Layu Fusarium

    OpenAIRE

    Nasution, Rima Yunisa

    2015-01-01

    Rima Yunisa Nasution. 2014. Effectiveness test on antagonist fungi Trichoderma sp. and Gliocladium Sp. to control fusarium disease (Fusarium oxysporum F.sp capsici) for chilli crop (Capsicum Annuum L.) at screen house. Supervised by Lahmuddin Lubis and Hassanuddin. The goal of the research is to examine antagonist fungi Trichoderma and Gliocladium virens towards F. Oxysporum that lead the fusarium disease for chilli crop at screen house. The research was conducted at Plant Dise...

  16. Molecular Identification of Trichoderma spp. in Garlic and Onion Fields and In Vitro Antagonism Trials on Sclerotium cepivorum

    OpenAIRE

    Luis Alvarado-Marchena and; William Rivera-Méndez

    2016-01-01

    ABSTRACT Trichoderma species are non-pathogenic microorganisms that protect against fungal diseases and contribute to increased crop yields. However, not all Trichoderma species have the same effects on crop or a pathogen, whereby the characterization and identification of strains at the species level is the first step in the use of a microorganism. The aim of this study was the identification – at species level – of five strains of Trichoderma isolated from soil samples obtained from garlic ...

  17. Tolerance to chitosan by Trichoderma species is associated with low membrane fluidity.

    Science.gov (United States)

    Zavala-González, Ernesto A; Lopez-Moya, Federico; Aranda-Martinez, Almudena; Cruz-Valerio, Mayra; Lopez-Llorca, Luis Vicente; Ramírez-Lepe, Mario

    2016-07-01

    The effect of chitosan on growth of Trichoderma spp., a cosmopolitan genus widely exploited for their biocontrol properties was evaluated. Based on genotypic (ITS of 18S rDNA) characters, four isolates of Trichoderma were identified as T. pseudokoningii FLM16, T. citrinoviride FLM17, T. harzianum EZG47, and T. koningiopsis VSL185. Chitosan reduces radial growth of Trichoderma isolates in concentration-wise manner. T. koningiopsis VSL185 was the most chitosan tolerant isolate in all culture media amended with chitosan (0.5-2.0 mg ml(-1) ). Minimal Inhibitory Concentration (MIC) and Minimal Fungicidal Concentration (MFC) were determined showing that T. koningiopsis VSL185 displays higher chitosan tolerance with MIC value >2000 μg ml(-1) while for other Trichoderma isolates MIC values were around 10 μg ml(-1) . Finally, free fatty acid composition reveals that T. koningiopsis VSL185, chitosan tolerant isolate, displays lower linolenic acid (C18:3) content than chitosan sensitive Trichoderma isolates. Our findings suggest that low membrane fluidity is associated with chitosan tolerance in Trichoderma spp. PMID:27213758

  18. Screening of different Trichoderma species against agriculturally important foliar plant pathogens.

    Science.gov (United States)

    Prabhakaran, Narayanasamy; Prameeladevi, Thokala; Sathiyabama, Muthukrishnan; Kamil, Deeba

    2015-01-01

    Different isolates of Trichoderma were isolated from soil samples which were collected from different part of India. These isolates were grouped into four Trichoderma species viz., Trichoderma asperellum (Ta), T. harzianum (Th), T. pseudokoningii (Tp) and T. longibrachiatum (Tl) based on their morphological characters. Identification of the above isolates was also confirmed through ITS region analysis. These Trichoderma isolates were tested for in vitro biological control of Alternaria solani, Bipolaris oryzae, Pyricularia oryzae and Sclerotinia scierotiorum which cause serious diseases like early blight (target spot) of tomato and potato, brown leaf spot disease in rice, rice blast disease, and white mold disease in different plants. Under in vitro conditions, all the four species of Trichoderma (10 isolates) proved 100% potential inhibition against rice blast pathogen Pyracularia oryzae. T. harzianum (Th-01) and T. asperellum (Ta-10) were effective with 86.6% and 97.7%, growth inhibition of B. oryzae, respectively. Among others, T. pseudokoningii (Tp-08) and T. Iongibrachiatum (Tl-09) species were particularly efficient in inhibiting growth of S. sclerotiorum by 97.8% and 93.3%. T. Iongibrachiatum (TI-06 and TI-07) inhibited maximum mycelial growth of A. solani by 87.6% and 84.75. However, all the T. harzianum isolates showed significantly higher inhibition against S. sclerotiorum (CD value 9.430), causing white mold disease. This study led to the selection of potential Trichoderma isolates against rice blast, early blight, brown leaf spot in rice and white mold disease in different crops.

  19. Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan.

    Science.gov (United States)

    Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz Ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

    2015-01-01

    This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract.

  20. Isolation and expression of two polyketide synthase genes from Trichoderma harzianum 88 during mycoparasitism.

    Science.gov (United States)

    Yao, Lin; Tan, Chong; Song, Jinzhu; Yang, Qian; Yu, Lijie; Li, Xinling

    2016-01-01

    Metabolites of mycoparasitic fungal species such as Trichoderma harzianum 88 have important biological roles. In this study, two new ketoacyl synthase (KS) fragments were isolated from cultured Trichoderma harzianum 88 mycelia using degenerate primers and analysed using a phylogenetic tree. The gene fragments were determined to be present as single copies in Trichoderma harzianum 88 through southern blot analysis using digoxigenin-labelled KS gene fragments as probes. The complete sequence analysis in formation of pksT-1 (5669bp) and pksT-2 (7901bp) suggests that pksT-1 exhibited features of a non-reducing type I fungal PKS, whereas pksT-2 exhibited features of a highly reducing type I fungal PKS. Reverse transcription polymerase chain reaction indicated that the isolated genes are differentially regulated in Trichoderma harzianum 88 during challenge with three fungal plant pathogens, which suggests that they participate in the response of Trichoderma harzianum 88 to fungal plant pathogens. Furthermore, disruption of the pksT-2 encoding ketosynthase-acyltransferase domains through Agrobacterium-mediated gene transformation indicated that pksT-2 is a key factor for conidial pigmentation in Trichoderma harzianum 88. PMID:26991299

  1. Screening of different Trichoderma species against agriculturally important foliar plant pathogens.

    Science.gov (United States)

    Prabhakaran, Narayanasamy; Prameeladevi, Thokala; Sathiyabama, Muthukrishnan; Kamil, Deeba

    2015-01-01

    Different isolates of Trichoderma were isolated from soil samples which were collected from different part of India. These isolates were grouped into four Trichoderma species viz., Trichoderma asperellum (Ta), T. harzianum (Th), T. pseudokoningii (Tp) and T. longibrachiatum (Tl) based on their morphological characters. Identification of the above isolates was also confirmed through ITS region analysis. These Trichoderma isolates were tested for in vitro biological control of Alternaria solani, Bipolaris oryzae, Pyricularia oryzae and Sclerotinia scierotiorum which cause serious diseases like early blight (target spot) of tomato and potato, brown leaf spot disease in rice, rice blast disease, and white mold disease in different plants. Under in vitro conditions, all the four species of Trichoderma (10 isolates) proved 100% potential inhibition against rice blast pathogen Pyracularia oryzae. T. harzianum (Th-01) and T. asperellum (Ta-10) were effective with 86.6% and 97.7%, growth inhibition of B. oryzae, respectively. Among others, T. pseudokoningii (Tp-08) and T. Iongibrachiatum (Tl-09) species were particularly efficient in inhibiting growth of S. sclerotiorum by 97.8% and 93.3%. T. Iongibrachiatum (TI-06 and TI-07) inhibited maximum mycelial growth of A. solani by 87.6% and 84.75. However, all the T. harzianum isolates showed significantly higher inhibition against S. sclerotiorum (CD value 9.430), causing white mold disease. This study led to the selection of potential Trichoderma isolates against rice blast, early blight, brown leaf spot in rice and white mold disease in different crops. PMID:26536792

  2. Biological control of rice brown spot with native isolates of three Trichoderma species

    Directory of Open Access Journals (Sweden)

    Elham Khalili

    2012-03-01

    Full Text Available Brown spot caused by Bipolaris oryzae is an important rice disease in Southern coast of Caspian Sea, the major rice growing region in Iran. A total of 45 Trichoderma isolates were obtained from rice paddy fields in Golestan and Mazandaran provinces which belonged to Trichoderma harzianum, T. virens and T. atroviride species. Initially, they were screened against B. oryzae by antagonism tests including dual culture, volatile and nonvolatile metabolites and hyperparasitism. Results showed that Trichoderma isolates can significantly inhibit mycelium growth of pathogen in vitro by producing volatile and nonvolatile metabolites Light microscopic observations showed no evidence of mycoparasitic behaviour of the tested isolates of Trichoderma spp. such as coiling around the B. oryzae. According to in vitro experiments, Trichoderma isolates were selected in order to evaluate their efficacy in controlling brown spot in glasshouse using seed treatment and foliar spray methods. Concerning the glasshouse tests, two strains of T. harzianum significantly controlled the disease and one strain of T. atroviride increased the seedling growth. It is the first time that the biological control of rice brown spot and increase of seedling growth with Trichoderma species have been studied in Iran.

  3. Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan.

    Science.gov (United States)

    Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz Ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

    2015-01-01

    This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract. PMID:26691463

  4. KADAR GLUKOSA DARI HIDROLISIS SELULOSA PADA ECENG GONDOK MENGGUNAKAN Trichoderma viride DENGAN VARIASI TEMPERATUR DAN WAKTU FERMENTASI

    Directory of Open Access Journals (Sweden)

    Purbowatiningrum R Sarjono

    2012-11-01

    Full Text Available Telah kita ketahui bahwa eceng gondok merupakan salah satu sumber selulosa yang melimpah di alam dan dapat dimanfaatkan sebagai sumber karbon bagi jamur Trichoderma viride. Eceng gondok memiliki bobot kering selulosa 21,5%, hemiselulosa 33,9% dan lignin 7,01%. Trichoderma viride adalah jamur saprofit yang berpotensi memproduksi selulase yang mampu mendegradasi ikatan β-1,4-glikosida pada rantai selulosa untuk menghasilkan glukosa. Glukosa dapat dimanfaatkan dalam produksi sirup gula, asam organik dan bioetanol. Penelitian ini bertujuan untuk mendapatkan Trichoderma viride yang mampu tumbuh pada media pertumbuhan hasil modifikasi eceng gondok serta memperoleh temperatur optimum aktivitas Trichoderma viride dalam menghidrolisis eceng gondok dan waktu fermentasi terbaik dalam menghasilkan glukosa. Proses pertama adalah persiapan sampel enceng gondok meliputi delignifikasi, kurva pertumbuhan Trichoderma viride dibuat dalam media modifikasi eceng gondok untuk mengetahui waktu optimum pertumbuhan Trichoderma viride. Penentuan temperatur optimum dan waktu fermentasi terbaik dari aktivitas Trichoderma viride didasarkan pada glukosa yang dihasilkan dari hidrolisis selulosa pada eceng gondok menggunakan metode Nelson Somogyi. Berdasarkan penelitian diperoleh hasil bahwa Trichoderma viride mampu tumbuh pada media modifikasi eceng gondok. Temperatur optimum aktivitas Trichoderma viride dalam menghidrolisis selulosa pada eceng gondok adalah 35oC dan waktu fermentasi terbaik dihasilkannya glukosa pada jam ke-96 yaitu sebesar 1,3864 mg/L.

  5. In vitro sensitivity of antagonistic Trichoderma atroviride to herbicides

    Directory of Open Access Journals (Sweden)

    Patricia Helena Santoro

    2014-04-01

    Full Text Available Trichoderma atroviride was tested in vitro for its sensitivity to different herbicides. The dosages tested were recommended dosage (RD, half dosage (½RD, and double dosage (2RD. Germination, colony-forming units (CFU, radial growth, and spore production were evaluated. Carfentrazone-ethyl and sulfentrazone inhibited the germination at RD and 2RD. A reduction in the CFU was observed for glufosinate-ammonium, atrazine, carfentrazone-ethyl, diuron + paraquat dichloride, imazapyr, oxyfluorfen, and sulfentrazone at each of the tested dosages. Radial growth was influenced by ametryn, atrazine, carfentrazone-ethyl, oxyfluorfen, and sulfentrazone herbicides, with an 80% reduction of the colonial area. Spore production was affected by carfentrazone-ethyl, oxyfluorfen, and sulfentrazone with colonial area reductions of over 70%. It was concluded that 2,4 D, clomazone, and imazapyr herbicides showed the least toxicity to T. atroviride and should be used in the crops where the fungus has been applied for phytopathogen control.

  6. A new cytotoxic cytochalasin from the endophytic fungus Trichoderma harzianum.

    Science.gov (United States)

    Chen, Huiqin; Daletos, Georgios; Okoye, Festus; Lai, Daowan; Dai, Haofu; Proksch, Peter

    2015-04-01

    The new natural product 4]-hydroxy-deacetyl-18-deoxycytochalasin H (1), together with the known deacetyl-18-deoxycytochalasin H (2) and 18-deoxycytochalasin H (3) were obtained from the endophytic fungus Trichoderma harzianum isolated from leaves of Cola nitida. The structure of the new compound was unambiguously determined by 1D and 2D NMR spectroscopy, and by HRESIMS measurements, as well as by comparison with the literature. Compounds 1-3 showed potent cytotoxic activity against the murine lymphoma (L5178Y) cell line and against human ovarian cancer (A2780 sens and A2780 CisR) cell lines (IC50 0.19-6.97 µM). The A2780 cell lines included cisplatin-sensitive (sens) and -resistant (R) cells. PMID:25973482

  7. [Antimicrobial Activity of Fungi Strains of Trichoderma from Middle Siberia].

    Science.gov (United States)

    Sadykova, V S; Kurakov, A V; Kuvarina, A E; Rogozhin, E A

    2015-01-01

    The antibiotic activity in 42 strains of 8 species of the Trichoderma genus (T. asperellum, T. viride, T. hamatum, T. koningii, T. atroviride, T. harzianum, T. Citrinoviride, and T. longibrachiatum) isolated from different Siberian ecotops was studied. It was shown that these species differ in the degree of their antibacterial and antifungal activity. The chosen strain, T. citrinoviride TV4-1, exhibited high activity and a wide range of actions against the opportunistic and pathogenic fungi of the Aspergillus and Candida albicans genus; bacteria, including methicillin-resistant Staphylococcus aureus; and cancer cells. According to mass and I R spectrometry data and the spectrum of biological action, peptaibols are probably the most active compounds in the strain culture extracts. PMID:26204779

  8. BIOTECHNOLOGY OF TRICHODERMA-BASED FEED ADDITIVE WITH CELLULOLYTIC PROPERTIES

    Directory of Open Access Journals (Sweden)

    Koshchayev A. G.

    2013-11-01

    Full Text Available In the work, we have presented the information of elaboration of a manufacturing process of Mycocel feed additive with the cellulolytic activity for poul-try industry. Manufacturing process includes follow-ing steps: stock culture maintenance and storage of Trichoderma lignorum 81-17, growing fluid culture of microscopic fungus in sucrose yeast extract me-dium, feed additive with cellulolytic properties out-put and quality control, packaging, storage and disposal of waste. We have shown that the Mycocel is non-toxic feed additive for protozoa and warm-blooded animals (laboratory mice and quails. This study demonstrated total population livability in the experimental group with feed additive. Quail body-weight of experimental group was higher by 6% as compared to the control and feed consumption per 1 kg of live weight of bird was 3,58 kg, 7,5% lower than the control

  9. 40 CFR 180.1102 - Trichoderma harzianum KRL-AG2 (ATCC #20847) strain T-22; exemption from requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Trichoderma harzianum KRL-AG2 (ATCC... PESTICIDE CHEMICAL RESIDUES IN FOOD Exemptions From Tolerances § 180.1102 Trichoderma harzianum KRL-AG2... of a tolerance is established for residues of the biofungicide Trichoderma harzianum KRL-AG2...

  10. CHARACTERIZATION OF SOIL TRICHODERMA ISOLATES FOR POTENTIAL BIOCONTROL OF PLANT PATHOGENS

    Directory of Open Access Journals (Sweden)

    S. Matei

    2011-12-01

    Full Text Available Various fungal strains belonging to genus Trichoderma act as biological control agents for soil born plant pathogens. Two new strains of Trichoderma harzianum (T.h. and Trichoderma viride (T.v. were isolated from forest soils in Ilfov county and their morphological aspects, enzymatic and antagonistic activity were examined. Current chemical fungicides had constantly, in time, less influence on pathogens due to their diversity, adaptability and increasing resistance.The paper present the morphological characterization of two strains of Trichoderma isolated from forest soils. Growth rate was higher in strain T.v.SP456 (0,675mm/h than in strain T.h.P8 (0,505mm/h when fungi were grown on Czapek culture medium.Morphological description is completed with photographs of colonies in Petri plates and microscopical aspects of fungal structures belonging to Trichoderma strains SP456 and P8.Comparative aspects concerning the level of main enzymes released by T.h. isolate P8 and T.v.SP456 in liquid culture media showed differences as a function of genetic structure of each fungal isolate. The optimum culture media for inducing peroxidase, polyphenol-oxidase, β-1,3-glucanase activity in T.v.SP456 isolate was Czapek and PDA for phenil-alanin-ammonium-oxidase and chitinase. T.v.SP456 was more efficient than T.h.P8 concerning enzymes activity.The interaction between Trichoderma fungal strains SP456 and P8 and strawberry plant pathogen strains, three belonging to Botrytis cinerea (S1, P1, P2 and one to Phytophtora spp. were examined, also. Both Trichoderma strains act as mycoparasites for plant pathogens. The inhibition percent of radial growth was higher for T.v.SP456 when compared with T.h.P8 for almost all pathogenic isolates.

  11. Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species.

    Science.gov (United States)

    Jiang, Yuan; Wang, Jin-Liang; Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

    2016-01-01

    We surveyed the Trichoderma (Hypocreales, Ascomycota) biodiversity in agricultural fields in four major agricultural provinces of East China. Trichoderma strains were identified based on molecular approaches and morphological characteristics. In three sampled seasons (spring, summer and autumn), 2078 strains were isolated and identified to 17 known species: T. harzianum (429 isolates), T. asperellum (425), T. hamatum (397), T. virens (340), T. koningiopsis (248), T. brevicompactum (73), T. atroviride (73), T. fertile (26), T. longibrachiatum (22), T. pleuroticola (16), T. erinaceum (16), T. oblongisporum (2), T. polysporum (2), T. spirale (2), T. capillare (2), T. velutinum (2), and T. saturnisporum (1). T. harzianum, T. asperellum, T. hamatum, and T. virens were identified as the dominant species with dominance (Y) values of 0.057, 0.052, 0.048, and 0.039, respectively. The species amount, isolate numbers and the dominant species of Trichoderma varied between provinces. Zhejiang Province has shown the highest diversity, which was reflected in the highest species amount (14) and the highest Shannon-Wiener diversity index of Trichoderma haplotypes (1.46). We observed that relative frequencies of T. hamatum and T. koningiopsis under rice soil were higher than those under wheat and maize soil, indicating the preference of Trichoderma to different crops. Remarkable seasonal variation was shown, with summer exhibiting the highest biodiversity of the studied seasons. These results show that Trichoderma biodiversity in agricultural fields varies by region, crop, and season. Zhejiang Province (the southernmost province in the investigated area) had more T. hamatum than Shandong Province (the northernmost province), not only in isolate amounts but also in haplotype amounts. Furthermore, at haplotype level, only T. hamatum showed a gradient distribution from south to north in correspondence analysis among the four dominant species. The above results would contribute to the

  12. Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species

    Science.gov (United States)

    Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

    2016-01-01

    We surveyed the Trichoderma (Hypocreales, Ascomycota) biodiversity in agricultural fields in four major agricultural provinces of East China. Trichoderma strains were identified based on molecular approaches and morphological characteristics. In three sampled seasons (spring, summer and autumn), 2078 strains were isolated and identified to 17 known species: T. harzianum (429 isolates), T. asperellum (425), T. hamatum (397), T. virens (340), T. koningiopsis (248), T. brevicompactum (73), T. atroviride (73), T. fertile (26), T. longibrachiatum (22), T. pleuroticola (16), T. erinaceum (16), T. oblongisporum (2), T. polysporum (2), T. spirale (2), T. capillare (2), T. velutinum (2), and T. saturnisporum (1). T. harzianum, T. asperellum, T. hamatum, and T. virens were identified as the dominant species with dominance (Y) values of 0.057, 0.052, 0.048, and 0.039, respectively. The species amount, isolate numbers and the dominant species of Trichoderma varied between provinces. Zhejiang Province has shown the highest diversity, which was reflected in the highest species amount (14) and the highest Shannon–Wiener diversity index of Trichoderma haplotypes (1.46). We observed that relative frequencies of T. hamatum and T. koningiopsis under rice soil were higher than those under wheat and maize soil, indicating the preference of Trichoderma to different crops. Remarkable seasonal variation was shown, with summer exhibiting the highest biodiversity of the studied seasons. These results show that Trichoderma biodiversity in agricultural fields varies by region, crop, and season. Zhejiang Province (the southernmost province in the investigated area) had more T. hamatum than Shandong Province (the northernmost province), not only in isolate amounts but also in haplotype amounts. Furthermore, at haplotype level, only T. hamatum showed a gradient distribution from south to north in correspondence analysis among the four dominant species. The above results would contribute to the

  13. DNA barcoding survey of Trichoderma diversity in soil and litter of the Colombian lowland Amazonian rainforest reveals Trichoderma strigosellum sp. nov. and other species

    NARCIS (Netherlands)

    Lopez-Quintero, C.A.; Atanasova, L.; Franco-Molano, A.E.; Gams, W.; Komon-Zelazowska, M.; Theelen, B.; Muller, W.H.; Boekhout, T.; Druzhinina, I.

    2013-01-01

    The diversity of Trichoderma (Hypocreales, Ascomycota) colonizing leaf litter as well as the rhizosphere of Garcinia macrophylla (Clusiaceae) was investigated in primary and secondary rain forests in Colombian Amazonia. DNA barcoding of 107 strains based on the internal transcribed spacers 1 and 2 (

  14. Phytohormone profiles induced by Trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants

    NARCIS (Netherlands)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A.; van Wees, Saskia C M

    2014-01-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the ph

  15. Using of green fluorescent reporter gene (GFP) to monitor the fate of Fusarium moniliforme mycoparasitized by Trichoderma viride

    Institute of Scientific and Technical Information of China (English)

    ZHU Ting-heng; WANG Wei-xia; WANG Chang-chun; YANG Rui-qin; CAI Xin-zhong

    2004-01-01

    @@ Fusarium moniliforme Sheld. is a rice pathogenic fungus and causes the disease called Bakanae,which has increasingly damaged rice production in the recent years. Trichoderma spp. has been one of the most widely used biological control agent of plant disease. By geneticaly labelling F. moniliforme with the GFP reporter gene, we have studied the antagonistic action of Trichoderma viride against this pathogenic fungus.

  16. The influence of formulation on Trichoderma biological activity and frosty pod rot disease management in Theobroma cacao

    Science.gov (United States)

    Frosty pod rot (FPR), caused by Moniliophthora roreri is responsible for significant losses in Theobroma cacao. Due to the limited options for FPR management, biological control methods using Trichoderma are being studied. Combinations of three formulations and two Trichoderma isolates were studied ...

  17. Identification and characterization of Trichoderma species aggressive to Pleurotus in Italy

    Institute of Scientific and Technical Information of China (English)

    Woo S L; Di Benedetto P; Senatore M; Abadi K; Gigante S; Soriente I; Ferraioli S; Scala F; Lorito M

    2004-01-01

    @@ In the late 1980's the development of a severe epidemic of green mold caused by Trichoderma spp.was noted in the commercial production of Agaricus bisporus (champignon) in the United Kingdom,North America, Spain and Holland, which caused extensive economic losses. The parasitic fungi isolated from the edible mushroom belonged to four biotypes, Thl, Th2, Th3 and Th4 of T.harzianum. However, among these biotypes, only Th2 (since classified as T. aggressivum f.europaeum) and Th4 (T. aggressivum f. aggressivum) were identified as the fungi causing problems in Agaricus production. In general, mushroom compost hosts both aggressive and innocuous isolates of Trichoderma, which are not morphologically distinguishable. About four years ago, a problem with green mold became apparent in the production of Pleurotus ostreatus in Northern Italy,which eventually developed to a crisis situation in the South two years later and threatened to seriously compromise the Pleurotus market. This study was initiated to: isolate and identify the aggressive fungi, then morphologically, physiologically and genetically characterize the isolates, determine the source and phases of infection, and study methods of control. Samples were obtained from different phases of compost preparation at the locality of a major producer and supplier of compost to the mushroom industry in Southern Italy, and microbial counts were conducted. Although the presence of Trichoderma was detected in the initial stages of composting, this value was reduced to zero from the phase of pasteurization to seeding with Pleurotus. Trichoderma infestations were noted in the packaged Pleurotus bales at various times during the incubation phase (7-15 days after seeding) and after shipping to the mushroom greenhouses, where the pathogen infestations greatly reduced the quality and quantity of the mushroom yield, as well as the number of potential harvest cycles.Preliminary results from the morphological and genetic

  18. Studies on strains of Trichoderma spp. from high altitude of Garhwal Himalayan region.

    Science.gov (United States)

    Joshi, B B; Vishwakarma, M P; Bahukhandi, D; Bhatt, R P

    2012-09-01

    Two species of Trichoderma i.e. T. harzianum and T. viride have been isolated from the soil samples collected from the higher altitude (2000-3500 m) of Garhwal Himalayan region in India. The two species were grown in Petri plates on TSM agar media and it was also observed that the optimum temperature and pH for Trichoderma growth was 30 degrees C and 5.5 respectively. When incubated on TSM agar medium at 4 degrees C, the fungus grew normally with heavy induced sporulation within three weeks of incubation. Induction of sporulation on exposure to low temperature appeared to be strategies for survival of these species in extreme cold environment temperature 4 to 5 degrees C. Antifungal activities of the two species of Trichoderma were demonstrated with phytopathogenic fungi in dual cultures. The antifungal metabolites produced by Trichoderma spp., diffusible as well as volatile, caused abnormalities in pathogenic fungi. Plant growth promotion of Trichoderma spp. was also shown through plant analysis in greenhouse.

  19. Arabidopsis thaliana polyamine content is modified by the interaction with different Trichoderma species.

    Science.gov (United States)

    Salazar-Badillo, Fatima Berenice; Sánchez-Rangel, Diana; Becerra-Flora, Alicia; López-Gómez, Miguel; Nieto-Jacobo, Fernanda; Mendoza-Mendoza, Artemio; Jiménez-Bremont, Juan Francisco

    2015-10-01

    Plants are associated with a wide range of microorganisms throughout their life cycle, and some interactions result on plant benefits. Trichoderma species are plant beneficial fungi that enhance plant growth and development, contribute to plant nutrition and induce defense responses. Nevertheless, the molecules involved in these beneficial effects still need to be identify. Polyamines are ubiquitous molecules implicated in plant growth and development, and in the establishment of plant microbe interactions. In this study, we assessed the polyamine profile in Arabidopsis plants during the interaction with Trichoderma virens and Trichoderma atroviride, using a system that allows direct plant-fungal contact or avoids their physical interaction (split system). The plantlets that grew in the split system exhibited higher biomass than the ones in direct contact with Trichoderma species. After 3 days of interaction, a significant decrease in Arabidopsis polyamine levels was observed in both systems (direct contact and split). After 5 days of interaction polyamine levels were increased. The highest levels were observed with T. atroviride (split system), and with T. virens (direct contact). The expression levels of Arabidopsis ADC1 and ADC2 genes during the interaction with the fungi were also assessed. We observed a time dependent regulation of ADC1 and ADC2 genes, which correlates with polyamine levels. Our data show an evident change in polyamine profile during Arabidopsis - Trichoderma interaction, accompanied by evident alterations in plant root architecture. Polyamines could be involved in the changes undergone by plant during the interaction with this beneficial fungus.

  20. Two new hyaline-ascospored species of Trichoderma and their phylogenetic positions.

    Science.gov (United States)

    Qin, W T; Zhuang, W Y

    2016-01-01

    Collections of hypocrealean fungi found on decaying wood in subtropical regions of China were examined. Two new species, Trichoderma confluens and T. hubeiense, were discovered and are described. Trichoderma confluens is characterized by its widely effuse to rarely pulvinate, yellow stromata with densely disposed yellowish brown ostioles, simple acremonium- to verticillium-like conidiophores, hyaline conidia and multiform chlamydospores. Trichoderma hubeiense has pulvinate, grayish yellow stromata with brownish ostioles, trichoderma- to verticillium-like conidiophores and hyaline conidia. The phylogenetic positions of the two fungi were investigated based on sequence analyses of RNA polymerase II subunit b and translation elongation factor 1-α genes. The results indicate that T. confluens belongs to the Hypocreanum clade and is associated with but clearly separated from T. applanatum and T. decipiens. Trichoderma hubeiense belongs to the Polysporum clade and related to T. bavaricum but obviously differs from other members of the clade in sequence data. Morphological distinctions between the new species and their close relatives are noted and discussed.

  1. Salicylic acid prevents Trichoderma harzianum from entering the vascular system of roots.

    Science.gov (United States)

    Alonso-Ramírez, Ana; Poveda, Jorge; Martín, Ignacio; Hermosa, Rosa; Monte, Enrique; Nicolás, Carlos

    2014-10-01

    Trichoderma is a soil-borne fungal genus that includes species with a significant impact on agriculture and industrial processes. Some Trichoderma strains exert beneficial effects in plants through root colonization, although little is known about how this interaction takes place. To better understand this process, the root colonization of wild-type Arabidopsis and the salicylic acid (SA)-impaired mutant sid2 by a green fluorescent protein (GFP)-marked Trichoderma harzianum strain was followed under confocal microscopy. Trichoderma harzianum GFP22 was able to penetrate the vascular tissue of the sid2 mutant because of the absence of callose deposition in the cell wall of root cells. In addition, a higher colonization of sid2 roots by GFP22 compared with that in Arabidopsis wild-type roots was detected by real-time polymerase chain reaction. These results, together with differences in the expression levels of plant defence genes in the roots of both interactions, support a key role for SA in Trichoderma early root colonization stages. We observed that, without the support of SA, plants were unable to prevent the arrival of the fungus in the vascular system and its spread into aerial parts, leading to later collapse.

  2. Use of Trichoderma spp.in remediation of polluted soils and waters

    Institute of Scientific and Technical Information of China (English)

    Gary E Harmant; James Lynch; Matteo Lorito

    2004-01-01

    @@ Trichoderma spp. probably have a role in remediation of polluted soils and waters. Highly rhizosphere competent strains persist on roots for an extended period of time (at least months) and continuously interact with the plants. They can increase general plant and root growth and increase uptake of a variety of materials. This makes the Trichoderma-plant interaction highly attractive for use in phytoextraction technologies. Moreover, Trichoderma spp. are resistant to a wide range of toxic compounds and can degrade some of these. One highly attractive target for remediation are soils that are polluted with cyanide and metallocyanides. Shrub willows (genus Salix) have been shown to take up and degrade these compounds by unknown mechanisms. Thus, they remove these compounds from soil but there are no cyanide residues in the plants. Similarly, Trichoderma spp. degrade free cyanide via production of extracellular enzymes and take up and then degrade metallocyanides such as Prussian blue. The willow-Trichoderma system therefore provides a plant-microbe system for degradation of these toxic compounds. The fungi also can be used directly in remediation strategies; for example,they degrade polyphenols such as those found in large quantities in waste water from production of olive oil. Thus, the abilities of the fungi to interact and enhance plant growth, their ability to grow in the presence of toxicants and their enzymatic abilities to degrade polluting substances provide a number of opportunities for either plant-microbe or pure fungal systems to remove pollutants from lands and waters.

  3. Diversity and effect of Trichoderma spp. associated with green mold disease on Lentinula edodes in China.

    Science.gov (United States)

    Wang, Gangzheng; Cao, Xiantao; Ma, Xiaolong; Guo, Mengpei; Liu, Changhao; Yan, Lianlian; Bian, Yinbing

    2016-08-01

    Lentinula edodes, one of the most important edible mushrooms in China, is affected heavily by the infection of green mold that overgrows mushroom mycelia. We collected the diseased samples from main L. edodes cultivation regions in China to characterize the pathogen and to study the effect of Trichoderma spp. on L. edodes species. We identified six Trichoderma species, that is, T. harzianum, T. atroviride, T. viride, T. pleuroticola, T. longibrachiatum, and T. oblongisporum based on the internal transcribed spacer or tef1-α sequences and morphology characteristics. In confrontation cultures on Petri plates or in tubes, and in L. edodes cultures in a medium containing Trichoderma metabolites, L. edodes mycelia were not only distorted and swollen, but also inhibited by Trichoderma isolates. It is not possible that adjusting pH value or temperature is used for controlling L. edodes green disease, because the growth of most of Trichoderma isolates and L. edodes shared similar pH and temperature conditions. PMID:27147196

  4. Use of Trichoderma spp.for biological control of the livestock feed contaminant fungus Fusarium proliferatum

    Institute of Scientific and Technical Information of China (English)

    Ruocco M; Ferraioli S; Scala F; Lorito M; Pane F; Ritieni A; Lanzuise S; Ambrosino P; Marra R; Woo S L; Ciliento R; Soriente I

    2004-01-01

    @@ Fusarium spp. are pathogens of many important agricultural crops, and are often strong mycotoxin producers. Fusarium proliferatum, in particular, causes disease in cereals and secretes the toxin Beauvaricin that contaminates livestock feed and cereals, producing a variety of toxicity symptoms ranging from poor weight gain to mortality. Beauvaricin is a cyclodepsipeptide and acts as a potent mycotoxin known to have insecticidal properties. This compound is highly toxic to human cell lines,where it induces apoptosis and specifically inhibits cholesterol acetyltransferase. Nothing is known about the role of this mycotoxin during the interaction of F. proliferatum with other microorganisms, including the fungal antagonists Trichoderma spp. In vitro tests have demonstrated that the antagonistic and mycoparasitic activity of Trichoderma is not inhibited by the presence of Beauvaricin at concentrations up to 10 mg/kg in the substrate. In vivo biocontrol assays on barley and wheat with Trichoderma against F. proliferatum isolates, producing and non-producing Beauvaricin, confirmed the ability of the antagonist to control this pathogen in all cases. Also Trichoderma culture filtrates obtained in conditions that promote _Cell Wall _Degrading Enzyme (CWDE) secretion, were able to inhibit spore germination of different F. proliferatum isolates.These results suggest the possibility of using Trichoderma and/or its metabolites to control contaminants of livestock feed by mycotoxin-producing Fusarium.

  5. Phytohormone profiles induced by trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants.

    Science.gov (United States)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A; Van Wees, Saskia C M

    2014-07-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the phytohormonal network of their host plant, thus leading to an improvement of plant growth and stress tolerance. In this study, we tested whether alterations in the phytohormone signature induced by different Trichoderma isolates correspond with their ability for biocontrol and growth promotion. Four Trichoderma isolates were collected from agricultural soils and were identified as the species Trichoderma harzianum (two isolates), Trichoderma ghanense, and Trichoderma hamatum. Their antagonistic activity against the plant pathogen Fusarium oxysporum f. sp. melonis was tested in vitro, and their plant growth-promoting and biocontrol activity against Fusarium wilt on melon plants was examined in vivo, and compared to that of the commercial strain T. harzianum T-22. Several growth- and defense-related phytohormones were analyzed in the shoots of plants that were root-colonized by the different Trichoderma isolates. An increase in auxin and a decrease in cytokinins and abscisic acid content were induced by the isolates that promoted the plant growth. Principal component analysis (PCA) was used to evaluate the relationship between the plant phenotypic and hormonal variables. PCA pointed to a strong association of auxin induction with plant growth stimulation by Trichoderma. Furthermore, the disease-protectant ability of the Trichoderma strains against F. oxysporum infection seems to be more related to their induced alterations in the content of the hormones abscisic acid, ethylene, and the cytokinin trans-zeatin riboside than to the in vitro antagonism activity against F. oxysporum.

  6. Biochemical and metabolic profiles of Trichoderma strains isolated from common bean crops in the Brazilian Cerrado, and potential antagonism against Sclerotinia sclerotiorum.

    Science.gov (United States)

    Lopes, Fabyano Alvares Cardoso; Steindorff, Andrei Stecca; Geraldine, Alaerson Maia; Brandão, Renata Silva; Monteiro, Valdirene Neves; Lobo, Murillo; Coelho, Alexandre Siqueira Guedes; Ulhoa, Cirano José; Silva, Roberto Nascimento

    2012-07-01

    Some species of Trichoderma have successfully been used in the commercial biological control of fungal pathogens, e.g., Sclerotinia sclerotiorum, an economically important pathogen of common beans (Phaseolus vulgaris L.). The objectives of the present study were (1) to provide molecular characterization of Trichoderma strains isolated from the Brazilian Cerrado; (2) to assess the metabolic profile of each strain by means of Biolog FF Microplates; and (3) to evaluate the ability of each strain to antagonize S. sclerotiorum via the production of cell wall-degrading enzymes (CWDEs), volatile antibiotics, and dual-culture tests. Among 21 isolates, we identified 42.86% as Trichoderma asperellum, 33.33% as Trichoderma harzianum, 14.29% as Trichoderma tomentosum, 4.76% as Trichoderma koningiopsis, and 4.76% as Trichoderma erinaceum. Trichoderma asperellum showed the highest CWDE activity. However, no species secreted a specific group of CWDEs. Trichoderma asperellum 364/01, T. asperellum 483/02, and T. asperellum 356/02 exhibited high and medium specific activities for key enzymes in the mycoparasitic process, but a low capacity for antagonism. We observed no significant correlation between CWDE and antagonism, or between metabolic profile and antagonism. The diversity of Trichoderma species, and in particular of T. harzianum, was clearly reflected in their metabolic profiles. Our findings indicate that the selection of Trichoderma candidates for biological control should be based primarily on the environmental fitness of competitive isolates and the target pathogen.

  7. Are mycoparasitism and chitinase production species or isolate dependent in Trichoderma ?

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Nagy V; Kovacs K

    2004-01-01

    @@ The relationship between taxonomic status of Trichoderma spp., chitinase production in solid substrate fermentation (SSF) on four media and mycoparasitism in dual culture (confrontation assay)against four plant pathogenic fungi was studied. Seventy five Trichoderma isolates belonging to 35species have been screened. The plant pathogenic fungi used in confrontation assay were Botrytis cinerea , Fusarium oxysporum f. sp. dianthi , Rhizoctonia solani and Sclerotinia sclerotiorum . The SSF media contained wheat bran, crude chitin (from crab shells, SIGMA) and salt solutions. The best performing isolates in mycoparasitism tests were Trichoderma flavofuscum, T. harzianum, T.inhamatum, T. koningii and T. strigosum. Some isolates exhibiting good mycoparasitism produced chitinase in SSF only at low or medium level. In contrary there were isolates with excellent extracellular chitinase production but their biocontrol potential did not belong to the leading group.Statistical methods have been used to evaluate the data.

  8. Population dynamics of Trichoderma species in the rhizosphere of tobacco and four species form China

    Institute of Scientific and Technical Information of China (English)

    YU Ze-fen; ZHANG Ke-qin

    2004-01-01

    @@ To study the effect of tobacco growth on Trichoderma population, we investigated the occurrence of Trichoderma species in the rhizosphere of tobacco plant during the period from transplanting (June) to harvesting (October) and measured relative environmental factors. Eleven species of Trichoderma were isolated, among which T. harzianum, T. viride, T. hamatum, T. atroviride, T.longibrachiatum, T. virens, T. koningii were identified, other four species Ty1, Ty2, Ty3, Ty4are new species. Of the species which occurred at high frequencies, T. harzianum and T. hamatum were most abundant in the July and T. viride in the August. The occurrence of the above three abundant species correlates significantly with the developmental phase of tobacco and correlates apparently with the soil moisture content, but not with the temperature. For the other eight species,no obvious correlation was found between the above factors and them.

  9. [Endocarditis caused by Trichoderma longibrachiatumin a patient receiving home parenteral nutrition].

    Science.gov (United States)

    Rodríguez Peralta, Laura I; Mañas Vera, Ma Reyes; García Delgado, Manuel J; Pérez de la Cruz, Antonio J

    2013-01-01

    Home parenteral nutrition (HPN) improves the quality of life of the patients although it has complications. Catheter-related infections and mechanical complications are the most frequent ones. We report the case of endocarditis over catheter in a man suffering from short bowel and receiving HPN. The special features of the case are firstly the catheter was a remaining fragment on the right atrial and secondly the infection was caused by Trichoderma longibrachiatum, an isolated fact regarding this pathology so far. Conventional surgery was applied to take the catheter out. Staphylococcus epidermidis, Ochrobactrum anthropi and Trichoderma longibrachiatum were isolated from the surgical specimen. The extraction of the infected catheter along with antibiotic therapy led to the complete recovery of the subject. Ochrobactrum anthropi and Trichoderma longibrachiatum are unusual microorganisms but they are acquiring more relevance. Although there is no agreement about intravascular retained catheter management, the most recommended approach consists on monitoring them and removing the device in case of complications.

  10. Isolation and Characterization of Trichoderma spp. for Antagonistic Activity Against Root Rot and Foliar Pathogens.

    Science.gov (United States)

    Kumar, Krishna; Amaresan, N; Bhagat, S; Madhuri, K; Srivastava, R C

    2012-06-01

    Trichoderma, soil-borne filamentous fungi, are capable of parasitising several plant pathogenic fungi. Twelve isolates of Trichoderma spp. isolated from different locations of South Andaman were characterized for their cultural, morphological and antagonistic activity against soil borne and foliar borne pathogens. The sequencing of these isolates showed seven different species. The isolates revealed differential reaction patterns against the test pathogens viz., Sclerotium rolfsii, Colletotrichum gloeosporioides and C. capsici. However, the isolates, TND1, TWN1, TWC1, TGD1 and TSD1 were most effective in percentage inhibition of mycelial growth of test pathogens. Significant chitinase and β-1,3-glucanase activities of all Trichoderma isolates has been recorded in growth medium. T. viride was found with highest chitinase whereas T. harzianum was recorded with highest β-1,3-glucanase activities. PMID:23729873

  11. Antagonisme in vitro de trichoderma spp. vis-a-vis de rhizoctonia solani kuhn

    OpenAIRE

    Camporota, P.

    1985-01-01

    Cet article présente les résultats obtenus lors de la réalisation de la première étape d’un programme de sélection de souches de Trichoderma spp. utilisables pour la lutte biologique contre Rhizoctonia solani dans le sol : 28 souches de Trichoderma ont été confrontées in vitro à 3 souches de R. solani appartenant à des groupes d’anastomose différents. On a mesuré, pour chaque souche de Trichoderma, la capacité à envahir les colonies de l’agent pathogène ainsi que les 3 modes d’action : my...

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

    Institute of Scientific and Technical Information of China (English)

    Sanz L; Hermosa M R; González F J; Monte E

    2004-01-01

    @@ Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE).Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderna.Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique.Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities.Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.

  13. The Trichoderma-plant interaction is mediated by avirulence proteins produced by this fungus

    Institute of Scientific and Technical Information of China (English)

    Ruocco M; Kip N; P J G M de Wit; Lorito M; Lanzuise S; Woo S L; Ambrosino P; Marra R; Turrà D; Gigante S; Formisno E; Scala F

    2004-01-01

    @@ The molecular basis of Trichoderma -plant interaction is very complex and still not completely understood. The colonization of the root system by rhizosphere competent strains of Trichoderma results in increased development of root/aerial systems, in improved yields and in plant disease control.Other beneficial effects, such as the induction of plant systemic resistance, have also been described.To understand the mechanisms involved we are using different approaches, including the making of transformants expressing genes that encode for compounds able to affect plant response to pathogens.Trichoderma transformants carrying the avirulence gene Avr4 from Cladosporium fulvum under the control of constitutive and inducible promoters were obtained and tested on tomato plants having the Cf4 resistance gene. Necrosis and suberification zones, similar to the symptoms appearing during Cladosporium-tomato interaction, were found when the roots of the Cf4 plants were treated with Avr4-Trichoderma. This demonstrates that selected Trichoderma strains are able to transfer to the plant molecules that may deeply affect metabolism, disease resistance etc. Therefore, these beneficial fungi can be regarded as biotechnological tools to provide a variety of crops with useful compounds.Moreover, in in vitro competition assays the transformants were found to be more effective as antagonists against Alternaria alternata than the wild type. Trichoderma sends a variety of biochemical signals to the plants including avirulence molecules; therefore the presence of avr-like proteins in the fungus proteome was investigated. Proteome analysis has permitted us to isolate and sequence many proteins potentially having this function. From the extraeellular protein extracts, we have purified and sequenced a protein with structural characteristics similar to Avr4 of C. fulvum.The protein, Hytra1, was found to be a hydrophobin with chitin binding activity, the typical 8cysteine residues, and 4

  14. In Vitro Antagonist Action of Trichoderma Strains Against Sclerotinia sclerotiorum and Sclerotium cepivorum

    Directory of Open Access Journals (Sweden)

    Francisco D.H. Castillo

    2011-01-01

    Full Text Available Problem statement: Sclerotinia sclerotiorum and Sclerotium cepivorum are soil pathogens which have generated resistance to synthetic fungicides. One biological alternative to this problem is to select novel strains of Trichoderma with different mechanisms of action against these plant pathogens to ensure efficient control. Approach: In the present research was determined the antagonism effect of Mexicans Trichoderma strains on S. sclerotiorum and S. cepivorum in vitro. It was used dual culture technique by determining the percentage of mycelia growth inhibition, days to contact and antagonism levels proposed by Bell. It was also determined the effect of volatile compounds and metabolites compound from Trichoderma produced in solutions. Results: The mycelia growth inhibition of S. sclerotiorum and S. cepivorum were 45-63.8 and 50.9-81.5% respectively by the effect of Trichoderma strains. The T. ghanense and T. longibrachiatum (T15 and T10 inhibited in higher proportion to S. sclerotiorum. On the other hand T. inhamatum and T. asperellum (T32 and T11 inhibited in higher proportion to S. cepivorum. The days to contact between the Trichoderma and phytopathogen species were between two to three days. The levels of antagonism according to the Bells scale were different between the two species of phytopathogen and for Trichoderma strains were grouped into I and II class. The maximum inhibition effect by volatiles compound was occasioned by T. longibrachiatum with 31.5 (T3 and 59.2% (T10 on S. sclerotiorum and S. cepivorum respectively. Respect to metabolites, these shown high effects on phytopathogen growth, where T. asperellum is the more outstanding specie which produce totals mycelia growth inhibition of two phytopathogen species. Conclusion: T. longibrachiatum (T3 and T10 and T. asperellum (T1 and T11 were the most efficient species with the highest antagonist effects against S. sclerotiorum and S. cepivorum.

  15. Experimental Evolution of Trichoderma citrinoviride for Faster Deconstruction of Cellulose.

    Science.gov (United States)

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J

    2016-01-01

    Engineering faster cellulose deconstruction is difficult because it is a complex, cooperative, multi-enzyme process. Here we use experimental evolution to select for populations of Trichoderma citrinoviride that deconstruct up to five-fold more cellulose. Ten replicate populations of T. citrinoviride were selected for growth on filter paper by serial culture. After 125 periods of growth and transfer to fresh media, the filter paper deconstruction increased an average of 2.5 fold. Two populations were examined in more detail. The activity of the secreted cellulase mixtures increased more than two-fold relative to the ancestor and the largest increase was in the extracellular β-glucosidase activity. qPCR showed at least 16-fold more transcribed RNA for egl4 (endoglucanase IV gene), cbh1 (cellobiohydrolase I gene) and bgl1 (extracellular β-glucosidase I gene) in selected populations as compared to the ancestor, and earlier peak expressions of these genes. Deep sequencing shows that the regulatory strategies used to alter cellulase secretion differ in the two strains. The improvements in cellulose deconstruction come from earlier expression of all cellulases and increased relative amount of β-glucosidase, but with small increases in the total secreted protein and therefore little increase in metabolic cost. PMID:26820897

  16. The Fungus Trichoderma Regulates Submerged Conidiation Using the Steroid Pregnenolone.

    Science.gov (United States)

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J

    2016-09-16

    In previous work, we evolved a population of Trichoderma citrinoviride in liquid cultures to speed up its asexual development cycle. The evolved population, called T-6, formed conidia 3 times sooner and in >1000-fold greater numbers. Here, we identify the steroid pregnenolone as a molecular signal for this different behavior. Media in which the ancestral T. citrinoviride population was grown (called ancestral spent media) contained a submerged conidiation inhibitor. Growing the evolved population T-6 in ancestral spent media eliminated the abundant formation of conidia. This inhibition depended on the amount and age of the ancestral spent medium and the time that the ancestral spent medium was added to the T-6 culture. Fractionation of the ancestral spent medium identified a hydrophobic inhibiting compound with a molecular weight less than 2000 g/mol. A combination of GC-MS, ELISA, and reaction with cholesterol oxidase identified it as pregnenolone. The addition of pregnenolone to cultures of T-6 inhibited submerged conidiation by inhibiting formation of conidiophores, while 10 other analogous steroids did not. Pregnenolone also inhibited submerged conidiation of Fusarium graminearum PH-1, a plant pathogen that causes head blight in wheat and barley. This identification of steroids as signal molecules in fungi creates opportunities to disrupt this signaling to control fungal behavior. PMID:27413801

  17. Genetic characterization of somatic recombination in Trichoderma pseudokoningii

    Directory of Open Access Journals (Sweden)

    Barcellos Fernando Gomes

    2003-01-01

    Full Text Available Crossing experiments via hyphal anastomosis between two strains contrasting for auxotrophic markers of Trichoderma pseudokoningii were conducted to characterize the somatic recombination process in this specie. Four crossings were made and a total of 1052 colonies obtained from conidial suspensions of the heterokaryotic colonies were analyzed. Sixty-eight recombinant colonies, from four growing generations, were analyzed for the auxotrophic markers. Of the 68 colonies analyzed, 58 were stable after four generations and the remainders were unstable, reverting to one of the parentals. Most of the recombinant colonies were unstable through subculture and after four growing generations they showed the leu ino met markers (auxotrophic for leucin, inositol and metionin respectively. The unstable recombinant colonies showed irregular growing borders, sparse sporulation and frequent sector formation. The results suggest the occurrence of recombination mechanisms in the heterokaryon (somatic recombination, different from those described for the parasexual cycle or parameiosis. Therefore, we proposed the ocurrence of nuclei degradation from one parental (non prevalent parental in the heterokaryon and that the resulting chromosomal fragments may be incorporated into whole nuclei of the another parental (prevalent parental. However the parameiosis as originally described cannot be excluded.

  18. Nutrient activation of Trichoderma fungal spores for improved biocontrol activity

    Institute of Scientific and Technical Information of China (English)

    Linda Gordon Hjeljord; Arne Tronsmo

    2004-01-01

    @@ The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum P1 (ThP1) to suppress Botrytis cinerea was investigated in laboratory, greenhouse and field trials. Preliminary nutrient activation at 21 ℃ accelerated subsequent germination of the antagonist at temperatures from 9 ℃ to 21 ℃; at ≥ 18 ℃ the germination time of preactivated ThP1conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea,concentrated inocula of preactivated but ungerminated ThP1 conidia reduced in vitro germination of the pathogen by ≥ 87 % at 12 ℃ to 25 ℃; initially-quiescent conidia achieved this level of suppression only at 25 ℃. Application of quiescent ThP1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by ≥85 % at 24 ℃ , but only by 35 % at 12 ℃. Preactivated conidia reduced infection by ≥60% at 12 ℃. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperture of 19 ℃, while only preactivated conidia were effective in the field at a mean temperature of 14 ℃ on the day of treatment application.

  19. Biosolubilization gamma irradiate ion result coal by mould trichoderma sp

    International Nuclear Information System (INIS)

    Biosolubilization of coal is process of converting solid coal to liquid fuel/chemicals by mean of microorganism. The aim of this research was to study the effect of gamma rays irradiation with varian doses of irradiation into solubilization of subbituminous coal by Trichoderma sp. The dosage used was 5, 10, and 20 kGy and unirradiated coal as control. The method was submerged culture in MSS+ medium and incubated at room temperature and agitated at 150 rpm for 21th days. The parameters observed were colonization, pH and biosolubilization product based on absorbance value at λ250nm and λ450nm and GC/MS analysis for the best treatment. The results showed that coal biosolubilization could be increased by gamma irradiation. The mould could growth well in medium containing irradiated coal and the medium of pH was decreased after incubation. The biosolubilization was increased but the irradiation dosage of coal didn't affect significantly. The best dose was 20 kGy with product biosolubilization similar to gasoline and solar. Based on the result, the pre-treatment of gamma irradiation on coal has potency to increased biosolubilization. (author)

  20. Isolation, purification and identification of three peptaibols from Trichoderma koningii with antibiotic activity against Ralstonia solancearum

    Institute of Scientific and Technical Information of China (English)

    SHEN Qing-tao; CHEN Xiu-lan; SUN Cai-yun; ZHANG Yu-zhong

    2004-01-01

    @@ The use of microorganisms for biological purposes has become an effective alternative to control plant pathogens. Trichoderma koningii Smf2 was chosen from eight Trichoderma strains for its thermostatic metabolites with antibiotic activity against Ralstonia solancearum Smith. Exclusion chromatography (LH20) was used twice to partially purify targeted metabolites combined with biological test. LC/ESI-MS, a powerful tool for rapid identification and sequence determination of peptides, identified these metabolites as three peptaibols named Trichokonin Ⅵ, Ⅶ and Ⅷ, and their sequences were confirmed with NMR.

  1. Role of Bradyrhizobium japonicum and Trichoderma spp. in the control of root rot disease of soybean

    Directory of Open Access Journals (Sweden)

    Syed Ehteshamul-Haque

    2014-08-01

    Full Text Available Seed treatment of soybean with Bndyrhizobium japonicum, Trichoderma harzianum, T. viride, T. hamatum, T. koningii and T. pseudokoningii significantly controlled the infection of 30-day-old seedlingsby Maerophomina phaseolina, Rhizoctonia solani and Fusarium spp. In 60-day-old plants Trichoderma spp.. and B. japonicum inhibited the grouth of R. solani and Fusarium spp., whereas the use of B. japonicum (TAL-102 with T. harzianum. T. viride, T. koningii and T. pseudokoningii controlled the infection by M. phaseolina. Greater grain yield was recorded when B. japonium (TAI-102 was used with T. hamatum.

  2. Antagonism of Trichoderma harzianum NF9 and Trichoderma virens TY against Three Plant Fungous Pathogens%木霉菌对植物病原真菌拮抗作用的研究

    Institute of Scientific and Technical Information of China (English)

    姚艳平; 李友莲; 王建明; 张作刚

    2013-01-01

    木霉菌是重要的植物病害生防菌.通过对峙培养法,测定了哈茨木霉(Trichoderma harzianum)菌株NF9和绿木霉(Tric hode rmavire ns)菌株TY对3种土传植物病原真菌的体外拮抗作用.结果表明,这2株木霉菌对黄瓜枯萎病菌、西瓜枯萎病菌及茄子白绢病菌均具有一定的拮抗作用,不同木霉菌株间拮抗效果存在差异,哈茨木霉菌株NF9优于绿木霉菌株TY.%The fungous Trichoderma is an important microorganism in the biological control of plant diseases. The antagonism of Trichoderma harzianum NF9 and Trichoderma virens TY against three soil-borne plant fungous pathogens, Fusarium oxysporum Schl. f. sp. cucumerin Owen, Sclerotium rolfsii and Fusarium oxysporum Schl. f. sp. niveum, were tested in vitro by dual—culture. The result showed that two Trichoderma isolates had antagonism effect on the tested pathogens, and Trichoderma harzianum NF9 was better than Trichoderma virens TY in antagonism effect.

  3. Trichoderma-plant root colonization: escaping early plant defense responses and activation of the antioxidant machinery for saline stress tolerance.

    Science.gov (United States)

    Brotman, Yariv; Landau, Udi; Cuadros-Inostroza, Álvaro; Tohge, Takayuki; Takayuki, Tohge; Fernie, Alisdair R; Chet, Ilan; Viterbo, Ada; Willmitzer, Lothar

    2013-03-01

    Trichoderma spp. are versatile opportunistic plant symbionts which can colonize the apoplast of plant roots. Microarrays analysis of Arabidopsis thaliana roots inoculated with Trichoderma asperelloides T203, coupled with qPCR analysis of 137 stress responsive genes and transcription factors, revealed wide gene transcript reprogramming, proceeded by a transient repression of the plant immune responses supposedly to allow root colonization. Enhancement in the expression of WRKY18 and WRKY40, which stimulate JA-signaling via suppression of JAZ repressors and negatively regulate the expression of the defense genes FMO1, PAD3 and CYP71A13, was detected in Arabidopsis roots upon Trichoderma colonization. Reduced root colonization was observed in the wrky18/wrky40 double mutant line, while partial phenotypic complementation was achieved by over-expressing WRKY40 in the wrky18 wrky40 background. On the other hand increased colonization rate was found in roots of the FMO1 knockout mutant. Trichoderma spp. stimulate plant growth and resistance to a wide range of adverse environmental conditions. Arabidopsis and cucumber (Cucumis sativus L.) plants treated with Trichoderma prior to salt stress imposition show significantly improved seed germination. In addition, Trichoderma treatment affects the expression of several genes related to osmo-protection and general oxidative stress in roots of both plants. The MDAR gene coding for monodehydroascorbate reductase is significantly up-regulated and, accordingly, the pool of reduced ascorbic acid was found to be increased in Trichoderma treated plants. 1-Aminocyclopropane-1-carboxylate (ACC)-deaminase silenced Trichoderma mutants were less effective in providing tolerance to salt stress, suggesting that Trichoderma, similarly to ACC deaminase producing bacteria, can ameliorate plant growth under conditions of abiotic stress, by lowering ameliorating increases in ethylene levels as well as promoting an elevated antioxidative capacity.

  4. Comparison of the effect of L-lysine-. cap alpha. -oxidase from Trichoderma harzianum Rifai and Trichoderma viride on nucleic acid synthesis in human tumor cells in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Khaduev, S.K.; Zhukova, O.S.; Dobrynin, Ya.V.; Soda, K.; Berezov, T.T.

    1986-10-01

    This paper gives comparative data on the effect of the new antitumor enzyme LO from a Soviet strain Trichoderma harzianum Rifai and from Trichoderma viride from Japan, on DNA and RNA synthesis in human ovarian carcinoma cells (CaOv strain) in culture, and also the results of the action of LO from T. harzianum Rifai on protein synthesis. Specific precursors were added before the end of the incubation time to the samples; /sup 3/H-thymidine as precursor for DNA synthesis, /sup 3/H-uridine as RNA precursor, and /sup 3/H-leucine as protein precursor. The final values of inhibition of DNA and RNA synthesis in the presence of enzyme from both sources are shown to depend in a virtually linear manner on enzyme concentration.

  5. Xylanase production by Trichoderma strains in solid substrate fermentation

    Institute of Scientific and Technical Information of China (English)

    Krisztina Kovacs; George Szakacs; Lew Christopher

    2004-01-01

    @@ The importance of microbial enzymes in pulp and paper manufacturing has grown significantly in the last two decades. Solid substrate fermentation (SSF) holds tremendous potential for the production of microbial enzymes of commercial interest. SSF can be of special interest in those processes where the crude fermented product (whole SSF culture, in situ enzyme) may be used directly as the enzyme source. Xylanase preparations practically free of cellulase activity are especially useful for biobleaching of crude cellulose pulps. Thirty-nine Trichoderma isolates have been screened in SSF for xylanase production on hardwood oxygen-delignified soda-aq pulp as carbon source and enzyme inducer.Xylanase activities varied between 0 and 2200 IU/g dry matter (DM) of initial substrate. In most instances, the simultaneously produced cellulase levels were below 1.0 Filter Paper Unit (FPU) /g DM. The xylanase to cellulase activity ratio varied in the range of 5 to 3500. The three most promising isolates (TUB F-1647, TUB F-1658 and TUB F-1684) yielded xylanase activity of 2040,1300 and 1500 IU/g DM xylanase, respectively, and 0.64, 0.43 and 0.43 FPU/g DM cellulase with a xylanase to cellulase activity ratio of 3200, 3000 and 3500, respectively. Wild strains F-1647, F-1658 and F-1684 were isolated from tree bark of Maldives, soils of Peru (last two), respectively.Medium optimization experiments to enhance the xylanase yield and to increase the xylanase to cellulase ratio have also been performed.

  6. Application of Trichoderma harzianum (T22) and Trichoderma atroviride (P1) as plant growth promoters, and their compatibility with copper oxychloride

    Institute of Scientific and Technical Information of China (English)

    Francesco Vinale; Gaetano D' Ambrosio; Khalid Abadi; Felice Scala; Roberta Marra; David Turrà; Sheridan L Woo; Matteo Lorito

    2004-01-01

    @@ Trichoderma strains are used in agriculture because they provide to the plants the following benefits:i) are rhizosphere competence and establish stable rhizosphere microbial communities; ii) control plant disease caused by pathogenic and competitive microflora, by using a variety of mechanisms; iii)improve vegetative growth, root development and yield; iv) make nutrients more available to the plant. In this work we have investigated the ability of T. harzianum T22 and T. atroviride P1 to improve plant growth of locally important horticultural crops: lettuce, tomatoes and peppers and to prevent disease in the greenhouse and field. The effect of the Trichoderma treatment was evaluated by determining the weight of fresh and dry roots and above ground plant biomass, measuring plants height, counting the number of emerged leaves (lettuce, tomatoes and peppers) and quantifying production (tomatoes and peppers). No disease symptoms were found during production, although Fusarium sp. strains pathogenic to tomato were detected in the soil. Compounds containing copper oxychloride are frequently used for fungal disease control in agriculture. In order to investigate the compatibility of T. harzianum T22 and T. atroviride P1 with copper oxychloride applications, the effect on mycelia growth was monitored in both liquid and solid medium. In general, the tests indicated a high level of tolerance of the Trichoderma strains to concentrations of copper oxychloride varying from 0.1 to 5 mmol/L.

  7. Efficacy of soil solarization, Trichoderma harzianum, and coffee pulp amendment against Armillaria sp

    NARCIS (Netherlands)

    Otieno, W.; Termorshuizen, A.J.; Jeger, M.J.

    2003-01-01

    Soil solarization was evaluated singly or in combination with Trichoderma harzianum infestation or coffee pulp amendment for its effect on wood-borne inoculum of an Armillaria sp. pathogenic on tea. Solarization increased maximum soil temperatures at 10 cm depth by 9-12degreesC and reduced viability

  8. Detection of peptaibols and their hydrolysis products in cultures of Trichoderma species.

    Science.gov (United States)

    Solfrizzo, M; Altomare, C; Visconti, A; Bottalico, A; Perrone, G

    1994-01-01

    Fifteen strains of Trichoderma belonging to 12 different species for potential use in biological control were examined for their ability to produce polypeptide antiobitics (peptaibols). Gas chromatography was used to detect the hydrolysis products of peptaibols after esterification with n-propanol and derivatisation with heptafluorobutyric anhydride. In particular, amino acids (alpha-aminoisobutyric acid, and isovalin) and amino alcohols (phenylalaninol, leucinol, and valinol) were used as markers for the detection of different peptaibols. alpha-Aminoisobutyric acid, an amino acid characterising all peptaibols, was detected in all cultures examined, indicating that the production of peptaibols is a common feature of the various Trichoderma species. Different amino acid/amino alcohol combinations suggested specific peptaibol production. On this basis, the peptaibol paracelsin was confirmed by HPTLC in 11 out of 15 Trichoderma strains cultures. Valinol was not found in any culture extract, indicating the lack of trichotoxins production. Culture extracts of the 6 tested Trichoderma strains were extremely toxic to Artemia salina larvae; for 5 of them this toxicity could be attributed to paracelsin (calculated LD50 = 2.2 microM). PMID:7704449

  9. A paralog of the proteinaceous elicitor sm1 affects colonization of maize roots by Trichoderma virens

    Science.gov (United States)

    The biocontrol agent, Trichoderma virens, has the ability to protect plants from pathogens by eliciting plant defense responses, involvement in mycoparasitism, or secreting antagonistic secondary metabolites. SM1, an elicitor of induced systemic resistance (ISR), was found to have three paralogs wi...

  10. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea.

  11. Relevance of trichothecenes in fungal physiology: Disruption of tri5 in Trichoderma arundinaceum

    Science.gov (United States)

    Trichothecenes are sesquiterpenoid mycotoxins produced mainly by Fusarium species. Harzianum A (HA), a non-phytotoxic trichothecene produced by Trichoderma arundinaceum, has recently been found to have antagonistic activity against fungal plant pathogens and to induce plant genes involved in defense...

  12. Trichoderma species mediated differential tolerance against biotic stress of phytopathogens in Cicer arietinum L.

    Science.gov (United States)

    Saxena, Amrita; Raghuwanshi, Richa; Singh, Harikesh Bahadur

    2015-02-01

    Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, β-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation.

  13. Promoção do crescimento do feijoeiro e controle da antracnose por Trichoderma spp

    Directory of Open Access Journals (Sweden)

    Erica Aparecida de Souza Pedro

    2012-11-01

    Full Text Available O objetivo deste trabalho foi avaliar a capacidade de Trichoderma spp. em promover o crescimento de plantas de feijão e reduzir a severidade da antracnose do feijoeiro (Colletotrichum lindemuthianum, bem como identificar os isolados mais eficientes. Sessenta isolados de Trichoderma spp. foram avaliados quanto à capacidade de promoção do crescimento nas plantas. Os sete isolados que mais se destacaram foram adicionados ao substrato de cultivo e avaliados quanto à redução na severidade da antracnose em plantas de feijão tratadas com conídios de C. lindemuthianum. Os mais eficientes no controle da doença foram identificados por sequenciamento de DNA. O isolado IB 28/07 foi avaliado nas concentrações 0,5, 1, 1,5 e 2% (peso:volume, que reduziram a severidade da doença em 41,51, 55,15, 81,82 e 96,06%, respectivamente. Os isolados mais eficientes de Trichoderma spp. podem proporcionar aumentos superiores a 30% na produção de matéria seca da parte aérea das plantas e reduzir a severidade da doença entre 63 e 98%. Esses isolados foram identificados como pertencentes às espécies Trichoderma harzianum, T. strigosum e T. theobromicola.

  14. 75 FR 43072 - Trichoderma Hamatum Isolate 382; Exemption from the Requirement of a Tolerance

    Science.gov (United States)

    2010-07-23

    ... 180.1102 (64 FR 16856, April 7, 1999 FRL-6070-3) and Trichoderma harzianum strain T-39 under 40 CFR...) 305-5805. II. Background and Statutory Findings In the Federal Register of July 22, 2009 (74 FR 36200... FR 28580, June 5, 1996, FRL-5371-2). This temporary exemption expired on March 1, 1998....

  15. Study on the effect of different nutrients on the growth of a Trichoderma aureoviride mutant

    Institute of Scientific and Technical Information of China (English)

    CHEN Jian-ai; WANG Wei-ming

    2004-01-01

    @@ Trichoderma aureoviride mutant T1010 was cultured in media containing different nutrients.Statistics showed significant differences and the results described below: The C/N tested were 3:1,6:1, 12: 1, 24: 1,48: 1. The best C/N was 24: 1. The amount of spores was 4.0×109 cfu/plate. The best carbon source among glucose, sucrose, maltose, lactose, cellulose and starch was the last one.With a starch-containing medium, T1010 produced 5.3 × 109 cfu/plate. The asparagine was the best nitrogen source. T1010 absorbed it easily and produced 9.6 × 109 cfu/plate. K, Mg, P, S, Fe were important to support T1010 growth and sporulation. In the absence of K, P, and Fe, Trichoderma grew poorly, while Mg didn' t seem to help Trichoderma colony development. A little amount of vitamins allwed T1010 to grow better. This data allowed us to improve the cultivation of Trichoderma at industrial level.

  16. On-Site Enzyme Production by Trichoderma asperellum for the Degradation of Duckweed

    DEFF Research Database (Denmark)

    Bech, Lasse; Herbst, Florian-Alexander; Grell, Morten Nedergaard;

    2015-01-01

    The on-site production of cell wall degrading enzymes is an important strategy for the development of sustainable bio-refinery processes. This study concerns the optimization of production of plant cell wall-degrading enzymes produced by Trichoderma asperellum. A comparative secretome analysis...

  17. Integrated genomic and transcriptomic analysis reveals mycoparasitism as the ancestoral life style of Trichoderma

    Energy Technology Data Exchange (ETDEWEB)

    Kubicek, Christian P.; Herrera-Estrella, Alfredo; Seidl, Verena; Crom, St& #233; phane Le; Martinez, Diego A.; Druzhinina, Irina S.; Zeilinger, Susanne; Casas-Flores, Sergio; Horwitz, Benjamin A.; Mukherjee, Prasun K.; Mukherjee, Mala; Kredics, L& #225; szlo; Alcaraz, Luis David; Aerts, Andrea; Antal, Zsuzsanna; Atanasova, Lea; Cervantes-Badillo, Mayte Guadalupe; Challacombe, Jean; Chertkov, Olga; McCluskey, Kevin; Coulpier, Fanny; Deshpande, Nandan; D& #246; hren, Hans von; Ebbole, Daniel J.; Esquivel-Naranjo, Edgardo Ulises; Fekete, Erzs& #233; bet; Flipphi, Michel; Glaser, Fabian; Gomez-Rodriguez, Elida Yazmin; Gruber, Sabine; Han, Cliff; Henrissat, Bernard; Hermosa, Rosa; Hern& #225; ndez-O?ate, Miguel; Karaffa, Levente; Kosti, Idit; Lindquist, Erika; Lucas, Susan; L& #252; beck, Mette; L& #252; beck, Peter Stephensen; Margeot, Antoine; Metz, Benjamin; Misra, Monica; Nevalainen, Helena; Omann, Markus; Packer, Nicolle; Perrone, Giancarlo; Uresti-Rivera, Edith Elena; Salamov, Asaf; Schmoll, Monika; Seiboth, Bernhard; Shapiro, Harris; Sukno, Serenella; Tamayo-Ramos, Juan Antonio; Thon, Michael; Tisch, Doris; Wiest, Aric; Wilkinson, Heather H.; Zhang, Michael; Coutinho, Pedro M.; Kenerley, Charles M.; Monte, Enrique; Baker, Scott E.; Grigoriev, Igor V.

    2011-04-29

    Mycoparasitism, a lifestyle where one fungus is parasitic on another fungus has special relevance when the prey is a plant pathogen, providing a strategy for biological control of pests for plant protection. Probably, the most studied biocontrol agents are species of the genus Hypocrea/Trichoderma.

  18. Study on the Biocontrol Activities of Trichoderma species in Greengram with Infected Fungal Pathogens

    International Nuclear Information System (INIS)

    Seven species of Trichoderma were isolated from rhizospheric soil sources and studied by cultural morphology and microscopic examinations. In dual plate assay, antifungal effects of seven Trichoderma strains were screened against three plant pathogenic fungi (Fusarium oxysporum, Rhizoctonia solani and Pythium sp.) on PDA medium and T-5 isolate showed a wide percentage of inhibitory effects on target pathogens with PIRG value. All Trichoderma strains exhibited a clear zone formation on minimal synthetic medium supplemented with 1% colloidal chitin. T-2 and T-5 were the best chitinase producer strains. In vitro screening for protease activity, the highest protease producing activity of Trichoderma isolate (T-2) were observed in pH indicator medium after 7 days incubation. In pot trial experiment, only T-5 strain exhibited more fungal suppression efficiency on green gram plant than commercial fungicide, Trisan and the other strains. So, it can be said that the effective strain was T-5 strain only which have been more antifungal producing power on three fungal pathogens than Trisan and the resting strains.

  19. Microencapsuling aerial conidia of Trichoderma harzianum through spray drying at elevated temperatures

    Science.gov (United States)

    Trichoderma conidia are mostly produced by solid fermentation systems. Inoculum is produced by liquid culturing, and then transferred to solid substrate for aerial conidial production. Aerial conidia of T. harzianum are hydrophilic in nature, and it is difficult to separate them from the solid subst...

  20. Draft Whole-Genome Sequence of the Biocontrol Agent Trichoderma harzianum T6776.

    Science.gov (United States)

    Baroncelli, Riccardo; Piaggeschi, Giulia; Fiorini, Lisa; Bertolini, Edoardo; Zapparata, Antonio; Pè, Mario Enrico; Sarrocco, Sabrina; Vannacci, Giovanni

    2015-01-01

    Trichoderma harzianum T6776 is a promising beneficial isolate whose effects consist of growth promotion, positive response of photosynthetic activity, hormonal signaling, and carbon partitioning in tomato, coupled with biocontrol of plant pathogens. Here, we present the first genome assembly of T6776, providing a useful platform for the scientific community. PMID:26067977

  1. Trichoderma L-Lysine-α-Oxidase Producer Strain Culture Fluid Inhibits Impatiens Necrotic Spot Virus.

    Science.gov (United States)

    Smirnova, I P; Shneider, Yu A; Karimova, E V

    2016-01-01

    A method for PCR diagnosis of impatiens necrotic spot virus is developed. Concentrated culture fluid with active L-lysine-α-oxidase (0.54 U/ml) from Trichoderma harzianum Rifai fungus producer strain F-180 inhibits vitally hazardous impatiens necrotic spot phytovirus. PMID:26742733

  2. Effect of some Trichoderma spp. isolates on promoting growth of cucumber seedlings under greenhouse conditions

    Directory of Open Access Journals (Sweden)

    M. Taghinasab Darzi

    2012-12-01

    Full Text Available This experiment was performed to investigate the effect of some Trichoderma spp. isolates as growth promoters of cucumber (Cucumis sativus L. seedlings under greenhouse conditions. Inoculai of 19 Thrichoderma spp. isolates were prepared from disinfected wheat grain. The upper half of the soil in pots (containing field soil and sand was mixed with these inoculai at 3% ratio and the pots were irrigated with tap water for 28 days. After four weeks, the seedlings were sampled for growth comparison on stem length, root length and total fresh weight. The results showed that some isolates improved significantly the cucumber seedlings’ growth and others had inhibitory effect. Application of Trichoderma spp. 17 and T. longibraciatum increased stem length more than 74% as compared to control. Also, these isolates increased significantly P<0.05 the total fresh weight about 40% and 25%, respectively, as compared to control. Furthermore, Trichoderma sp. 19 decreased significantly the stem length, root length and total fresh weight as compared to control. These results show the ability of Persian Trichoderma spp. isolates in promoting cucumber growth and its potential for other plants.

  3. Antagonistic and plant growth activity of Trichoderma isolates of Western Himalayas.

    Science.gov (United States)

    Joshi, B B; Bhatt, R P; Bahukhandi, D

    2010-11-01

    The genus Trichoderma is rapidly growing colonies bearing tufted or postulate, repeatedly branched conidiophores with lageniform phialides and hyaline or green conidia born in slimy heads. 62 isolates of Trichoderma species were isolated from different rhizospheric soil samples collected from different places located in Western Himalayas region. Out of these only two species were found i.e. Trichoderma hazianum and Trichoderma viride. Their efficacy against soil borne plant pathogens like Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotiorum revealed that only three isolates amounting to 5% of the total collected isolates of this region were found highly antagonist. Among them 5% isolates were found against S. rolfsii, 13% isolates against R. solani, 10% against sclerotium caused above 80% inhibition of mycelial growth respectively. 6% isolates out of twenty seven utilized chitin by more than 80 and 16% isolates consumed cellulose by above 80% and therefore are producers of chitinase and cellulases. 58% isolates produced colonies having cottony texture and 41% produced dark green colonies. Pigmentation as observed from reverse side of the colony revealed that 70% of them did not produced pigment in the medium. Plant growth promotion measured as root and shoot lengths were significantly higher than in control. The maximum root length and shoot length were recorded when seeds were treated with isolates were recorded at Srinagar Garhwal was 4.70 and 4.75 cm out of all the isolates in which isolate recorded from Srinagar no 3 caused maximum percent seed germination which was significantly higher 79.49%. PMID:21506476

  4. Trichoderma species mediated differential tolerance against biotic stress of phytopathogens in Cicer arietinum L.

    Science.gov (United States)

    Saxena, Amrita; Raghuwanshi, Richa; Singh, Harikesh Bahadur

    2015-02-01

    Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, β-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation. PMID:25205162

  5. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea. PMID:25171761

  6. Randomly Amplified Polymorphic DNA of Trichoderma isolates and antagonism against Rhizoctonia solani

    Directory of Open Access Journals (Sweden)

    Larissa Brandão Góes

    2002-06-01

    Full Text Available Random Amplified Polymorphic DNA (RAPD procedure was used to examine the genetic variability among fourteen isolates of Trichoderma and their ability to antagonize Rhizoctonia solani using a dual-culture assay for correlation among RAPD products and their hardness to R. solani. Seven oligodeoxynucleotide primers were selected for the RAPD assays which resulted in 197 bands for 14 isolates of Trichoderma. The data were entered into a binary matrix and a similarity matrix was constructed using DICE similarity (SD index. A UPGMA cluster based on SD values was generated using NTSYS (Numerical Taxonomy System, Applied Biostatistics computer program. A mean coefficient of similarity obtained for pairwise comparisons among the most antagonics isolates was around 40%. The results presented here showed that the variability among the isolates of Trichoderma was very high. No relationship was found between the polymorphism showed by the isolates and their hardness, origin and substrata.A técnica de RAPD (Random Amplified Polymorphic DNA foi utilizada para examinar a variabilidade genética em quatorze isolados de Trichoderma além de sua capacidade de antagonizar o fungo fitopatogênico Rhizoctonia solani usando pareamento in vitro, e a possível relação entre perfís de RAPD e agressividade dos isolados de Trichoderma a R. solani. Foram selecionados sete primers para os ensaios de RAPD, os quais produziram 197 bandas. Os dados foram introduzidos no programa de computador NTSYS (Numerical Taxonomy System, Applied Biostatisticsna forma de uma matrix binária, sendo construída uma matriz de similaridade utilizando-se o coeficiente de similaridade de DICE (SD e baseado nos valores SD, pelo método de agrupamento UPGMA um dendrograma. Observou-se que o grau de similaridade das amostras que apresentaram melhor desempenho antagônico foi bastante baixo, em torno de 40%. Os resultados demonstraram que a variabilidade entre os isolados de Trichoderma é muito

  7. Trichoderma sp Native from Chili Region of Poanas, Durango, Mexico Antagonist against Phytopathogen Fungi

    Directory of Open Access Journals (Sweden)

    Gabriela B. Valencia

    2011-01-01

    Full Text Available Problem statement: Presence of Trichoderma spp. in agricultural soils decrease incidence of diseases by phytopathogen fungi. Sanity diagnostic require to know if exist beneficial microorganism and what agricultural practices help to their propagation. Approach: Samples (30 were taken from soils and sick plants of ten sites in four localities of Valley of Poanas. Phytophthora capsici Leo, Rhizoctonia solani Kuhn and Trichoderma sp were isolated in agar V8 and were identified by microscopy. Results: In the 30 samples analyzed the presence of Phytophthora capsici Leo and Rhizoctonia solani Kuhn was determined. Two isolations of Trichoderma sp were obtained from soil, they had antagonist activity against to P. capsici and R. solani on agar-V8 medium and showed chitinase activity. Sugar production in chitinase (10 mg.mL-1 by crude extract of Trichoderma growth in basal medium more chitin was determined. The average of sugar production from strains were 0.1175 and 0.1125 mg.mL-1 and standard deviations were 0.0567 and 0.0567 in four repetition. Interviews were applied to fifty farmers about cultivars and cultivation practices. At least seven types of chili were cultivated in the region of the Valley of Poanas, inorganic fertilization, irrigation systems by channel, gates and pumps were used. One hundred percent of farmers reported diseases of Damping off and Phytophthora root. Biocides were not used to control these diseases. Conclusion: The natural presence of Trichoderma spp was detected in Valley of Poanas, but some practices as inorganic fertilization and irrigation system can be contributing to propagation of phytopathogen fungi.

  8. Variabilidade entre isolados de Trichoderma harzianum: I - Aspectos citológicos Variability among Trichoderma harzianum isolates: I - Cytological aspects

    Directory of Open Access Journals (Sweden)

    E. Peres

    1995-04-01

    Full Text Available Objetivou-se neste trabalho estudar a variabilidade de isolados selvagens de Trichoderma harzianum baseado nas características culturais e citológicas. Observaram-se o tamanho dos fialosporos, número de núcleos por fialosporos e crescimento e esporulação em meio de malte-ágar. Pelos resultados aqui encontrados foi possível reconhecer que há variação entre os isolados selvagens da espécie T. harzianum. Com relação ao número de núcleos, verificou-se uma variação de 1 a 3 núcleos por fialosporos. Também observou-se padrões diferenciais de crescimento e morfologia da colônia. Mais de 50% dos isolados atingiram o máximo de crescimento em 48 horas.This study is based largely on morphological and cultural characters of Tríchoderma harzianum isolates. It were observed the size of phialospores, mycelial growth and sporulation on malt extract agar and nuclei number per phialospores, stained with Giemsa. A x 100 oil immersion len was used in examining and in measuring phialospores. Based on the size of phialospores, it was possible recognize that there is variaton among the wild isolates for the specie T. harzianum. Also, with relation to mycelial growth and sporulation can itself distinguish from one another different pattern. Up to 50% of isolates had maximum growth in 48 hours. The nuclei number of 1 to 3 per phialospore was observed.

  9. Biocontrol de la pudrición de raíz de nochebuena de interior con Trichoderma spp. Root rot biocontrol for indoor poinsettia with Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Felipe de Jesús Osuna-Canizalez

    2012-06-01

    Full Text Available En Morelos, la pudrición de la raíz causada por Fusarium spp., es una de las principales enfermedades de la nochebuena de interior. Por su efecto devastador, en su prevención o control se realizan aplicaciones frecuentes de productos químicos, con los riesgos inherentes a la salud humana y al ambiente. En la búsqueda de alternativas bioracionales al manejo de esta enfermedad, se realizó un ensayo en el que se evaluaron tres cepas comerciales de Trichoderma spp., en tres diferentes sustratos: S1= "tierra de hoja" (70% v/v+tezontle grueso (15% v/v+tezontle fino (10% v/v+agrolita (5% v/v; S2= turba (80% v/v+ fibra de coco (20% v/v; S3= "tierra de hoja" (70% + "tepojal" (30%, en las variedades comerciales más comunes, Freedom Red y Prestige Red. Se utilizó un diseño factorial de tratamientos 4 x 3 x 2 y los tratamientos resultantes se evaluaron en un diseño completamente al azar con seis repeticiones. Respecto a la incidencia de pudrición de la raíz, las cepas comerciales de Trichoderma spp., no mostraron diferencias entre sí ni con el testigo químico. La pudrición de la raíz estuvo asociada con S2, debido a una baja capacidad de aireación, y sólo se presentó en Prestige Red. La población (UFC g-¹ de Trichoderma spp., en el sustrato al término del ciclo, fue igual (pIn Morelos, root rot caused by Fusarium spp., is one of the main diseases of indoor poinsettia. In order to prevent or control its devastating effect, frequent applications of chemical products are performed, with inherent risks to human health and environment. In quest for alternative biorational options, an essay in which three commercial strains of Trichoderma spp., was done, in three different substrates: S1= "organic soil" (70% v/v+thick tezontle (15% v/v+thin tezontle (10% v/v+agrolita (5% v/v; S2= peat moss (80% v/v+ coconut fiber (20% v/v; S3= "organic soil" (70%+"tepojal" (30%, in most common commercial varieties, Freedom Red and Prestige Red. A factorial

  10. KINETICS AND EQUILIBRIUM PARAMETERS OF BIOSORPTION AND BIOACCUMULATION OF LEAD IONS FROM AQUEOUS SOLUTIONS BY TRICHODERMA LONGIBRACHIATUM

    OpenAIRE

    Enitan S. Balogun; John B. Durosanya; Sarafadeen O. Kareem; Abideen I. Adeogun

    2012-01-01

    Biosorption and bioaccumulation of Lead ions (Pb(II)) by Trichoderma longibrachiatum were investigated in a batch system. The effects of some important parameters such as pH, initial metal concentration, temperature and inoculum concerntration on biosorption capacity were also studied. The maximum biosorption capacity of Trichoderma longibrachiatum was at 25 ppm of lead, showed 100 % removal at pH 7 and 25 oC after fifteen days. Biosorption equilibrium was established in 150 minutes. The proc...

  11. Cara Aplikasi Trichoderma spp. untuk Menekan Infeksi Busuk Pangkal Batang (Athelia rolfsii (Curzi)) pada Beberapa Varietas Kedelai di Rumah Kassa

    OpenAIRE

    Husna, Rafika

    2016-01-01

    Athelia rolfsii (Curzi) is soil borne disease which has wide host range such as rice, mungbean, peanut, soybean, sweet potato, banana, wheat, and potato. Trichoderma spp. is antagonistic fungi that can suppress of some pathogens. This experiment aimed to test the applications of Trichoderma spp. with seed treatment and direct application to Athelia rolfsii (Curzi) with three different soybean varieties including Anjasmoro, Grobogan, and Willis. The experiment was conducted in Plant Disease La...

  12. Adaptation of Trichoderma Species to Pesticide Confidor and Evaluation of their Growth Ability in the Media Containing Confidor

    OpenAIRE

    Farnaz Ershadfath; Hossein Banejad; Fariba Mohsenzadeh

    2015-01-01

    Introduction: Contamination caused by pesticides is considered as one of the environmental problems. Bioremediation is exploiting the ability of microorganisms to remove pollutants. Trichoderma species are free-living fungi that exist naturally in the environment. These fungi have the ability to uptake some contaminants biologically. The aim of this study is to evaluate the effect of Confidor, as an environmental contaminant, on the growth ability of Trichoderma sp. as a contaminant absorber....

  13. In Vitro Inhibition of Cellulolytic Enzymes of Fusarium Oxysporum by Trichoderma spp and Pseudomonas Fluorescens on Arachis Hypogaea L

    OpenAIRE

    P.Rajeswari

    2015-01-01

    In an attempt to develop biocontrol system for management of Fusarium wilt in groundnut, Trichoderma viride, Trichoderma harzianum,and Pseudomonas fluorescens were evaluated for their antagonistic activity against Fusarium oxysporum in vitro. .Fusarium wilt diseasescaused by the fungus Fusarium oxysporum lead to significant yield losses of crops. Experiments were conducted on the effect of culture filtratesof T.viride (1%), T. harzianum (1.5%), and P. fluorescens (2%) on the in vitro inhibiti...

  14. Isolation, screening, and molecular characterization of plant growth promoting rhizobacteria isolates of Azotobacter and Trichoderma and their beneficial activities

    OpenAIRE

    Kasa, Parameswari; Modugapalem, Hemalatha; Battini, Kishori

    2015-01-01

    Objectives: The present study was conducted for isolation, screening, and identification of Azotobacter and Trichoderma from different soil samples. Methods: A total of 10 isolates of Azotobacter and Trichoderma were isolated from rhizospheric soils. The test isolates were biochemically characterized and screened in in-vitro conditions for their plant growth promoting properties. DNA polymorphism of isolates was studied using randomly amplified polymorphic DNA analysis. Results: A total of 41...

  15. The Epl1 and Sm1 proteins from Trichoderma atroviride and Trichoderma virens differentially modulate systemic disease resistance against different life style pathogens in Solanum lycopersicum

    Directory of Open Access Journals (Sweden)

    Miguel Angel eSalas-Marina

    2015-02-01

    Full Text Available Fungi belonging to the genus Trichoderma, commonly found in soil or colonizing plant roots, exert beneficial effects on plants, including the promotion of growth and the induction of resistance to disease. T. virens and T. atroviride secrete the proteins Sm1 and Epl1, respectively, which elicit local and systemic disease resistance in plants. In this work, we show that these fungi promote growth in tomato (Solanum lycopersicum plants. T. virens was more effective than T. atroviride in promoting biomass gain, and both fungi were capable of inducing systemic protection in tomato against Alternaria solani, Botrytis cinerea, and Pseudomonas syringae pv. tomato (Pst DC3000. Deletion (KO of epl1 in T. atroviride resulted in diminished systemic protection against A. solani and B. cinerea, whereas the T. virens sm1 KO strain was less effective in protecting tomato against Pst DC3000 and B. cinerea. Importantly, over-expression (OE of epl1 and sm1 led to an increase in disease resistance against all tested pathogens. Although the Trichoderma WT strains induced both systemic acquired resistance (SAR- and induced systemic resistance (ISR-related genes in tomato, inoculation of plants with OE and KO strains revealed that Epl1 and Sm1 play a minor role in the induction of these genes. However, we found that Epl1 and Sm1 induce the expression of a peroxidase and an α-dioxygenase encoding genes, respectively, which could be important for tomato protection by Trichoderma spp. Altogether, these observations indicate that colonization by beneficial and/or infection by pathogenic microorganisms dictates many of the outcomes in plants, which are more complex than previously thought.

  16. Inoculação de sementes de milho utilizando o Trichoderma harzianum como promotor de crescimento Corn seed inoculation using Trichoderma harzianum as a growth promoter

    Directory of Open Access Journals (Sweden)

    Maria de Lourdes Resende

    2004-08-01

    Full Text Available Conduziu-se este trabalho com o objetivo de estudar o efeito do fungo Trichoderma harzianum como promotor de crescimento de sementes de milho tratadas com diferentes fungicidas. O experimento foi realizado em vasos em casa-de-vegetação, adotando o delineamento experimental inteiramente casualizado em esquema fatorial (3x2, utilizando os tratamentos com fungicidas: Captan®, Maxim® e testemunha e inoculação (com e sem inóculo em quatro repetições. As características agronômicas avaliadas foram: altura de planta, peso da matéria seca da parte aérea e de raízes e também a técnica de isolamento do fungo nas raízes aos 45 dias após semeadura. Observou-se diferença significativa somente para o fator inóculo na característica peso de matéria seca de raízes. Pelos resultados obtidos, verificou-se que o fungo Trichoderma harzianum estimulou maior acúmulo de matéria seca nas raízes das plantas de milho e também estava presente nas raízes das plantas oriundas de sementes inoculadas, independentemente do tratamento fungicida das sementes.The objective of this work was to study the effect of the fungus Trichoderma harzianum as a growth promoter on the corn seeds treated with different fungicides. This experiment was carried out in pots in a greenhouse by adopting the experimental completely randomized design in (3 X 2 factorial scheme, by utilizing the treatments with fungicides: Captan®, Maximim® and control and inoculation (with and without an inoculum with four replicates. The agronomic characteristics evaluated were: plant height, shoot and root dry matter weight and also the inoculum isolation technique on the rots after sowing. Significant differences were found only for the factor inoculum in the characteristic root dry matter weight. The results obtained revealed that the fungus Trichoderma harzianum stimulated greatest dry matter accumulation in the roots of corn plants and also it was present in the roots of plants from

  17. Biocatalytic and antimicrobial activities of gold nanoparticles synthesized by Trichoderma sp.

    Science.gov (United States)

    Mishra, Aradhana; Kumari, Madhuree; Pandey, Shipra; Chaudhry, Vasvi; Gupta, K C; Nautiyal, C S

    2014-08-01

    The aim of this work was to synthesize gold nanoparticles by Trichoderma viride and Hypocrea lixii. The biosynthesis of the nanoparticles was very rapid and took 10 min at 30 °C when cell-free extract of the T. viride was used, which was similar by H. lixii but at 100 °C. Biomolecules present in cell free extracts of both fungi were capable to synthesize and stabilize the formed particles. Synthesis procedure was very quick and environment friendly which did not require subsequent processing. The biosynthesized nanoparticles served as an efficient biocatalyst which reduced 4-nitrophenol to 4-aminophenol in the presence of NaBH₄ and had antimicrobial activity against pathogenic bacteria. To the best of our knowledge, this is the first report of such rapid biosynthesis of gold nanoparticles within 10 min by Trichoderma having plant growth promoting and plant pathogen control abilities, which served both, as an efficient biocatalyst, and a potent antimicrobial agent.

  18. Identification of Trichoderma Species on Mushrooms%食用菌木霉种类鉴定

    Institute of Scientific and Technical Information of China (English)

    贺字典; 孙焕顷; 高玉峰

    2008-01-01

    从不同食用菌上分离到57株木霉,按Rifai和Bissett的分类系统共鉴定出6种木霉:康氏木霉(Tri-choderma koningii)、拟康木霉(Trichoderma pseudokoningii)、哈茨木霉(Trichodema harzianum)、桔绿木霉(Tri-choderma citrinviride)、长枝木霉(Trichoderma longibrachiatum)和非钩木霉(Trichodema inhamatum).首次从白灵菇的培养基质上分离到了非钩木霉、康氏木霉和拟康木霉,后两种是食用菌上木霉的优势种,分别占总菌株数的50.87%和26.32%.

  19. Trichoderma harzianum as a biocontrol agent against Alternaria alternata on tobacco

    Directory of Open Access Journals (Sweden)

    Jugoslav Ziberoski

    2012-06-01

    Full Text Available Trichoderma fungi are the most popular agents used in a biological control. Therefore, our aim was to determine an impact of Trichoderma harzianum on the fungus Alternaria alternata - a causing agent of the brown spot disease on tobacco. In vitro analyses were made in several variants of double culture, in order to study the effect of difusible and volatile metabolites. There was strong reducing effect on the development of A.alternata with various mechanisms of antagonistic influence. The volatile metabolites have also shown reducing effect. Some abnormalities were observed in the pathogen's morphology both in difusible and volatile metabolites. The strong reducing effect of T.harzianum towards A. alternata can be applied in biological control of this pathogen.

  20. Trichoderma koningii assisted biogenic synthesis of silver nanoparticles and evaluation of their antibacterial activity

    Science.gov (United States)

    Tripathi, R. M.; Gupta, Rohit Kumar; Shrivastav, Archana; Singh, M. P.; Shrivastav, B. R.; Singh, Priti

    2013-09-01

    The present study demonstrates the biosynthesis of silver nanoparticles using Trichoderma koningii and evaluation of their antibacterial activity. Trichoderma koningii secretes proteins and enzymes that act as reducing and capping agent. The biosynthesized silver nanoparticles (AgNPs) were characterized by UV-Vis spectroscopy, dynamic light scattering (DLS), transmission electron microscopy (TEM) and x-ray diffraction (XRD). UV-Vis spectra showed absorbance peak at 413 nm corresponding to the surface plasmon resonance of silver nanoparticles. DLS was used to find out the size distribution profile. The size and morphology of the AgNPs was determined by TEM, which shows the formation of spherical nanoparticles in the size range of 8-24 nm. X-ray diffraction showed intense peaks corresponding to the crystalline silver. The antibacterial activity of biosynthesized AgNPs was evaluated by growth curve and inhibition zone and it was found that the AgNPs show potential effective antibacterial activity.

  1. Exposing of Trichoderma spp. to gamma radiation for stimulating its pesticide biodegradation activity

    International Nuclear Information System (INIS)

    This work has been conducted to study the possibility of making use of fungi for degrading insecticide-carbofuran. Trichoderma spp. were showed highly potentiality to metabolize carbofuran (200 mg/ kg) to 3-ketocarbofuran in soil as a sole carbon and energy source within 14 days. Carbofuran and its main metabolite were analyzed by high performance liquid chromatography (HPLC). Studies on biodegradation in the soil showed that 81.5 % and 86 % of carbofuran degraded within 14 days of incubation by T. harzianum and T. viride strains, respectively. The lowest dose of gamma irradiation 0.25 KGy enhanced the mycelial dry weight by 22.8 % and 16.2 % for T. harzianum and T. viride strains, respectively. This indicated that the isolates of Trichoderma spp. were potentially useful for carbofuran bioremediation.

  2. Effect of microelement and chemical fungicides on biocontrol effect of Trichoderma T23

    Institute of Scientific and Technical Information of China (English)

    ZHUANG Jing-hua; GAO Zeng-gui; YANG Chang-cheng; LIU Xian; CHEN Jie

    2004-01-01

    @@ Recently there have been many reports about soil diseases controlled by Trichoderma, but few could be applied on agriculture production in large areas. T23 isolated from soil around plant roots in the field by Biopesticide Engineering Center of Shenyang Agricultural University could control effectively Cucurbits Fusarium Wilt. The effects of 9 microelements which include copper, zinc, iron, boron,molybdenum, calcium, manganese, magnesium, potassium and frequently-used chemical fungicides,such as-carbendazim, thiram, thiophanate-methyl, chlorothalonil and hymexazol on the growth and the amounts of spores of Fusarium oxysporum FJ and Trichoderma T23 were studied. The effects of those factors on control effect of T23 to melon diseases were discussed and gave basis for the screening of synergistic agents and fungicides in controlling synergistically the pathogen.

  3. Primary Study on Biological Control Potential of Trichoderma harzianum TL-1

    Institute of Scientific and Technical Information of China (English)

    Su; Zhenyu; Xiao; Man; Gao; Xinzheng; Tang; Libo; Li; Li

    2014-01-01

    Trichoderma harzianum is a widely used biocontrol fungus. The growth promoting effect of strain Trichoderma harzianum TL-1 on tomato and pepper and its biological control effects against tomato seedling damping-off and pepper blight were investigated through pot experiments. The results showed that the stain TL-1 had significant promotion effect on growth of pepper and tomato in sterilized and natural soils. With the application dose of 3. 0 and 0. 5g/ pot,their dry weight were increased up to 46% and 150% compared with control,respectively. In addition,TL-1 had good control effects against tomato seedling damping-off and pepper blight. Compared with fungicide treatment,TL-1 treatment could control diseases for long term,without repeat occurrence of diseases.

  4. Genome-scale investigation of phenotypically distinct but nearly clonal Trichoderma strains.

    Science.gov (United States)

    Lange, Claudia; Weld, Richard J; Cox, Murray P; Bradshaw, Rosie E; McLean, Kirstin L; Stewart, Alison; Steyaert, Johanna M

    2016-01-01

    Biological control agents (BCA) are beneficial organisms that are applied to protect plants from pests. Many fungi of the genus Trichoderma are successful BCAs but the underlying mechanisms are not yet fully understood. Trichoderma cf. atroviride strain LU132 is a remarkably effective BCA compared to T. cf. atroviride strain LU140 but these strains were found to be highly similar at the DNA sequence level. This unusual combination of phenotypic variability and high DNA sequence similarity between separately isolated strains prompted us to undertake a genome comparison study in order to identify DNA polymorphisms. We further investigated if the polymorphisms had functional effects on the phenotypes. The two strains were clearly identified as individuals, exhibiting different growth rates, conidiation and metabolism. Superior pathogen control demonstrated by LU132 depended on its faster growth, which is a prerequisite for successful distribution and competition. Genome sequencing identified only one non-synonymous single nucleotide polymorphism (SNP) between the strains. Based on this SNP, we successfully designed and validated an RFLP protocol that can be used to differentiate LU132 from LU140 and other Trichoderma strains. This SNP changed the amino acid sequence of SERF, encoded by the previously undescribed single copy gene "small EDRK-rich factor" (serf). A deletion of serf in the two strains did not lead to identical phenotypes, suggesting that, in addition to the single functional SNP between the nearly clonal Trichoderma cf. atroviride strains, other non-genomic factors contribute to their phenotypic variation. This finding is significant as it shows that genomics is an extremely useful but not exhaustive tool for the study of biocontrol complexity and for strain typing. PMID:27190719

  5. Virulence and Experimental Treatment of Trichoderma longibrachiatum, a Fungus Refractory to Treatment.

    Science.gov (United States)

    Paredes, Katihuska; Capilla, Javier; Mayayo, Emilio; Guarro, Josep

    2016-08-01

    Different inocula of Trichoderma longibrachiatum were tested in a murine model, and only the highest one (1 × 10(7) CFU/animal) killed all of the mice at day 15 postinfection, with spleen and liver the most affected organs. The efficacies of amphotericin B deoxycholate, liposomal amphotericin B, voriconazole, and micafungin were evaluated in the same model, with very poor results. Our study demonstrated the low virulence but high resistance to antifungal compounds of this fungus. PMID:27216056

  6. Trichoderma harzianum transformant has high extracellular alkaline proteinase expression during specific mycoparasitic interactions

    Directory of Open Access Journals (Sweden)

    Goldman Maria Helena S.

    1998-01-01

    Full Text Available The mycoparasite Trichoderma harzianum produces an alkaline proteinase that may be specifically involved in mycoparasitism. We have constructed transformant strains of this fungus that overexpress this alkaline proteinase. Some of the transformants were assessed for alkaline proteinase activity, and those with higher activity than the wild type were selected for further studies. One of these transformant strains produced an elevated and constitutive pbr1 mRNA level during mycoparasitic interactions with Rhizoctonia solani.

  7. Phylogeny and biodiversity of Trichoderma and Hypocrea and its implications on taxonomy

    Institute of Scientific and Technical Information of China (English)

    Christian P Kubicek

    2004-01-01

    @@ Safe strain identification and species recognition is an important issue for Trichoderma and Hypocrea,because members of the genus are economically important producers of industrial enzymes and antibiotics, have application as biocontrol agents against plant pathogens, whereas some have become known as opportunistic pathogens of immunocompromised mammals and humans. However, classical approaches based on the use of morphological and phenetic characters have been difficult to apply, due to the plasticity of characters and the discordance of morphological and molecular evolution.

  8. Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

    Science.gov (United States)

    Nor, Nurafifah Mohd; Hadibarata, Tony; Zubir, Meor Mohd Fikri Ahmad; Lazim, Zainab Mat; Adnan, Liyana Amalina; Fulazzaky, Mohamad Ali

    2015-11-01

    Cresol Red belongs to the triphenylmethane (TPM) class of dyes which are potentially carcinogenic or mutagenic. However, very few studies on biodegradation of Cresol Red were investigated as compared to other type dyes such as azo and anthraquinone dye. The aim of this work is to evaluate triphenylmethane dye Cresol Red degradation by fungal strain isolated from the decayed wood in Johor Bahru, Malaysia. Detailed taxonomic studies identified the organisms as Trichoderma species and designated as strain Trichoderma harzianum M06. In this study, Cresol Red was decolorized up to 88% within 30 days under agitation condition by Trichoderma harzianum M06. Data analysis revealed that a pH value of 3 yielded a highest degradation rate among pH concentrations (73%), salinity concentrations of 100 g/L (73%), and a volume of 0.1 mL of Tween 80 (79%). Induction in the enzyme activities of manganese peroxidase, lignin peroxidase, laccase, 1,2- and 2,3-dioxygenase indicates their involvement in Cresol Red removal. Various analytical studies such as Thin-Layer Chromatography (TLC), UV-Vis spectrophotometer, and Gas chromatography mass spectrometry (GC-MS) confirmed the biotransformation of Cresol Red by the fungus. Two metabolites were identified in the treated medium: 2,4-dihydroxybenzoic acid (t R 7.3 min and m/z 355) and 2-hydroxybenzoic acid (t R 8.6 min and m/z 267). Based on these products, a probable pathway has been proposed for the degradation of Cresol Red by Trichoderma harzianum M06.

  9. Antifungal activity of six plant essential oils from Serbia against Trichoderma aggressivum f. europaeum

    OpenAIRE

    Rada Đurović-Pejčev; Ivana Potočnik; Svetlana Milijašević-Marčić; Biljana Todorović; Emil Rekanović; Miloš Stepanović

    2014-01-01

    Six essential oils (EOs) extracted from plants originating in Serbia were assayed for inhibitory and fungicidal activity against a major fungal pathogen of button mushroom causing green mould disease, Trichoderma agressivum f. europaeum. The strongest activity was demonstrated by the oils of basil (Ocimum basilicum L.) and peppermint (Mentha piperita L.). Medium antifungal activity of St. John's wort (Hypericum perforatum L.) and walnut [Juglans regia (F)] ...

  10. Construction of new GFP-tagged fusants for Trichoderma harzianum with enhanced biocontrol activity

    Directory of Open Access Journals (Sweden)

    Kowsari Mojegan

    2014-07-01

    Full Text Available Trichoderma is one of the most exploited biocontrol agents for the management of plant diseases. In biocontrol ecology, the critical factors are detection, and the monitoring and recovery of specific biocontrol agents either naturally present or deliberately released into the environment. Protoplast fusion is an appropriate tool for the improvement of biocontrol Trichoderma strains. Protoplast isolation from Trichoderma harzianum was achieved using 24 h culture age, 6.6 mg/ml Novazym L 1412 at 30°C which resulted the maximum protoplast yield of 5 × 108/ml. The self-fused protoplasts were regenerated and 12 fusants were selected based on their growth rate on 2% colloidal chitin medium. Next, a comparison was done for chitinase and antagonistic activity. Transcriptomic analysis based on quantitative real-time RT-PCR, demonstrated that T8-05 fusant expressed 1.5 fold of chit42 transcript as compared with the parental line. This fusant with 7.02±0.15U chitinase activity showed a higher growth inhibition rate (100% than the parent strain, against Rhizoctonia solani. To obtain a genetically marked fusant that can be used as a biomonitor, the fusant was cotransformed with the gfp and amdS genes. The morphology and viability of selected cotransformant (FT8-7MK-05-2 was similar to the parent. Green fluorescing conidia were observed within the first 2 days of incubation in the soil, and this was followed by the formation of chlamydopores after 60 days. The colonisation of the gfp-tagged fusant was also monitored visually on R. solani sclerotia by scanning electron microscopy. Production of gfp-tagged fusant of Trichoderma spp. provides a potentially useful tool for monitoring hyphal growth patterns and the population of biocontrol agent isolates introduced into environmental systems.

  11. Novel plant bio-protectants based on Trichoderma spp. strains with superior characteristics

    OpenAIRE

    Abadi, Khalid M.

    2008-01-01

    Global warming caused by the greenhouse effect will alter the geographical distribution of host and pathogen populations, thus affecting the natural physiology of their interaction and reducing the efficacy of chemical and biological control strategies presently used in agriculture. In perspective to this scenario, new management practices will be required. The main task of this thesis was to isolate and characterize new biocontrol agents of the genus Trichoderma originating from Libya, where...

  12. Cellulase Activity in Solid State Fermentation of Palm Kernel Cake with Trichoderma sp.

    Directory of Open Access Journals (Sweden)

    Massaud, M. B. N.

    2012-01-01

    Full Text Available Aims: The effect of different types of fungal inocula to the cellulase activity measured on palm kernel cake (PKC was studied. Methodology and Results: Isolate Pro-A1 which was identified as Trichoderma sp. was selected as a potential producer of cellulase via solid state fermentation technique (SSF. Two types of PKCs were used; raw PKC (containing residual oil and defatted PKC. The PKCs were inoculated with different concentrations of conidia and varying amounts (g of solid mycelia plugs (SMP for SSF. The effect of ultrafiltered crude fungal filtrate (CFF as inocula was also being tested. The highest cellulase activity of 2.454 FPU/mL was detected with 60% (wt/wt SMP applied to the raw PKC. Conversely, 2.059 FPU/mL of cellulase activity was measured when 80% (wt/wt of SMP was applied to the defatted PKC which is 62.3% higher than the untreated defatted PKC; and more than 100% increase in enzymatic activity compared to raw PKC. The cellulase activity in the SSF inoculated with 8 x 106 conidia /mL and 12 x 106 conidia /mL were 1.704 FPU/mL for raw PKC and 1.856 FPU/mL for defatted PKC, an enhancement of about 46% from uninoculated batch. Inoculation with CFF bears corresponding maximum improvement of the cellulase activity on both PKCs of 13.58% (raw and 2.86% (defatted. Conclusion, significance and impact of study: The current study proves that Trichoderma sp. in the form of SMP can enhance the cellulase activity on PKCs effectively with more than 100% increment. Fungal conidia are also a better choice in enhancing cellulase activity of Trichoderma sp. permitted that the PKC used is devoid of oil. From this study, Trichoderma sp. holds the potential of converting lignocellulosic materials into products of commercial and industrial values such as glucose and other biofuels.

  13. A Selective Medium for Quantitative Reisolation of Trichoderma harzianum from Agaricus bisporus Compost

    OpenAIRE

    Williams, Josie; Clarkson, John M.; Mills, Peter R.; Cooper, Richard M

    2003-01-01

    We adapted a selective medium, previously developed for reisolation of Trichoderma spp. from soil, for quantitative determination of growth of T. harzianum from commercial Agaricus bisporus composts. This medium enables comparisons of aggressive (sensu inhibition of A. bisporus yield) with nonaggressive T. harzianum groups. The resulting medium contains the antimicrobials chloramphenicol, streptomycin, quintozene, and propamocarb and was highly selective, allowing the recovery of T. harzianum...

  14. Optimum Concentrations of Trichoderma longibrachiatum and Cadusafos for Controlling Meloidogyne javanica on Zucchini Plants

    OpenAIRE

    Sokhandani, Zahra; Moosavi, Mohammad Reza; Basirnia, Tahereh

    2016-01-01

    A factorial experiment was established in a completely randomized design to verify the effect of different inoculum levels of an Iranian isolate of Trichoderma longibrachiatum separately and in combination with various concentrations of cadusafos against Meloidogyne javanica in the greenhouse. Zucchini seeds were soaked for 12 hr in five densities (0, 105, 106, 107, and 108 spores/ml suspension) of the fungus prior to planting in pots containing four concentrations of cadusafos (0, 0.5, 1, an...

  15. Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride

    OpenAIRE

    Goyal, Meenakshi; Kalra, K. L.; V.K. Sareen; G. Soni

    2008-01-01

    In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing l...

  16. Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

    Science.gov (United States)

    Nor, Nurafifah Mohd; Hadibarata, Tony; Zubir, Meor Mohd Fikri Ahmad; Lazim, Zainab Mat; Adnan, Liyana Amalina; Fulazzaky, Mohamad Ali

    2015-11-01

    Cresol Red belongs to the triphenylmethane (TPM) class of dyes which are potentially carcinogenic or mutagenic. However, very few studies on biodegradation of Cresol Red were investigated as compared to other type dyes such as azo and anthraquinone dye. The aim of this work is to evaluate triphenylmethane dye Cresol Red degradation by fungal strain isolated from the decayed wood in Johor Bahru, Malaysia. Detailed taxonomic studies identified the organisms as Trichoderma species and designated as strain Trichoderma harzianum M06. In this study, Cresol Red was decolorized up to 88% within 30 days under agitation condition by Trichoderma harzianum M06. Data analysis revealed that a pH value of 3 yielded a highest degradation rate among pH concentrations (73%), salinity concentrations of 100 g/L (73%), and a volume of 0.1 mL of Tween 80 (79%). Induction in the enzyme activities of manganese peroxidase, lignin peroxidase, laccase, 1,2- and 2,3-dioxygenase indicates their involvement in Cresol Red removal. Various analytical studies such as Thin-Layer Chromatography (TLC), UV-Vis spectrophotometer, and Gas chromatography mass spectrometry (GC-MS) confirmed the biotransformation of Cresol Red by the fungus. Two metabolites were identified in the treated medium: 2,4-dihydroxybenzoic acid (t R 7.3 min and m/z 355) and 2-hydroxybenzoic acid (t R 8.6 min and m/z 267). Based on these products, a probable pathway has been proposed for the degradation of Cresol Red by Trichoderma harzianum M06. PMID:26275435

  17. Filtration enrichment method for isolation of auxotrophic mutants of Trichoderma harzianum rifai

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    Cassiolato Ana Maria R.

    1999-01-01

    Full Text Available The isolation of genetic markers, like drug resistance and auxotrophy, is a laborious but important step in genetic research. The isolation of auxotrophic mutants of Trichoderma harzianum using the filtration enrichment technique was more effective than using the total isolation technique. Most of 12 auxotrophic mutants exhibited similar growth rate and higher sporulation when compared with the wild type, but only two mutants (TWS-410 and TW5-523 could grow in 500µg/L of benomyl.

  18. Efficient leaching of cellulases produced by Trichoderma harzianum in solid state fermentation

    OpenAIRE

    Roussos, Sevastianos; Raimbault, Maurice; Saucedo-Castaneda, G.; Lonsane, B.K.

    1992-01-01

    Recovery of cellulases from solid state culture of #Trichoderma harzianum$ was efficiently achieved by hydraulic pressing. Pressing of fermented solids yielded carboxymethyl-cellulase (CMCase) extraction efficiency of 71% and a ratio of leachate to fermented solids of 0.58 (v/w). Addition of water to pressed solids and second pressing improved the efficiency (95%) with simultaneous increase in the ratio to 1.16 (v/w). The overall extraction of filter paper activity was lower (85%) than that o...

  19. Eradication of Sclerotinia sclerotiorum Sclerotia from Soil Using Organic Waste Materials as Trichoderma Fungi Carriers

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    Smolińska Urszula

    2016-06-01

    Full Text Available Sclerotinia sclerotiorum (Lib. de Bary is considered as one of the most harmful soilborne pathogens, which reduces productivity of horticultural crops. Currently used chemical or biological methods for the eradication of S. sclerotiorum from a soil are not very effective. The aim of this study was to evaluate the possibility of eradication of S. sclerotiorum sclerotia from a soil using the Trichoderma isolates, which were multiplied on the organic carriers prepared from agro-industrial wastes and by-products: WsA (wheat straw + apple pomaces, WsP (wheat straw + potato pulp and T-GRAN (dry onion rind, apples and strawberry pomaces, rapeseed meal. The results showed that soil amendment with organic materials overgrown with the Trichoderma fungi had a significant reducing effect on S. sclerotiorum. Especially effective was the carrier WsA overgrown with T. virens TRS114, which completely prevented the survival of sclerotia of S. sclerotiorum regardless of the dose of application. Less effective was the WsP carrier. However, addition WsP overgrown with T. atroviride TRS40 at the 5% w/v, resulted in survival only 6.7% of sclerotia. In the greenhouse experiments with lettuce, the application of granulates T-GRAN into the soil had different impact on S. sclerotiorum depending on the conditions to the pathogen development. In conducive conditions, an addition of the organic substances without Trichoderma significantly decreased the yield of lettuce plants. A positive effect on the growth of plants was observed after the application of T-GRAN overgrown with Trichoderma.

  20. Molecular Identification Of Trichoderma Strains Collected To Develop Plant Growth-Promoting And Biocontrol Agents

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    Oskiera Michał

    2015-06-01

    Full Text Available Trichoderma strains that are beneficial to both the growth and health of plants can be used as plant growth-promoting fungi (PGPF or biological control agents (BCA in agricultural and horticultural practices. In order to select PGPF or BCA strains, their biological properties and taxonomy must be carefully studied. In this study, 104 strains of Trichoderma collected at geographically different locations in Poland for selection as PGPF or BCA were identified by DNA barcoding, based on the sequences of internal transcribed spacers 1 and 2 (ITS1 and 2 of the ribosomal RNA gene cluster and on the sequences of translation elongation factor 1 alpha (tef1, chitinase 18-5 (chi18-5, and RNA polymerase II subunit (rpb2 gene fragments. Most of the strains were classified as: T. atroviride (38%, T. harzianum (21%, T. lentiforme (9%, T. virens (9%, and T. simmonsii (6%. Single strains belonging to T. atrobrunneum, T. citrinoviride, T. crassum, T. gamsii, T. hamatum, T. spirale, T. tomentosum, and T. viridescens were identified. Three strains that are potentially pathogenic to cultivated mushrooms belonging to T. pleuroticola and T. aggressivum f. europaeum were also identified. Four strains: TRS4, TRS29, TRS33, and TRS73 were classified to Trichoderma spp. and molecular identification was inconclusive at the species level. Phylogeny analysis showed that three of these strains TRS4, TRS29, and TRS33 belong to Trichoderma species that is not yet taxonomically established and strain TRS73 belongs to the T. harzianum complex, however, the species could not be identified with certainty.

  1. Efectos del Trichoderma sp. sobre el crecimiento y desarrollo de la arveja (Pisum sativum L.

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    David Fernando Camargo-Cepeda

    2015-04-01

    Full Text Available Se estima que el cultivo de arveja en Colombia genera alrededor de 2,3 millones de jornales y unos 15.000 empleos directos; de él dependen más de 26.000 productores (1. Ante la ausencia de alternativas de producción, el agricultor ha recurrido tradicionalmente a la aplicación de productos de síntesis química, práctica que cada vez se encuentra más restringida por razones económicas y ecológicas (2; por esto, se hace necesario encontrar nuevos modelos que contribuyan a mejorar la calidad de vida de los productores. El trabajo determinó los efectos de la aplicación de Trichoderma sp. sobre el crecimiento y desarrollo de la arveja. Se realizó el aislamiento de la cepa nativa de Trichoderma sp. a partir de suelo proveniente de cultivos de arveja; luego se procedió a hacer las diluciones tanto de Trichoderma sp. nativa como de la comercial; se inocularon las plantas y se realizó la medición de las variables de crecimiento y desarrollo. La aplicación de Trichoderma sp. comercial en el cultivo de arveja mejora notablemente su crecimiento y desarrollo, influyendo en variables fisiológicas como germinación, área foliar, peso seco y fresco de la raíz, peso seco y fresco de la parte aérea, y longitud de raíz.

  2. Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease

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    Felix Carlos

    2002-01-01

    Full Text Available Abstract Background Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes has been considered as the main mechanism involved in the antagonistic process. However, although Trichoderma strains were found to impair development of Crinipellis perniciosa, the causal agent of cocoa plant witches' broom disease, no fungal strain is available for effective control of this disease. We have then undertaken a program of construction of hydrolytic enzyme-overproducing Trichoderma strains aiming improvement of the fungal antagonistic capacity. The protease of an indian Trichoderma isolate showing antagonistic activity against C. perniciosa was purified to homogeneity and characterized for its kinetic properties and action on the phytopathogen cell wall. Results A protease produced by the Trichoderma harzianum isolate 1051 was purified to homogeneity by precipitation with ammonium sulfate followed by hydrophobic chromatography. The molecular mass of this protease as determined by SDS-polyacrylamide gel electrophoresis was about 18.8 kDa. Its N-terminal amino acid sequence shares no homology with any other protease. The purified enzyme substantially affected the cell wall of the phytopathogen C. perniciosa. Western-blotting analysis showed that the enzyme was present in the culture supernatant 24 h after the Trichoderma started to grow in casein-containing liquid medium. Conclusions The capacity of the Trichoderma harzianum protease to hydrolyze the cell wall of C. perniciosa indicates that this enzyme may be actually involved in the antagonistic process between the two fungi. This fact strongly suggest that hydrolytic enzyme over-producing transgenic fungi may show superior biocontrol capacity.

  3. Screening of Trichoderma isolates for their potential of biosorption of nickel and cadmium.

    Science.gov (United States)

    Nongmaithem, Nabakishor; Roy, Ayon; Bhattacharya, Prateek Madhab

    2016-01-01

    Fourteen Trichoderma isolates were evaluated for their tolerance to two heavy metals, nickel and cadmium. Three isolates, MT-4, UBT-18, and IBT-I, showed high levels of nickel tolerance, whereas MT-4, UBT-18, and IBT-II showed better tolerance of cadmium than the other isolates. Under nickel stress, biomass production increased up to a Ni concentration of 60ppm in all strains but then decreased as the concentrations of nickel were further increased. Among the nickel-tolerant isolates, UBT-18 produced significantly higher biomass upon exposure to nickel (up to 150ppm); however, the minimum concentration of nickel required to inhibit 50% of growth (MIC50) was highest in IBT-I. Among the cadmium-tolerant isolates, IBT-II showed both maximum biomass production and a maximum MIC50 value in cadmium stress. As the biomass of the Trichoderma isolates increased, a higher percentage of nickel removal was observed up to a concentration of 40ppm, followed by an increase in residual nickel and a decrease in biomass production at higher nickel concentrations in the medium. The increase in cadmium concentrations resulted in a decrease in biomass production and positively correlated with an increase in residual cadmium in the culture broth. Nickel and cadmium stress also influenced the sensitivity of the Trichoderma isolates to soil fungistasis. Isolates IBT-I and UBT-18 were most tolerant to fungistasis under nickel and cadmium stress, respectively. PMID:26991295

  4. Biological Action of Plant Extracts on a Fungal Plant Biostimulant Strain of Trichoderma Viride

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    Şesan Tatiana Eugenia

    2015-11-01

    Full Text Available The antifungal activity of nine plant extracts manufactured by Hofigal Import Export S.A. Romania against the biocontrol fungal agent Trichoderma viride Pers. (isolate Tv 82 was assessed in vitro for the first time in Romania. In general, the development (mycelial growth and sporulation was not inhibited by the six plant extracts (Satureja hortensis, Achillea millefolium, Allium sativum, Mentha sp., Hyssopus officinalis, Artemisia dracunculus 'Sativa', with three exceptions (Rosmarinus officinalis, Valeriana officinalis, Tagetes patula, applied in a concentration of 20%. Among these, the extract of Tagetes patula has inhibited the Tv 82 development, applied as lower concentrations (10% and 5%, efficacy being 54.3% and 50%, respectively. In addition, the tested plant extracts of Satureja hortensis, Achillea millefolium, Mentha sp. proved stimulative effect on Tv 82 development. This approach add to the early studies on the selectivity of Trichoderma spp. to chemicals used in plant protection, new data referring to the use of antagonistic fungi, like Trichoderma spp., as a protective mean against phytopathogens. Also, these data sustain the possibility of applying plant extracts as an alternative in plant protection or to apply together chemical (pesticides and biological means (plant extracts especially to protect ecological crops, as vegetables, medicinal plants a.o.

  5. Trichoderma spp. alleviate phytotoxicity in lettuce plants (Lactuca sativa L.) irrigated with arsenic-contaminated water.

    Science.gov (United States)

    Caporale, Antonio G; Sommella, Alessia; Lorito, Matteo; Lombardi, Nadia; Azam, Shah M G G; Pigna, Massimo; Ruocco, Michelina

    2014-09-15

    The influence of two strains of Trichoderma (T. harzianum strain T22 and T. atroviride strain P1) on the growth of lettuce plants (Lactuca sativa L.) irrigated with As-contaminated water, and their effect on the uptake and accumulation of the contaminant in the plant roots and leaves, were studied. Accumulation of this non-essential element occurred mainly into the root system and reduced both biomass development and net photosynthesis rate (while altering the plant P status). Plant growth-promoting fungi (PGPF) of both Trichoderma species alleviated, at least in part, the phytotoxicity of As, essentially by decreasing its accumulation in the tissues and enhancing plant growth, P status and net photosynthesis rate. Our results indicate that inoculation of lettuce with selected Trichoderma strains may be helpful, beside the classical biocontrol application, in alleviating abiotic stresses such as that caused by irrigation with As-contaminated water, and in reducing the concentration of this metalloid in the edible part of the plant.

  6. Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains.

    Science.gov (United States)

    Chaverri, Priscila; Branco-Rocha, Fabiano; Jaklitsch, Walter; Gazis, Romina; Degenkolb, Thomas; Samuels, Gary J

    2015-01-01

    Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture 'T. harzianum T22' was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex.

  7. The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens

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    Rashmi Singh

    2016-03-01

    Full Text Available Abstract The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2% and Trichoderma atroviride (40.3% were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69 U/mL and β-1,3-glucanase activity (61.9 U/mL. Th-m1 also showed high competitive saprophytic ability (CSA among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%.

  8. Diversity of Trichoderma spp. causing Pleurotus green mould diseases in Central Europe.

    Science.gov (United States)

    Błaszczyk, Lidia; Siwulski, Marek; Sobieralski, Krzysztof; Frużyńska-Jóźwiak, Dorota

    2013-07-01

    The present study includes the molecular characteristics of Trichoderma pleurotum and Trichoderma pleuroticola isolates collected from green moulded cereal straw substrates at 47 oyster mushroom farms in Poland. The screening of the 80 Trichoderma isolates was performed by morphological observation and by using the multiplex PCR assay. This approach enabled specific detection of 47 strains of T. pleurotum and 2 strains of T. pleuroticola. Initial identifications were confirmed by sequencing the fragment of internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of the rRNA gene cluster and the fragment including the fourth and fifth introns and the last long exon of the translation-elongation factor 1-alpha (tef1) gene. ITS and tef1 sequence information was also used to establish the intra- and interspecies relationship of T. pleurotum and T. pleuroticola originating from the oyster mushroom farms in Poland and from other countries. Comparative analysis of the ITS sequences showed that all T. pleurotum isolates from Poland represent one haplotype, identical to that of T. pleurotum strains from Hungary and Romania. Sequence analysis of the tef1 locus revealed two haplotypes ("T" and "N") of Polish T. pleurotum isolates. The "T" type isolates of T. pleurotum were identical to those of strains from Hungary and Romania. The "N" type isolates possessed a unique tef1 allele. Detailed analysis of the ITS and tef1 sequences of two T. pleuroticola isolates showed their identicalness to Italian strain C.P.K. 1540.

  9. Biological control of Rhizoctonia solani on potato by using indigenous Trichoderma spp.

    Science.gov (United States)

    Durak, Emre Demirer

    2016-04-01

    At this study, it was aimed to determine the effect of Trichoderma isolates that was isolated from the soil samples taken from the different regions on black scurf and stem canker disease caused by Rhizoctonia solani Kühn that has been one of the biggest problems of the potato cultivation. At the end of the soil isolations, totally 81 Trichoderma isolates were obtained and their species were identified. Of these isolates, T. harzianum (42%), T. virens (31%), T. asperellum (15%) and T. viride (12%). All of the isolates were tested in vitro for their antagonistic activity against the R. solani isolate. The isolates that show high inhibition rate was selected and tested against R. solani in vitro. Potato plants were grown in a greenhouse for about 10 weeks. Then the plants were evaluated according to the scale, plant height, shoot fresh and dry weights, root fresh and dry weights were noted. The experiment was conducted two times in three replications. At the in vitro tests, generally, it was determined that Trichoderma isolates have inhibited to R. solani and in vivo, they were reduced the effects of the disease and they were raised the development of the plant. In particular, it was determined that some isolates of the T. harzianum and T. virens have reduced the severity of the disease. It was determined that both in vitro and in vivo isolates have shown different efficiency against R. solani.

  10. Fungicide sensitivity of Trichoderma spp. from Agaricus bisporus farms in Serbia.

    Science.gov (United States)

    Kosanović, Dejana; Potočnik, Ivana; Vukojević, Jelena; Stajić, Mirjana; Rekanović, Emil; Stepanović, Miloš; Todorović, Biljana

    2015-01-01

    Trichoderma species, the causal agents of green mould disease, induce great losses in Agaricus bisporus farms. Fungicides are widely used to control mushroom diseases although green mould control is encumbered with difficulties. The aims of this study were, therefore, to research in vitro toxicity of several commercial fungicides to Trichoderma isolates originating from Serbian and Bosnia-Herzegovina farms, and to evaluate the effects of pH and light on their growth. The majority of isolates demonstrated optimal growth at pH 5.0, and the rest at pH 6.0. A few isolates also grew well at pH 7. The weakest mycelial growth was noted at pH 8.0-9.0. Generally, light had an inhibitory effect on the growth of tested isolates. The isolates showed the highest susceptibility to chlorothalonil and carbendazim (ED50 less than 1 mg L(-1)), and were less sensitive to iprodione (ED50 ranged 0.84-6.72 mg L(-1)), weakly resistant to thiophanate-methyl (ED50 = 3.75-24.13 mg L(-1)), and resistant to trifloxystrobin (ED50 = 10.25-178.23 mg L(-1)). Considering the toxicity of fungicides to A. bisporus, carbendazim showed the best selective toxicity (0.02), iprodione and chlorothalonil moderate (0.16), and thiophanate-methyl the lowest (1.24), while trifloxystrobin toxicity to A. bisporus was not tested because of its inefficiency against Trichoderma isolates.

  11. Involvement of Trichoderma asperellum strain T6 in regulating iron acquisition in plants.

    Science.gov (United States)

    Zhao, Lei; Wang, Fei; Zhang, Yaqing; Zhang, Jiaojiao

    2014-07-01

    Iron (Fe) deficiency is a major plant nutritional disorder in many parts of the world, particularly in areas with saline soils. Among the numerous root-associated microbes that are beneficial for plant nutrient uptake, Trichoderma spp. are the most effective rhizosphere fungi for enhancing plant growth and plant resistance to biotic and abiotic stresses. To investigate the potential mechanisms of action of Trichoderma on insoluble Fe in the soil, which is difficult for plants to absorb and utilize, a high siderophore-producing strain of Trichoderma T6, was isolated from the rhizosphere of cucumber plants. The strain was identified as T. asperellum based on the morphological features and molecular phylogenetic analyses. Applying strain T6 to sterile soil could increase soil levels of Fe(2+) and siderophores, as well as increase Fe(2+) and Fe(3+)-chelate reductase (FCR) activity in cucumber tissues. Purified siderophore eluent (PSE) increased plant growth, thus confirming its role in plant growth promotion. Moreover, extracellular Fe(3+) reducing activity and three kinds of organic acids were detected in the culture filtrate of strain T6. These results indicate that strain T6 influences plant Fe absorption in several ways. Siderophore-based Fe chelation is effective in providing Fe to plants, organic acids, and Fe(3+) reducing enzymes may participate in the solubilization and reduction of insoluble Fe(3+) to Fe(2+).

  12. Antibiosis of Trichoderma spp strains native to northeastern Mexico against the pathogenic fungus Macrophomina phaseolina

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    José Luis Hernández Mendoza

    2015-12-01

    Full Text Available Abstract Sampling of agricultural soils from the Mexican northeastern region was performed to detect Trichoderma spp., genetically characterize it, and assess its potential use as a biologic control agent against Macrophomina phaseolina. M. phaseolina is a phytopathogen that attacks over 500 species of cultivated plants and causes heavy losses in the regional sorghum crop. Sampling was performed immediately after sorghum or corn harvest in an area that was approximately 170 km from the Mexico-USA border. Sixteen isolates were obtained in total. Using colony morphology and sequencing the internal transcribed spacers (ITS 1 and 4 of 18S rDNA, 14 strains were identified as Trichoderma harzianum, T. koningiopsis and T. virens. Subsequently, their antagonistic activity against M. phaseolina was evaluated in vitro, and 11 isolates showed antagonism by competition and stopped M. phaseolina growth. In 4 of these isolates, the antibiosis phenomenon was observed through the formation of an intermediate band without growth between colonies. One strain, HTE808, was identified as Trichoderma koningiopsis and grew rapidly; when it came into contact with the M. phaseolina colony, it continued to grow and sporulated until it covered the entire petri dish. Microscopic examination confirmed that it has a high level of hyperparasitism and is thus considered to have high potential for use in the control of this phytopathogen.

  13. Optimum Concentrations of Trichoderma longibrachiatum and Cadusafos for Controlling Meloidogyne javanica on Zucchini Plants.

    Science.gov (United States)

    Sokhandani, Zahra; Moosavi, Mohammad Reza; Basirnia, Tahereh

    2016-03-01

    A factorial experiment was established in a completely randomized design to verify the effect of different inoculum levels of an Iranian isolate of Trichoderma longibrachiatum separately and in combination with various concentrations of cadusafos against Meloidogyne javanica in the greenhouse. Zucchini seeds were soaked for 12 hr in five densities (0, 10(5), 10(6), 10(7), and 10(8) spores/ml suspension) of the fungus prior to planting in pots containing four concentrations of cadusafos (0, 0.5, 1, and 2 mg a.i./kg soil). The data were analyzed using a custom response surface regression model and the response surface curve and contour plots were drawn. Reliability of the model was examined by comparing the result of new experimental treatments with the predicted results. The optimal levels of these two variables also were calculated. The interactive effects of concentrations of Trichoderma and cadusafos were insignificant for several responses such as the total number of eggs per gram soil, the number of intact eggs per gram soil, nematode reproduction factor, and control percent. Closeness of experimental mean values with the expected values proved the validity of the model. The optimal levels of the cadusafos concentration and Trichoderma concentration that caused the best plant growth and lowest nematode reproduction were 1.7 mg a.i./kg soil and 10(8) conidia/ml suspension, respectively. PMID:27168653

  14. The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens.

    Science.gov (United States)

    Rashmi, Singh; Maurya, Sudarshan; Upadhyay, Ram Sanmukh

    2016-01-01

    The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2%) and Trichoderma atroviride (40.3%) were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69U/mL) and β-1,3-glucanase activity (61.9U/mL). Th-m1 also showed high competitive saprophytic ability (CSA) among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%).

  15. Endocarditis por Trichoderma longibrachiatum en paciente con nutrición parenteral domiciliaria

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    Laura I. Rodríguez Peralta

    2013-06-01

    Full Text Available La modalidad domiciliaria de la nutrición parenteral (NPD mejora la calidad de vida de los pacientes, pero tiene complicaciones como infecciones asociadas a catéter (IAC y complicaciones mecánicas. Presentamos el caso de un paciente con NPD por intestino corto que desarrolla una endocarditis sobre catéter con matices especiales: asentar sobre un catéter abandonado en aurícula derecha y tratarse de una infección con participación de un microorganismo no descrito hasta el momento en esta patología, Trichoderma longibrachiatum. El catéter se extrajo mediante cirugía convencional. En la pieza quirúrgica se aislaron Staphylococcus epidermidis, Ochrobactrum anthropi y Trichoderma longibrachiatum. Combinando el tratamiento antibiótico y la eliminación del foco infeccioso se consiguió la recuperación completa. Ochrobactrum anthropi y Trichoderma longibrachiatum son microorganismos poco habituales, pero que cada vez adquieren mayor relevancia. Aunque no existe acuerdo en el manejo de los catéteres intravasculares "abandonados", es recomendable el seguimiento y eliminarlos en caso de complicación.

  16. Isolation and identification of Trichoderma harzianum from groundwater: An effective biosorbent for defluoridation of groundwater.

    Science.gov (United States)

    Koshle, Shalini; Mahesh, S; Swamy, S Nanjunda

    2016-01-01

    The ability of non-viable form of Trichoderma harzianum, isolated from fluoride rich groundwater, was investigated as biosorbent for defluoridation of groundwater. Biosorption experiments were carried out at laboratory scale for removal of fluoride from groundwater. Significant effect of operational parameters on fluoride biosorption using Trichoderma harzianum as biosorbent was evaluated by varying operational parameters such as: initial fluoride concentration (2-8 mgl(-1)), biosorbent dose (0.4-1.6g/100ml), groundwater pH (6-10), temperature (30-50 degrees C) and biosorption time (30-120 min). The fluoride adsorption isotherms were modeled by Langmuir and Freundlich isotherms. Our result showed that fluoride biosorption, significantly increased with increase in groundwater pH, biosorbent dose, temperature and biosorption time, whereas increase in initial fluoride concentration reduced fluoride removal. The fluoride biosorption was rapid and maximum fluoride uptake was attained with 1.6g 100ml(-1) biosorbent within 60 min. Optimal pH 10 and temperature 50 degrees C gave maximum defluoridation efficiency. Freundlich isotherm fits well for defluoridation of groundwater using Trichoderma harzianum as biosorbent which indicated that biosorbent surface sites were heterogeneous in nature and fitted into heterogeneous site binding model. PMID:26930871

  17. Biocontrol mechanisms of Trichoderma harzianum against soybean charcoal rot caused by Macrophomina phaseolina

    Directory of Open Access Journals (Sweden)

    Khaledi Nima

    2016-01-01

    Full Text Available Throughout the world, charcoal rot, caused by Macrophomina phaseolina, is one of the most destructive and widespread diseases of crop plants such as soybean. In this study, the biological control capability of 11 Trichoderma spp. isolates against M. phaseolina was investigated using screening tests. Among all the tested Trichoderma spp. isolates, inhibition varied from 20.22 to 58.67% in dual culture tests. Dual culture, volatile and non-volatile tests revealed that two isolates of Trichoderma harzianum (including the isolates T7 and T14 best inhibited the growth of M. phaseolina in vitro. Therefore, these isolates were selected for biocontrol of M. phaseolina in vivo. The results of greenhouse experiments revealed that disease severity in the seed treatment with T. harzianum isolates was significantly lower than that of the soil treatment. In most of the cases, though, soil treatment with T. harzianum resulted in higher plant growth parameters, such as root and shoot weight. The effects of T. harzianum isolates on the activity of peroxidase enzyme and phenolic contents of the soybean root in the presence and absence of M. phaseolina were determined in greenhouse conditions. Our results suggested that a part of the inhibitory effect of T. harzianum isolates on soybean charcoal rot might be related to the indirect influence on M. phaseolina. Plant defense responses were activated as an elicitor in addition to the direct effect on the pathogen growth.

  18. The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens.

    Science.gov (United States)

    Rashmi, Singh; Maurya, Sudarshan; Upadhyay, Ram Sanmukh

    2016-01-01

    The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2%) and Trichoderma atroviride (40.3%) were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69U/mL) and β-1,3-glucanase activity (61.9U/mL). Th-m1 also showed high competitive saprophytic ability (CSA) among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%). PMID:26887221

  19. Antibiosis of Trichoderma spp strains native to northeastern Mexico against the pathogenic fungus Macrophomina phaseolina.

    Science.gov (United States)

    Mendoza, José Luis Hernández; Pérez, María Isabel Sánchez; Prieto, Juan Manuel González; Velásquez, Jesús DiCarlo Quiroz; Olivares, Jesús Gerardo García; Langarica, Homar Rene Gill

    2015-01-01

    Sampling of agricultural soils from the Mexican northeastern region was performed to detect Trichoderma spp., genetically characterize it, and assess its potential use as a biologic control agent against Macrophomina phaseolina. M. phaseolina is a phytopathogen that attacks over 500 species of cultivated plants and causes heavy losses in the regional sorghum crop. Sampling was performed immediately after sorghum or corn harvest in an area that was approximately 170 km from the Mexico-USA border. Sixteen isolates were obtained in total. Using colony morphology and sequencing the internal transcribed spacers (ITS) 1 and 4 of 18S rDNA, 14 strains were identified as Trichoderma harzianum, T. koningiopsis and T. virens. Subsequently, their antagonistic activity against M. phaseolina was evaluated in vitro, and 11 isolates showed antagonism by competition and stopped M. phaseolina growth. In 4 of these isolates, the antibiosis phenomenon was observed through the formation of an intermediate band without growth between colonies. One strain, HTE808, was identified as Trichoderma koningiopsis and grew rapidly; when it came into contact with the M. phaseolina colony, it continued to grow and sporulated until it covered the entire petri dish. Microscopic examination confirmed that it has a high level of hyperparasitism and is thus considered to have high potential for use in the control of this phytopathogen. PMID:26691467

  20. Identification and Characterization of Trichoderma Species Damaging Shiitake Mushroom Bed-Logs Infested by Camptomyia Pest.

    Science.gov (United States)

    Kim, Jun Young; Kwon, Hyuk Woo; Yun, Yeo Hong; Kim, Seong Hwan

    2016-05-28

    The shiitake mushroom industry has suffered from Camptomyia (gall midges) pest, which feeds on the mycelium of shiitake mushroom during its cultivation. It has been postulated that fungal damage of shiitake bed-logs is associated with infestation by the insect pest, but this is not well understood. To understand the fungal damage associated with Camptomyia pest, various Trichoderma species were isolated, identified, and characterized. In addition to two previously known Trichoderma species, T. citrinoviride and T. deliquescens, two other Trichoderma species, T. harzianum and T. atroviride, were newly identified from the pestinfested bed-log samples obtained at three mushroom farms in Cheonan, Korea. Among these four species, T. harzianum was the most evident. The results of a chromogenic media-based assay for extracellular enzymes showed that these four species have the ability to produce amylase, carboxyl-methyl cellulase, avicelase, pectinase, and β-glucosidase, thus indicating that they can degrade wood components. A dual culture assay on PDA indicated that T. harzianum, T. atroviride, and T. citrinoviride were antagonistic against the mycelial growth of a shiitake strain (Lentinula edodes). Inoculation tests on shiitake bed-logs revealed that all four species were able to damage the wood of bed-logs. Our results provide evidence that the four green mold species are the causal agents involved in fungal damage of shiitake bed-logs infested by Camptomyia pest. PMID:26930351

  1. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

    Directory of Open Access Journals (Sweden)

    M Irfan

    2012-03-01

    Full Text Available Summary: In the present study xylanase enzyme was produced from Trichoderma viride in solid state fermentation using sugarcane bagasse as a substrate. The whole fermentation process was carried out in 250ml Erlenmeyer flask at 30oC for seven days of fermentation period. The enzyme was partially purified by ammonium sulphate (60% fractionation followed by dialysis. The partially purified enzyme was further characterized showing optimum pH and temperature of 5.0 and 50oC respectively. Metal profile of the enzyme showed that it was stimulated by FeSO4 (134%, CaCl2 (129%, BaCl2 (105%, MgSO4 (113%, MnCl2 (102% or AgCl (107% and it was strongly inhibited by EDTA (26% or HgSO4 (32%. Industrial Relevance: In the present study, xylanase enzyme was produced and characterized from Trichoderma viride in solid state fermentation using cheap substrate. This enzyme is very helpful in industrial sector especially in pulp and paper industry, food industry and also in bioethanol production. Pilot scale production of this enzyme in industries can reduce the import cost of the enzyme and make the whole process cost effective. Keywords: Partial purification; Characterization; Xylanase; Trichoderma viride; SSF

  2. Characterisation of a Trichoderma hamatum monooxygenase gene involved in antagonistic activity against fungal plant pathogens.

    Science.gov (United States)

    Carpenter, Margaret A; Ridgway, Hayley J; Stringer, Alison M; Hay, Amanda J; Stewart, Alison

    2008-04-01

    A monooxygenase gene was isolated from a biocontrol strain of Trichoderma hamatum and its role in biocontrol was investigated. The gene had homologues in other fungal genomes, but was not closely related to any fully characterised gene. The T. hamatum monooxygenase gene was expressed specifically in response to the plant pathogens Sclerotinia sclerotiorum, Sclerotinia minor and Sclerotium cepivorum, but not in response to Botrytis cinerea or T. hamatum. Expression of the gene did not occur until contact had been made between the two fungal species. Homologues in T. atroviride and T. virens showed similar expression patterns. Expression of the gene in response to S. sclerotiorum was influenced by pH, with a peak of expression at pH 4, and was subject to nitrogen catabolite repression. Disruption of the monooxygenase gene did not affect the growth or morphology of T. hamatum, but caused a decrease in its ability to inhibit the growth and sclerotial production of S. sclerotiorum. The monooxygenase gene had a role in the antagonistic activity of Trichoderma species against specific fungal plant pathogens and is therefore a potentially important factor in biocontrol by Trichoderma species. PMID:18231791

  3. 卓越的画质,专为发烧级玩家而设计 Schneider(施耐德)CINE-DIGITAR ANAMORPHIC 1.33×XL

    Institute of Scientific and Technical Information of China (English)

    王韦

    2010-01-01

    为了满足想消除投影画面上那两条粗大的黑边、追求更广阔视野又不愿意牺牲接近1/3投影机像素的挑剔玩家,2.35:1的变形镜头出现了。但是许多玩家发现使用了变形镜头之后,对画面的亮度、锐度、清晰度的损失实在是非常严重,于是进一步催生了高价高品质的顶级变形镜头的诞生。其中由德国著名光学大厂Schneider Optics(施耐德光学)所推出的CINE-DIGITARANAMORPHIC 1.33×系列就是高素质变形镜头的代表,绝佳的画面畸变的控制,优秀的色彩还原能力,都是普通变形镜头难以望其项背的。而来到编辑部视听室的则是最高端的CINE-DIGITAR ANAMORPHIC 1.33×XL。

  4. Chinese new record Trichoderma capillare and its functional assessment%中国新记录种毛细木霉Trichoderma capillare及功能评价

    Institute of Scientific and Technical Information of China (English)

    陈凯; 李纪顺; 张广志; 王贻莲; 赵晓燕; 吴晓青; 扈进冬; 杨合同

    2016-01-01

    木霉菌株TW22166分离自湖南省洞庭湖滩涂湿地,该菌株分生孢子梗具有长而发达的主轴,二次分枝丰富,不规则,一般单生,分生孢子梗向顶方向着生单个瓶梗.利用ITS、tef1及rpb2序列分析,发现该菌株同Trichoderma capiUare同源性达到99%以上;在以rpb2构建的系统发育进化树中,TW22166与Trichoderma capillare strain GJS 06-66[JN175530.1]位于同一分支.该菌株在PDA平板上对9种植物病原真菌均有抑制作用,其中对Fusarium.oxysporum的抑制率为100.0%.菌株具有蛋白酶、几丁质酶及纤维素酶活性,其中蛋白酶活性较高,透明圈宽度达16.0 mm.该菌株鉴定为毛细木霉Trichoderma capiUare,是木霉属中国新记录种,也是一株较有潜力的生物防治菌株.

  5. Biocontrol potential of salinity tolerant mutants of Trichoderma harzianum against Fusarium oxysporum Potencial de biocontrole de mutantes sal-tolerantes de Trichoderma harzianum contra Fusarium oxysporum

    Directory of Open Access Journals (Sweden)

    Hassan Abdel-Latif A. Mohamed

    2006-06-01

    Full Text Available Exposing a wild-type culture of Trichoderma harzianum to gamma irradiation induced two stable salt-tolerant mutants (Th50M6 and Th50M11. Under saline conditions, both mutants greatly surpassed their wild type strain in growth rate, sporulation and biological proficiency against Fusarium oxysporum, the causal agent of tomato wilt disease. Tolerant T. harzianum mutants detained a capability to grow and convinced sporulation in growth media containing up to 69 mM NaCl. In comparison with their parent strain, characterization of both mutants confirmed that they have reinforced contents of proline and hydroxyproline, relatively higher sodium content compared to potassium, calcium or magnesium contents, higher level of total phenols. Electrophoretic analysis of total soluble proteins in the salt tolerance mutant Th50M6 showed different bands accumulated in response to 69 mM NaCl. Data also showed that mutants produce certain active metabolites, such as chitinases, cellulases, beta-galactosidases, as well as, some antibiotics i.e., trichodermin, gliotoxin and gliovirin. Trichoderma mutants significantly reduced wilt disease incidence and improved yield and mineral contents of tomato plants under both saline and non-saline soil conditions, as well as, under infested and natural conditions. T. harzianum mutants were also more efficient in dropping the F. oxysporum growth in rhizosphere compared to the wild type strain. Population density of both mutants in rhizosphere far exceeded that of T. harzianum wild type strain.A exposição de uma cepa selvagem de Trichoderma harzianum à irradiação gama induziu dois mutantes tolerantes a sal (Th50M6 e Th50M11. Em condições salinas, os dois mutantes foram muito superiores à cepa selvagem em relação à velocidade de multiplicação, esporulação e eficiência contra Fusarium oxysporum, o agente causador da doença wilt do tomate. Os mutantes tolerantes foram capazes de multiplicação e esporulação em

  6. Structural and mechanistic analysis of engineered trichodiene synthase enzymes from Trichoderma harzianum: towards higher catalytic activities empowering sustainable agriculture.

    Science.gov (United States)

    Kumari, Indu; Chaudhary, Nitika; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

    2016-06-01

    Trichoderma spp. are well-known bioagents for the plant growth promotion and pathogen suppression. The beneficial activities of the fungus Trichoderma spp. are attributed to their ability to produce and secrete certain secondary metabolites such as trichodermin that belongs to trichothecene family of molecules. The initial steps of trichodermin biosynthetic pathway in Trichoderma are similar to the trichothecenes from Fusarium sporotrichioides. Trichodiene synthase (TS) encoded by tri5 gene in Trichoderma catalyses the conversion of farnesyl pyrophosphate to trichodiene as reported earlier. In this study, we have carried out a comprehensive comparative sequence and structural analysis of the TS, which revealed the conserved residues involved in catalytic activity of the protein. In silico, modelled tertiary structure of TS protein showed stable structural behaviour during simulations. Two single-substitution mutants, i.e. D109E, D248Y and one double-substitution mutant (D109E and D248Y) of TS with potentially higher activities are screened out. The mutant proteins showed more stability than the wild type, an increased number of electrostatic interactions and better binding energies with the ligand, which further elucidates the amino acid residues involved in the reaction mechanism. These results will lead to devise strategies for higher TS activity to ultimately enhance the trichodermin production by Trichoderma spp. for its better exploitation in the sustainable agricultural practices.

  7. Structural and mechanistic analysis of engineered trichodiene synthase enzymes from Trichoderma harzianum: towards higher catalytic activities empowering sustainable agriculture.

    Science.gov (United States)

    Kumari, Indu; Chaudhary, Nitika; Sandhu, Padmani; Ahmed, Mushtaq; Akhter, Yusuf

    2016-06-01

    Trichoderma spp. are well-known bioagents for the plant growth promotion and pathogen suppression. The beneficial activities of the fungus Trichoderma spp. are attributed to their ability to produce and secrete certain secondary metabolites such as trichodermin that belongs to trichothecene family of molecules. The initial steps of trichodermin biosynthetic pathway in Trichoderma are similar to the trichothecenes from Fusarium sporotrichioides. Trichodiene synthase (TS) encoded by tri5 gene in Trichoderma catalyses the conversion of farnesyl pyrophosphate to trichodiene as reported earlier. In this study, we have carried out a comprehensive comparative sequence and structural analysis of the TS, which revealed the conserved residues involved in catalytic activity of the protein. In silico, modelled tertiary structure of TS protein showed stable structural behaviour during simulations. Two single-substitution mutants, i.e. D109E, D248Y and one double-substitution mutant (D109E and D248Y) of TS with potentially higher activities are screened out. The mutant proteins showed more stability than the wild type, an increased number of electrostatic interactions and better binding energies with the ligand, which further elucidates the amino acid residues involved in the reaction mechanism. These results will lead to devise strategies for higher TS activity to ultimately enhance the trichodermin production by Trichoderma spp. for its better exploitation in the sustainable agricultural practices. PMID:26207800

  8. Adaptation of Trichoderma Species to Pesticide Confidor and Evaluation of their Growth Ability in the Media Containing Confidor

    Directory of Open Access Journals (Sweden)

    Farnaz Ershadfath

    2015-12-01

    Full Text Available Introduction: Contamination caused by pesticides is considered as one of the environmental problems. Bioremediation is exploiting the ability of microorganisms to remove pollutants. Trichoderma species are free-living fungi that exist naturally in the environment. These fungi have the ability to uptake some contaminants biologically. The aim of this study is to evaluate the effect of Confidor, as an environmental contaminant, on the growth ability of Trichoderma sp. as a contaminant absorber. Materials and methods: Five species of Trichoderma fungi were cultured in PDA media. Then the fungi were adapted with 3 different concentrations of Confidor gradually (5, 10 and 20 mg/l. The diameter of the fungal colonies growing in different concentrations of the toxin, were measured after 24 hr and were compared with the control samples (medium without toxin. Results: Results showed that in all species of fungi the colony diameters increased significantly with increasing toxin concentrations. The largest colony diameter was related to T.tomentosum, T.asperellum and T.harzianum (88.88, 87.5 and 86.95%, respectively at the concentration of 20 mg of toxic. Also, in all studied fungal species, in the medium containing 20 (mg/ l of toxic, the aerial hyphae expanded much thicker and faster than other concentrations. Discussion and conclusion: The results indicate a significant increase in the growth ability of Trichoderma strains with increasing Confidor concentration. Therefore it could be concluded that Trichoderma fungi have a high potentiality for biodegradation of Confidor.

  9. 平菇代料栽培中污染木霉菌的鉴定%Identification of Trichoderma Species from Contaminated Substrate During Cultivation of Pleurotus ostreatus

    Institute of Scientific and Technical Information of China (English)

    高苇; 李宝聚; 孙军德; 石延霞

    2007-01-01

    从北京平菇(Pleurotus ostreatus)产区收集的木霉病害样本,通过分离纯化及显微观察,确定了5种引起袋栽平菇发病的木霉菌,分别是绿色木霉(Trichoderma viride)、康氏木霉(Trichoderma koningii)、拟康氏木霉(Trichoderma pseudokoningii)、哈茨木霉(Trichoderma harzianum)和桔绿木霉(Trichoderma citrinoviride),并发现了平菇栽培中的优势污染种--绿色木霉和康氏木霉.

  10. EFEITO ANTAGÔNICO DE Trichoderma sp. NO DESENVOLVIMENTO DE Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok ANTAGONISTIC EFFECT OF Trichoderma sp. ON THE DEVELOPMENT OF Beauveria bassiana (Bals. Vuill. AND Metarhizium anisopliae (Metsch. SOROK

    Directory of Open Access Journals (Sweden)

    Alcides Moino Jr.

    1999-01-01

    Full Text Available Este trabalho teve por objetivo avaliar o efeito de Trichoderma sp. no desenvolvimento de Beauveria bassiana e Metarhizium anisopliae. Trichoderma sp., B. bassiana (isolado 634 e M. anisopliae (isolado E-9 foram inoculados em meio BDA, com intervalos de 0, 48, 120 e 168 horas entre a inoculação de Trichoderma sp. e dos entomopatógenos. Avaliou-se o crescimento radial das colônias nos períodos de 48 e 120 horas após a inoculação de Trichoderma sp., sendo que este afetou o desenvolvimento dos entomopatógenos quando inoculado simultaneamente ou após 48 horas. B. bassiana e M. anisopliae desenvolveram-se normalmente quando inoculados 168 horas antes de Trichoderma sp.. Também foi avaliado o efeito de um extrato de Trichoderma sp. sobre os entomopatógenos, com a adição de 0,1; 0,5; 1,0 e 5,0 ml de extrato/100,0 ml de meio, onde foram inoculados os entomopatógenos. Foram medidos os diâmetros de colônias e o número de conídios produzidos por B. bassiana e M. anisopliae na presença do extrato. A concentração de 5,0 ml de extrato/100,0 ml de meio alterou o crescimento e a conidiogênese de B. bassiana. O fungo M. anisopliae foi afetado a partir da adição de 1,0 ml de extrato/100,0 ml de meio.The objective of this work was to evaluate the effect of Trichoderma sp. on the development of Beauveria bassiana and Metarhizium anisopliae. The fungus Trichoderma sp. was inoculated on PDA culture medium, 0, 48, 120 and 168 hours after inoculation of the same plates with either B. bassiana (isolate 634 or M. anisopliae (isolate E-9. The radial growth of fungal colonies was measured 48 and 120 hours after Trichoderma sp. inoculation. Trichoderma sp. affected the development of both entomopathogenic fungi when inoculated simultaneously or 48 hours later. B. bassiana and M. anisopliae had normal development when inoculated 168 hours before Trichoderma sp. The effect of a toxic extract from Trichoderma sp. on the entomopathogenic fungi was also

  11. Peptaibol, Secondary‐Metabolite, and Hydrophobin Pattern of Commercial Biocontrol Agents Formulated with Species of the Trichoderma harzianum Complex

    DEFF Research Database (Denmark)

    Degenkolb, Thomas; Nielsen, Kristian Fog; Dieckmann, Ralf;

    2015-01-01

    The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics...... in plant‐protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna®, Trichosan......®, Vitalin®, Promot® WP, and TrichoMax®, formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well‐established, HPLC/MS‐based peptaibiomics approach, it could unequivocally be demonstrated that all...

  12. Eficiencia biológica de cepas nativas de Trichoderma spp., en el control de Sclerotium rolfsii Sacc., en cacahuate

    OpenAIRE

    A. C. Michel-Aceves; M. A. Otero-Sánchez; R. Ariza-Flores; A. Barrios-Ayala; N. Alarcón-Cruz

    2013-01-01

    En el control de Sclerotium rolfsii Sacc., se evaluó el efecto antagónico de aislados nativos de Trichoderma spp., bajo condiciones in vitro y de invernadero, en la zona Norte del estado de Guerrero. Se obtuvieron doce aislados nativos de Trichoderma spp. (dos de Santa Teresa y diez de Tlaxmalac, Gro.), de los cuales se seleccionaron seis (Tcn-4, Tcn-5, Tcn-6, Tcn-7, Tcn-8 y Tcn-11) mediante el método del papel celofán. En condiciones in vitro, el porcentaje de inhibición varió desde 10 a 94....

  13. KINETICS AND EQUILIBRIUM PARAMETERS OF BIOSORPTION AND BIOACCUMULATION OF LEAD IONS FROM AQUEOUS SOLUTIONS BY TRICHODERMA LONGIBRACHIATUM

    Directory of Open Access Journals (Sweden)

    Enitan S. Balogun

    2012-04-01

    Full Text Available Biosorption and bioaccumulation of Lead ions (Pb(II by Trichoderma longibrachiatum were investigated in a batch system. The effects of some important parameters such as pH, initial metal concentration, temperature and inoculum concerntration on biosorption capacity were also studied. The maximum biosorption capacity of Trichoderma longibrachiatum was at 25 ppm of lead, showed 100 % removal at pH 7 and 25 oC after fifteen days. Biosorption equilibrium was established in 150 minutes. The process fitted well into pseudo second order kinetic model and was best explained by Langmuir isotherm.

  14. Limitations in controlling white mold on common beans with Trichoderma spp. at the fall-winter season

    Directory of Open Access Journals (Sweden)

    Trazilbo José de Paula Júnior

    2012-12-01

    Full Text Available We studied the effectiveness of application of Trichoderma spp. in controlling white mold on common beans at the fall-winter crop in the Zona da Mata region of the State of Minas Gerais, Brazil. There was no effect of the antagonist in reducing the disease severity, which could be explained by the low temperatures and the high inoculum pressure in the field. We concluded that Trichoderma applications are not recommended for control of white mold on common beans at the fall-winter season in regions with average temperature bellow 20 °C, since this condition favor more the pathogen than the antagonist.

  15. Purificación y caracterización de hidrolasas implicadas en el microparasitismo de Trichoderma harzianum

    OpenAIRE

    Cruz Díaz, Jesús de la

    1994-01-01

    Los hongos del genero Trichoderma son buenos agentes de control biológico. Entre los mecanismos que estos utilizan para combatir las enfermedades de plantas producidas por hongos se encuentra el fenómeno denominado micoparasitismo. Se sugiere que este proceso requiere la producción de hidrolasas de pared celular. En este trabajo se ha estudiado la producción de quitinasas y BETA-1,3 y BETA-1-6-GLUCANASAS en la estirpe micoparasitaria Trichoderma harzianum CECT 2413. Se ha analizado la regulac...

  16. Diversidad de Anamorfos de Ascomycota en bosques nativos de Celtis tala (Ulmaceae en la Provincia de Buenos Aires, Argentina Diversity of anamorphic fungi in Celtis tala (Ulmaceae native forest from Buenos Aires province, Argentina

    Directory of Open Access Journals (Sweden)

    Natalia Allegrucci

    2007-07-01

    Full Text Available En el presente trabajo se analizó la diversidad de microhongos que constituyen la comunidad fúngica saprótrofa (anamorfos de Ascomycota presente en hojarasca y suelo en bosques nativos de Celtis tala (tala en el partido de Magdalena, provincia de Buenos Aires. Se realizaron muestreos estacionales durante dos años (2004-2005 y se aislaron e identificaron los hongos presentes. Se calculó la frecuencia relativa porcentual de cada taxón; estos datos fueron utilizados para evaluar la diversidad fúngica mediante el cálculo del Indice de Diversidad de Shanon y Weaver ( H '. Para discriminar las comunidades fúngicas se utilizó el coeficiente de similitud de Sorensen ( S '. Se identificaron 104 taxones de anamorfos de Ascomycota, de los cuales 54 fueron aislados de hojarasca y 58 de suelo, registrándose 8 especies en común para ambos tipos de muestra. De las especies compartidas, las que presentaron frecuencias más altas para hojarasca fueron las menos representadas en suelo y viceversa. El resultado del cálculo del coeficiente de similitud de Sorensen fue de 0.14, indicando que la comunidad de hongos saprótrofos que crece en la hojarasca de tala está integrada por diferentes especies a las que caracterizan la micobiota del suelo de la misma área. No se encontraron diferencias significativas en el índice de diversidad.In this paper we analyze the diversity of species that compose the saprotrophic (anamorphic Ascomycota fungi community in the leaf litter and soil in Celtis tala forest in Magdalena , located in the province of Buenos Aires . Seasonal samples were taken during two years (2004-2005, and fungi were isolated and identified. The relative frequencies of fungi were calculated. To compare the similarity of the fungi composition between different habitats, Sorensen's index of similarity (S´ was applied. The frequencies of occurrence of these fungi were recorded and Shannon Weaver index (H´ was applied to evaluate fungal

  17. Bioremediation potential of a copper-resistant Trichoderma strain for copper-containing wastewater%一株抗铜木霉对含铜废水的修复潜力研究

    Institute of Scientific and Technical Information of China (English)

    田晔; 滕应; 赵静; 骆永明; 李振高; 张明旭

    2012-01-01

    基于真菌作为含铜废水生物修复剂的优点,探讨了一株里氏木霉FS10-C对含铜废水的修复潜力.采用摇瓶试验研究了不同Cu2+含量溶液下木霉FS10-C的生长发育和铜积累率;扫描电子显微镜(SEM)和X射线光散射能谱(EDX)观察成熟FS10-C细胞吸附Cu2+后形态和各元素变化;同时进行了该株木霉的生物安全性初步评价.结果表明:FS10-C孢子在pH为(4.0±0.2)~(5.6±0.2),Cu2+的质量浓度在不大于600mg/L的溶液中均可生长发育,且在质量浓度为100,200和400mg/L的Cu2+溶液中其铜积累率分别达50.6%,30.3%和4.7%.SEM显示,在质量浓度为100mg/L的Cu2+溶液中,成熟FS10-C细胞形态无改变,400mg/L时菌体结构仍完整;EDX元素分析显示,在质量浓度为100,200和400mg/L的Cu2+溶液中,FS10-C细胞铜的平均质量分数分别为18.50%和30.40%,且菌丝大于孢子.生物安全性初步评价表明FS10-C及代谢产物对人畜均无害.里氏木霉FS10-C是一株颇具研究潜力的含铜废水修复菌.%Based on the advantages of fungi as a bioremediation agent for copper-containing wastewater, bioremediation potential of Trichoderma reesei FS10-C for copper-containing wastewater was investigated. Flask tests were conducted on analyzing the development, growth and accumulation rate of FS10-C under copper stress in aqueous solutions. Scanning e- lectron microscope (SEM) and energy-dispersive X-ray analysis (EDX) were used to observe the morphology and element variations of FSI0-C, and bio-safety assessment of the strain was also performed. The results show that the spore of FS10-C can grow and develop in aqueous solutions with pH of (4.0±0. 2)- (5.6±0. 2) and Cu( Ⅱ ) concentration below 600 mg/L.Copper accumulation rates of FS10-C under 100, 200 and 400 mg/L Cu( Ⅱ ) concentrations were 50.60/oo, 30.3% and 4.70%, respectively. The SEM showed no morphological variation in FS10-C at 100 mg/L Cu

  18. Características de Trichoderma harzianum, como agente limitante en el cultivo de hongos comestibles

    Directory of Open Access Journals (Sweden)

    Omar Romero Arenas

    2009-10-01

    Full Text Available El incremento dramático de incidencia y severidad de los “mohos verdes” en la producción de hongos comestibles se refleja en la aparición de formas altamente agresivas de éstos patógenos, como es el caso de los biotipos de Trichoderma harzianum (Th1, Th2., Th3 y Th4, que han sido encontrados en Europa y Norte América, donde la importancia de la patogenicidad de dicho “moho” se comprobó en 1995 con las pérdidas del 30-100% en las plantas de hongos comestibles en Chester, Pennsylvania. En México se han identificado diversos “mohos contaminantes”, entre los cuales Trichoderma spp., se encuentra frecuentemente en la producción de hongos comestibles (Agaricus bisporus, Pleurotus ostreatus y Lentinula edodes, en el 2004, un grupo de investigación detectó la presencia de cepas altamente agresivas de T. aggressivum f. aggressivum, identificadas con técnicas clásicas y moleculares, en muestras de substrato (compost contaminado, proporcionado por la principal planta de hongos de México. Actualmente se desconoce la situación sobre la distribución de Trichoderma harzianum y los problemas de contaminación en la producción de hongos comestibles, tanto de zonas rurales, como de zonas industrializadas en México, puede causar serias disminuciones en la producción de hongos comestibles y presentar pérdidas económicas para los productores de la región. Palabras Claves: T. harzianum; T. aggressivum; patogenicidad; producción de hongos comestibles. Abstract The dramatic increase of green mould incidence and severity in edible mushroom production has been reflected in the emergence of highly aggressive forms of these pathogens, such as Trichoderma harzianum biotypes (Th1, Th2, Th3 and Th4. These have been found in Europe and North America where the importance of the mould’s pathogenicity was discovered in 1995 leading to 30%-100% losses from Th4 in edible mushrooms from farms in Chester, Pennsylvania. Several contaminating

  19. Harzianolide, a novel plant growth regulator and systemic resistance elicitor from Trichoderma harzianum.

    Science.gov (United States)

    Cai, Feng; Yu, Guanghui; Wang, Ping; Wei, Zhong; Fu, Lin; Shen, Qirong; Chen, Wei

    2013-12-01

    A detailed understanding of the effect of natural products on plant growth and protection will underpin new product development for plant production. The isolation and characterization of a known secondary metabolite named harzianolide from Trichoderma harzianum strain SQR-T037 were described, and the bioactivity of the purified compound as well as the crude metabolite extract in plant growth promotion and systemic resistance induction was investigated in this study. The results showed that harzianolide significantly promoted tomato seedling growth by up to 2.5-fold (dry weight) at a concentration of 0.1 ppm compared with the control. The result of root scan suggested that Trichoderma secondary metabolites may influence the early stages of plant growth through better root development for the enhancement of root length and tips. Both of the purified harzianolide and crude metabolite extract increased the activity of some defense-related enzymes to response to oxidative stress. Examination of six defense-related gene expression by real-time reverse transcription-PCR analysis revealed that harzianolide induces the expression of genes involved in the salicylic acid (PR1 and GLU) and jasmonate/ethylene (JERF3) signaling pathways while crude metabolite extract inhibited some gene expression (CHI-II and PGIP) related to basal defense in tomato plants. Further experiment showed that a subsequent challenge of harzianolide-pretreated plants with the pathogen Sclerotinia sclerotiorum resulted in higher systemic resistance by the reduction of lesion size. These results indicate that secondary metabolites of Trichoderma spp., like harzianolide, may play a novel role in both plant growth regulation and plant defense responses.

  20. G protein signalling involved in host recognition and mycoparasitismrelated chitinase expression in Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

    Susanne Zeilinger; Barbara Reithner; Kurt Brunner; Valeria Scala; Isabel Peiβl; Matteo Lorito; Robert L Mach

    2004-01-01

    @@ Mycoparasitic species of Trichoderma are commercially applied as biological control agents against various fungal pathogens. The mycoparasitic interaction is host specific and includes recognition,attack and subsequent penetration and killing of the host. Investigations on the underlying events revealed that Trichoderma responds to multiple signals from the host (e. g. lectins or other ligands such as low molecular weight components released from the host's cell wall) and host attack is accompanied by morphological changes and the secretion of hydrolytic enzymes and antibiotics.Degradation of the cell wall of the host fungus is-besides glucanases and proteases-mainly achieved by chitinases. In vivo studies showed that the ech42 gene (encoding endochitinase 42) is expressed before physical contact of Trichoderma with its host, probably representing one of the earliest events in mycoparasitism, whereas Nag1 (N-acetylglucosaminidase) plays a key role in the general induction of the chitinolytic enzyme system of T. atroviride . Investigations on the responsible signal transduction pathways of T. atroviride led to the isolation of several genes encoding key components of the cAMP and MAP kinase signaling pathways, as alpha and β subunits of heterotrimeric G proteins, the regulatory subunit of cAMP-dependent protein kinase,adenylate cyclase, and three MAP kinases. Analysis of knockout mutants, generated by Agrobacterium-mediated transformation, revealed that at least two alpha-subunits of heterotrimeric G proteins are participating in mycoparasitism-related signal transduction. The Tga1 G alpha subunit was shown to be involved in mycoparasitism-related processes such as chitinase expression and overproduction of toxic secondary metabolites, whereas Tga3 was found to be completely avirulent showing defects in chitinase formation and host recognition.

  1. Deciphering the hormonal signalling network behind the systemic resistance induced by Trichoderma harzianum in tomato

    Directory of Open Access Journals (Sweden)

    Ainhoa eMartinez-Medina

    2013-06-01

    Full Text Available Root colonization by selected Trichoderma isolates can activate in the plant a systemic defence response that is effective against a broad spectrum of plant pathogens. Diverse plant hormones play pivotal roles in the regulation of the defence signalling network that leads to the induction of systemic resistance triggered by beneficial organisms (ISR. Among them, jasmonic acid (JA and ethylene (ET signalling pathways are generally essential for ISR. However, Trichoderma ISR (TISR is believed to involve a wider variety of signalling routes, interconnected in a complex network of cross-communicating hormone pathways. Using tomato as a model, an integrative analysis of the main mechanisms involved in the systemic resistance induced by Trichoderma harzianum against the necrotrophic leaf pathogen Botrytis cinerea was performed. Root colonization by T. harzianum rendered the leaves more resistant to B. cinerea independently of major effects on plant nutrition. The analysis of disease development in shoots of tomato mutant lines impaired in the synthesis of the key defence related hormones JA, ET, salicylic acid (SA and abscisic acid (ABA and the peptide prosystemin (PS evidenced the requirement of intact JA, SA and ABA signalling pathways for a functional TISR. Expression analysis of several hormone related marker genes point to the role of priming for enhanced JA-dependent defence responses upon pathogen infection. Together, our results indicate that although TISR induced in tomato against the necrotrophs is mainly based on boosted JA-dependent responses, the pathways regulated by the plant hormones SA- and ABA are also required for successful TISR development

  2. Phylogeny and taxonomical investigation of Trichoderma spp. from Indian region of Indo-Burma Biodiversity hot spot region with special reference to Manipur.

    Science.gov (United States)

    Kamala, Th; Devi, S Indira; Sharma, K Chandradev; Kennedy, K

    2015-01-01

    Towards assessing the genetic diversity and occurrence of Trichoderma species from the Indian region of Indo-Burma Biodiversity hotspot, a total of 193 Trichoderma strains were isolated from cultivated soils of nine different districts of Manipur comprising 4 different agroclimatic zones. The isolates were grouped based on the morphological characteristics. ITS-RFLP of the rDNA region using three restriction digestion enzymes: Mob1, Taq1, and Hinf1, showed interspecific variations among 65 isolates of Trichoderma. Based on ITS sequence data, a total of 22 different types of representative Trichoderma species were reported and phylogenetic analysis showed 4 well-separated main clades in which T. harzianum was found to be the most prevalent spp. among all the Trichoderma spp. Combined molecular and phenotypic data leads to the development of a taxonomy of all the 22 different Trichoderma spp., which was reported for the first time from this unique region. All these species were found to produce different extrolites and enzymes responsible for the biocontrol activities against the harmful fungal phytopathogens that hamper in food production. This potential indigenous Trichoderma spp. can be targeted for the development of suitable bioformulation against soil and seedborne pathogens in sustainable agricultural practice.

  3. Biosorption Potential of Trichoderma gamsii Biomass for Removal of Cr(VI) from Electroplating Industrial Effluent

    OpenAIRE

    Haresh Keharia; Kavita, B.

    2012-01-01

    The potential use of acid-treated biomass of Trichoderma gamsii to remove hexavalent chromium ions from electroplating industrial effluent was evaluated. Electroplating industrial effluent contaminated with 5000 mg/L of Cr(VI) ions, collected from industrial estate of Gujarat, India, was mixed with acid-treated biomass of T. gamsii at biomass dose of 10 mg/mL. Effect of contact time and initial Cr(VI) ions was studied. The biosorption of Cr(VI) ions attained equilibrium at time interval of 24...

  4. Effective Biosorption of Nickel(II) from Aqueous Solutions Using Trichoderma viride

    OpenAIRE

    B. Naresh Kumar; Kalarani, V.; P.SUJATHA

    2013-01-01

    The primary objective of the present study is to evaluate the optimization conditions such as kinetic and equilibrium isotherm models involved in the removal of Ni(II) from the aqueous solutions by Trichoderma viride. The biosorbent was characterized by FTIR and SEM. The optimum biosorption conditions were determined as a function of pH, biomass dosage, contact time, initial metal ion concentration, and temperature. The maximum Ni(II) biosorption was obtained at pH 4.5. The equilibrium data w...

  5. PRODUCCIÓN MASIVA DE Trichoderma harzianum Rifai EN DIFERENTES SUSTRATOS ORGÁNICOS

    OpenAIRE

    A. C. Michel-Aceves; M. A. Otero-Sánchez; R. D. Martínez-Rojero; N. L. Rodríguez-Morán; R. Ariza-Flores; A. Barrios-Ayala

    2008-01-01

    Uno de los sustratos utilizados para reproducir a Trichoderma spp., es el grano entero de arroz, el cual tiene costo relativamente alto. Con el propósito de encontrar un sustrato orgánico, económico y de fácil adquisición en la región, en el cual este hongo tenga un buen desarrollo y una alta producción de esporas viables, se estableció esta investigación que tuvo como objetivos evaluar 15 sustratos orgánicos en la reproducción masiva y viabilidad de esporas de T. harzianum y asociarlo con la...

  6. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

    OpenAIRE

    Irfan, M.; Q. Syed

    2012-01-01

    Summary: In the present study xylanase enzyme was produced from Trichoderma viride in solid state fermentation using sugarcane bagasse as a substrate. The whole fermentation process was carried out in 250ml Erlenmeyer flask at 30oC for seven days of fermentation period. The enzyme was partially purified by ammonium sulphate (60%) fractionation followed by dialysis. The partially purified enzyme was further characterized showing optimum pH and temperature of 5.0 and 50oC respectively. Metal pr...

  7. New C13 lipids from the marine-derived fungus Trichoderma harzianum.

    Science.gov (United States)

    Li, Bo; Huang, Qi-Xi; Gao, Du; Liu, Dong; Ji, Yu-Bin; Liu, Hua-Gang; Lin, Wen-Han

    2015-05-01

    Chemical examination of the fermentation broth of a sponge-associated fungus Trichoderma harzinum HMS-15-3 led to the isolation of four pairs of new C13 lipid enantiomers namely harzianumols A-H (1a-4b). Their structures were elucidated on the basis of extensive spectroscopic (IR, MS, 1D, and 2D NMR) data analysis, including the modified Mosher's method for the assignment of their absolute configurations. The new compounds were evaluated for antihyperlipidemic effects in HepG2 cells. PMID:26031203

  8. Production of xylan-degrading enzymes by a Trichoderma harzianum strain

    Directory of Open Access Journals (Sweden)

    Cacais André O.Guerreiro

    2001-01-01

    Full Text Available Trichoderma harzianum strain 4 produced extracellular xylan-degrading enzymes, namely beta-xylanase, beta-xylosidase and alpha-arabinofuranosidase, when grown in liquid medium cultures containing oat spelt xylan as inducer. Cellulase activity was not detected. The pattern of xylan-degrading enzymes induction was influenced by the form of xylan present in the medium. They were detected in different incubation periods. Electrophoretic separation of the proteins from liquid culture filtrates by SDS-PAGE showed a variety of bands with high and low molecular weights.

  9. Immobilization of Trichoderma harzianum α-Amylase on Treated Wool: Optimization and Characterization

    OpenAIRE

    Saleh A. Mohamed; Khan, Jalaluddin A; Omar A.M. Al-Bar; El-Shishtawy, Reda M.

    2014-01-01

    α-Amylase from Trichoderma harzianum was covalently immobilized on activated wool by cyanuric chloride. Immobilized α-amylase exhibited 75% of its initial activity after 10 runs. The soluble and immobilized α-amylases exhibited maximum activity at pH values 6.0 and 6.5, respectively. The immobilized enzyme was more thermally stable than the soluble one. Various substrates were hydrolyzed by immobilized α-amylase with high efficiencies compared to those of soluble α-amylase. The inhibition of ...

  10. L-glutaminase production by Trichoderma koningii under solid-state fermentation

    OpenAIRE

    El-Sayed, Ashraf S. A.

    2009-01-01

    Solid state fermentation was conducted for the production of L-glutaminase by Trichoderma koningii Oud.aggr. using different agro-industrial byproducts inlcuding wheat bran, groundnut residues, rice hulls, soya bean meal, corn steep, sesamum oil cake, cotton seed residues and lentil industrial residues as solid substrates. Wheat bran was the best substrate for induction of L-glutaminase (12.1 U/mg protein) by T. koningii. The maximum productivity (23.2 U/mg protein) and yield (45.0 U/gds) of ...

  11. Bioconversions of Palm Kernel Cake and Rice Bran Mixtures by Trichoderma viride Toward Nutritional Contents

    OpenAIRE

    Yana Sukaryana; Umi Atmomarsono; Vitus D. Yunianto; Ejeng Supriyatna

    2010-01-01

    The objective of the research is to examine the mixtures of palm kernel cake and rice bran of fermented by Trichoderma viride. Completely randomized design in factorial pattern 4 x 4 was used in this experiment. factor I is the doses of inoculums; D1 = 0%, D2 =  0,1% , D3 =  0,2%, D4 =  0,3%, and  complement factor II is mixtures of palm kernel cake and rice bran : T1=20:80% ; T2=40:60% ; T3=60:40% ; T4=80:20%. The treatment each of three replicate. Fermentation was conduc...

  12. Características de Trichoderma harzianum, como agente limitante en el cultivo de hongos comestibles

    OpenAIRE

    Omar Romero-Arenas; Manuel Huerta Lara; Miguel Angel Damián Huato; Francisco Domínguez Hernández; Daniel Alfonso Arellano Victoria

    2009-01-01

    El incremento dramático de incidencia y severidad de los mohos verdes en la producción de hongos comestibles se refleja en la aparición de formas altamente agresivas de éstos patógenos, como es el caso de los biotipos de Trichoderma harzianum (Th1, Th2., Th3 y Th4), que han sido encontrados en Europa y Norte América, donde la importancia de la patogenicidad de dicho moho se comprobó en 1995 con las pérdidas del 30-100% en las plantas de hongos comestibles en Chester, Pennsylvania. En Méxi...

  13. Cellulase production by two mutant strain of Trichoderma longibrachiatum Qm9414 and Rut C30

    International Nuclear Information System (INIS)

    Native or pretreated biomass from Onopordum nervosum boiss, has been examined as candidate feedstock for cellulase production by two mutant strain of trichoderma longibrachiatum QM9414 and Rut C30. Batch cultivation methods were evaluated and compared with previous experiments using ball-milled, crystalline cellulose (Solka floc). Batch cultivation of T. longibrachiatum Rut C30 on 55% (W/V) acid pretreated O. nervosum biomass yielded enzyme productivities and activities comparable to those obtained on Solka floc. However, the overall enzyme production performance was lower than on Solka floc at comparable cellulose concentrations. This fact may be due to the accumulation of pretreated by products and lignin in the fermentor.(author)

  14. Effect of Trichoderma spp. On Three Pathogens Causing Vegetable Diseases in Greenhouse%木霉(Trichoderma spp.)对三种引起大棚蔬菜病害病原菌的影响

    Institute of Scientific and Technical Information of China (English)

    宋瑞清; 周秀华; Sirajul HASAH

    2004-01-01

    通过木霉属(Trichoderma)3菌株与双鸭山蔬菜大棚中的黄瓜枯萎病菌(Fusarium oxysporum Schlecht.f.cucumerinum)、黄瓜果腐病菌(Phytophthora capsici Leonian)、菜豆叶枯病菌(Cladosporium sp.)的对峙培养试验,结果表明:绿色木霉1(Trichoderma viride Pers.ex Gray 1)可作为双鸭山蔬菜大棚中的黄瓜枯萎病、黄瓜果腐病、菜豆叶枯病3种病害的生物防治拮抗菌加以利用,该拮抗菌对菜豆叶枯病菌抑制效果最好;绿色木霉2(Trichoderma viride 2)对黄瓜果腐病菌抑制效果最好;而哈茨木霉(Trichoderma harzianum Rifai)对以上3种病原菌都有抑制效果,对菜豆叶枯病菌抑制效果最好.从试验结果还可看出,绿色木霉2对黄瓜枯萎病菌和菜豆叶枯病菌的生长有促进作用.

  15. Antagonism of Trichoderma species on Cladosporium herbarum and their enzimatic characterization Antagonismo de espécies de Trichoderma sobre Cladosporium herbarum e suas caracterização enzimática

    Directory of Open Access Journals (Sweden)

    Maria Angélica G. Barbosa

    2001-06-01

    Full Text Available The verrucose caused by Cladosporium herbarum reduces production and quality of Passion fruit (Passiflora edulis Sims., a largely consumed tropical fruit. This work aimed to investigate the antagonism of Trichoderma species (T. polysporum, T. koningii, T. viride and T. harzianum against Cladosporium herbarum, and to study the production of extracellular hydrolytic enzymes by the pathogen and the antagonists. The results showed considerable antagonistic potential for the biocontrol of C. herbarum isolates by all Trichoderma species, except T. koningii. The most distinguished effect was observed for T. polysporum. In relation to the pattern of esterase obtained by electrophoresis in poliacrylamyde gel, the major activity presented by the isolates was observed after five days of incubation. The C. herbarum isolates produced extracellular enzymes, lipase, pectinase, cellulase, and protease, all possibly related to the infection process. Amylase excretion, not known to be associated with phytopathogens, was detected in Trichoderma species, but not in C. herbarum. In addition to amylase, all Trichoderma species tested produced also extracellular cellulase and pectinase, except T. koningii in relation to the latter enzyme. The demonstration of various esterase isoenzymes in zymograms of the Trichoderma species and C. herbarum isolates was markedly improved by washing the mycelia with detergents or EDTA. This fact suggested that a major fraction of extracelular enzymes may remain attached to outside fungal cell wall after being excreted.O maracujá (Passiflora edulis Sims., um fruto tropical amplamente consumido, tem sua produção e a qualidade dos seus frutos reduzidos pela verrugose causada por Cladosporium herbarum. Este trabalho objetivou investigar o antagonismo de espécies de Trichoderma (T. polysporum, T.koningii, T. viride e T. harzianum contra C. herbarum, e estudar a produção de enzimas hidrolíticas extracelulares pelo fitopatógeno e

  16. Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi

    Science.gov (United States)

    Trichothecenes are phytotoxic sesquiterpenoid compounds of fungal origin which can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in the biosynthesis of HA is the conversion of farnesyl diphosphate t...

  17. Trichothecenes and aspinolides produced by Trichoderma arundinaceum regulate expression of Botrytis cinerea genes involved in virulence and growth

    Science.gov (United States)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea (B05.10) produce the sesquiterpenoid compounds harzianum A (HA) and botrydial (BOT), respectively. T. arundinaceum Ta(delta)Tri5, a mutant that does not produce HA, produces high levels of the polyketide compounds aspinolide (Asp) B and C. We anal...

  18. Botrydial and botcinins produced by Botrytis cinerea regulate expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis

    Science.gov (United States)

    Trichoderma arundinaceum (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides (Asp) and botcinines (Botc), respectively. In the present work, we analyzed the role of BOT and Botcs in the T. arundinaceum-B. cin...

  19. Novel aspinolide production by Trichoderma arundinaceum with a potential role in Botrytis cinerea antagonistic activity and plant defense priming

    Science.gov (United States)

    Harzianum A (HA), a trichothecene produced by Trichoderma arundinaceum, has recently been described to have antagonistic activity against fungal plant pathogens and to induce plant defence genes. In the present work, we have shown that a tri5 genedisrupted mutant that lacks HA production overproduce...

  20. Regulation of Botrytis cinerea virulence genes in interaction with Trichoderma arundinaceum is mediated by the sesquiterpene harzianum A

    Science.gov (United States)

    Trichoderma includes a great diversity of species, some of them with the ability to control the growth of fungal phytopathogens. Many of these strains produce secondary metabolites that are able to inhibit the growth of their fungal preys. However, pathogens can also produce metabolites that in some...