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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. Sulphur metabolism and cellulase gene expression are connected processes in the filamentous fungus Hypocrea jecorina (anamorph Trichoderma reesei

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    Schmoll Monika

    2008-10-01

    Full Text Available Abstract Background Sulphur compounds like cysteine, methionine and S-adenosylmethionine are essential for the viability of most cells. Thus many organisms have developed a complex regulatory circuit that governs the expression of enzymes involved in sulphur assimilation and metabolism. In the filamentous fungus Hypocrea jecorina (anamorph Trichoderma reesei little is known about the participants in this circuit. Results Analyses of proteins binding to the cellulase activating element (CAE within the promotor of the cellobiohydrolase cbh2 gene led to the identification of a putative E3 ubiquitin ligase protein named LIMPET (LIM1, which is an orthologue of the sulphur regulators SCON-2 of Neurospora crassa and Met30p of Saccharomyces cerevisiae. Transcription of lim1 is specifically up-regulated upon sulphur limitation and responds to cellulase inducing conditions. In addition, light dependent stimulation/shut down of cellulase gene transcription by methionine in the presence of sulphate was observed. Further, lim1 transcriptionally reacts to a switch from constant darkness to constant light and is subject to regulation by the light regulatory protein ENVOY. Thus lim1, despite its function in sulphur metabolite repression, responds both to light as well as sulphur- and carbon source. Upon growth on cellulose, the uptake of sulphate is dependent on the light status and essential for growth in light. Unlike other fungi, growth of H. jecorina is not inhibited by selenate under low sulphur conditions, suggesting altered regulation of sulphur metabolism. Phylogenetic analysis of the five sulphate permeases found in the genome of H. jecorina revealed that the predominantly mycelial sulphate permease is lacking, thus supporting this hypothesis. Conclusion Our data indicate that the significance of the sulphate/methionine-related signal with respect to cellulase gene expression is dependent on the light status and reaches beyond detection of sulphur

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

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    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

  5. Synergy in Trichoderma Reesei cellulases

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    Jepsen Kudal, Eva Rose; Ejaz, Rooshanie Nadia; Poniewierska, Julia Alicja

    2016-01-01

    Synergy has been observed between the various cellulolytic enzymes of the fungus Trichoderma Reesei. Termed cellulases, these enzymes come together to form a potent cocktail of biomass degrading enzymes. The biofuel industry has seen a recent surge partly due to the advances in attaining a more applicable understanding of the mechanisms of synergy that occur when said enzymes are used to degrade biomass into fermentable sugar. To that end, this review summarizes the various theories postulati...

  6. Immobilization of Trichoderma reesei by radiation polymerization

    International Nuclear Information System (INIS)

    Immobilization of Trichoderma reesei was carried out by radiation polymerization. It was found that the activity of fixed cells increased with increasing surface area of the carrier and was affected by the concentration of monomer tetraethylenglycol dimethacrylate and the shape of the substrate composition and structure of cotton textile fabrics. (author)

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

    OpenAIRE

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

    2012-01-01

    Abstract Background 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 i...

  8. Molecular evidence that the asexual industrial fungus Trichoderma reesei is a clonal derivative of the ascomycete Hypocrea jecorina.

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    Kuhls, K; Lieckfeldt, E; Samuels, G. J.; Kovacs, W; Meyer, W.; Petrini, O; Gams, W.; T. Börner; Kubicek, C P

    1996-01-01

    The relationship of the important cellulase producing asexual fungus Trichoderma reesei to its putative teleomorphic (sexual) ancestor Hypocrea jecorina and other species of the Trichoderma sect. Longibrachiatum was studied by PCR-fingerprinting and sequence analyses of the nuclear ribosomal DNA region containing the internal transcribed spacers (ITS-1 and ITS-2) and the 5.8S rRNA gene. The differences in the corresponding ITS sequences allowed a grouping of anamorphic (asexual) species of Tr...

  9. Space mutagenic effect of Trichoderma reesei

    International Nuclear Information System (INIS)

    The slant mycelia cultured with or without mutagenic agent diethyl sulfate (DS) and spores of Trichoderma reesei were loaded in the 18th returning satellite. Systematical screening showed that the life cycle and morphology of some strains had changed after space flight. After selection and domestication, 3 mutant strains with high cellulose enzyme activity were isolated. The cellulose enzyme productivities of the mutants were significantly increased more than 50%, and the mutant were generically stable. (authors)

  10. Expression of Barley Endopeptidase B in Trichoderma reesei

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    Saarelainen, R.; Mantyla, A.; Nevalainen, H.; Suominen, P.

    1997-01-01

    The gene for barley endopeptidase B (EPB) has been expressed in the filamentous fungus Trichoderma reesei from the cbh1 promoter. The EPB signal sequence allowed secretion of over 90% of the recombinant protein. Yields reached about 500 mg of immunoreactive protein per liter and exceeded values for any other protein derived from a higher eukaryotic organism produced in T. reesei.

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

    Directory of Open Access Journals (Sweden)

    Schuster André

    2012-01-01

    Full Text Available Abstract Background 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  13. Improved production of Trichoderma harzianum endochitinase by expression in Trichoderma reesei.

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    Margolles-Clark, E.; Hayes, C K; Harman, G. E.; Penttilä, M

    1996-01-01

    The chromosomal endochitinase gene (ThEn-42) of the mycoparasite fungus Trichoderma harzianum P1 was isolated and overexpressed in the filamentous fungus Trichoderma reesei under the promoter of the major cellulase gene cbhl1. The host strain RutC-30 did not produce any endogenous endochitinase activity. The prepro region of the T harzianum endochitinase was correctly processed in T. reesei. No differences in expression were observed when the prepro region was replaced with the CBHI signal se...

  14. Studies on Laccase Activity in the Filamentous Fungus Trichoderma reesei

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    Sekme, Semra; Atacı, Neşe; Arısan, İnci

    2014-01-01

    Laccase have been mainly studied in wood rot fungal species of the basidiomycetes family especiallyin white rot fungi. Studies in other fungal families are largely lacking. This study has evaluated laccaseactivity fromTrichoderma reesei in catechol based medium. Results showed that laccase enzyme from T.reesei was active in acidic pH range and that optimum pH was 4.5. The optimum temperature oflaccase from T. reesei was also 270C. Laccase activity in medium containing 10 gL-1 catechol was 1.2...

  15. Genome Sequence and Annotation of Trichoderma parareesei, the Ancestor of the Cellulase Producer Trichoderma reesei

    OpenAIRE

    Yang, Dongqing; Pomraning, Kyle; Kopchinskiy, Alexey; Karimi Aghcheh, Razieh; Atanasova, Lea; Chenthamara, Komal; Baker, Scott E.; Zhang, Ruifu; Shen, Qirong; Freitag, Michael; Kubicek, Christian P.; Druzhinina, Irina S.

    2015-01-01

    The filamentous fungus Trichoderma parareesei is the asexually reproducing ancestor of Trichoderma reesei, the holomorphic industrial producer of cellulase and hemicellulase. Here, we present the genome sequence of the T. parareesei type strain CBS 125925, which contains genes for 9,318 proteins.

  16. Genome Sequence and Annotation of Trichoderma parareesei, the Ancestor of the Cellulase Producer Trichoderma reesei.

    Science.gov (United States)

    Yang, Dongqing; Pomraning, Kyle; Kopchinskiy, Alexey; Karimi Aghcheh, Razieh; Atanasova, Lea; Chenthamara, Komal; Baker, Scott E; Zhang, Ruifu; Shen, Qirong; Freitag, Michael; Kubicek, Christian P; Druzhinina, Irina S

    2015-01-01

    The filamentous fungus Trichoderma parareesei is the asexually reproducing ancestor of Trichoderma reesei, the holomorphic industrial producer of cellulase and hemicellulase. Here, we present the genome sequence of the T. parareesei type strain CBS 125925, which contains genes for 9,318 proteins. PMID:26272569

  17. Expression of the mammalian peptide hormone obestatin in Trichoderma reesei.

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    Sun, Angela; Peterson, Robyn; Te'o, Junior; Nevalainen, Helena

    2016-01-25

    The filamentous fungus Trichoderma reesei is an expression host widely exploited for the production of recombinant proteins. However, its capacity for expressing small peptides (reesei cellobiohydrolase I core (CBHI) or xylanase 2 (XYN2) pro-region as a carrier and the cbh1 promoter for gene expression, in high protein-low protease producing mutant strains T. reesei Rut-C30 and HEPI. The yield of obestatin was improved from about 300 ng/ml to up to 5.5 μg/ml through adaptive laboratory evolution and modifications to the cultivation strategy, which included adjustments of the type and ratio of carbon and nitrogen sources used in the medium. The successful expression of Obe-HFBI demonstrated the potential of T. reesei as an expression host for small peptides and further enhancement of the recombinant yield through modification of culture conditions. PMID:26341165

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

    OpenAIRE

    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...

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

    OpenAIRE

    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...

  20. Ethanol Perturbs Glycosylation and Inhibits Hypersecretion in Trichoderma reesei

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    Merivuori, Hannele; Montenecourt, Bland S.; Sands, Jeffrey A.

    1987-01-01

    The effects of ethanol and phenylethanol on the growth of and glycoprotein secretion by Trichoderma reesei were studied. Low levels (1.5%, vol/vol) of ethanol perturbed the glycosylation process, as shown by alterations in the isoelectric profile of the secreted proteins and a reduction in the rate of incorporation of mannose into oligosaccharides. In addition to these effects on posttranslational modification, ethanol drastically lowered the protein secretion level of a hypersecretory strain.

  1. Systems biological approaches towards understanding cellulase production by Trichoderma reesei

    OpenAIRE

    Kubicek, Christian P.

    2013-01-01

    Recent progress and improvement in “-omics” technologies has made it possible to study the physiology of organisms by integrated and genome-wide approaches. This bears the advantage that the global response, rather than isolated pathways and circuits within an organism, can be investigated (“systems biology”). The sequencing of the genome of Trichoderma reesei (teleomorph Hypocrea jecorina), a fungus that serves as a major producer of biomass-degrading enzymes for the use of renewable lignoce...

  2. Pectinase from Trichoderma reesei QM 9414

    Energy Technology Data Exchange (ETDEWEB)

    Haltmeier, T.; Leisola, M.; Ulmer, D.; Waldner, R.; Fiechter, A.

    1983-06-01

    The present study was undertaken to determine whether T. reesei produces pectinolytic enzymes and, if so, to determine their action pattern and specificity. The aim was also to find out the practical importance of pectinolytic activity in the hydrolysis of cellulosic materials. It was found that the presence of pectinase does not appear to facilitate the hydrolysis of cellulose in plant material through any cell-separating mechanism. It is concluded that cellulase and xylanase activities are the important activities in the practical hydrolysis of cellulosic materials and that lignin is the real limiting factor. (Refs. 12).

  3. Production of cellulase by Trichoderma reesei from dairy manure.

    Science.gov (United States)

    Wen, Zhiyou; Liao, Wei; Chen, Shulin

    2005-03-01

    Cellulase production by the fungi Trichoderma reesei was studied using dairy manure as a substrate. Data showed that T. reesei RUT-C30 had higher cellulase production than T. reesei QM 9414 and that a homogenized manure, treated by a blender to reduce fiber size, led to higher cellulase production. The cellulase production was further optimized by growing T. reesei RUT-C30 on homogenized manure. The effects of manure concentration, pH, and temperature on cellulase production were investigated with optimal parameter values determined to be 10 g/l manure (dry basis), 25.5 degrees C, and pH 5.7, respectively. Elimination of CaCl2, MgSO4, nitrogen sources (NH4+ and urea) and trace elements (Fe2+, Zn2+, Co2+ and Mn2+) from the original salt solution had no negative influence on the cellulase production, while phosphate elimination did reduce cellulase production. Based on above results, the final medium composition was simplified with manure additives being KH2PO4, tween-80 and CoCl2 only. Using this medium composition and a reaction time of 6-8 days, a maximum cellulase production activity of 1.74 IU/ml of filter paper activity, 12.22 IU/ml of CMCase activity, and 0.0978 IU/ml of beta-glucosidase was obtained. This filter paper activity is the highest ever reported in cellulase production from agricultural wastes. PMID:15491832

  4. Clonal species Trichoderma parareesei sp. nov. likely resembles the ancestor of the cellulase producer Hypocrea jecorina/T. reesei.

    Science.gov (United States)

    Atanasova, Lea; Jaklitsch, Walter M; Komoń-Zelazowska, Monika; Kubicek, Christian P; Druzhinina, Irina S

    2010-11-01

    We have previously reported that the prominent industrial enzyme producer Trichoderma reesei (teleomorph Hypocrea jecorina; Hypocreales, Ascomycota, Dikarya) has a genetically isolated, sympatric sister species devoid of sexual reproduction and which is constituted by the majority of anamorphic strains previously attributed to H. jecorina/T. reesei. In this paper we present the formal taxonomic description of this new species, T. parareesei, complemented by multivariate phenotype profiling and molecular evolutionary examination. A phylogenetic analysis of relatively conserved loci, such as coding fragments of the RNA polymerase B subunit II (rpb2) and GH18 chitinase (chi18-5), showed that T. parareesei is genetically invariable and likely resembles the ancestor which gave raise to H. jecorina. This and the fact that at least one mating type gene of T. parareesei has previously been found to be essentially altered compared to the sequence of H. jecorina/T. reesei indicate that divergence probably occurred due to the impaired functionality of the mating system in the hypothetical ancestor of both species. In contrast, we show that the sexually reproducing and correspondingly more polymorphic H. jecorina/T. reesei is essentially evolutionarily derived. Phenotype microarray analyses performed at seven temperature regimens support our previous speculations that T. parareesei possesses a relatively high opportunistic potential, which probably ensured the survival of this species in ancient and sustainable environment such as tropical forests. PMID:20817800

  5. Production of Trichoderma reesei cellulases on glucose-containing media.

    OpenAIRE

    Nakari-Setälä, T; Penttilä, M

    1995-01-01

    The filamentous fungus Trichoderma reesei was shown to secrete active cellobiohydrolase I and the endoglucanase I catalytic core domain into the culture medium when the fungus was grown on glucose-containing medium. The expression of the proteins was driven by the promoters of the elongation factor 1 alpha, tef1, and the unidentified gene for cDNA1. The cDNA1 promoter gave the best yields. The highest amounts of cellobiohydrolase I and the endoglucanase I core, being 50 to 100 mg/liter, accou...

  6. Method To Estimate Growth of Trichoderma reesei and Aspergillus wentii in Mixed Culture on Cellulosic Substrates

    OpenAIRE

    Panda, T.; Bisaria, V. S.; Ghose, T. K.

    1989-01-01

    A simple differential method based on measurement of an intracellular pigment of Aspergillus wentii was developed for estimation of the individual growths of two fungi, Trichoderma reesei and A. wentii, in mixed-culture fermentation of an insoluble cellulosic substrate.

  7. Hydrolysis of Palm Kernel Cake (Elaeis guineensis Jacq) by Fungi Trichoderma reesei that Degrades Mannan Polysaccharides

    OpenAIRE

    Achmad Jaelani; Wiranda Gentini Piliang; Suryahadi; Iman Rahayu

    2008-01-01

    The research was conducted to study the growth characterization of Trichoderma reesei, the degradability of mannan polysaccharides from Palm Kernel Cake (PKC) with different concentrations of fungi and thickness of the PKC medium, as well as the improvement of the nutritive value of palm kernel cake. Experiment 1 was to study the growth characterization of Trichoderma reesei, and Experiment 2 was conducted using concentrations of fungi (104, 105, and 106 CFU per cc) and thickness of the PKC m...

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

    Directory of Open Access Journals (Sweden)

    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.

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

    OpenAIRE

    Jørgensen, Mikael Skaanning; Skovlund, Dominique Aubert; Johannesen, Pia Francke; Mortensen, Uffe Hasbro

    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 for genetic engineering. RESULTS: In this study we present new genetic tools for T. reesei to further expand its use in industrial production. We have developed an expression platform where genes are ...

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

    DEFF Research Database (Denmark)

    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.

    Science.gov (United States)

    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

  13. Linking hydrolysis performance to Trichoderma reesei cellulolytic enzyme profile.

    Science.gov (United States)

    Lehmann, Linda; Rønnest, Nanna P; Jørgensen, Christian I; Olsson, Lisbeth; Stocks, Stuart M; Jørgensen, Henrik S; Hobley, Timothy

    2016-05-01

    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, for example, by spiking with single enzymes and monitoring hydrolysis performance. In this study, a multivariate approach, partial least squares regression, was used to see whether it could help explain the correlation between enzyme profile and hydrolysis performance. Diverse enzyme mixtures were produced by T. reesei Rut-C30 by exploiting various fermentation conditions and used for hydrolysis of washed pretreated corn stover as a measure of enzyme performance. In addition, the enzyme mixtures were analyzed by liquid chromatography-tandem mass spectrometry to identify and quantify the different proteins. A multivariate model was applied for the prediction of enzyme performance based on the combination of different proteins present in an enzyme mixture. The multivariate model was used for identification of candidate proteins that are correlated to enzyme performance on pretreated corn stover. A very large variation in hydrolysis performance was observed and this was clearly caused by the difference in fermentation conditions. Besides β-glucosidase, the multivariate model identified several xylanases, Cip1 and Cip2, as relevant proteins to study further. Biotechnol. Bioeng. 2016;113: 1001-1010. © 2015 Wiley Periodicals, Inc. PMID:26524197

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

    Science.gov (United States)

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

    OpenAIRE

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

    2010-01-01

    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 gen...

  16. Comparison of Extracellular Cellulase Activities of Clostridium thermocellum LQRI and Trichoderma reesei QM9414

    OpenAIRE

    Ng, Thomas K.; Zeikus, J. G.

    1981-01-01

    The crude extracellular cellulase of Clostridium thermocellum LQRI (virgin strain) was very active and solubilized microcrystalline cellulose at one-half the rate observed for the extracellular cellulase of Trichoderma reesei QM9414 (mutant strain). C. thermocellum cellulase activity differed considerably from that of T. reesei as follows: higher endoglucanase/exoglucanase activity ratio; absence of extracellular cellobiase or β-xylosidase activity; long-chain oligosaccharides instead of shor...

  17. A homologous production system for Trichoderma reesei secreted proteins in a cellulase-free background

    OpenAIRE

    Uzbaş, Fatma; Uzbas, Fatma; Sezerman, Uğur; Sezerman, Ugur; Hartl, Lukas; Kubicek, Christian P.; Seiboth, Bernhard

    2012-01-01

    Recent demands for the production of biofuels from lignocellulose led to an increased interest in engineered cellulases from Trichoderma reesei or other fungal sources. While the methods to generate such mutant cellulases on DNA level are straightforward, there is often a bottleneck in their production since a correct posttranslational processing of these enzymes is needed to obtain highly active enzymes. Their production and subsequent enzymatic analysis in the homologous host T. reesei is, ...

  18. Achieving efficient protein expression in Trichoderma reesei by using strong constitutive promoters

    OpenAIRE

    Li Junxin; Wang Juan; Wang Shaowen; Xing Miao; Yu Shaowen; Liu Gang

    2012-01-01

    Abstract Backgrounds The fungus Trichoderma reesei is an important workhorse for expression of homologous or heterologous genes, and the inducible cbh1 promoter is generally used. However, constitutive expression is more preferable in some cases than inducible expression that leads to production of unwanted cellulase components. In this work, constitutive promoters of T. reesei were screened and successfully used for high level homologous expression of xylanase II. Results The transcriptional...

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

    Science.gov (United States)

    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

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

    OpenAIRE

    Denton Jai A; Kelly Joan M

    2011-01-01

    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 functi...

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

    OpenAIRE

    Maritje Aleonor Hilakore; Suryahadi; Komang Wiryawan; Djumali Mangunwijaya

    2013-01-01

    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  tr...

  2. Identification of microRNA-Like RNAs in the Filamentous Fungus Trichoderma reesei by Solexa Sequencing

    OpenAIRE

    Kang, Kang; Zhong, Jiasheng; Jiang, Liang; Liu, Gang; Gou, Christine Yuan; Wu, Qiong; Wang, You; Luo, Jun; Gou, Deming

    2013-01-01

    microRNAs (miRNAs) are non-coding small RNAs (sRNAs) capable of negatively regulating gene expression. Recently, microRNA-like small RNAs (milRNAs) were discovered in several filamentous fungi but not yet in Trichoderma reesei, an industrial filamentous fungus that can secrete abundant hydrolases. To explore the presence of milRNA in T. reesei and evaluate their expression under induction of cellulose, two T. reesei sRNA libraries of cellulose induction (IN) and non-induction (CON) were gener...

  3. The Polyketide Synthase Gene pks4 of Trichoderma reesei Provides Pigmentation and Stress Resistance

    OpenAIRE

    Atanasova, Lea; Knox, Benjamin P.; Kubicek, Christian P.; Druzhinina, Irina S.; Baker, Scott E.

    2013-01-01

    Species of the fungal genus Trichoderma (Hypocreales, Ascomycota) are well-known for their production of various secondary metabolites. Nonribosomal peptides and polyketides represent a major portion of these products. In a recent phylogenomic investigation of Trichoderma polyketide synthase (PKS)-encoding genes, the pks4 from T. reesei was shown to be an orthologue of pigment-forming PKSs involved in synthesis of aurofusarin and bikaverin in Fusarium spp. In this study, we show that deletion...

  4. Production of cellulase-free xylanase by Trichoderma reesei SAF3 Produção de xilanase livre de celulase por Trichoderma reesei SAF3

    OpenAIRE

    Sanjay Kar; Asish Mandal; Das Mohapatra, Pradeep K.; Mondal, Keshab C.; Bikash R. Pati

    2006-01-01

    A xylanase producing fungi has been isolated from soil and identified as Trichoderma reesei SAF3. Maximum growth of the organism was found at 48 h under submerged condition in xylan containing enriched medium, whereas highest enzyme production (4.75U/mL) was recorded at 72 h. No detectable cellulase activity was noted during whole cultivation period. The partially purified enzyme hydrolyzed xylan into xylopentose and xylose. All these properties of xylanase highlighten its promising uses in i...

  5. Ultrastructural features of the early secretory pathway in Trichoderma reesei.

    Science.gov (United States)

    Nykänen, Marko; Birch, Debra; Peterson, Robyn; Yu, Hong; Kautto, Liisa; Gryshyna, Anna; Te'o, Junior; Nevalainen, Helena

    2016-05-01

    We have systematically analysed the ultrastructure of the early secretory pathway in the Trichoderma reesei hyphae in the wild-type QM6a, cellulase-overexpressing Rut-C30 strain and a Rut-C30 transformant BV47 overexpressing a recombinant BiP1-VenusYFP fusion protein with an endoplasmic reticulum (ER) retention signal. The hyphae were studied after 24 h of growth using transmission electron microscopy, confocal microscopy and quantitative stereological techniques. All three strains exhibited different spatial organisation of the ER at 24 h in both a cellulase-inducing medium and a minimal medium containing glycerol as a carbon source (non-cellulase-inducing medium). The wild-type displayed a number of ER subdomains including parallel tubular/cisternal ER, ER whorls, ER-isolation membrane complexes with abundant autophagy vacuoles and dense bodies. Rut-C30 and its transformant BV47 overexpressing the BiP1-VenusYFP fusion protein also contained parallel tubular/cisternal ER, but no ER whorls; also, there were very few autophagy vacuoles and an increasing amount of punctate bodies where particularly the recombinant BiP1-VenusYFP fusion protein was localised. The early presence of distinct strain-specific features such as the dominance of ER whorls in the wild type and tub/cis ER in Rut-C30 suggests that these are inherent traits and not solely a result of cellular response mechanisms by the high secreting mutant to protein overload. PMID:26699139

  6. 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...

  7. 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...

  8. 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 in...

  9. 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...

  10. 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. To...

  11. 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...

  12. 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...

  13. PROTEIN EXPRESSION AND SECRETION BY TRICHODERMA REESEI UNDER LOW ENDOGENOUS PROTEIN BACKGROUND

    Science.gov (United States)

    Trichoderma reesei (Hypocrea jecorina) is one of the most commonly used fungi for the manufacturing of industrial enzyme products. The fungus is capable of secreting proteins in levels up to 100 grams per liter. A number of homologous and heterologous proteins have been successfully over-expressed...

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

    DEFF Research Database (Denmark)

    Rosgaard, Lisa; Pedersen, Sven; Langston, J; Akerhielm, D; Cherry, JR; Meyer, Anne S.

    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...... cellobiohydrolases (CBHs) and endo-1,4-beta-glucanases (EGs). Since the original T. reesei strain was isolated from decaying canvas, the T reesei CBH and EG activities might be present in suboptimal ratios for hydrolysis of pretreated lignocellulosic substrates. We employed statistically designed combinations of the...... cellulases secreted by T. reesei. Hence, the data indicate the feasibility of designing minimal enzyme mixtures for pretreated lignocellulosic biomass by careful combination of monocomponent enzymes. This strategy can promote both a more efficient enzymatic hydrolysis of (ligno)cellulose and a more rational...

  15. Understanding the Role of the Master Regulator XYR1 in Trichoderma reesei by Global Transcriptional Analysis

    OpenAIRE

    dos Santos Castro, Lilian; de Paula, Renato G.; Antoniêto, Amanda C. C.; Persinoti, Gabriela F.; Silva-Rocha, Rafael; Silva, Roberto N.

    2016-01-01

    We defined the role of the transcriptional factor—XYR1—in the filamentous fungus Trichoderma reesei during cellulosic material degradation. In this regard, we performed a global transcriptome analysis using RNA-Seq of the Δxyr1 mutant strain of T. reesei compared with the parental strain QM9414 grown in the presence of cellulose, sophorose, and glucose as sole carbon sources. We found that 5885 genes were expressed differentially under the three tested carbon sources. Of these, 322 genes were...

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

    Directory of Open Access Journals (Sweden)

    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.

  17. The cargo and the transport system: secreted proteins and protein secretion in Trichoderma reesei (Hypocrea jecorina).

    Science.gov (United States)

    Saloheimo, Markku; Pakula, Tiina M

    2012-01-01

    Trichoderma reesei (Hypocrea jecorina) is an efficient cell factory for protein production that is exploited by the enzyme industry. Yields of over 100 g secreted protein l(-1) from industrial fermentations have been reported. In this review we discuss the spectrum of proteins secreted by T. reesei and the studies carried out on its protein secretion system. The major enzymes secreted by T. reesei under production conditions are those degrading plant polysaccharides, the most dominant ones being the major cellulases, as demonstrated by the 2D gel analysis of the secretome. According to genome analysis, T. reesei has fewer genes encoding enzymes involved in plant biomass degradation compared with other fungi with sequenced genomes. We also discuss other T. reesei secreted enzymes and proteins that have been studied, such as proteases, laccase, tyrosinase and hydrophobins. Investigation of the T. reesei secretion pathway has included molecular characterization of the pathway components functioning at different stages of the secretion process as well as analysis of the stress responses caused by impaired folding or trafficking in the pathway or by expression of heterologous proteins. Studies on the transcriptional regulation of the secretory pathway have revealed similarities, but also interesting differences, with other organisms, such as a different induction mechanism of the unfolded protein response and the repression of genes encoding secreted proteins under secretion stress conditions. PMID:22053009

  18. 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.

  19. 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

  20. 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

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

    Science.gov (United States)

    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

  2. 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.

  3. 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...

  4. 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

    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...

  5. 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...

  6. Optimization of the production of trichoderma reesei endoglucanase I in pichia pastoris

    OpenAIRE

    Çalık, Aslı; Calik, Asli

    2010-01-01

    The aim of this study is to produce Trichoderma reesei endoglucanase I in Pichia pastoris. Pichia pastoris is the host of the production since it has many advantages like high level production of foreign proteins and possession of a tightly regulated promoter, alcohol oxidase I. Pichia cells expressing heterologous endoglucanase I was grown in fermenter with different strategies. The effects of temperature and methanol concentration on EG I production were tried to be understood. Especially t...

  7. Precipitation of Trichoderma reesei commercial cellulase preparations under standard enzymatic hydrolysis conditions for lignocelluloses

    OpenAIRE

    Chylenski, Piotr; Felby, Claus; Haven, Mai Østergaard; Gama, F. M.; Selig, Michael J

    2012-01-01

    Comparative studies between commercial Trichoderma reesei cellulase preparations show that, depending on the preparation and loading, total protein precipitation can be as high as 30 % under standard hydrolysis conditions used for lignocellulosic materials. ATR-IR and SDS-PAGE data verify precipitates are protein-based and contain key cell wall hydrolyzing enzymes. Precipitation increased considerably with incubation temperature; roughly 50–150 % increase from 40 to 50 °C and 800 % greater at...

  8. Enzymatic hydrolyzing performance of Acremonium cellulolyticus and Trichoderma reesei against three lignocellulosic materials

    OpenAIRE

    Murakami Katsuji; Inoue Hiroyuki; Fang Xu; Fujii Tatsuya; Sawayama Shigeki

    2009-01-01

    Abstract Background Bioethanol isolated from lignocellulosic biomass represents one of the most promising renewable and carbon neutral alternative liquid fuel sources. Enzymatic saccharification using cellulase has proven to be a useful method in the production of bioethanol. The filamentous fungi Acremonium cellulolyticus and Trichoderma reesei are known to be potential cellulase producers. In this study, we aimed to reveal the advantages and disadvantages of the cellulase enzymes derived fr...

  9. Regulation of genes encoding enzymes involved in plant cell wall deconstruction in Trichoderma reesei

    OpenAIRE

    Ries, Laure Nicolas Annick

    2013-01-01

    This study describes the regulation of genes encoding plant cell wall-degrading enzymes in the presence of different carbon sources from the biotechnologically important fungus Trichoderma reesei. It was shown that different carbon sources influence fungal growth rate, biomass production and subsequent enzyme secretion. Several genes were identified and suggested to play a role in the development of conidia and in maintaining polarised growth. RNA-sequencing studies showed an increase in t...

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

    OpenAIRE

    Viruthagiri, T.; Muthuvelayudham, R.; Saravanan, P.

    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, KH2PO4, and CoCl2 ·6H2O were selected based on their positive influence on cellulase production. The com...

  11. Exo and endo-glucanolytic activity of cellulases purified from Trichoderma reesei

    OpenAIRE

    Gama, F. M.; Vilanova, Manuel; Mota, M

    1998-01-01

    Four cellulases, produced by Trichoderma reesei, have been purified by preparative isoelectric focusing (Rotofor), size exclusion (Sephacryl 100 HR), anionic (Mono Q) and cationic (Mono S) chromatography and chromatofocusing (Mono P). Enzymatic activity with a large number of substrates allowed the proteins to be classified as: cellobiohydrolase I, cellobiohydrolase II, endoglucanase I and endoglucanase II. The exo- or endo- glucanase character of these enzymes was analysed by using a techniq...

  12. Enzyme production in immobilized Trichoderma reesei cells with hydrophobic polymers prepared by radiation polymerization method

    International Nuclear Information System (INIS)

    Trichoderma reesei cells were immobilized on paper covered with hydrophobic monomer, trimethylpropane triacrylate by radiation polymerization. The effect of immobilization condition on enzyme productivity was studied by measuring filter paper and cellobiose activity. The cells were adhered and grew on the surface of the carrier with the polymer giving high enzyme productivity in the immobilized cells in comparison with the free cells. Optimum concentration and volume of the coating monomer for the preparation of the immobilized cells were obtained. (author)

  13. Mechanism by which cellulose triggers cellobiohydrolase I gene expression in Trichoderma reesei.

    OpenAIRE

    el-Gogary, S; Leite, A; Crivellaro, O; Eveleigh, D E; el-Dorry, H

    1989-01-01

    The expression of cellobiohydrolase I mRNA from Trichoderma reesei, measured by Northern blot hybridization, is controlled by the nature of carbon sources used in the culture medium. Cellulose and the soluble disaccharide sophorose, but not glycerol or glucose, act as inducers. Cellobiohydrolase I mRNA was undetectable when antibodies to the major members of the cellulolytic system were present in the culture medium prior to the addition of cellulose. These antibodies had no repressive effect...

  14. Comparative secretome analyses of two Trichoderma reesei RUT-C30 and CL847 hypersecretory strains

    OpenAIRE

    Lignon Sabrina; Mollé Daniel; Jan Gwénaël; Dolla Alain; Margeot Antoine; Herpoël-Gimbert Isabelle; Mathis Hughes; Sigoillot Jean-Claude; Monot Frédéric; Asther Marcel

    2008-01-01

    Abstract Background Due to its capacity to produce large amounts of cellulases, Trichoderma reesei is increasingly been researched in various fields of white biotechnology, especially in biofuel production from lignocellulosic biomass. The commercial enzyme mixtures produced at industrial scales are not well characterized, and their proteinaceous components are poorly identified and quantified. The development of proteomic methods has made it possible to comprehensively overview the enzymes i...

  15. Effects of gamma-ray irradiation on cellulase secretion of Trichoderma reesei

    International Nuclear Information System (INIS)

    Trichoderma reesei was irradiated with gamma rays to investigate the effects of different dosages on cellulase production. Doses above 0.7 kGy induced cell lysis. Cell growth began to be obstructed at 2.0 kGy. As a result, the cells irradiated at 2.0 kGy secreted 1.8 times as much cellulase as the untreated cells

  16. Effect of Colloidal Materials on Cellulase Production by Trichoderma reesei Rut-C30

    OpenAIRE

    Duff, Sheldon J.B.; Cooper, David G.; Fuller, O. Maynard

    1985-01-01

    The addition of positively charged colloidal materials to the growth medium markedly increased the concentration of cellulase enzymes produced by Trichoderma reesei Rut-C30. Filter paper activities of up to 4 and 13 IU/ml have been achieved by the addition of colloidal materials, using 3% lactose and 3% cellulose, respectively, as a substrate. The particles exert their effect by binding soluble sugars and slowing their uptake by the organism.

  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.... reesei has been shown to produce a secondary metabolite known as paracelsin, which is a peptaibol... secondary metabolite known as paracelsin, which is associated with a variety of toxic effects to...

  18. Production of cellulase-free xylanase by Trichoderma reesei SAF3 Produção de xilanase livre de celulase por Trichoderma reesei SAF3

    Directory of Open Access Journals (Sweden)

    Sanjay Kar

    2006-12-01

    Full Text Available A xylanase producing fungi has been isolated from soil and identified as Trichoderma reesei SAF3. Maximum growth of the organism was found at 48 h under submerged condition in xylan containing enriched medium, whereas highest enzyme production (4.75U/mL was recorded at 72 h. No detectable cellulase activity was noted during whole cultivation period. The partially purified enzyme hydrolyzed xylan into xylopentose and xylose. All these properties of xylanase highlighten its promising uses in industrial scale.A partir de solo, isolou-se uma cepa de fungos produtos de xilanase, posteriormente identificado como Trichoderma reesei SAF3. O crescimento máximo do fungo foi obtido após 48h em condições submersas em meio de cultura contendo xilano, enquanto produção máxima de enzima (4,75U/mL ocorreu em 72h. Durante o período de cultivo, não foi detectada atividade celulásica. A enzima parcialmente purificada hidrolizou xilano a xilopentose e xilose. Essas propriedades da xilanase destacam seu uso promissor em escala industrial.

  19. Regulation of xyn3 gene expression in Trichoderma reesei PC-3-7.

    Science.gov (United States)

    Xu, J; Nogawa, M; Okada, H; Morikawa, Y

    2000-09-01

    The characteristics of regulation of the gene encoding the third xylanase (Xyn III) of a filamentous fungus, Trichoderma reesei PC-3-7, were studied by Northern blot analysis. A partial DNA sequence (185 bp) of the xyn3 gene was obtained by PCR amplification of genomic DNA of T. reesei PC-3-7 and sequenced. This xyn3 gene fragment was used as a probe for Southern and Northern blot analysis. The expression of the xyn3 gene was regulated at the transcriptional level. The xyn3 mRNA was expressed in mycelia of T. reesei PC-3-7 induced by Avicel, L-sorbose and sophorose, but not by xylose, xylooligosaccharides and birchwood xylan. Furthermore, it was observed that xyn3 was synchronously expressed with egll but not with xyn1 and xyn2 by L-sorbose, indicating that the xyn3 gene may be coordinately expressed with cellulase genes. By Southern blot analysis, the xyn3 gene was confirmed to exist as a single copy in both strains of T. reesei PC-3-7 and QM9414. However, no xyn3 mRNA appeared in the mycelia induced by any kind of inducers in T. reesei QM9414 even when total RNA was used in large excess, suggesting that the xyn3 gene in T. reesei QM9414 is in the dormant state and cannot be expressed. Therefore, T. reesei PC-3-7 may be a very useful strain for elucidating the induction mechanism of xylanase biosynthesis by cellulosic and xylanosic substrates, and also the regulatory correlation between cellulase and xylanase induction. PMID:11030574

  20. Enzymatic hydrolyzing performance of Acremonium cellulolyticus and Trichoderma reesei against three lignocellulosic materials

    Directory of Open Access Journals (Sweden)

    Murakami Katsuji

    2009-10-01

    Full Text Available Abstract Background Bioethanol isolated from lignocellulosic biomass represents one of the most promising renewable and carbon neutral alternative liquid fuel sources. Enzymatic saccharification using cellulase has proven to be a useful method in the production of bioethanol. The filamentous fungi Acremonium cellulolyticus and Trichoderma reesei are known to be potential cellulase producers. In this study, we aimed to reveal the advantages and disadvantages of the cellulase enzymes derived from these fungi. Results We compared A. cellulolyticus and T. reesei cellulase activity against the three lignocellulosic materials: eucalyptus, Douglas fir and rice straw. Saccharification analysis using the supernatant from each culture demonstrated that the enzyme mixture derived from A. cellulolyticus exhibited 2-fold and 16-fold increases in Filter Paper enzyme and β-glucosidase specific activities, respectively, compared with that derived from T. reesei. In addition, culture supernatant from A. cellulolyticus produced glucose more rapidly from the lignocellulosic materials. Meanwhile, culture supernatant derived from T. reesei exhibited a 2-fold higher xylan-hydrolyzing activity and produced more xylose from eucalyptus (72% yield and rice straw (43% yield. Although the commercial enzymes Acremonium cellulase (derived from A. cellulolyticus, Meiji Seika Co. demonstrated a slightly lower cellulase specific activity than Accellerase 1000 (derived from T. reesei, Genencor, the glucose yield (over 65% from lignocellulosic materials by Acremonium cellulase was higher than that of Accellerase 1000 (less than 60%. In addition, the mannan-hydrolyzing activity of Acremonium cellulase was 16-fold higher than that of Accellerase 1000, and the conversion of mannan to mannobiose and mannose by Acremonium cellulase was more efficient. Conclusion We investigated the hydrolysis of lignocellulosic materials by cellulase derived from two types of filamentous fungi. We

  1. Achieving efficient protein expression in Trichoderma reesei by using strong constitutive promoters

    Directory of Open Access Journals (Sweden)

    Li Junxin

    2012-06-01

    Full Text Available Abstract Backgrounds The fungus Trichoderma reesei is an important workhorse for expression of homologous or heterologous genes, and the inducible cbh1 promoter is generally used. However, constitutive expression is more preferable in some cases than inducible expression that leads to production of unwanted cellulase components. In this work, constitutive promoters of T. reesei were screened and successfully used for high level homologous expression of xylanase II. Results The transcriptional profiles of 13 key genes that participate in glucose metabolism in T. reesei were analyzed by quantitative real-time reverse-transcription polymerase chain reaction (RT-qPCR. The results indicated that the mRNA levels of pdc (encoding pyruvate decarboxylase and eno (encoding enolase genes were much higher than other genes under high glucose conditions. Recombinant T. reesei strains that homologously expressed xylanase II were constructed by using the promoters of the pdc and eno genes, and they respectively produced 9266 IU/ml and 8866 IU/ml of xylanase activities in the cultivation supernatant in a medium with high glucose concentration. The productivities of xylanase II were 1.61 g/L (with the pdc promoter and 1.52 g/L (with the eno promoter, approximately accounted for 83% and 82% of the total protein secreted by T. reesei, respectively. Conclusions This work demonstrates the screening of constitutive promoters by using RT-qPCR in T. reesei, and has obtained the highest expression of recombinant xylanase II to date by using these promoters.

  2. Exploring laccase-like multicopper oxidase genes from the ascomycete Trichoderma reesei: a functional, phylogenetic and evolutionary study

    OpenAIRE

    Levasseur Anthony; Saloheimo Markku; Navarro David; Andberg Martina; Pontarotti Pierre; Kruus Kristiina; Record Eric

    2010-01-01

    Abstract Background The diversity and function of ligninolytic genes in soil-inhabiting ascomycetes has not yet been elucidated, despite their possible role in plant litter decay processes. Among ascomycetes, Trichoderma reesei is a model organism of cellulose and hemicellulose degradation, used for its unique secretion ability especially for cellulase production. T. reesei has only been reported as a cellulolytic and hemicellulolytic organism although genome annotation revealed 6 laccase-lik...

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

    OpenAIRE

    Jun Huang; Dong Chen; Yutuo Wei; Qingyan Wang; Zhenchong Li; Ying Chen; Ribo Huang

    2014-01-01

    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...

  4. Systems Analysis of Lactose Metabolism in Trichoderma reesei Identifies a Lactose Permease That Is Essential for Cellulase Induction

    OpenAIRE

    Ivanova, Christa; Bååth, Jenny A.; Seiboth, Bernhard; Kubicek, Christian P.

    2013-01-01

    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...

  5. Glycoprotein Hypersecretion Alters the Cell Wall in Trichoderma reesei Strains Expressing the Saccharomyces cerevisiae Dolichylphosphate Mannose Synthase Gene▿

    OpenAIRE

    Perlińska-Lenart, Urszula; Orłowski, Jacek; Laudy, Agnieszka E.; Zdebska, Ewa; Palamarczyk, Grażyna; Kruszewska, Joanna S.

    2006-01-01

    Expression of the Saccharomyces cerevisiae DPM1 gene (coding for dolichylphosphate mannose synthase) in Trichoderma reesei (Hypocrea jecorina) increases the intensity of protein glycosylation and secretion and causes ultrastructural changes in the fungal cell wall. In the present work, we undertook further biochemical and morphological characterization of the DPM1-expressing T. reesei strains. We established that the carbohydrate composition of the fungal cell wall was altered with an increas...

  6. The VELVET A orthologue VEL1 of Trichoderma reesei regulates fungal development and is essential for cellulase gene expression

    OpenAIRE

    Karimi Aghcheh, Razieh; Németh Zoltán (1987-) (biomérnök); Atanasova, Lea; Fekete Erzsébet (1975-) (biotechnológus); Paholcsek Melinda (1984-) (molekuláris biológus, genetikus); Sándor Erzsébet; Aquino, Benigno; Druzhinina, Irina S.; Karaffa Levente; Kubicek, Christian P.

    2014-01-01

    Trichoderma reesei is the industrial producer of cellulases and hemicellulases for biorefinery processes. Their expression is obligatorily dependent on the function of the protein methyltransferase LAE1. The Aspergillus nidulans orthologue of LAE1 - LaeA - is part of the VELVET protein complex consisting of LaeA, VeA and VelB that regulates secondary metabolism and sexual as well as asexual reproduction. Here we have therefore investigated the function of VEL1, the T. reesei orthologue of A. ...

  7. 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

  8. Cloning and expression of Trichoderma reesei cellobiohydrolase I in Pichia pastoris

    Energy Technology Data Exchange (ETDEWEB)

    Godbole, S.; Decker, S.R.; Nieves, R.A.; Adney, W.S.; Vinzant, T.B.; Baker, J.O.; Thomas, S.R.; Himmel, M.E.

    1999-10-01

    Pichia pastoris was transformed with the Trichoderma reesei cbh1 gene, and the recombinant enzyme was purified and analyzed kinetically and by circular dichroism. The P. pastoris rCBH I was recognized by MoAb raised to T. reesei CBH I but was found in multiple molecular weight species on SDS-PAGE gels. Carbohydrate content determination and SDS-PAGE western analysis indicated that the recombinant protein was hyperglycosylated, although a species very similar in molecular weight to the T. reesei enzyme could be isolated chromatographically. The P. pastoris rCBH I also demonstrated activity toward soluble and insoluble substrates (i.e., pNPL and Sigmacell), although at a level significantly lower than the wild-type enzyme. More seriously, the yeast-expressed enzyme showed non-wild-type secondary structure by circular dichroism. The authors conclude that P. pastoris may not serve as an adequate host for the site-directed mutagenesis of T. reesei CBH I.

  9. 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.

  10. The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation

    Directory of Open Access Journals (Sweden)

    Seiboth Bernhard

    2011-05-01

    Full Text Available Abstract Background The identification and characterization of the transcriptional regulatory networks governing the physiology and adaptation of microbial cells is a key step in understanding their behaviour. One such wide-domain regulatory circuit, essential to all cells, is carbon catabolite repression (CCR: it allows the cell to prefer some carbon sources, whose assimilation is of high nutritional value, over less profitable ones. In lower multicellular fungi, the C2H2 zinc finger CreA/CRE1 protein has been shown to act as the transcriptional repressor in this process. However, the complete list of its gene targets is not known. Results Here, we deciphered the CRE1 regulatory range in the model cellulose and hemicellulose-degrading fungus Trichoderma reesei (anamorph of Hypocrea jecorina by profiling transcription in a wild-type and a delta-cre1 mutant strain on glucose at constant growth rates known to repress and de-repress CCR-affected genes. Analysis of genome-wide microarrays reveals 2.8% of transcripts whose expression was regulated in at least one of the four experimental conditions: 47.3% of which were repressed by CRE1, whereas 29.0% were actually induced by CRE1, and 17.2% only affected by the growth rate but CRE1 independent. Among CRE1 repressed transcripts, genes encoding unknown proteins and transport proteins were overrepresented. In addition, we found CRE1-repression of nitrogenous substances uptake, components of chromatin remodeling and the transcriptional mediator complex, as well as developmental processes. Conclusions Our study provides the first global insight into the molecular physiological response of a multicellular fungus to carbon catabolite regulation and identifies several not yet known targets in a growth-controlled environment.

  11. Crystallization and preliminary diffraction analysis of a β-galactosidase from Trichoderma reesei

    International Nuclear Information System (INIS)

    The crystallization and preliminary diffraction analysis of a β-galactosidase from T. reesei is described. The crystals diffracted to 1.2 Å resolution. An extracellular β-galactosidase from Trichoderma reesei was crystallized from sodium cacodylate buffer using polyethylene glycol (PEG) as a precipant. Crystals grown by homogenous streak-seeding belonged to space group P1, with unit-cell parameters a = 67.3, b = 69.1, c = 81.5 Å, α = 109.1, β = 97.3, γ = 114.5°. The crystals diffracted to 1.8 Å resolution using a rotating-anode generator and to 1.2 Å resolution using a synchrotron source. On the basis of the Matthews coefficient (VM = 3.16 Å3 Da−1), one molecule is estimated to be present in the asymmetric unit. The aim of the determination of the crystal structure is to increase the understanding of this industrially significant enzyme

  12. Trichoderma reesei FS10-C enhances phytoremediation of Cd-contaminated soil by Sedum plumbizincicola and associated soil microbial activities

    OpenAIRE

    Teng, Ying; Luo, Yang; Ma, Wenting; Zhu, Lingjia; Ren, Wenjie; Luo, Yongming; Christie, Peter; Li, Zhengao

    2015-01-01

    This study aimed to explore the effects of Trichoderma reesei FS10-C on the phytoremediation of Cd-contaminated soil by the hyperaccumulator Sedum plumbizincicola and on soil fertility. The Cd tolerance of T. reesei FS10-C was characterized and then a pot experiment was conducted to investigate the growth and Cd uptake of S. plumbizincicola with the addition of inoculation agents in the presence and absence of T. reesei FS10-C. The results indicated that FS10-C possessed high Cd resistance (u...

  13. Characterization of the pyruvate kinase-encoding gene (pki1) of Trichoderma reesei.

    Science.gov (United States)

    Schindler, M; Mach, R L; Vollenhofer, S K; Hodits, R; Gruber, F; Visser, J; De Graaff, L; Kubicek, C P

    1993-08-25

    The pyruvate kinase-encoding gene (pki1) from Trichoderma reesei was isolated by hybridization to the corresponding Aspergillus nidulans pkiA gene. The 1614-bp nucleotide (nt) sequence of the cloned gene codes for a 538-amino-acid protein. The coding sequence contains a single intron of 246 nt at a position identical to that of intron E in the A. nidulans gene. The PKI protein shows extensive homology to the PKIs of A. nidulans and A. niger (67%) and Saccharomyces cerevisiae (59%). The 5' non-coding sequence contains a number of motifs typical for yeast glycolytic genes, but so far only rarely found in filamentous fungi. PMID:8359694

  14. 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

  15. Functional analysis of Trichoderma reesei CKIIα2, a catalytic subunit of casein kinase II.

    Science.gov (United States)

    Wang, Mingyu; Yang, Hui; Zhang, Meiling; Liu, Kuimei; Wang, Hanbin; Luo, Yi; Fang, Xu

    2015-07-01

    Trichoderma reesei is the most important industrial cellulase-producing filamentous fungus. Although its molecular physiology has been investigated, the signal transduction pathways are not fully understood. In particular, the role of casein kinase II (CKII) is not yet clear. In this work, we carried out functional investigations on a catalytic subunit of CKII, CKIIα2. Comparison of the phenotypic features of T. reesei parent and Δck2α2 strains showed significant changes following ck2α2 disruption. T. reesei Δck2α2 form significantly smaller mycelial pellets in glucose-containing liquid minimum media, have shorter and fewer branch hyphae, produce smaller amounts of chitinases, produce more spores, show more robust growth on glucose-containing agar plates, and consume glucose at a significantly higher rate. Suggestions can be made that CKIIα2 governs chitinase expression, and the disruption of ck2α2 results in lower levels of chitinase production, leading to a weaker cell wall disruption capability, further resulting in weaker hyphal branching, which eventually leads to smaller mycelial pellets in liquid media. Further conclusions can be made that CKIIα2 is involved in repression of sporulation and glucose metabolism, which is consistent with the proposal that CKIIα2 represses global metabolism. These observations make the deletion of ck2α2 a potentially beneficial genetic disruption for T. reesei during industrial applications, as smaller mycelial pellets, more spores and more robust glucose metabolism are all desired traits for industrial fermentation. This work reports novel unique functions of a CKII catalytic subunit and is also the first genetic and physiological investigation on CKII in T. reesei. PMID:25833183

  16. 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

  17. 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...

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

    OpenAIRE

    Wan, Qun; Kovalevsky, Andrey; Zhang, Qiu; Hamilton-Brehm, Scott; Upton, Rosalynd; Weiss, Kevin L.; Mustyakimov, Marat; Graham, David; Coates, Leighton; Langan, Paul

    2013-01-01

    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.

  19. Regulation of the cellulolytic system in Trichoderma reesei by sophorose: induction of cellulase and repression of beta-glucosidase.

    OpenAIRE

    Sternberg, D; Mandels, G. R.

    1980-01-01

    Sophorose has two regulatory roles in the production of cellulase enzymes in Trichoderma reesei: beta-glucosidase repression and cellulase induction. Sophorose also is hydrolyzed by the mycelial-associated beta-glucosidase. Repression of beta-glucosidase reduces sophorose hydrolysis and thus may increase cellulase induction.

  20. 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. PMID:26319683

  1. Cellophane based mini-prep method for DNA extraction from the filamentous fungus Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    Henrique-Silva Flavio

    2002-06-01

    Full Text Available Abstract Background Methods for the extraction of DNA from filamentous fungi are frequently laborious and time consuming because most of the available protocols include maceration in liquid nitrogen after the mycelium has been grown in a liquid culture. This paper describes a new method to replace those steps, which involves the growth of the mycelium on cellophane disks overlaid on solid medium and the use of glass beads for cell wall disruption. Results Extractions carried out by this method provided approximately 2 μg of total DNA per cellophane disk for the filamentous fungus Trichoderma reesei. To assess the DNA's quality, we made a PCR (Polymerase Chain Reaction amplification of a gene introduced by a transformation in this fungus's genome (hph gene, with successful results. We also confirmed the quality of the DNA by the use of Southern blotting to analyze the presence of the same gene, which was easily detected, resulting in a sharply defined and strong band. Conclusions The use of this method enabled us to obtain pure DNA from Trichoderma reesei, dispensing with the laborious and time-consuming steps involved in most protocols. The DNA obtained was found to be suitable for PCR and Southern blot analyses. Another advantage of this method is the fact that several samples can be processed simultaneously, growing the fungus on multiple well cell culture plates. In addition, the absence of maceration also reduces sample handling, minimizing the risks of contamination, a particularly important factor in work involving PCR.

  2. Cellulase induction in Trichoderma reesei by cellulose requires its own basal expression.

    Science.gov (United States)

    Carle-Urioste, J C; Escobar-Vera, J; El-Gogary, S; Henrique-Silva, F; Torigoi, E; Crivellaro, O; Herrera-Estrella, A; El-Dorry, H

    1997-04-11

    The induction of cellulases by cellulose, an insoluble polymer, in the filamentous fungus Trichoderma reesei is puzzling. We previously proposed a mechanism that is based on the presence of low levels of cellulase in the uninduced fungus; this basal cellulase activity would digest cellulose-releasing oligosaccharides that could enter the cell and trigger expression of cellulases. We now present experiments that lend further support to this model. We show here that transcripts of two members of the cellulase system, cbh1 and egl1, are present in uninduced T. reesei cells. These transcripts are induced at least 1100-fold in the presence of cellulose. We also show that a construct containing the hygromycin B resistance-encoding gene driven by the cbh1 promoter confers hygromycin B resistance to T. reesei cells grown in the absence of cellulose. Moreover, cellulose-induced production of the cbh1 transcript was suppressed when antisense RNA against three members of the cellulase system was expressed in vivo. Experiments are presented indicating that extracellular cellulase activity is the rate-limiting event in induction of synthesis of the cellulase transcripts by cellulose. The results reveal a critical requirement for basal expression of the cellulase system for induction of synthesis of its own transcripts by cellulose. PMID:9092563

  3. 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

  4. Understanding the Role of the Master Regulator XYR1 in Trichoderma reesei by Global Transcriptional Analysis

    Science.gov (United States)

    dos Santos Castro, Lilian; de Paula, Renato G.; Antoniêto, Amanda C. C.; Persinoti, Gabriela F.; Silva-Rocha, Rafael; Silva, Roberto N.

    2016-01-01

    We defined the role of the transcriptional factor—XYR1—in the filamentous fungus Trichoderma reesei during cellulosic material degradation. In this regard, we performed a global transcriptome analysis using RNA-Seq of the Δxyr1 mutant strain of T. reesei compared with the parental strain QM9414 grown in the presence of cellulose, sophorose, and glucose as sole carbon sources. We found that 5885 genes were expressed differentially under the three tested carbon sources. Of these, 322 genes were upregulated in the presence of cellulose, while 367 and 188 were upregulated in sophorose and glucose, respectively. With respect to genes under the direct regulation of XYR1, 30 and 33 are exclusive to cellulose and sophorose, respectively. The most modulated genes in the Δxyr1 belong to Carbohydrate-Active Enzymes (CAZymes), transcription factors, and transporters families. Moreover, we highlight the downregulation of transporters belonging to the MFS and ABC transporter families. Of these, MFS members were mostly downregulated in the presence of cellulose. In sophorose and glucose, the expression of these transporters was mainly upregulated. Our results revealed that MFS and ABC transporters could be new players in cellulose degradation and their role was shown to be carbon source-dependent. Our findings contribute to a better understanding of the regulatory mechanisms of XYR1 to control cellulase gene expression in T. reesei in the presence of cellulosic material, thereby potentially enhancing its application in several biotechnology fields. PMID:26909077

  5. Heterologous Expression of an Alkali and Thermotolerant Lipase from Talaromyces thermophilus in Trichoderma reesei.

    Science.gov (United States)

    Zhang, Xu; Li, Xueqi; Xia, Liming

    2015-07-01

    To heterologously express a Talaromyces thermophilus lipase gene in Trichoderma reesei, an efficient binary vector pChph-pCBH1sigpro-ttl which includes a newly designed cbh1 promoter and hygromycin-resistant marker was constructed. This plasmid was then transformed into T. reesei via improved Agrobacterium EHA 105-mediated transformation. After modification of co-culture conditions and enzymolysis treatment of conidia, 258 transformants were produced. A two-step screening method based on antibiotic resistance and capacity to utilize lactose and tributyrin was introduced to further select promising candidates, which would be additionally verified by PCR analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and lipase activity assay. Lipase production was carried out in shaking flasks, and the activity reached 241 IU/mL (7415.4 IU/mg) after 84-h fermentation. It was found that this lipase performed high alkali and thermostable tolerance with the optimal pH 9.5 and temperature 60 °C, and it could retain more than 70 % activity after being disposed in pH 11 or 70 °C for 1 h. This study herein would benefit the genetic engineering of T. reesei and the industrial application of this important fungal lipase. PMID:26077681

  6. Purification and Characterization of β-1,3-Glucanase from the Antagonistic Fungus Trichoderma reesei

    Directory of Open Access Journals (Sweden)

    SRI WAHYUNI BUDIARTI

    2009-09-01

    Full Text Available Trichoderma enzymes that inhibit fungal cell walls have been suggested to play an important role in mycoparasitic action against fungal root rot pathogen Ganoderma philippii. This experiment was aimed to purify and characterize the β-1,3-glucanase of T. reesei. Extracellular β-1,3-glucanase was produced by growing mycoparasite T. reesei isolate T13 in colloidal chitin and sucrose as carbon sources. The enzyme was then purified to its homogeneity by precipitation with ammonium sulfate, followed by gel filtration chromatography and chromatofocusing. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE 12% was used to confirm the purity of enzyme at each stage of preparation and to characterize purified protein. The results showed that T. reesei produced at least three extracellular β-1,3-glucanases. Estimation of molecular weight based on SDS-PAGE 12% have three isoform of β-1,3-glucanase were 90 kDa for β-1,3-glucanase-I, 75 kDa for β-1,3-glucanase-II, and 64 kDa for β-1,3-glucanase-III. Their optimum pH and temperature were 5 and 50 oC, respectively.

  7. 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

    2016-03-01

    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. PMID:26540299

  8. Understanding the Role of the Master Regulator XYR1 in Trichoderma reesei by Global Transcriptional Analysis.

    Science.gov (United States)

    Dos Santos Castro, Lilian; de Paula, Renato G; Antoniêto, Amanda C C; Persinoti, Gabriela F; Silva-Rocha, Rafael; Silva, Roberto N

    2016-01-01

    We defined the role of the transcriptional factor-XYR1-in the filamentous fungus Trichoderma reesei during cellulosic material degradation. In this regard, we performed a global transcriptome analysis using RNA-Seq of the Δxyr1 mutant strain of T. reesei compared with the parental strain QM9414 grown in the presence of cellulose, sophorose, and glucose as sole carbon sources. We found that 5885 genes were expressed differentially under the three tested carbon sources. Of these, 322 genes were upregulated in the presence of cellulose, while 367 and 188 were upregulated in sophorose and glucose, respectively. With respect to genes under the direct regulation of XYR1, 30 and 33 are exclusive to cellulose and sophorose, respectively. The most modulated genes in the Δxyr1 belong to Carbohydrate-Active Enzymes (CAZymes), transcription factors, and transporters families. Moreover, we highlight the downregulation of transporters belonging to the MFS and ABC transporter families. Of these, MFS members were mostly downregulated in the presence of cellulose. In sophorose and glucose, the expression of these transporters was mainly upregulated. Our results revealed that MFS and ABC transporters could be new players in cellulose degradation and their role was shown to be carbon source-dependent. Our findings contribute to a better understanding of the regulatory mechanisms of XYR1 to control cellulase gene expression in T. reesei in the presence of cellulosic material, thereby potentially enhancing its application in several biotechnology fields. PMID:26909077

  9. Comparative secretome analyses of two Trichoderma reesei RUT-C30 and CL847 hypersecretory strains

    Directory of Open Access Journals (Sweden)

    Lignon Sabrina

    2008-12-01

    Full Text Available Abstract Background Due to its capacity to produce large amounts of cellulases, Trichoderma reesei is increasingly been researched in various fields of white biotechnology, especially in biofuel production from lignocellulosic biomass. The commercial enzyme mixtures produced at industrial scales are not well characterized, and their proteinaceous components are poorly identified and quantified. The development of proteomic methods has made it possible to comprehensively overview the enzymes involved in lignocellulosic biomass degradation which are secreted under various environmental conditions. Results The protein composition of the secretome produced by industrial T. reesei (strain CL847 grown on a medium promoting the production of both cellulases and hemicellulases was explored using two-dimensional electrophoresis and MALDI-TOF or LC-MS/MS protein identification. A total of 22 protein species were identified. As expected, most of them are potentially involved in biomass degradation. The 2D map obtained was then used to compare the secretomes produced by CL847 and another efficient cellulolytic T. reesei strain, Rut-C30, the reference cellulase-overproducing strain using lactose as carbon source and inducer of cellulases. Conclusion This study provides the most complete mapping of the proteins secreted by T. reesei to date. We report on the first use of proteomics to compare secretome composition between two cellulase-overproducing strains Rut-C30 and CL847 grown under similar conditions. Comparison of protein patterns in both strains highlighted many unexpected differences between cellulase cocktails. The results demonstrate that 2D electrophoresis is a promising tool for studying cellulase production profiles, whether for industrial characterization of an entire secretome or for a more fundamental study on cellulase expression at genome-wide scale.

  10. [Use of Endoglucanase IV from Trichoderma reesei to Enhance the Hydrolytic Activity of a Cellulase Complex from the Fungus Penicillium verruculosum].

    Science.gov (United States)

    Proskurina, O V; Korotkova, O G; Rozhkova, A M; Kondrat'eva, E G; Matys, V Yu; Zorov, I N; Koshelev, A V; Okunev, O N; Nemashkalov, V A; Bubnova, T V; Sinitsyn, A P

    2015-01-01

    The effect of polysaccharide monooxygenase (endoglucanase IV) from the fungus Trichoderma reesei on the hydrolysis of polysaccharide substrates by cellulases secreted by the fungus Penicillium verruculosum has been investigated. Supplementation of the enzyme complex from P. verruculosum by endoglucanase IV from T. reesei has been shown to elevate the efficiency of cellulose hydrolysis by 45%. PMID:26859961

  11. 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.

  12. 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

    OpenAIRE

    M Iyan Sofyan

    2004-01-01

    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 l...

  13. The Hypocrea jecorina (Trichoderma reesei hypercellulolytic mutant RUT C30 lacks a 85 kb (29 gene-encoding region of the wild-type genome

    Directory of Open Access Journals (Sweden)

    Hartl Lukas

    2008-07-01

    Full Text Available Abstract Background The hypercellulolytic mutant Hypocrea jecorina (anamorph Trichoderma reesei RUT C30 is the H. jecorina strain most frequently used for cellulase fermentations and has also often been employed for basic research on cellulase regulation. This strain has been reported to contain a truncated carbon catabolite repressor gene cre1 and is consequently carbon catabolite derepressed. To date this and an additional frame-shift mutation in the glycoprotein-processing β-glucosidase II encoding gene are the only known genetic differences in strain RUT C30. Results In the present paper we show that H. jecorina RUT C30 lacks an 85 kb genomic fragment, and consequently misses additional 29 genes comprising transcription factors, enzymes of the primary metabolism and transport proteins. This loss is already present in the ancestor of RUT C30 – NG 14 – and seems to have occurred in a palindromic AT-rich repeat (PATRR typically inducing chromosomal translocations, and is not linked to the cre1 locus. The mutation of the cre1 locus has specifically occurred in RUT C30. Some of the genes that are lacking in RUT C30 could be correlated with pronounced alterations in its phenotype, such as poor growth on α-linked oligo- and polyglucosides (loss of maltose permease, or disturbance of osmotic homeostasis. Conclusion Our data place a general caveat on the use of H. jecorina RUT C30 for further basic research.

  14. Small-Angle Neutron Scattering Reveals pH-Dependent Conformational Changes in Trichoderma reesei Cellobiohydrolase I: Implications for Enzymatic Activity

    Energy Technology Data Exchange (ETDEWEB)

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

    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 solutions having pH values of 7.0, 6.0, 5.3, and 4.2. As the pH decreases from 7.0 to 5.3, the enzyme structure remains well defined, possessing a spatial differentiation between the cellulose binding domain and the catalytic core that only changes subtly. At pH 4.2, the solution conformation of the enzyme changes to a structure that is intermediate between a properly folded enzyme and a denatured, unfolded state, yet the secondary structure of the enzyme is essentially unaltered. The results indicate that at the pH of optimal activity, the catalytic core of the enzyme adopts a structure in which the compact packing typical of a fully folded polypeptide chain is disrupted and suggest that the increased range of structures afforded by this disordered state plays an important role in the increased activity of Cel7A through conformational selection.

  15. 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.

  16. 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.

  17. Immobilization of trichoderma REESEI (QM 9414) cells with paper covered with ionic copolymer by radiation polymerization

    International Nuclear Information System (INIS)

    Cationic-hydrophobic copolymer and anionic-hydrophobic copolymer was covered onto surface of paper by radiation polymerization. The paper covered with ionic copolymer was used as carrier of immobilizing Trichoderma reesei cells. Results showed that the cells were immobilized firmly on the carriers and not dislocated from the carriers by shaking. All of FPA of the cells immobilized with the carriers covered with cationic copolymer were higher than that of un-immobilized free cells. The carriers covered with anionic copolymer showed good effect on immobilization of the cells. The weight of immobilized cells increase as increasing the component of DEAEMA in poly (DEAEMA-ATMPT) or decreasing the component of AA in poly (AA-ATMPT). It also increase with the increase of water absorption in poly (DEAEMA-ATMPT) or decrease of water absorption in poly (AA-ATMPT). It shows the static interaction play an important role in the immobilization of cells with ionic copolymer materials

  18. 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.

  19. 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. PMID:26672443

  20. Dose-dependency of radiation on enzyme production in Trichoderma reesei

    International Nuclear Information System (INIS)

    Effect of irradiation dose on the production of cellulase and amylase related enzymes in Trichoderma reesei was studied in which post-irradiation time response pattern was measured. The damage of the cells irradiated with certain irradiation doses (1.40±0.20x105, 2.20±0.10x105, 3.00±0.50x105 and 3.50±0.20x105 rad) was rapidly recovered. The increased enzyme production in the culture of the irradiated cells resulted from the recovery of radiation damage after irradiation. The function of cell growth was not affected by irradiation below dose of 5x105 rad, though the function of enzyme synthesis was drastically affected. (orig.)

  1. Morphological characterization and viability assessment of Trichoderma reesei by image analysis.

    Science.gov (United States)

    Lecault, Véronique; Patel, Nilesh; Thibault, Jules

    2007-01-01

    The production of cellulase from the filamentous fungus Trichoderma reesei is a critical step in the industrial process leading to cellulose ethanol. As a result of the lack of quantitative analysis tools, the intimate relationship that exists between the morphological and physiological states of the microorganism, the shear field in the bioreactor, and the process performance is not yet fully understood. A semiautomatic image analysis protocol was developed to characterize the mycelium morphology and to estimate its percentage viability during the fermentation process based on four morphological types (unbranched, branched, entangled, and clumped microorganisms). Pictures taken under bright field microscopy combined with images of fluorescein diacetate stained fungi were used to assess the morphological parameters and the percentage viability of microorganisms simultaneously. The method was tested during the course of fed-batch fermentation in a reciprocating plate bioreactor. The use of the image analysis protocol was found to be successful in quantifying the variations in the morphology and the viability of T. reesei throughout the fermentation. PMID:17373824

  2. 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

  3. 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

  4. New-to-nature sophorose analog: a potent inducer for gene expression in Trichoderma reesei.

    Science.gov (United States)

    Huang, Tom Tao; Wages, John M

    2016-04-01

    Controlled hydrolysis of lactonic sophorolipids from Starmerella bombicola yields a previously undescribed sophorose analog that potently induces cellulase in Trichoderma reesei Rut-C30. Acid treatment of natural sophorolipids results in a mixture of monoacetylated, deacetylated, and diacetylated sophorolipids in acidic and lactonic forms. Isolation of the active components of the mixture, followed by structure determination by MS and NMR, reveals a new chemical entity, in which the lactone ring has been opened at the C-1' rather than at the C-4″ position of the sophorose moiety. This sophorose ester is resistant to degradation by the host and is at least 28 times more powerful an inducer than sophorose in shake-flask culture. Even at low concentrations (0.05mM), the chemically modified sophorolipid effectively induces cellulase. With further improvements, this highly enabling technology can potentially reduce the cost of enzymes produced in T. reesei and can facilitate the rapid deployment of enzyme plants to support the nascent cellulosic biofuels and biochemicals industries. PMID:26920480

  5. Novel Strategies for Genomic Manipulation of Trichoderma reesei with the Purpose of Strain Engineering.

    Science.gov (United States)

    Derntl, Christian; Kiesenhofer, Daniel P; Mach, Robert L; Mach-Aigner, Astrid R

    2015-09-01

    The state-of-the-art procedure for gene insertions into Trichoderma reesei is a cotransformation of two plasmids, one bearing the gene of interest and the other a marker gene. This procedure yields up to 80% transformation efficiency, but both the number of integrated copies and the loci of insertion are unpredictable. This can lead to tremendous pleiotropic effects. This study describes the development of a novel transformation system for site-directed gene insertion based on auxotrophic markers. For this purpose, we tested the applicability of the genes asl1 (encoding an enzyme of the l-arginine biosynthesis pathway), the hah1 (encoding an enzyme of the l-lysine biosynthesis pathway), and the pyr4 (encoding an enzyme of the uridine biosynthesis pathway). The developed transformation system yields strains with an additional gene at a defined locus that are prototrophic and ostensibly isogenic compared to their parental strain. A positive transformation rate of 100% was achieved due to the developed split-marker system. Additionally, a double-auxotrophic strain that allows multiple genomic manipulations was constructed, which facilitates metabolic engineering purposes in T. reesei. By employing goxA of Aspergillus niger as a reporter system, the influence on the expression of an inserted gene caused by the orientation of the insertion and the transformation strategy used could be demonstrated. Both are important aspects to be considered during strain engineering. PMID:26150462

  6. 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. PMID:26481618

  7. 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. PMID:26293912

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

    OpenAIRE

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

    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 decre...

  9. 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 ...

  10. 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...

  11. Common features and interesting differences in transcriptional responses to secretion stress in the fungi Trichoderma reesei and Saccharomyces cerevisiae

    OpenAIRE

    Suortti Tapani; Valkonen Mari; Saloheimo Markku; Lanthaler Karin; Pakula Tiina; Arvas Mikko; Robson Geoff; Penttilä Merja

    2006-01-01

    Abstract Background Secretion stress is caused by compromised folding, modification or transport of proteins in the secretory pathway. In fungi, induction of genes in response to secretion stress is mediated mainly by the unfolded protein response (UPR) pathway. This study aims at uncovering transcriptional responses occurring in the filamentous fungi Trichoderma reesei exposed to secretion stress and comparing these to those found in the yeast Saccharomyces cerevisiae. Results Chemostat cult...

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

    OpenAIRE

    Tisch Doris; Kubicek Christian P; Schmoll Monika

    2011-01-01

    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 fo...

  13. Identification of residues important for substrate uptake in a glucose transporter from the filamentous fungus Trichoderma reesei

    OpenAIRE

    Weixin Zhang; Yanli Cao; Jing Gong; Xiaoming Bao; Guanjun Chen; Weifeng Liu

    2015-01-01

    The glucose transporter is an important player in cell metabolism that mediates the intracellular uptake of glucose. Here, we characterized the glucose transporter Stp1 from the filamentous fungus Trichoderma reesei. The individual substitution of several conserved residues for Ala in Stp1 corresponding to those interacting with D-glucose in the xylose/H+ symporter XylE inflicted contrasting effects on its ability to support the growth of an hxt-null yeast on glucose. The targeted change of P...

  14. Praperlakuan Fisik Dan Biologi Terhadap Biomassa Eceng Gondok Untuk Produksi Enzim Selulase Oleh Aspergillus Niger Dan Trichoderma Reesei

    OpenAIRE

    Amriani, Feni

    2015-01-01

    The Overgrowth of water hyacinth leads to eutrophication of water bodies as it exhausts nutrient and oxygen contents in water. However, it can be potentially used as lignocellulose biomass for cellulase production by several types of microbes such as Aspergillus niger and Trichoderma reesei. Physical pretreatment is conducted by size reduction of biomass and biological pretreatment by relying white rot fungus in which used to degrade lignin and improve accessibility of microbes to the cellulo...

  15. 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. PMID:26672453

  16. Array comparative genomic hybridization analysis of Trichoderma reesei strains with enhanced cellulase production properties

    Directory of Open Access Journals (Sweden)

    Penttilä Merja

    2010-07-01

    Full Text Available Abstract Background Trichoderma reesei is the main industrial producer of cellulases and hemicellulases that are used to depolymerize biomass in a variety of biotechnical applications. Many of the production strains currently in use have been generated by classical mutagenesis. In this study we characterized genomic alterations in high-producing mutants of T. reesei by high-resolution array comparative genomic hybridization (aCGH. Our aim was to obtain genome-wide information which could be utilized for better understanding of the mechanisms underlying efficient cellulase production, and would enable targeted genetic engineering for improved production of proteins in general. Results We carried out an aCGH analysis of four high-producing strains (QM9123, QM9414, NG14 and Rut-C30 using the natural isolate QM6a as a reference. In QM9123 and QM9414 we detected a total of 44 previously undocumented mutation sites including deletions, chromosomal translocation breakpoints and single nucleotide mutations. In NG14 and Rut-C30 we detected 126 mutations of which 17 were new mutations not documented previously. Among these new mutations are the first chromosomal translocation breakpoints identified in NG14 and Rut-C30. We studied the effects of two deletions identified in Rut-C30 (a deletion of 85 kb in the scaffold 15 and a deletion in a gene encoding a transcription factor on cellulase production by constructing knock-out strains in the QM6a background. Neither the 85 kb deletion nor the deletion of the transcription factor affected cellulase production. Conclusions aCGH analysis identified dozens of mutations in each strain analyzed. The resolution was at the level of single nucleotide mutation. High-density aCGH is a powerful tool for genome-wide analysis of organisms with small genomes e.g. fungi, especially in studies where a large set of interesting strains is analyzed.

  17. Overexpression of the Gene Encoding GTP:Mannose-1-Phosphate Guanyltransferase, mpg1, Increases Cellular GDP-Mannose Levels and Protein Mannosylation in Trichoderma reesei

    OpenAIRE

    Zakrzewska, Anna; Palamarczyk, Grazyna; Krotkiewski, Hubert; Zdebska, Ewa; Saloheimo, Markku; Penttilä, Merja; Kruszewska, Joanna S.

    2003-01-01

    To elucidate the regulation and limiting factors in the glycosylation of secreted proteins, the mpg1 and dpm1 genes from Trichoderma reesei (Hypocrea jecorina) encoding GTP:α-d-mannose-1-phosphate guanyltransferase and dolichyl phosphate mannose synthase (DPMS), respectively, were overexpressed in T. reesei. No significant increases were observed in DPMS activity or protein secretion in dpm1-overexpressing transformants, whereas overexpression of mpg1 led to a twofold increase in GDP-mannose ...

  18. 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.

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. Transcriptional profiling of biomass degradation-related genes during Trichoderma reesei growth on different carbon sources.

    Science.gov (United States)

    Chen, Xiuzhen; Luo, Yingfeng; Yu, Hongtao; Sun, Yuhui; Wu, Hong; Song, Shuhui; Hu, Songnian; Dong, Zhiyang

    2014-03-10

    To identify all the gene products involved in cellulosic biomass degradation, we employed RNA sequencing technology to perform a genome-wide comparison of gene expression during growth of Trichoderma reesei QM9414 on cellulose or glucose. Due to their important role in lignocellulose decomposition, we focused on CAZymes and other secreted proteins. In total, 122 CAZymes showed at least a two-fold change in mRNA abundance, and 97 of those were highly induced by cellulose. Compared to the well-characterized cellulases and hemicellulases, a majority of the other upregulated CAZymes showed lower transcriptional levels. In addition, 64 secreted proteins, including oxidoreductases, exhibited at least two-fold upregulation on cellulose medium. To better understand the potential roles of low-abundance CAZymes in cellulose breakdown, we compared the expression patterns of 25 glycoside hydrolase genes under different conditions via real-time PCR. Substantial differences for the 25 genes were observed for individual strains grown on different carbon sources, and between QM9414 and RUTC30 when grown on the same carbon source. Moreover, we identified 3 genes that are coregulated with known cellulases. Collectively, this study highlights a comprehensive transcriptional profile for biomass degradation-related proteins and provides a first step toward the identification of candidates to construct optimized enzyme cocktails. PMID:24445169

  5. Effect of culture medium composition on Trichoderma reesei's morphology and cellulase production.

    Science.gov (United States)

    Ahamed, Aftab; Vermette, Patrick

    2009-12-01

    The objective of this study was to determine how fungal morphology influences the volumetric cellulase productivity of Trichoderma reesei cultured in four media with lactose and lactobionic acid as fed-batch in a 7 L stirred tank bioreactor. The use of a cellulose-yeast extract culture medium yielded the highest enzyme production with a volumetric enzyme activity of 69.8 U L(-1) h(-1), and a maximum fungal biomass of 14.7 g L(-1). These findings were associated with the following morphological characteristics of the fungus: total mycelia was 98% of total mean projected area, mean hyphae length of 10 mm, mean hyphae volume of 45.1 mm(3), mean hyphae diameter of 7.9 microm, number of branches 9, and number of tips per hypha 29. A positive correlation was found between the total mycelia, the number of tips and the volumetric enzyme productivity, indicating the weight of these variables on the enzyme productivity. PMID:19592237

  6. New polygalacturonases from Trichoderma reesei: characterization and their specificities to partially methylated and acetylated pectins.

    Science.gov (United States)

    Mohamed, Saleh A; Christensen, Tove M I E; Mikkelsen, Jorn Dalgaard

    2003-03-14

    Two extracellular isoenzymes of polygalacturonases PG1 and PG2 were isolated from 3-day-old culture filtrates of Trichoderma reesei. The two enzymes were purified to homogeneity by ion-exchange, gel filtration and hydrophobic interaction chromatographies. PG1 and PG2 exhibit similar molecular weights from gel filtration and SDS-PAGE. Their properties, including optimal pH and temperature, thermal stability and Km were compared. Characterization of substrate specificity showed that the two enzymes had higher affinity toward PGA (B0100) derived from sugar beet pectin (SBP) than PGA from lime pectin. A series of SBPs with different distribution patterns of methyl and acetyl groups, produced by treatment with either plant pectin methylesterase (P-series) or fungal pectin methylesterase (F-series) or base catalysis (B-series), was used as substrates for PG1 and PG2. Substrates with a low degree of esterification were preferred substrates. The activities of PG1 and PG2 were strongly correlated to the degree of methylation and very little effect from acetylation. The products generated by digestion of selected lime and SBPs were analysed using matrix assisted laser desorption ionisation time of flight (MALDI TOF) MS. A mode of action revealed a random cleavage pattern for PG1 and PG2, confirming that these enzymes are endopolygalacturonases. PMID:12668107

  7. Modulating the thermostability of Endoglucanase I from Trichoderma reesei using computational approaches.

    Science.gov (United States)

    Bayram Akcapinar, Gunseli; Venturini, Alessandro; Martelli, Pier Luigi; Casadio, Rita; Sezerman, Ugur O

    2015-05-01

    In the last decades, effective cellulose degradation became a major point of interest due to the properties of cellulose as a renewable energy source and the widespread application of cellulases (the cellulose degrading enzymes) in many industrial processes. Effective bioconversion of lignocellulosic biomass into soluble sugars for ethanol production requires use of thermostable and highly active cellulases. The library of current cellulases includes enzymes that can work at acidic and neutral pH in a wide temperature range. However, only few cellulases are reported to be thermostable. In order to alleviate this, we have performed a hybrid approach for the thermostabilization of a key cellulase, Endoglucanase I (EGI) from Trichoderma reesei. We combined in silico and in vitro experiments to modulate the thermostability of EGI. Four different predictive algorithms were used to set up a library of mutations. Three thermostabilizer mutations (Q126F, K272F, Q274V) were selected and molecular dynamics simulations at room temperature and high temperatures were performed to analyze the effect of the mutations on enzyme structure and stability. The mutations were then introduced into the endoglucanase 1 gene, using site-directed mutagenesis, and the effect of the mutations on enzyme structure and stability were determined. MD simulations supported the fact that Q126F, K272F and Q274V mutations have a thermostabilizing effect on the protein structure. Experimental studies validated that all of the mutants exhibited higher thermostability compared with native EGI albeit with a decrease in specific activity. PMID:25784767

  8. 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.

  9. 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.

  10. Physiological evaluation of the filamentous fungus Trichoderma reesei in production processes by marker gene expression analysis

    Directory of Open Access Journals (Sweden)

    Penttilä Merja

    2007-05-01

    Full Text Available Abstract Background Biologically relevant molecular markers can be used in evaluation of the physiological state of an organism in biotechnical processes. We monitored at high frequency the expression of 34 marker genes in batch, fed-batch and continuous cultures of the filamentous fungus Trichoderma reesei by the transcriptional analysis method TRAC (TRanscript analysis with the aid of Affinity Capture. Expression of specific genes was normalised either with respect to biomass or to overall polyA RNA concentration. Expressional variation of the genes involved in various process relevant cellular functions, such as protein production, growth and stress responses, was related to process parameters such as specific growth and production rates and substrate and dissolved oxygen concentrations. Results Gene expression of secreted cellulases and recombinant Melanocarpus albomyces laccase predicted the trends in the corresponding extracellular enzyme production rates and was highest in a narrow "physiological window" in the specific growth rate (μ range of 0.03 – 0.05 h-1. Expression of ribosomal protein mRNAs was consistent with the changes in μ. Nine starvation-related genes were found as potential markers for detection of insufficient substrate feed for maintaining optimal protein production. For two genes induced in anaerobic conditions, increasing transcript levels were measured as dissolved oxygen decreased. Conclusion The data obtained by TRAC supported the usefulness of focused and intensive transcriptional analysis in monitoring of biotechnical processes providing thus tools for process optimisation purposes.

  11. 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

  12. Folding and stability of endoglucanase III, a single-domain cellulase from Trichoderma reesei.

    Science.gov (United States)

    Arunachalam, U; Kellis, J T

    1996-09-01

    The reversible folding of an endoglucanase (EGIII) from the filamentous fungus Trichoderma reesei was investigated by activity, tryptophan fluorescence, and peptide CD measurements. Equilibrium stability was determined by urea denaturation at various pH and temperature values. Unfolding and refolding rates were measured over a range of urea concentrations. The data from the equilibrium and kinetic studies fit a simple two-state model, except at lower urea concentrations, where the folding kinetics indicate a transient intermediate. Unfolding is very slow, with a half-life of about 2 h in 8 M urea at pH 5.5 and 25 degrees C. Comparison of the urea dependence of the folding kinetics and equilibrium indicates the protein undergoes 93% of its total change in solvent exposure on going from the unfolded state to the transition state. Thus, the transition state is quite compact. The presence of dithiothreitol destabilized the protein by 7 kcal/mol, indicating the presence of an unusually strong disulfide linkage between the two cysteines in the molecule. Protein stability is dramatically reduced at alkaline pH values; this can be attributed to a titratable shift (pKa = 7.8) in the slope of the urea dependence of unfolding. PMID:8784193

  13. Characterization of the Dielectric Constant in the Trichoderma reesei Cel7B Active Site.

    Science.gov (United States)

    Song, Xiangfei; Wang, Yefei; Zhang, Shujun; Yan, Shihai; Li, Tong; Yao, Lishan

    2015-07-27

    An attempt is made to evaluate the dielectric constant of the Trichoderma reesei Cel7B active site. Through kinetic measurements, the pKa value of the catalytic acid E201 is determined. Mutations (away from E201) with net charge changes are introduced to perturb the E201 pKa. It is shown that the mutation with a +1 charge change (including G225R, G230R, and A335R) decreases the pKa of E201, whereas the mutation with a -1 charge change (including Q149E, A222D, G225D, and G230D) increases the pKa. This effect is consistent with the electrostatic interaction between the changed charge and the E201 side chain. The fitting of the experimental data yields an apparent dielectric constant of 25-80. Molecular dynamics simulations with explicit water molecules indicate that the high solvent accessibility of the active site contributes largely to the high dielectric constant. ONIOM calculations show that high dielectric constant benefits the catalysis through decreasing the energy of the transition state relative to that of the enzyme substrate complex. PMID:26114648

  14. 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.

  15. 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...

  16. Isolation and Partial Purification of Extracellular Enzyme (1, 3)-β-D Glucanase from Trichoderma reesei (3929)

    OpenAIRE

    R. Saravanan; V. Pavani Devi; Shanmugam, A.; D. Sathish Kumar

    2007-01-01

    The fungus Trichoderma reesei produces extracellular lytic enzymes such as β-1, 4 glucanases, β-1, 6 glucanases and β-1, 3 D glucanases. In the present study the PDA medium was used for the production of β-1, 3 glucanases from the fungal strain. Extra cellular protein (β-1, 3 glucanase) was partially purified by ammonium sulphate precipitation and dialysis. The amount of reducing sugar (126 mg/100 mL) liberated by the action of enzyme was determined by using DNS method. The molecular weight o...

  17. From in silico to in vitro: modelling and production of trichoderma reesei endoglucanase 1 and its mutant in pichia pastoris

    OpenAIRE

    Bayram Akçapınar, Günseli; Bayram Akcapinar, Gunseli; Gül, Özgür; Gul, Ozgur; Sezerman, Uğur; Sezerman, Ugur

    2011-01-01

    In this study, a major cellulase, namely endoglucanase 1 (EGI) from Trichoderma reesei was mutated by the introduction of four different Lysine and Glycine rich loops to create a hotspot for directed crosslinking of EGI away from the active site. The impact of the inserted loops on the stability of the enzyme was analyzed using Molecular Dynamics (MD) and the effect on the active site was studied using Molecular Mechanics (MM) simulations. The best loop mutation predicted in silico (EGI_L5) w...

  18. Cloning and expression in Saccharomyces cerevisiae of a Trichoderma reesei beta-mannanase gene containing a cellulose binding domain.

    OpenAIRE

    Stålbrand, H; Saloheimo, A; Vehmaanperä, J; HENRISSAT, B.; Penttilä, M

    1995-01-01

    beta-Mannanase (endo-1,4-beta-mannanase; mannan endo-1,4-beta-mannosidase; EC 3.2.1.78) catalyzes endo-wise hydrolysis of the backbone of mannan and heteromannans, including hemicellulose polysaccharides, which are among the major components of plant cell walls. The gene man1, which encodes beta-mannanase, of the filamentous fungus Trichoderma reesei was isolated from an expression library by using antiserum raised towards the earlier-purified beta-mannanase protein. The deduced beta-mannanas...

  19. Enhancing saccharification of wheat straw by mixing enzymes from genetically-modified Trichoderma reesei and Aspergillus niger

    OpenAIRE

    Jiang, Yanping; Duarte, Alexandra Vivas; van den Brink, Joost; Wiebenga, Ad; Zou, Gen; Wang, Chengshu; De Vries, Ronald P; Zhou, Zhihua; Benoit, Isabelle

    2015-01-01

    Objectives To increase the efficiency of enzymatic hydrolysis for plant biomass conversion into renewable biofuel and chemicals. Results By overexpressing the point mutation A824 V transcriptional activator Xyr1 in Trichoderma reesei, carboxymethyl cellulase, cellobiosidase and β-d-glucosidase activities of the best mutant were increased from 1.8 IU/ml, 0.1 IU/ml and 0.05 IU/ml to 4.8 IU/ml, 0.4 IU/ml and 0.3 IU/ml, respectively. The sugar yield of wheat straw saccharification by combining en...

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

    OpenAIRE

    Gloria López; Pilar Estrada

    2014-01-01

    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 calorime...

  1. 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

  2. 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

  3. Regulation of cellulase gene expression in the filamentous fungus Trichoderma reesei.

    Science.gov (United States)

    Ilmén, M; Saloheimo, A; Onnela, M L; Penttilä, M E

    1997-04-01

    Basic features of regulation of expression of the genes encoding the cellulases of the filamentous fungus Trichoderma reesei QM9414, the genes cbh1 and cbh2 encoding cellobiohydrolases and the genes egl1, egl2 and egl5 encoding endoglucanases, were studied at the mRNA level. The cellulase genes were coordinately expressed under all conditions studied, with the steady-state mRNA levels of cbh1 being the highest. Solka floc cellulose and the disaccharide sophorose induced expression to almost the same level. Moderate expression was observed when cellobiose or lactose was used as the carbon source. It was found that glycerol and sorbitol do not promote expression but, unlike glucose, do not inhibit it either, because the addition of 1 to 2 mM sophorose to glycerol or sorbitol cultures provokes high cellulase expression levels. These carbon sources thus provide a useful means to study cellulase regulation without significantly affecting the growth of the fungus. RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose. The results clearly show that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus. However, derepression of cellulase expression occurs without apparent addition of an inducer once glucose has been depleted from the medium. This expression seems not to arise simply from starvation, since the lack of carbon or nitrogen as such is not sufficient to trigger significant expression. PMID:9097427

  4. 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 at...

  5. 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.

  6. 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)

  7. 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.

  8. 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.

  9. Swollenin, a Trichoderma reesei protein with sequence similarity to the plant expansins, exhibits disruption activity on cellulosic materials.

    Science.gov (United States)

    Saloheimo, Markku; Paloheimo, Marja; Hakola, Satu; Pere, Jaakko; Swanson, Barbara; Nyyssönen, Eini; Bhatia, Amit; Ward, Michael; Penttilä, Merja

    2002-09-01

    Plant cell wall proteins called expansins are thought to disrupt hydrogen bonding between cell wall polysaccharides without hydrolyzing them. We describe here a novel gene with sequence similarity to plant expansins, isolated from the cellulolytic fungus Trichoderma reesei. The protein named swollenin has an N-terminal fungal type cellulose binding domain connected by a linker region to the expansin-like domain. The protein also contains regions similar to mammalian fibronectin type III repeats, found for the first time in a fungal protein. The swollenin gene is regulated in a largely similar manner as the T. reesei cellulase genes. The biological role of SWOI was studied by disrupting the swo1 gene from T. reesei. The disruption had no apparent effect on the growth rate on glucose or on different cellulosic carbon sources. Non-stringent Southern hybridization of Trichoderma genomic DNA with swo1 showed the presence of other swollenin-like genes, which could substitute for the loss of SWOI in the disruptant. The swollenin gene was expressed in yeast and Aspergillus niger var. awamori. Activity assays on cotton fibers and filter paper were performed with concentrated SWOI-containing yeast supernatant that disrupted the structure of the cotton fibers without detectable formation of reducing sugars. It also weakened filter paper as assayed by an extensometer. The SWOI protein was purified from A. niger var. awamori culture supernatant and used in an activity assay with Valonia cell walls. It disrupted the structure of the cell walls without producing detectable amounts of reducing sugars. PMID:12199698

  10. Common features and interesting differences in transcriptional responses to secretion stress in the fungi Trichoderma reesei and Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Suortti Tapani

    2006-02-01

    Full Text Available Abstract Background Secretion stress is caused by compromised folding, modification or transport of proteins in the secretory pathway. In fungi, induction of genes in response to secretion stress is mediated mainly by the unfolded protein response (UPR pathway. This study aims at uncovering transcriptional responses occurring in the filamentous fungi Trichoderma reesei exposed to secretion stress and comparing these to those found in the yeast Saccharomyces cerevisiae. Results Chemostat cultures of T. reesei expressing human tissue plasminogen activator (tPA and batch bioreactor cultures treated with dithiothreitol (DTT to prevent correct protein folding were analysed with cDNA subtraction and cDNA-amplified fragment length polymorphism (AFLP experiments. ESTs corresponding to 457 unique genes putatively induced under secretion stress were isolated and the expression pattern of 60 genes was confirmed by Northern analysis. Expression of these genes was also studied in a strain over-expressing inositol-requiring enzyme 1 (IREI protein, a sensor for the UPR pathway. To compare the data with that of S. cerevisiae, published transcriptome profiling data on various stress responses in S. cerevisiae was reanalysed. The genes up-regulated in response to secretion stress included a large number of secretion related genes in both organisms. In addition, analysis of T. reesei revealed up regulation of the cpc1 transcription factor gene and nucleosomal genes. The induction of the cpcA and histone gene H4 were shown to be induced also in cultures of Aspergillus nidulans treated with DTT. Conclusion Analysis of the genes induced under secretion stress has revealed novel features in the stress response in T. reesei and in filamentous fungi. We have demonstrated that in addition to the previously rather well characterised induction of genes for many ER proteins or secretion related proteins also other types of responses exist.

  11. 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

  12. Characterization of Secretory Genes ypt1/yptA and nsf1/nsfA from Two Filamentous Fungi: Induction of Secretory Pathway Genes of Trichoderma reesei under Secretion Stress Conditions

    OpenAIRE

    Saloheimo, Markku; Wang, Huaming; Valkonen, Mari; Vasara, Tuija; Huuskonen, Anne; Riikonen, Marjukka; Pakula, Tiina; Ward, Michael; Penttilä, Merja

    2004-01-01

    Two genes involved in protein secretion, encoding the Rab protein YPT1/YPTA and the general fusion factor NSFI/NSFA, were characterized from two filamentous fungi, Trichoderma reesei and Aspergillus niger var. awamori. The isolated genes showed a high level of conservation with their Saccharomyces cerevisiae and mammalian counterparts, and T. reesei ypt1 was shown to complement yeast Ypt1p depletion. The transcriptional regulation of the T. reesei ypt1, nsf1, and sar1 genes, involved in prote...

  13. 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

  14. 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.

  15. Exploring laccase-like multicopper oxidase genes from the ascomycete Trichoderma reesei: a functional, phylogenetic and evolutionary study

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    Levasseur Anthony

    2010-08-01

    Full Text Available Abstract Background The diversity and function of ligninolytic genes in soil-inhabiting ascomycetes has not yet been elucidated, despite their possible role in plant litter decay processes. Among ascomycetes, Trichoderma reesei is a model organism of cellulose and hemicellulose degradation, used for its unique secretion ability especially for cellulase production. T. reesei has only been reported as a cellulolytic and hemicellulolytic organism although genome annotation revealed 6 laccase-like multicopper oxidase (LMCO genes. The purpose of this work was i to validate the function of a candidate LMCO gene from T. reesei, and ii to reconstruct LMCO phylogeny and perform evolutionary analysis testing for positive selection. Results After homologous overproduction of a candidate LMCO gene, extracellular laccase activity was detected when ABTS or SRG were used as substrates, and the recombinant protein was purified to homogeneity followed by biochemical characterization. The recombinant protein, called TrLAC1, has a molecular mass of 104 kDa. Optimal temperature and pH were respectively 40-45°C and 4, by using ABTS as substrate. TrLAC1 showed broad pH stability range of 3 to 7. Temperature stability revealed that TrLAC1 is not a thermostable enzyme, which was also confirmed by unfolding studies monitored by circular dichroism. Evolutionary studies were performed to shed light on the LMCO family, and the phylogenetic tree was reconstructed using maximum-likelihood method. LMCO and classical laccases were clearly divided into two distinct groups. Finally, Darwinian selection was tested, and the results showed that positive selection drove the evolution of sequences leading to well-known laccases involved in ligninolysis. Positively-selected sites were observed that could be used as targets for mutagenesis and functional studies between classical laccases and LMCO from T. reesei. Conclusions Homologous production and evolutionary studies of the first

  16. Re-annotation of the CAZy genes of Trichoderma reesei and transcription in the presence of lignocellulosic substrates

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    Häkkinen Mari

    2012-10-01

    Full Text Available Abstract Background Trichoderma reesei is a soft rot Ascomycota fungus utilised for industrial production of secreted enzymes, especially lignocellulose degrading enzymes. About 30 carbohydrate active enzymes (CAZymes of T. reesei have been biochemically characterised. Genome sequencing has revealed a large number of novel candidates for CAZymes, thus increasing the potential for identification of enzymes with novel activities and properties. Plenty of data exists on the carbon source dependent regulation of the characterised hydrolytic genes. However, information on the expression of the novel CAZyme genes, especially on complex biomass material, is very limited. Results In this study, the CAZyme gene content of the T. reesei genome was updated and the annotations of the genes refined using both computational and manual approaches. Phylogenetic analysis was done to assist the annotation and to identify functionally diversified CAZymes. The analyses identified 201 glycoside hydrolase genes, 22 carbohydrate esterase genes and five polysaccharide lyase genes. Updated or novel functional predictions were assigned to 44 genes, and the phylogenetic analysis indicated further functional diversification within enzyme families or groups of enzymes. GH3 β-glucosidases, GH27 α-galactosidases and GH18 chitinases were especially functionally diverse. The expression of the lignocellulose degrading enzyme system of T. reesei was studied by cultivating the fungus in the presence of different inducing substrates and by subjecting the cultures to transcriptional profiling. The substrates included both defined and complex lignocellulose related materials, such as pretreated bagasse, wheat straw, spruce, xylan, Avicel cellulose and sophorose. The analysis revealed co-regulated groups of CAZyme genes, such as genes induced in all the conditions studied and also genes induced preferentially by a certain set of substrates. Conclusions In this study, the CAZyme

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

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    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

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

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    Piotr Janas

    2014-08-01

    Full Text Available 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 increase in the level of xylanolytic enzymes produced by the four mutants. In addition. it effected the complex composition of cellulolytic enzymes secreted by VTT-D-79l 24 (i.c. increased and reduced excertion of (β-glucosidase and β-1,4-endoglucanase respectively.

  19. Preparative purification of Trichoderma reesei native and {open_quotes}core{close_quotes} cellobiohydrolase I by electrophoresis and chromatofocusing

    Energy Technology Data Exchange (ETDEWEB)

    Offord, D.A.; Lee, N.E.; Woodward, J. [Oak Ridge National Lab., TN (United States)

    1991-12-31

    The enzymes present in the cellulose complex produced by the fungus Trichoderma reesei have been the subject of considerable attention in consequence of their potential for converting cellulosic materials into glucose for further use in fermentation processes. Cellobiohydrolase I (CBH I) is the major component of crude commercial fungal cellulose preparations and catalyzes the conversion of insoluble cellulose into cellobiose. The primary structure of CBH I is known, and its tertiary structure, deduced from small-angle X-ray scattering studies, takes the shape of a tadpole with a catalytic head region known as {open_quotes}core{close_quotes} CBH I and a C-terminal cellulose-binding tail region. Removal of the tail can be accomplished with the protease papain, resulting in reduced activity towards insoluble substrates but unchanged activity towards soluble substrates.

  20. Preparative purification of Trichoderma reesei native and core'' Cellobiohydrolase I by electrophoresis and chromatofocussing

    Energy Technology Data Exchange (ETDEWEB)

    Offord, D.A.; Lee, N.E.; Woodward, J.

    1990-01-01

    The enzymes present in the cellulase complex produced by the fungus Trichoderma reesei have been the subject of considerable attention due to their potential for converting cellulosic materials into glucose for further use in fermentation processes. Cellobiohydrolase I (CBH I) is the major component of crude commercial fungal cellulase preparations and catalyzes the conversion of insoluble cellulose into cellobiose. We have been interested, recently, in the reduction of native and core'' CBH I (the primary structure of CBH I with a catalytic head region) and have needed to develop a method for their preparative purification. We now report that by using electrophoresis and chromatofocussing, preparative quantities of both native and core'' CBH I have been obtained. Since their pI values are different, milligram quantities of core'' CBH I can be generated and purified from the native enzyme by chromatofocussing within 2 hours. 13 refs., 7 figs., 2 tabs.

  1. Papain digestion of crude Trichoderma reesei cellulase: Purification and properties of cellobiohydrolase I and II core proteins

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J.; Brown, J.P.; Evans, B.R.; Affholter, K.A.

    1992-01-01

    Papain digestion of a crude Trichoderma reesei cellulose preparation followed by gel filtration on a Superdex column resulted in the separation of cellobiohydrolase (CBH) I and II core proteins (cp). They were further purified to apparent homogeneity by chromatofocusing. N-terminal protein sequencing of the CBH II cp preparation confirmed its identity. A comparison of the catalytic activity and cellulose-binding ability of these core proteins was made. The major differences between them were the findings that CBH II cp possessed a sixfold higher specific activity toward p-nitrophenylcellobioside than the native CBH II preparation and still bound to microcrystalline cellulose, unlike CBH I cp. Neither CBH I cp nor CBH II cp had activity toward carboxymethylcellulose, but both were able to hydrolyze barley b-glucan. These data suggest that removal of the cellulose-binding domain and hinge region from CBH I and II have different effects on their properties.

  2. Papain digestion of crude Trichoderma reesei cellulase: Purification and properties of cellobiohydrolase I and II core proteins

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J.; Brown, J.P.; Evans, B.R.; Affholter, K.A.

    1992-12-01

    Papain digestion of a crude Trichoderma reesei cellulose preparation followed by gel filtration on a Superdex column resulted in the separation of cellobiohydrolase (CBH) I and II core proteins (cp). They were further purified to apparent homogeneity by chromatofocusing. N-terminal protein sequencing of the CBH II cp preparation confirmed its identity. A comparison of the catalytic activity and cellulose-binding ability of these core proteins was made. The major differences between them were the findings that CBH II cp possessed a sixfold higher specific activity toward p-nitrophenylcellobioside than the native CBH II preparation and still bound to microcrystalline cellulose, unlike CBH I cp. Neither CBH I cp nor CBH II cp had activity toward carboxymethylcellulose, but both were able to hydrolyze barley b-glucan. These data suggest that removal of the cellulose-binding domain and hinge region from CBH I and II have different effects on their properties.

  3. Multivariable parameter optimization for the endoglucanase production by Trichoderma reesei Rut C30 from Ocimum gratissimum seed

    Directory of Open Access Journals (Sweden)

    Mithu Das

    2008-02-01

    Full Text Available The aim of this study was to evaluate the interaction effects of the physico-chemical parameters on the endoglucanase (CMCase production by Trichoderma reesei Rut C30 on a cellulosic agro-residue by the solid-state fermentation (SSF and to determine their optimum values by the EVOP factorial design technique. The best combination of physical parameters for the maximum production of the endoglucanase (CMCase was 28ºC temperature, 79% relative humidity and 4.8 pH of the medium. The best combination of the chemical parameters was (mg/L nicotinic acid 15, naphthalene acetic acid 7, ferric chloride 5 and Tween-80 6. With the application of this technique, the yield of the CMCase increased by ~ 2.3 fold.

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

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    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.

  5. Crystallization and preliminary X-ray crystallographic studies of an exo-β-d-glucosaminidase from Trichoderma reesei

    International Nuclear Information System (INIS)

    An exo-β-d-glucosaminidase from T. reesei (Gls93) has been crystallized by the hanging-drop vapour-diffusion method. Diffraction data have been collected using synchrotron radiation. Chitosan is degraded to glucosamine (GlcN) by chitosanase and exo-β-d-glucosaminidase (GlcNase). GlcNase from Trichoderma reesei (Gls93) is a 93 kDa extracellular protein composed of 892 amino acids. The enzyme liberates GlcN from the nonreducing end of the chitosan chain in an exo-type manner and belongs to glycoside hydrolase family 2. For crystallographic investigations, Gls93 was overexpressed in Pichia pastoris cells. The recombinant Gls93 had two molecular forms of ∼105 kDa (Gls93-F1) and ∼100 kDa (Gls93-F2), with the difference between them being caused by N-glycosylation. Both forms were crystallized by the hanging-drop vapour-diffusion method. Crystals of Gls93-F1 belonged to the orthorhombic space group P212121, with unit-cell parameters a = 98.27, b = 98.42, c = 108.28 Å, and diffracted to 1.8 Å resolution. Crystals of Gls93-F2 belonged to the orthorhombic space group P212121, with unit-cell parameters a = 67.84, b = 81.62, c = 183.14 Å, and diffracted to 2.4 Å resolution. Both crystal forms were suitable for X-ray structure analysis at high resolution

  6. I-SceI-mediated double-strand DNA breaks stimulate efficient gene targeting in the industrial fungus Trichoderma reesei.

    Science.gov (United States)

    Ouedraogo, Jean Paul; Arentshorst, Mark; Nikolaev, Igor; Barends, Sharief; Ram, Arthur F J

    2015-12-01

    Targeted integration of expression cassettes for enzyme production in industrial microorganisms is desirable especially when enzyme variants are screened for improved enzymatic properties. However, currently used methods for targeted integration are inefficient and result in low transformation frequencies. In this study, we expressed the Saccharomyces cerevisiae I-SceI meganuclease to generate double-strand breaks at a defined locus in the Trichoderma reesei genome. We showed that the double-strand DNA breaks mediated by I-SceI can be efficiently repaired when an exogenous DNA cassette flanked by regions homologous to the I-SceI landing locus was added during transformation. Transformation efficiencies increased approximately sixfold compared to control transformation. Analysis of the transformants obtained via I-SceI-mediated gene targeting showed that about two thirds of the transformants resulted from a homologous recombination event at the predetermined locus. Counter selection of the transformants for the loss of the pyrG marker upon integration of the DNA cassette showed that almost all of the clones contained the cassette at the predetermined locus. Analysis of independently obtained transformants using targeted integration of a glucoamylase expression cassette demonstrated that glucoamylase production among the transformants was high and showing limited variation. In conclusion, the gene targeting system developed in this study significantly increases transformation efficiency as well as homologous recombination efficiency and omits the use of Δku70 strains. It is also suitable for high-throughput screening of enzyme variants or gene libraries in T. reesei. PMID:26272087

  7. 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

  8. 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...

  9. UNIQUE ACETYLATION OF OLIGOSACCHARIDES BY TRICHODERMA REESEI ACETYL ESTERASE IN WATER - VINYL ACETATE MIXTURE

    Science.gov (United States)

    Purified T. reesei RUT C-30 acetyl esterase catalyzes acetyl transfer to a variety of carbohydrates in water in the presence of vinyl acetate as the acetyl group donor. The degree of conversion and the number of formed acetates depended on the acceptor used. With some acceptors, such as methyl or ...

  10. 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.

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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.

  16. 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

  17. I-SceI enzyme mediated integration (SEMI) for fast and efficient gene targeting in Trichoderma reesei.

    Science.gov (United States)

    Ouedraogo, Jean Paul; Arentshorst, Mark; Nikolaev, Igor; Barends, Sharief; Ram, Arthur F J

    2016-03-20

    We previously showed that creation of a double strand DNA break (DSB) by expressing I-SceI in an engineered Trichoderma reesei (Hypocrea jecorina) strain containing a I-SceI recognition site improved transformation and homologous integration efficiencies. In this study, we further improved homologous integration frequencies by combining I-SceI mediated double strand break with disruption of the tku70 gene. The inability of the tku70 mutant to repair a I-SceI mediated DSB via NHEJ was used to force integration of an expression cassette with homologous flanks surrounding the DSB site. Besides expressing I-SceI from a plasmid, we also show that adding I-SceI enzyme during transformation was successful to generate DSBs. The I-SceI enzyme mediated integration, or SEMI, in combination with a Δtku70 mutant has a synergistic effect on homologous recombination efficiencies as 90-100% of the transformants exhibited integration of the expression cassette at the homologous site. PMID:26860210

  18. Improvements in Glucose Sensitivity and Stability of Trichoderma reesei β-Glucosidase Using Site-Directed Mutagenesis.

    Science.gov (United States)

    Guo, Boyang; Amano, Yoshihiko; Nozaki, Kouichi

    2016-01-01

    Glucose sensitivity and pH and thermal stabilities of Trichoderma reesei Cel1A (Bgl II) were improved by site-directed mutagenesis of only two amino acid residues (L167W or P172L) at the entrance of the active site. The Cel1A mutant showed high glucose tolerance (50% of inhibitory concentration = 650 mM), glucose stimulation (2.0 fold at 50 mM glucose), and enhanced specific activity (2.4-fold) compared with those of the wild-type Cel1A. Furthermore, the mutant enzyme showed stability at a wide pH range of 4.5-9.0 and possessed high thermal stability up to 50 °C with 80% of the residual activities compared with the stability seen at the pH range of 6.5-7.0 and temperatures of up to 40 °C in the wild-type Cel1A. Kinetic studies for hydrolysis revealed that the Cel1A mutant was competitively inhibited by glucose at similar levels as the wild-type enzyme. Additionally, the mutant enzyme exhibited substrate inhibition, which gradually disappeared with an increasing glucose concentration. These data suggest that the glucose stimulation was caused by relieve the substrate inhibition in the presence of glucose. To conclude, all the properties improved by the mutagenesis would be great advantages in degradation of cellulosic biomass together with cellulases. PMID:26790148

  19. Synergistic action of recombinant accessory hemicellulolytic and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation.

    Science.gov (United States)

    Laothanachareon, Thanaporn; Bunterngsook, Benjarat; Suwannarangsee, Surisa; Eurwilaichitr, Lily; Champreda, Verawat

    2015-12-01

    Synergism between core cellulases and accessory hydrolytic/non-hydrolytic enzymes is the basis of efficient hydrolysis of lignocelluloses. In this study, the synergistic action of three recombinant accessory enzymes, namely GH62 α-l-arabinofuranosidase (ARA), CE8 pectin esterase (PET), and GH10 endo-1,4-beta-xylanase (XYL) from Aspergillus aculeatus expressed in Pichia pastoris to a commercial Trichoderma reesei cellulase (Accellerase® 1500; ACR) on hydrolysis of alkaline pretreated rice straw was studied using a mixture design approach. Applying the full cubic model, the optimal ratio of quaternary enzyme mixture was predicted to be ACR:ARA:PET:XYL of 0.171:0.079:0.100:0.150, which showed a glucose releasing efficiency of 0.173 gglc/FPU, higher than the binary ACR:XYL mixture (0.122 gglc/FPU) and ACR alone (0.081 gglc/FPU) leading to a 47.3% increase in glucose yield compared with that from ACR at the same cellulase dosage. The result demonstrates the varying degree of synergism of accessory enzymes to cellulases useful for developing tailor-made enzyme systems for bio-industry. PMID:26433794

  20. A new stoichiometric miniaturization strategy for screening of industrial microbial strains: application to cellulase hyper-producing Trichoderma reesei strains

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    Jourdier Etienne

    2012-05-01

    Full Text Available Abstract Background During bioprocess development, secondary screening is a key step at the boundary between laboratory and industrial conditions. To ensure an effective high-throughput screening, miniaturized laboratory conditions must mimic industrial conditions, especially for oxygen transfer, feeding capacity and pH stabilization. Results A feeding strategy has been applied to develop a simple screening procedure, in which a stoichiometric study is combined with a standard miniaturization procedure. Actually, the knowledge of all nutriments and base or acid requirements leads to a great simplification of pH stabilization issue of miniaturized fed-batch cultures. Applied to cellulase production by Trichoderma reesei, this strategy resulted in a stoichiometric mixed feed of carbon and nitrogen sources. While keeping the pH between shake flask and stirred bioreactor comparable, the developed shake flask protocol reproduced the strain behaviour under stirred bioreactor conditions. Compared to a an already existing miniaturized shake flasks protocol, the cellulase concentration was increased 5-fold, reaching about 10 g L-1. Applied to the secondary screening of several clones, the newly developed protocol succeeded in selecting a clone with a high industrial potential. Conclusions The understanding of a bioprocess stoichiometry contributed to define a simpler and more effective miniaturization. The suggested strategy can potentially be applied to other fed-batch processes, for the screening of either strain collections or experimental conditions.

  1. 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. PMID:26285758

  2. Identification of residues important for substrate uptake in a glucose transporter from the filamentous fungus Trichoderma reesei.

    Science.gov (United States)

    Zhang, Weixin; Cao, Yanli; Gong, Jing; Bao, Xiaoming; Chen, Guanjun; Liu, Weifeng

    2015-01-01

    The glucose transporter is an important player in cell metabolism that mediates the intracellular uptake of glucose. Here, we characterized the glucose transporter Stp1 from the filamentous fungus Trichoderma reesei. The individual substitution of several conserved residues for Ala in Stp1 corresponding to those interacting with D-glucose in the xylose/H(+) symporter XylE inflicted contrasting effects on its ability to support the growth of an hxt-null yeast on glucose. The targeted change of Phe 50, proximal to the substrate-binding site, was also found to exert a profound effect on the activity of Stp1. In contrast with the charged residues, the substitution of Phe 50 with either the hydrophilic residues Asn and Gln or the small residues Gly and Ala significantly enhanced the transport of glucose and its fluorescent analogue, 2-NBDG. On the other hand, a variant with the three substitutions I115F, F199I and P214L displayed remarkably improved activity on glucose and 2-NBDG transport. Further analysis indicated that the combined mutations of Ile 115 and Pro 214, positioned on the lateral surface of the Stp1 N-domain, fully accounted for the enhanced transport activity. These results provide insight into the structural basis for glucose uptake in fungal sugar transporters. PMID:26345619

  3. 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. PMID:26026380

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

    OpenAIRE

    Elena Ciornea; Tamara Barbaneagra; Alexandru Manoliu; Mihaela Cristica

    2010-01-01

    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 cat...

  5. 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

  6. Defining the genome-wide role of CRE1 during carbon catabolite repression in Trichoderma reesei using RNA-Seq analysis.

    Science.gov (United States)

    Antoniêto, Amanda Cristina Campos; dos Santos Castro, Lílian; Silva-Rocha, Rafael; Persinoti, Gabriela Felix; Silva, Roberto Nascimento

    2014-12-01

    The ascomycete Trichoderma reesei is one of the most well-studied cellulolytic fungi and is widely used by the biotechnology industry in the production of second generation bioethanol. The carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1. CCR represses genes related to cellulase production when a carbon source is readily available in the medium. Using RNA sequencing, we investigated CCR during the synthesis of cellulases, comparing the T. reesei Δcre1 mutant strain with its parental strain, QM9414. Of 9129 genes in the T. reesei genome, 268 genes were upregulated and 85 were downregulated in the presence of cellulose (Avicel). In addition, 251 genes were upregulated and 230 were downregulated in the presence of a high concentration of glucose. Genes encoding cellulolytic enzymes and transcription factors and genes related to the transport of nutrients and oxidative metabolism were also targets of CCR, mediated by CRE1 in a carbon source-dependent manner. Our results also suggested that CRE1 regulates the expression of genes related to the use of copper and iron as final electron acceptors or as cofactors of enzymes that participate in biomass degradation. As a result, the final effect of CRE1-mediated transcriptional regulation is to modulate the access of cellulolytic enzymes to cellulose polymers or blocks the entry of cellulase inducers into the cell, depending on the glucose content in the medium. These results will contribute to a better understanding of the mechanism of carbon catabolite repression in T. reesei, thereby enhancing its application in several biotechnology fields. PMID:25459535

  7. Subcellular fractionation of a hypercellulolytic mutant, Trichoderma reesei Rut-C30: localization of endoglucanase in microsomal fraction.

    Science.gov (United States)

    Glenn, M; Ghosh, A; Ghosh, B K

    1985-11-01

    The growing mycelia of Trichoderma reesei Rut-C30 are richly endowed with endoplasmic reticula and a variety of pleomorphic subcellular bodies. Mycelia of the culture growing in presence of avicel pH101 was fractionated in sucrose density gradients, and several morphologically and biochemically distinct fractions were isolated. Mycelia were homogenized in a Bead Beater, and the homogenate was freed of nucleus and wall fragments by low-speed centrifugation before fractionation. Organelle-free cytosol, which did not penetrate the gradient, contained (of the total) 72% of the vanadate-sensitive ATPase, 26% of carboxymethyl cellulase (CMCase), 2% of cytochrome c reductase, and 13% of the protein. Significant fractions separated on a gradient were light vesicles containing heavily stained material inside and ribosomes attached to the outside surface, intact vesicles resembling condensing vacuoles, large vesicles derived from the plasma membrane, and heavy vesicles containing crystalline material. The light-vesicle fraction contained a large portion of the cell-bound CMCase activity. The particle-bound ATPase and cytochrome c reductase activities were concentrated in heavy fractions. The fractionation in the presence of MgCl2 improved the preservation of subcellular bodies derived from the endoplasmic reticula. Although the CMCase activity of the light-vesicle fraction was 4 times higher than the activity in the heavy-vesicle fraction, the CMCase antibody-binding capacities of both fractions were about the same. This discrepancy between the catalytic activity and the antibody-binding capacity suggests that the heavy vesicles might have contained considerable amount of inactive CMCase compared with that present in the light vesicles. PMID:4091550

  8. 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

    Directory of Open Access Journals (Sweden)

    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

  9. The effect of denaturation and reduction on cellobiohydrolase 1 from Trichoderma reesei

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J. (Oak Ridge National Lab., TN (USA)); Herrmann, P.C. (Andrews Univ., Berrien Springs, MI (USA))

    1990-01-01

    The origin of these studies from our interest in the use of protein denaturants to elute cellulase enzyme components from residual cellulosic substrates. A high concentration of the denaturant guanidine hydrochloride, required for elution, was found to only partially unfold the major component of T. reesei cellulase, cellobiohydrolase I(CBH I), and that the reason for the loss in catalytic activity was due to the competitive inhibition of this enzyme and not, as expected, due to its unfolding or denaturation. Papain digestion of CBH I generated core'' enzyme which, apparently, completely broke up upon reduction. These data are discussed in relation to structure/function relationships of this key cellulase enzyme component. In this paper studies on the effect of denaturation and reduction of CBH I are described. The rationale for these studies is to further our understanding of the structure and function relationships of this key cellulase enzyme and to determine the methodology for its successful unfolding and refolding that may be necessary for its recovery. 18 refs., 5 figs.

  10. Characterization of the Ca(2+) -responsive signaling pathway in regulating the expression and secretion of cellulases in Trichoderma reesei Rut-C30.

    Science.gov (United States)

    Chen, Ling; Zou, Gen; Wang, Jingzhi; Wang, Jin; Liu, Rui; Jiang, Yanping; Zhao, Guoping; Zhou, Zhihua

    2016-05-01

    Calcium signaling plays pivotal roles in the hyphal growth, conidiation, and osmosis sensitivity of fungi through the Ca(2+) /calmodulin-calcineurin-dependent pathway. This study found that an appropriate extracellular Ca(2+) concentration markedly stimulated the hyphal growth, cellulase production, and total protein secretion of the cellulase hyper-producing strain, Trichoderma reesei Rut-C30. Transcription analysis revealed upregulation of not only encoding genes of cellulases and the transcriptional activator XYR1 but also several genes encoding endoplasmic reticulum-chaperones after Ca(2+) addition. The function of CRZ1, T. reesei calcineurin-responsive zinc finger transcription factor 1, was further characterized by gene disruption. Electrophoretic mobility shift assays (EMSAs) in combination with chromatin immunoprecipitation (ChIP) verified that CRZ1 could bind directly to the upstream regions of xyr1 and cbh1 (cellobiohydrolase I-encoding gene) in response to Ca(2+) . A DNase I footprinting assay identified its putative binding consensus site (5'-[T/G]GGCG-3' or 5'-GGGC[G/T]-3'). EMSAs confirmed that CRZ1 competed for occupancy of the xyr1 promoter with another transcription factor, ACE1. These results revealed putative signaling pathways downstream of calcineurin in response to extracellular Ca(2+) involved in upregulation of cellulose degradation-related genes, reflecting progress in the study of Ca(2+) signaling in filamentous fungi. This study also provides insight that will facilitate further improvement of (hemi-)cellulase production by T. reesei. PMID:27109892

  11. 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.

  12. 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

  13. Improvement of Napier Grass Silage Nutritive Value by Using Inoculant and Crude Enzyme from Trichoderma reesei and Its Effect on in Vitro Rumen Fermentation

    Directory of Open Access Journals (Sweden)

    D. J. Nurjana

    2016-04-01

    Full Text Available This research was aimed to evaluate the effect of inoculant and crude enzyme of Trichoderma reesei on fermentation quality and rumen digestibility of napier grass silage. This research consisted of two stages. The first stage was fermentation quality using a completely randomized design with three treatments and six replications. The second stage was rumen fermentability using a randomized block design with three treatments and six replications. The treatments of both stages were: T0= control of napier grass silage (Hi-fer+, T1= control + T. reesei inoculant at the level of 2.13 x 107 cfu/kg, and T2= control + crude enzyme from T. reesei at the level of 11.4 unit/kg. Napier grass silage were kept at room temperature and opened 21 d after fermentation. The result of the first stage experiment showed that T2 decreased (P<0.01 dry matter, organic matter, crude fiber, and neutral detergent fiber (NDF and also increased (P<0.05 crude protein, total digestible nutrient (TDN, pH, acetic acid, and propionic acid of napier grass silage while T1 decreased (P<0.01 NDF and increased (P<0.05 propionic acid. The result of the second stage experiment showed that there were no significant differences in rumen pH, in vitro dry matter and organic matter digestibility, total VFA, proportion of VFA, ratio of acetic to propionic acids, and also NH3. It is concluded that the addition of crude enzyme from T. reesei could improve fermentation quality of napier grass silage with decreasing NDF and increasing TDN but did not affect rumen fermentability.

  14. 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

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

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    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

  16. 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.

  17. 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.

  18. The N-acetylglucosamine catabolic gene cluster in Trichoderma reesei is controlled by the Ndt80-like transcription factor RON1.

    Science.gov (United States)

    Kappel, Lisa; Gaderer, Romana; Flipphi, Michel; Seidl-Seiboth, Verena

    2016-02-01

    Chitin is an important structural constituent of fungal cell walls composed of N-acetylglucosamine (GlcNAc) monosaccharides, but catabolism of GlcNAc has not been studied in filamentous fungi so far. In the yeast Candida albicans, the genes encoding the three enzymes responsible for stepwise conversion of GlcNAc to fructose-6-phosphate are clustered. In this work, we analysed GlcNAc catabolism in ascomycete filamentous fungi and found that the respective genes are also clustered in these fungi. In contrast to C. albicans, the cluster often contains a gene for an Ndt80-like transcription factor, which we named RON1 (regulator of N-acetylglucosamine catabolism 1). Further, a gene for a glycoside hydrolase 3 protein related to bacterial N-acetylglucosaminidases can be found in the GlcNAc gene cluster in filamentous fungi. Functional analysis in Trichoderma reesei showed that the transcription factor RON1 is a key activator of the GlcNAc gene cluster and essential for GlcNAc catabolism. Furthermore, we present an evolutionary analysis of Ndt80-like proteins in Ascomycota. All GlcNAc cluster genes, as well as the GlcNAc transporter gene ngt1, and an additional transcriptional regulator gene, csp2, encoding the homolog of Neurospora crassa CSP2/GRHL, were functionally characterised by gene expression analysis and phenotypic characterisation of knockout strains in T. reesei. PMID:26481444

  19. The effects of disruption of phosphoglucose isomerase gene on carbon utilisation and cellulase production in Trichoderma reesei Rut-C30

    Directory of Open Access Journals (Sweden)

    Pakula Tiina

    2011-05-01

    Full Text Available Abstract Background Cellulase and hemicellulase genes in the fungus Trichoderma reesei are repressed by glucose and induced by lactose. Regulation of the cellulase genes is mediated by the repressor CRE1 and the activator XYR1. T. reesei strain Rut-C30 is a hypercellulolytic mutant, obtained from the natural strain QM6a, that has a truncated version of the catabolite repressor gene, cre1. It has been previously shown that bacterial mutants lacking phosphoglucose isomerase (PGI produce more nucleotide precursors and amino acids. PGI catalyzes the second step of glycolysis, the formation of fructose-6-P from glucose-6-P. Results We deleted the gene pgi1, encoding PGI, in the T. reesei strain Rut-C30 and we introduced the cre1 gene in a Δpgi1 mutant. Both Δpgi1 and cre1+Δpgi1 mutants showed a pellet-like and growth as well as morphological alterations compared with Rut-C30. None of the mutants grew in media with fructose, galactose, xylose, glycerol or lactose but they grew in media with glucose, with fructose and glucose, with galactose and fructose or with lactose and fructose. No growth was observed in media with xylose and glucose. On glucose, Δpgi1 and cre1+Δpgi1 mutants showed higher cellulase activity than Rut-C30 and QM6a, respectively. But in media with lactose, none of the mutants improved the production of the reference strains. The increase in the activity did not correlate with the expression of mRNA of the xylanase regulator gene, xyr1. Δpgi1 mutants were also affected in the extracellular β-galactosidase activity. Levels of mRNA of the glucose 6-phosphate dehydrogenase did not increase in Δpgi1 during growth on glucose. Conclusions The ability to grow in media with glucose as the sole carbon source indicated that Trichoderma Δpgi1 mutants were able to use the pentose phosphate pathway. But, they did not increase the expression of gpdh. Morphological characteristics were the result of the pgi1 deletion. Deletion of pgi1 in

  20. 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.

  1. 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.

  2. 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.

  3. 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.

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

    International Nuclear Information System (INIS)

    Highlights: ► The CSLP and fusion enzyme were successfully expressed in the Pichia pastoris. ► The fusion enzyme was stable at 80 °C for 120-min. ► The fusion enzyme was responsible for cellulose-binding capacity. ► 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 (α 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 °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.

  5. 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.

  6. Overexpression of an endo-1,4-β-glucanase V gene (EGV) from Trichoderma reesei leads to the accumulation of cellulase activity in transgenic rice.

    Science.gov (United States)

    Li, X Y; Liu, F; Hu, Y F; Xia, M; Cheng, B J; Zhu, S W; Ma, Q

    2015-01-01

    The ectopic expression of cellulase in biomass can reduce the cost of biofuel conversion. This trait modification technique is highly beneficial for biofuel production. In this study, we isolated an endo-1,4-beta-glucanase gene (EGV) from Trichoderma reesei and inserted this gene downstream of a fragment encoding the signal peptide Apo-SP in a modified pCAMBIA1301 vector to obtain an Apo-SP and AsRed fusion protein. Transient expression of this fusion protein in onion epidermal cells showed that the Apo-SP signal was localized to the plastids. EGV transgenic rice plants that did not carry screening marker genes were obtained through overexpression of the pDTB double T-DNA vector. Western blotting showed that EGV was expressed in the dry straw of T0 generation transgenic rice plants and in fresh leaves of the T1 generation. More importantly, our results also showed that the peptide product of EGV in the transgenic plants folded correctly and was capable of digesting the cellulase substrate CMC. Additionally, cellulase activity remained stable in the straw that had been dried at room temperature for three months. This study presents an important technical approach for the development of transgenic rice straw that has stable cellulase activity and can be used for biofuel conversion. PMID:26782396

  7. 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.

  8. 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

  9. 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

  10. 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.

  11. Expression of endo-1, 4-beta-xylanase from Trichoderma reesei in Pichia pastoris and functional characterization of the produced enzyme

    Directory of Open Access Journals (Sweden)

    He Jun

    2009-06-01

    Full Text Available Abstract Background In recent years, xylanases have attracted considerable research interest because of their potential in various industrial applications. The yeast Pichia pastoris can neither utilize nor degrade xylan, but it possesses many attributes that render it an attractive host for the expression and production of industrial enzymes. Results The Xyn2 gene, which encodes the main Trichoderma reesei Rut C-30 endo-β-1, 4-xylanase was cloned into the pPICZαA vector and expressed in Pichia pastoris. The selected P. pastoris strains produced as 4,350 nkat/ml β-xylanase under the control of the methanol inducible alcohol oxidase 1 (AOX1 promoter. The secreted recombinant Xyn2 was estimated by SDS-PAGE to be 21 kDa. The activity of the recombinant Xyn2 was highest at 60°C and it was active over a broad range of pH (3.0–8.0 with maximal activity at pH 6.0. The enzyme was quite stable at 50°C and retained more than 94% of its activity after 30 mins incubation at this temperature. Using Birchwood xylan, the determined apparent Km and kcat values were 2.1 mg/ml and 219.2 S-1, respectively. The enzyme was highly specific towards xylan and analysis of xylan hydrolysis products confirmed as expected that the enzyme functions as endo-xylanase with xylotriose as the main hydrolysis products. The produced xylanase was practically free of cellulolytic activity. Conclusion The P. pastoris expression system allows a high level expression of xylanases. Xylanase was the main protein species in the culture supernatant, and the functional tests indicated that even the non-purified enzyme shows highly specific xylanase activity that is free of cellulolytic side acitivities. Therefore, P pastoris is a very useful expression system when the goal is highly specific and large scale production of glycosyl hydrolases.

  12. 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. PMID:26073313

  13. 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

  14. 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.

  15. 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

  16. 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.

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

    OpenAIRE

    Bambang Dwi Argo; Yusuf Hendrawan; Dewi Maya Maharani; Angky Wahyu Putranto; Sri Winarsih

    2016-01-01

    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 activit...

  18. Nonparaxial anamorphic diffractive lenses

    International Nuclear Information System (INIS)

    Applications of anamorphic diffractive lenses, or in other words elliptical zone plates, are growing in number and diversity. Following this progress, the manufacturing methods of anamorphic diffractive lenses are also a subject of continuous improvement. On the other hand, their design is still based mainly on the paraxial approximation. In the present paper nonparaxial elliptical zone plates are presented and their difference from the paraxial version, termed as the elliptical aberration is shown. A comparison of their focusing quality was performed based on results of numerical modelling as well as experiments. (paper)

  19. The efficacy of a new 6-phytase obtained from Buttiauxella spp. expressed in Trichoderma reesei on digestibility of amino acids, energy, and nutrients in pigs fed a diet based on corn, soybean meal, wheat middlings, and corn distillers' dried grains with solubles.

    Science.gov (United States)

    Adedokun, S A; Owusu-Asiedu, A; Ragland, D; Plumstead, P; Adeola, O

    2015-01-01

    Sixteen cannulated pigs were used to evaluate the effect of a new 6-phytase derived from Buttiauxella spp. and expressed in Trichoderma reesei on apparent ileal digestibility (AID) of AA and apparent total tract digestibility (ATTD) of DM, N, Ca, P, Na, Mg, K, Cl, and energy. Pigs were fed 4 diets for 2 periods in a crossover design. Within each period, there were 4 blocks of 4 pigs per block with each diet represented within each block. The average initial BW in periods 1 and 2 were 22 and 30 kg, respectively. Each period lasted 9 d with fecal collection on d 5 and 6 and a 12-h ileal digesta collection on d 7, 8, and 9. Pigs received a daily feed allowance of approximately 4.5% of their BW. The experimental diets were based on corn, soybean meal, wheat middlings, and corn distillers dried grain with solubles. Phytase was added at 0; 500; 1,000; or 2,000 phytase units/kg of diet to a basal diet that contained 205, 15, 5.4, and 10 g of CP, Lys, total P (1.6 g of nonphytate P), and Ca/kg diet, respectively. The addition of phytase improved (P pigs in a dose-dependent manner. PMID:25568365

  20. Biological Control of Sclerotium rolfsii Damping-off of Tropical Pine (Pinus merkusii) with Three Isolates of Trichoderma spp.

    OpenAIRE

    S. M. Widyastuti; Harjono; Sumardi .; D. Yuniarti

    2003-01-01

    Trichoderma koningii, T. reesei and T. harzianum were tested for their ability to control Sclerotium rolfsii damping-off of tropical pine seedlings. Light microscopic observation on dual culture assay showed that the hyphae of all Trichoderma isolates could grow parallel to the hypae of S. rolfsii. However only T. reesei and T. harzianum coiled around the hyphae of S. rolfsii and formed appresoria and hook-like structures. Both isolates indicated equally high colony growth inhibition toward ...

  1. 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.

  2. Biotransformation of Trichoderma spp. and Their Tolerance to Aromatic Amines, a Major Class of Pollutants

    OpenAIRE

    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

    2013-01-01

    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-me...

  3. [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

  4. 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

  5. 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)

  6. 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)

  7. Sexual development in the industrial workhorse Trichoderma reesei

    OpenAIRE

    Seidl, Verena; Seibel, Christian; Kubicek, Christian P.; Schmoll, Monika

    2009-01-01

    Filamentous fungi are indispensable biotechnological tools for the production of organic chemicals, enzymes, and antibiotics. Most of the strains used for industrial applications have been—and still are—screened and improved by classical mutagenesis. Sexual crossing approaches would yield considerable advantages for research and industrial strain improvement, but interestingly, industrially applied filamentous fungal species have so far been considered to be largely asexual. This is also true...

  8. 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.

  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. Biosynthesis of the enzymes of the cellulase system by T. Reesei QM 9414 in the presence of sophorose

    Science.gov (United States)

    Gritzali, M.

    1982-12-01

    As conventional, nonrenewable energy sources are rapidly depleted and it was necessary to search for alternative sources of energy. It was increasingly apparent that biomass and waste are alternatives well worth exploring. The sources of biomass and wastes that considered for conversion to useful products are quite diverse, but the most abundant constituent of almost every type is cellulose. Cellulose is cleanly converted to soluble fermentable sugars enzymatically, and cellulose enzymes were isolated from a number of microbial sources. It is generally agreed that the most effective system of enzymes for the conversion of cellulose to glucose is produced by species of the imperfect fungus Trichoderma. The mutant organism Trichoderma reesei QM 9414 is among the best producers of high levels of enzymes; these are extracellular and have carbonhydrate covalently bound to the peptide. Trichoderma produces three types of enzymes which, in a sequential and cooperative manner, convert cellulose to soluble oligosaccharides and glucose.

  11. Phylogeny and systematics of the anamorphic, entomopathogenic genus Beauveria

    Science.gov (United States)

    Beauveria is a cosmopolitan anamorph genus of arthropod pathogens that includes the agronomically important species B. bassiana and B. brongniartii, which are used as mycoinsecticides for the biological control of pest insects. Recent phylogenetic evidence demonstrates that Beauveria is monophyletic...

  12. 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.

  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. 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...

  15. КСИЛАНАЗЫ TRICHODERMA REESEI – БИОСИНТЕЗ И ПРИМЕНЕНИЕ ДЛЯ ГИДРОЛИЗА ЗЕРНОВЫХ КОРМОВ

    OpenAIRE

    Морозова, Юлия; Скворцов, Евгений; Алимова, Фарида

    2013-01-01

    Штамм Trichoderma reesei M18.2 способен к активному синтезу ксиланаз и целлюлаз на отходе спиртового производства – послеспиртовой барде. При глубинном культивировании на барде активность ксиланаз и целлюлаз достигала максимального значения на четвёртые сутки культивирования и составляла 530 и 3.3 МЕ/мл соответственно. Исследован состав углеводной фракции фуражных сортов зерновых культур Республики Татарстан. Содержание ксиланов в ней достигало 11.4%. Глубина ферментативного гидролиза зерново...

  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. 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.

  18. 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.

  19. Femtosecond micromachining in transparent bulk materials using an anamorphic lens.

    Science.gov (United States)

    Desautels, G Logan; Brewer, Chris D; Walker, Mark A; Juhl, Shane B; Finet, Marc A; Powers, Peter E

    2007-10-01

    A unique anamorphic lens design was applied to a circular 780nm femtosecond laser pulse to transform it into an elliptically shaped beam at focus. This lens was developed to give an alternative method of micromachining bulk transparent materials. The challenge for femtosecond laser processing is to control the nonlinear affect of self-focusing, which can occur when using a fast f-number lens. Once the focused spot is dominated by self-focusing the predicted focused beam becomes a filament inside the bulk, which is an undesirable effect. The anamorphic lens resolves this self-focusing by increasing the numerical aperture (NA) and employing an elliptical beam shape. The anamorphic lens was designed to furnish a 2.5mum by 190mum line at focus. Provided the pulse energy is high enough, transparent bulk material will be damaged with a single femtosecond laser pulse. Damage in this text refers to visual change in the index of refraction as observed under an optical microscope. Using this elliptical shape (or line), grating structures were micro-machined on the surface of SiC bulk transparent substrate. SiC was chosen because it is known for its micromachining difficulty and its crystalline structure. From the lack of self-focusing and using energy that is just above the damage threshold the focused line beam generated from the anamorphic lens grating structures produced a line shape nearly identical to the geometrical approximation. In this paper we discuss a new method of writing gratings (or other types of structures) in bulk transparent materials using a single femtosecond laser pulse. We will investigate the grating structures visually (inspected under an optical microscope) and also by use of an atomic force microscopy (AFM). In addition, we test the grating diffraction efficiency (DE) as a function of grating spacing, d. PMID:19550582

  20. Mathematical model for enzymatic hydrolysis and fermentation of cellulose by Trichoderma

    Energy Technology Data Exchange (ETDEWEB)

    Peitersen, N.; Ross, E.W. Jr.

    1979-06-01

    This paper describes a mathematical model for the enzymatic hydrolysis and fermentation of cellulose by Trichoderma reesei. The principal features of the model are the assumption of two forms of cellulose (crystalline and amorphous), two sugars (cellobiose and glucose), and two enzymes (cellulase and ..beta..-glucosidase). An inducer-repressor-messenger RNA mechanism is used to predict enzyme formation, and pH effects are included. The model consists of 12 ordinary differential equations for 12 state variables and contains 38 parameters. The parameters were estimated from four sets of experimental data by optimization. The results appear satisfactory, and the computer programs permit simulation of a variety of system changes.

  1. 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...... degrading enzymes and strong secretion ability of this genus. Here we report extensive transcriptome analysis of plant cell wall degrading enzymes in T. asperellum. The production of cell wall degrading enzymes by T. asperellum was tested on a range of cellulosic materials under various conditions. When T...... the theory that the glycoside hydrolases have evolved from a common ancestor, followed by a specialization in which saprotrophic fungi such as T. reesei and T. longibrachiatum lost a significant number of genes including several glycoside hydrolases....

  2. 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; Smith, Dale A.

    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...

  3. 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. PMID:25839184

  4. 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.

  5. Alternative management of a problematic weed of wheat Avena fatua L. by metabolites of Trichoderma Alternativas de manejo de una maleza problemática de trigo Avena fatua L. por metabolitos de Trichoderma

    Directory of Open Access Journals (Sweden)

    Arshad Javaid

    2011-06-01

    Full Text Available Wild oat (Avena fatua L. is a problematic weed of wheat (Triticum aestivum L. in Pakistan. The present study was designed to evaluate the herbicidal activity of culture filtrates of four Trichoderma spp., namely T. harzianum, T. pseudokoningii, T. reesei and T. viride, against this weed species. In a laboratory bioassay, original (100% and diluted (50% culture filtrates of T. harzianum and T. pseudokoningii significantly reduced shoot and root growth of A. fatua seedlings. Only original culture filtrates of T. reesei exhibited significant effects, while the effect of filtrates of T. viride was insignificant against shoot and root growth of the target weed species. Generally, original concentrations of culture filtrates of all Trichoderma spp., except T. harzianum, significantly reduced various parameters of root and shoot growth of wheat seedlings. In foliar spray bioassay, pot-grown 1-wk and 2-wk old A. fatua and wheat seedlings were sprayed with 100% culture filtrates of the four Trichoderma spp. thrice with 5 d interval each. Culture filtrates of all except T. viride significantly diminished root and shoot biomass of A. fatua plants in 1-wk old plants. The effect of foliar spray on root and shoot growth of wheat was insignificant. Present study concludes that metabolites of T. harzianum, T. reesei and T. pseudokoningii contain herbicidal constituents for the management of A. fatua.La avena silvestre (Avena fatua L. es una maleza problemática del trigo (Triticum aestivum L. en Paquistán. El presente estudio se diseñó para evaluar la actividad de filtrados de cultivos de cuatro Trichoderma spp.: T. harzianum, T. pseudokoningii, T. reesei y T. viride, contra esta especie. En un bioensayo de laboratorio, filtrados de cultivos originales (100% y diluidos (50% de T. harzianum and T. pseudokoningii redujeron significativamente el crecimiento de brotes y raíces de plántulas de A fatua. Sólo los filtrados de cultivos originales de T. reesei

  6. 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...

  7. In Situ Stability of Substrate-Associated Cellulases Studied by DSC

    DEFF Research Database (Denmark)

    Borch, Kim; Cruys-Bagger, Nicolaj; Badino, Silke Flindt; Sørensen, Trine Holst; Windahl, Michael Skovbo; Westh, Peter; Jensen, Kenneth; Alasepp, Kadri

    2014-01-01

    of the transition peak was used as a gauge of the population of native enzyme. Analogous measurements were made for enzymes in pure buffer. Investigations of two cellobiohydrolases, Cel6A and Cel7A, from Trichoderma reesei, which is an anamorph of the fungus Hypocrea jerorina, showed that these...

  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. 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.

  10. 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

  11. 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.

  12. 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

  13. 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.

  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. 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...

  16. Production of cellulase from kraft paper mill sludge by Trichoderma reesei rut C-30.

    Science.gov (United States)

    Wang, Wei; Kang, Li; Lee, Yoon Y

    2010-05-01

    Paper mill sludge is a solid waste material generated from pulping and papermaking operations. Because of high glucan content and its well-dispersed structure, paper mill sludges are well suited for bioconversion into value-added products. It also has high ash content originated from inorganic additives used in papermaking, which causes hindrance to bioconversion. In this study, paper mill sludges from Kraft process were de-ashed by a centrifugal cleaner and successive treatment by sulfuric acid and sodium hydroxide, and used as a substrate for cellulase production. The treated sludge was the only carbon source for cellulase production, and predominantly inorganic nutrients were used as the nitrogen source for this bioprocess. The cellulase enzyme produced from the de-ashed sludge exhibited cellulase activity of 8 filter paper unit (FPU)/mL, close to that obtainable from pure cellulosic substrates. The yield of cellulase enzyme was 307 FPU/g glucan of de-ashed sludge. Specific activity was 8.0 FPU/mg protein. In activity tests conducted against the corn stover and alpha-cellulose, the xylanse activity was found to be higher than that of a commercial cellulase. Relatively high xylan content in the sludge appears to have induced high xylanase production. Simultaneous saccharification and fermentation (SSF) was performed using partially de-ashed sludge as the feedstock for ethanol production using Sacharomyces cerevisiae and the cellulase produced in-house from the sludge. With 6% (w/v) glucan feed, ethanol yield of 72% of theoretical maximum and 24.4 g/L ethanol concentration were achieved. These results were identical to those of the SSF using commercial cellulases. PMID:19997787

  17. Cellulase activity mapping of Trichoderma reesei cultivated in sugar mixtures under fed-batch conditions

    OpenAIRE

    Jourdier, Etienne; Cohen, Céline; Poughon, Laurent; Larroche, Christian; Monot, Frédéric; Ben Chaabane, Fadhel

    2013-01-01

    Background: On-site cellulase production using locally available lignocellulosic biomass (LCB) is essential for cost-effective production of 2nd-generation biofuels. Cellulolytic enzymes (cellulases and hemicellulases) must be produced in fed-batch mode in order to obtain high productivity and yield. To date, the impact of the sugar composition of LCB hydrolysates on cellulolytic enzyme secretion has not been thoroughly investigated in industrial conditions. Results: The effect of sugar mixtu...

  18. Cell wall modification in tobacco by differential targeting of recombinant endoglucanase from Trichoderma reesei

    OpenAIRE

    Klose, Holger; Günl, Markus; Usadel, Björn; Fischer, Rainer; Commandeur, Ulrich

    2015-01-01

    Background The development of transgenic plants as a production platform for biomass-degrading enzymes is a promising tool for an economically feasible allocation of enzymes processing lignocellulose. Previous research has already identified a major limitation of in planta production such as interference with the structure and integrity of the plant cell wall resulting in a negative influence on plant growth and development. Results Here, we describe the in planta expression of endoglucanase ...

  19. Optimization of Cellulase Production by Trichoderma reesei HY07 Using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Shikai Wang

    2013-05-01

    Full Text Available Response Surface Methodology (RSM based on a three-level, three-variable Box and Behnken Factorial Design (BBFD was used to evaluate the interactive effects of corn stalk: bran ratio, tween80 and temperature on the cellulase production by solid fermentation. The optimum conditions derived via RSM were: corn stalk: bran ratio 1.29:1, Tween80 11.05 &muL and temperature 31 °C for carboxymethyl cellulase (CMCase and corn stalk: bran ratio 0.77:1, Tween80 12.54&muL and temperature 32 °C for Filter Paper Activity (FPA. The actual experimental yield was 406.42 U/g for CMCase and 93.62 U/g for FPA under optimum condition, which compared well to the maximum predicted value of 405.67 U/g and 91.29 U/g. The cellulase yield under optimal conditions was 1.45 fold for CMCase and 1.33 fold for FPA to the control.

  20. Phosphorylated N-Glycans of Glycoprotein CBH I from the Gungus Trichoderma Reesei

    Czech Academy of Sciences Publication Activity Database

    De Bruyn, A.; Van der Eycken, J.; Schraml, Jan; Busson, R.; Sandra, K.; Clayessens, M.; Van Beeumen, J.; Geysens, S.; Contreras, R.

    2002. s. PL 368. [International Conference on Magnetic Resonance in Biological Systems /20./. 25.08.2002-30.08.2002, Toronto] Institutional research plan: CEZ:AV0Z4072921 Subject RIV: CF - Physical ; Theoretical Chemistry

  1. 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.

  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. 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.

  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. 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.

  6. 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

  7. Isolation and analysis of lip2 gene from Trichoderma harzianum

    OpenAIRE

    Vaz, Madalena; Belo, Hélio; Jorge, Lurdes; Gonzalez, Francisco J.; Monte, Enrique; Choupina, Altino

    2011-01-01

    The genus Trichoderma is cosmopolitan in soils, wood decomposition and plant material. Species of Trichoderma are often dominant components of the soil microflora in various habitats. This is due to different metabolic capacity of the Trichoderma species and its aggressive competitiveness in nature. The genus Trichoderma are frequently used in biological control because of its antagonist ability of phytopathogenic fungi. The mechanisms employed by Trichoderma spp. to antagon...

  8. 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.

  9. 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.

  10. 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.

  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. Gene expression analysis of the biocontrol fungus Trichoderma harzianum in the presence of tomato plants, chitin, or glucose using a high-density oligonucleotide microarray

    Directory of Open Access Journals (Sweden)

    Suárez M Belén

    2009-10-01

    Full Text Available Abstract Background It has recently been shown that the Trichoderma fungal species used for biocontrol of plant diseases are capable of interacting with plant roots directly, behaving as symbiotic microorganisms. With a view to providing further information at transcriptomic level about the early response of Trichoderma to a host plant, we developed a high-density oligonucleotide (HDO microarray encompassing 14,081 Expressed Sequence Tag (EST-based transcripts from eight Trichoderma spp. and 9,121 genome-derived transcripts of T. reesei, and we have used this microarray to examine the gene expression of T. harzianum either alone or in the presence of tomato plants, chitin, or glucose. Results Global microarray analysis revealed 1,617 probe sets showing differential expression in T. harzianum mycelia under at least one of the culture conditions tested as compared with one another. Hierarchical clustering and heat map representation showed that the expression patterns obtained in glucose medium clustered separately from the expression patterns observed in the presence of tomato plants and chitin. Annotations using the Blast2GO suite identified 85 of the 257 transcripts whose probe sets afforded up-regulated expression in response to tomato plants. Some of these transcripts were predicted to encode proteins related to Trichoderma-host (fungus or plant associations, such as Sm1/Elp1 protein, proteases P6281 and PRA1, enchochitinase CHIT42, or QID74 protein, although previously uncharacterized genes were also identified, including those responsible for the possible biosynthesis of nitric oxide, xenobiotic detoxification, mycelium development, or those related to the formation of infection structures in plant tissues. Conclusion The effectiveness of the Trichoderma HDO microarray to detect different gene responses under different growth conditions in the fungus T. harzianum strongly indicates that this tool should be useful for further assays that

  13. Molecular Characterization and Identification of Biocontrol Isolates of Trichoderma spp.

    OpenAIRE

    Hermosa, M. R.; Grondona, I; Iturriaga, E A; Diaz-Minguez, J. M.; Castro, C.; Monte, E.; Garcia-Acha, I.

    2000-01-01

    The most common biological control agents (BCAs) of the genus Trichoderma have been reported to be strains of Trichoderma virens, T. harzianum, and T. viride. Since Trichoderma BCAs use different mechanisms of biocontrol, it is very important to explore the synergistic effects expressed by different genotypes for their practical use in agriculture. Characterization of 16 biocontrol strains, previously identified as “Trichoderma harzianum” Rifai and one biocontrol strain recognized as T. virid...

  14. 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.

  15. 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

  16. 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...

  17. 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...

  18. Visualization of Trichoderma reesei Cellobiohydrolase I and Endoglucanase I on Aspen Cellulose by Using Monoclonal Antibody-Colloidal Gold Conjugates

    OpenAIRE

    Nieves, Rafael A.; Robert P. Ellis; Todd, Roberta J.; Johnson, Timothy J. A.; Grohmann, Karel; Himmel, Michael E.

    1991-01-01

    Monoclonal antibodies (MAbs) specific for cellobiohydrolase I (CBH I) and endoglucanase I (EG I) were conjugated to 10- and 15-nm colloidal gold particles, respectively. The binding of CBH I and EG I was visualized by utilizing the MAb-colloidal gold probes. The visualization procedure involved immobilization of cellulose microfibrils on copper electron microscopy grids, incubation of the cellulose-coated grids with cellulase(s), binding of MAb-colloidal gold conjugates to cellulase(s), and v...

  19. A new stoichiometric miniaturization strategy for screening of industrial microbial strains: application to cellulase hyper-producing Trichoderma reesei strains

    OpenAIRE

    Jourdier Etienne; Poughon Laurent; Larroche Christian; Monot Frédéric; Chaabane Fadhel

    2012-01-01

    Abstract Background During bioprocess development, secondary screening is a key step at the boundary between laboratory and industrial conditions. To ensure an effective high-throughput screening, miniaturized laboratory conditions must mimic industrial conditions, especially for oxygen transfer, feeding capacity and pH stabilization. Results A feeding strategy has been applied to develop a simple screening procedure, in which a stoichiometric study is combined with a standard miniaturization...

  20. The CRE1 carbon catabolite repressor of the fungus Trichoderma reesei: a master regulator of carbon assimilation.

    OpenAIRE

    Seiboth Bernhard; Druzhinina Irina S; Karaffa Levente; Hartl Lukas; Sándor Erzsébet; Fekete Erzsébet (1975-) (biotechnológus); Atanasova Lea; Linke Rita; Margeot Antoine; Portnoy Thomas; Le Crom Stéphane; Kubicek Christian P

    2011-01-01

    Abstract Background The identification and characterization of the transcriptional regulatory networks governing the physiology and adaptation of microbial cells is a key step in understanding their behaviour. One such wide-domain regulatory circuit, essential to all cells, is carbon catabolite repression (CCR): it allows the cell to prefer some carbon sources, whose assimilation is of high nutritional value, over less profitable ones. In lower multicellular fungi, the C2H2 zinc finger CreA/C...

  1. 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.

  2. [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

  3. 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.

  4. 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

  5. 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...

  6. 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...

  7. Influence of Environmental Parameters on Trichoderma Strains with Biocontrol Potential

    OpenAIRE

    Kredics, László; Antal, Zsuzsanna; Manczinger, László; Szekeres, András; Kevei, Ferenc; Nagy, Erzsébet

    2003-01-01

    Several mycoparasitic strains belonging to the filamentous fungal genus Trichoderma are promising candidates for the biological control of plant pathogenic fungi. When planning the application of antagonistic Trichoderma strains for the purposes of biological control, it is very important to consider the environmental parameters affecting the biocontrol agents in the soil. A series of abiotic and biotic environmental parameters has an influence on the biocontrol efficacy of Trichoderma. Some ...

  8. 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...

  9. 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.

  10. Purifying selection and birth-and-death evolution in the class II hydrophobin gene families of the ascomycete Trichoderma/Hypocrea

    Directory of Open Access Journals (Sweden)

    Kenerley Charles M

    2008-01-01

    Full Text Available Abstract Background Hydrophobins are proteins containing eight conserved cysteine residues that occur uniquely in mycelial fungi. Their main function is to confer hydrophobicity to fungal surfaces in contact with air or during attachment of hyphae to hydrophobic surfaces of hosts, symbiotic partners or themselves resulting in morphogenetic signals. Based on their hydropathy patterns and solubility characteristics, hydrophobins are divided into two classes (I and II, the latter being found only in ascomycetes. Results 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 draft sequenced genomes (H. jecorina = T. reesei, H. atroviridis = T. atroviride; H. virens = T. virens an additional 14,000 ESTs from six other Trichoderma spp. (T. asperellum, H. lixii = T. harzianum, T. aggressivum var. europeae, T. longibrachiatum, T. cf. viride. The former three contained six, ten and nine members, respectively. Ten is the highest number found in any ascomycete so far. All the hydrophobins we examined had the conserved four beta-strands/one helix structure, which is stabilized by four disulfide bonds. In addition, a small number of these hydrophobins (HFBscontained an extended N-terminus rich in either proline and aspartate, or glycine-asparagine. Phylogenetic analysis reveals a mosaic of terminal clades containing 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 were from Sordariomycetes. A combined phylogeny of these sequences with those of Trichoderma showed that the Trichoderma HFBs mostly formed their own clades

  11. 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.

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

    Science.gov (United States)

    Ijjas, Anna; Steinhardt, Paul J.

    2016-02-01

    The results from Planck2015, when combined with earlier observations from the Wilkinson Microwave Anisotropy Probe, Atacama Cosmology Telescope, South Pole Telescope 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 cosmology, has been proposed that can produce distinctive predictions.

  13. 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...

  14. 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

  15. 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

  16. 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

  17. Pseudozyma vetiver sp. nov., a novel anamorphic ustilaginomycetous yeast species isolated from the phylloplane in Thailand.

    Science.gov (United States)

    Chamnanpa, Thunnicha; Limtong, Pitayakon; Srisuk, Nantana; Limtong, Savitree

    2013-11-01

    Three strains representing one novel yeast species were isolated from the phylloplanes of the vetiver grasses (DMKU-LV90 and DMKU-LV99(T)) and sugarcane (DMKU-SP260) collected in Thailand by leaf washing followed by a plating technique. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and the sequence analysis of the D1/D2 region of the large subunit (LSU) rRNA gene and the internal transcribed spacer region (ITS), the three strains were found to represent a single novel anamorphic ustilaginomycetous yeast species in the genus Pseudozyma. The name Pseudozyma vetiver sp. nov. is proposed for this novel species. The type strain is DMKU-LV99(T) (BCC 61021 = CBS 12824). The novel species showed phylogenetic relationships to the other members of the genus Pseudozyma and to teleomorphic fungal genera, namely Ustilago, Sporisorium and Anomalomyces in Ustilaginaceae, Ustilaginales. The three strains showed identical sequences both in the D1/D2 and ITS regions. The Pseudozyma species closest to the novel species in terms of pairwise sequence similarity in the D1/D2 region was Pseudozyma pruni but with 2.3 % nucleotide substitutions (14 nucleotide substitutions and no gaps out of 606 nt). The novel species and P. pruni differed by 10.9 % nucleotide substitutions (75 nucleotide substitutions and 31 gaps out of 691 nt) in the ITS region. The phylogenetic analysis based on the combined sequences of the ITS region and the D1/D2 region of the LSU rRNA gene showed that the novel species was found to be most closely related to Pseudozyma fusiformata but with 2.9 % nucleotide substitutions in the D1/D2 region and 7.4 % nucleotide substitutions in the ITS region. PMID:23877892

  18. Anamorphic preclinical SPECT imaging with high-resolution silicon double-sided strip detectors

    Science.gov (United States)

    Durko, Heather L.

    Preclinical single-photon emission computed tomography (SPECT) is an essential tool for studying progression, response to treatment, and physiological changes in small animal models of human disease. The wide range of imaging applications is often limited by the static design of many preclinical SPECT systems. We have developed a prototype imaging system that replaces the standard static pinhole aperture with two sets of movable, keel-edged copper-tungsten blades configured as crossed (skewed) slits. These apertures can be positioned independently between the object and detector, producing an anamorphic image in which the axial and transaxial magnications are not constrained to be equal. We incorporated a 60 mm x 60 mm, millimeter-thick megapixel silicon double-sided strip detector that permits ultrahigh-resolution imaging. While the stopping power of silicon is low for many common clinical radioisotopes, its performance is sufficient in the range of 20-60 keV to allow practical imaging experiments. The low-energy emissions of 125I fall within this energy window, and the 60-day half life provides an advantage for longitudinal studies. The flexible nature of this system allows the future application of adaptive imaging techniques. We have demonstrated ˜225-mum axial and ˜175-mum transaxial resolution across a 2.65 cm3 cylindrical field of view, as well as the capability for simultaneous multi-isotope acquisitions. We describe the key advancements that have made this system operational, including bringing up a new detector readout ASIC, development of detector control software and data-processing algorithms, and characterization of operating characteristics. We describe design and fabrication of the adjustable slit aperture platform, as well as the development of an accurate imaging forward model and its application in a novel geometric calibration technique and a GPU-based ultrahigh-resolution reconstruction code.

  19. 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

  20. 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...

  1. 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.

  2. 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.

  3. 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

  4. 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

    enzymes is susceptible to inhibition by compounds liberated by physico‐chemical pre‐treatment if the biomass is kept unwashed. Xylan and xylo‐oligosaccharides (XOS) have been proposed to play a key role in inhibition of cellobiohydrolases of glycoside hydrolase family 7. To elucidate the mechanism behind......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 of such...

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

    OpenAIRE

    Itamar Soares de Melo; Jane L. Faull

    2000-01-01

    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 v...

  6. 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

  7. 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.

  8. 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.

  9. 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

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

    OpenAIRE

    Susanne Zeilinger; Markus Omann

    2007-01-01

    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 pla...

  11. 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.

  12. BIOLOGICAL CONTROL OF SUGAR BEET DAMPING-OFF WITH TRICHODERMA SPP.

    Science.gov (United States)

    Biological control of damping-off in sugar beet seedlings with Trichoderma species. Isolates of Trichoderma virens and other Trichoderma species are effective biocontrol agents for diseases of several crops. Control of damping-off caused by Rhizoctonia solani has been observed in a number of c...

  13. 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

  14. 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

  15. 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.

  16. Unraveling Trichoderma species in the attine ant environment: description of three new taxa.

    Science.gov (United States)

    Montoya, Quimi Vidaurre; Meirelles, Lucas Andrade; Chaverri, Priscila; Rodrigues, Andre

    2016-05-01

    Fungus-growing "attine" ants forage diverse substrates to grow fungi for food. In addition to the mutualistic fungal partner, the colonies of these insects harbor a rich microbiome composed of bacteria, filamentous fungi and yeasts. Previous work reported some Trichoderma species in the fungus gardens of leafcutter ants. However, no studies systematically addressed the putative association of Trichoderma with attine ants, especially in non-leafcutter ants. Here, a total of 62 strains of Trichoderma were analyzed using three molecular markers (ITS, tef1 and rpb2). In addition, 30 out of 62 strains were also morphologically examined. The strains studied correspond to the largest sampling carried out so far for Trichoderma in the attine ant environment. Our results revealed the richness of Trichoderma in this environment, since we found 20 Trichoderma species, including three new taxa described in the present work (Trichoderma attinorum, Trichoderma texanum and Trichoderma longifialidicum spp. nov.) as well as a new phylogenetic taxon (LESF 545). Moreover, we show that all 62 strains grouped within different clades across the Trichoderma phylogeny, which are identical or closely related to strains derived from several other environments. This evidence supports the transient nature of the genus Trichoderma in the attine ant colonies. The discovery of three new species suggests that the dynamic foraging behavior of these insects might be responsible for accumulation of transient fungi into their colonies, which might hold additional fungal taxa still unknown to science. PMID:26885975

  17. 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.

  18. 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

  19. Competition in artifical plant growth media by Trichoderma spp

    DEFF Research Database (Denmark)

    Sarocco, Sabrina; Lübeck, Mette; Vannacci, Giovanni

    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...... isolates in a natural pot mix in order to select potential fungal antagonists to be employed in the biocontrol of Rhizoctonia solani damping-off of radish, and 2) to verify the hypothesis that competition for a food base plays a role in reducing pathogen activity. Fifteen Trichoderma spp., selected among...

  20. 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.

  1. 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

  2. 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...

  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. Production and Optimization of Cellulase Enzyme Using Aspergillus niger USM AI 1 and Comparison with Trichoderma reesei via Solid State Fermentation System

    OpenAIRE

    Ibrahim, C. O.; Darah, I.; Lee, C. K.

    2011-01-01

    Novel design solid state bioreactor, FERMSOSTAT, had been evaluated in cellulase production studies using local isolate Aspergillus niger USM AI 1 grown on sugarcane bagasse and palm kernel cake at 1 : 1 (w/w) ratio. Under optimised SSF conditions of 0.5 kg substrate; 70% (w/w) moisture content; 30°C; aeration at 4 L/h · g fermented substrate for 5 min and mixing at 0.5 rpm for 5 min, about 3.4 U/g of Filter paper activity (FPase) was obtained. At the same time, comparative studies of the enz...

  6. Trichoderma volatiles effecting Arabidopsis

    DEFF Research Database (Denmark)

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

    2015-01-01

    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...

  7. 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...

  8. 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.

  9. 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

  10. 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

  11. 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:

  12. 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...

  13. 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.

  14. Comparative genome sequence analysis underscores mycoparasitism as the ancestral life style of Trichoderma

    OpenAIRE

    Kubicek, Christian P.; Herrera-Estrella, Alfredo; Seidl-Seiboth, Verena; Martinez, Diego A.; Druzhinina, Irina S.; Thon, Michael; Zeilinger, Susanne; Casas-Flores, Sergio; Horwitz, Benjamin A.; Mukherjee, Prasun K; Mukherjee, Mala; Kredics László; Alcaraz, Luis D; Aerts, Andrea; Antal Zsuzsanna (Szeged)

    2011-01-01

    Background 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. Results Here we report an analysis of the genome sequences of the two biocontrol species Trichoderma atroviride (teleomorph Hypocrea atroviridis) and Trichoderma virens (formerly Gliocl...

  15. Comparative genome sequence analysis underscores mycoparasitism as the ancestral life style of Trichoderma

    OpenAIRE

    Kubichek, C.P.; Tamayo Ramos, J.A.

    2011-01-01

    Background: 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. Results: Here we report an analysis of the genome sequences of the two biocontrol species Trichoderma atroviride (teleomorph Hypocrea atroviridis) and Trichoderma virens (formerly Glio...

  16. 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...

  17. 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.

  18. Experimental Evolution of Trichoderma citrinoviride for Faster Deconstruction of Cellulose

    OpenAIRE

    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 populati...

  19. 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.

  20. 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...

  1. 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...

  2. 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...

  3. 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.

  4. 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.

  5. 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...

  6. 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, chi18-13 and...... Trichoderma clades are observed. These observations show that Trichoderma chitinases chi18-13 and chi18-15 evolve in a manner consistent with rapid co-evolutionary interactions and identifies putative target regions involved in determining substrate-specificity....

  7. Trichoderma spp.: a biocontrol agent for sustainable management of plant diseases

    International Nuclear Information System (INIS)

    Trichoderma spp. are mainly asexual fungi that are present in all types of agricultural soils and also in decaying wood. The antagonistic activity of Trichoderma species showed that it is parasitic on many soil-borne and foliage pathogens. The fungus is also a decomposer of cellulosic waste materials. Recent discoveries show that the fungi not only act as biocontrol agents, but also stimulate plant resistance, and plant growth and development resulting in an increase in crop production. The biocontrol activity involving mycoparasitism, antibiotics and competition for nutrients, also induces defence responses or systemic resistance responses in plants. These responses are an important part of Trichoderma in biocontrol program. Currently, Trichoderma spp., is being used to control plant diseases in sustainable diseases management systems. This paper reviews the published information on Trichoderma spp., and its biocontrol activity in sustainable disease management programs. (author)

  8. 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.

  9. 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.

  10. 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

  11. The biodegradation of tallow by Trichoderma harzianum rifai RP1

    OpenAIRE

    Fleming, John Patrick

    2002-01-01

    Wastewater, activated sludge and tallow were used as sources o f organisms in enrichment cultures to screen for species capable of degrading the hard fat, tallow. A total of 58 strains were isolated, o f which seven non-filamentous and two filamentous organisms removed greater than 20% of 20g/l tallow from batch cultures. Optimum fat removal of 83% by the strain F2, identified as the fungus Trichoderma harzianum Rifai RP1, was observed in cultures buffered to pH 6, incubated at 25°C, shak...

  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. 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

  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. 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 .

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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 ...

  8. 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 ...

  9. 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...

  10. Kecernaan in vitro bahan kering dan bahan organik jerami jagung yang diinokulasi dengan Trichoderma sp pada lama inkubasi yang berbeda

    OpenAIRE

    GANI K., RASUL

    2013-01-01

    ABSTRAK Tujuan penelitian ini adalah untuk mengetahui kecernaan in vitro bahan kering dan bahan organik jerami jagung yang diinokulasi dengan Trichoderma sp. pada lama inkubasi yang berbeda. Penelitian ini disusun berdasarkan Rancangan Acak Lengkap (RAL) dengan 4 perlakuan dan 3 ulangan yaitu: P0= jerami jagung tanpa inokulasi (kontrol), P1= Jerami jagung + 5% Trichoderma sp. dengan lama waktu inkubasi 1 minggu, P2= Jerami jagung + 5% Trichoderma sp. dengan lama waktu inkubasi 2 minggu, ...

  11. 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 ...

  12. Biological control of rice brown spot with native isolates of three Trichoderma species.

    Science.gov (United States)

    Khalili, Elham; Sadravi, Mehdi; Naeimi, Shahram; Khosravi, Vahid

    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 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. PMID:24031832

  13. 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

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. Effect of Trichoderma sp. on Sclerotium rolfsii, the Causative Agent of Collar Rot on Zamioculcas zamiifolia and an on Farm Method to Mass Produce Trichoderma species

    Directory of Open Access Journals (Sweden)

    R.L.C. Wijesundera

    2010-01-01

    Full Text Available The antagonistic effect of three local isolates of Trichoderma viride and one local isolate of Trichoderma harzianum were tested against the pathogenic fungus Sclerotium rolfsii. The latter organism is responsible for major loss due to collar rot of the ornamental crop Zamioculcas zamiifolia in Sri Lanka. The disease causes massive losses. The antagonistic potential of the local isolates against the phytopathogenic fungi Sclerotium rolfsii was investigated in dual culture, poison food technique, pot trials and field trials on Zamioculcas zamiifolia plants. All Trichoderma isolates tested under in-vitro conditions significantly inhibited the growth of S. rolfsii. Of these isolates, Trichoderma viride isolate Tv1, showed highest percentage inhibition and was thus selected for in vivo field trials. Data recorded from bi monthly field application of this organism over the two growing seasons, confirmed the success of the treatment in controlling collar rot disease at the economic threshold level. Field application of testing isolate T. viride Tv1 as a conidial suspension (1011 cfu mL-1 greatly reduced the disease incidence of Zamioculcas zamiifolia plants by a percentage of 75.54%. On farm mass production of this isolate was developed to help facilitate the establishment of an integrated eco-friendly disease management system for growers of Zamioculcas zamiifolia. Different media was also evaluated to mass produce the Trichoderma isolate. The media evaluated in this study included the solid substrates barley seeds, paddy, cow pea, maize and sorghum and semi solid, liquid substrates such as potato dextrose, rice extract, paddy extracts, respectively. Although mycelial growth was fastest in barley and paddy media. And the highest yield of spores of the Trichoderma isolate was observed 7 days after inoculation in Barley and Paddy media.

  19. Promoção do crescimento do feijoeiro e controle da antracnose por Trichoderma spp Plant growth promotion of common bean and anthracnose control by 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.The objective of this work was to evaluate the ability of Trichoderma spp. to promote growth of common bean plants and to reduce severity of anthracnose (Colletotrichum lindemuthianum, as well as to identify the best performing isolates. Sixty Trichoderma spp. isolates were evaluated as to their capacity to promote growth in common bean. The seven isolates that stood out were added to the culture substrate and assessed for reduction in severity of anthracnose in bean plants treated with C. lindemuthianum conidia. The most efficient isolates in controlling the disease were identified by DNA sequencing. The IB 28/07 isolate was evaluated in the concentrations 0.5, 1, 1.5, and 2% (weight:volume, which reduced disease severity in 41.51, 55.15, 81.82, and 96.06%, respectively. The most efficient Trichoderma spp

  20. Promoção do crescimento do feijoeiro e controle da antracnose por Trichoderma spp Plant growth promotion of common bean and anthracnose control by Trichoderma spp

    OpenAIRE

    Erica Aparecida de Souza Pedro; Ricardo Harakava; Cleusa Maria Mantovanello Lucon; Sylvia Dias Guzzo

    2012-01-01

    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 antr...

  1. Efficient production of Taka-amylase A by Trichoderma viride.

    Science.gov (United States)

    Cheng, C; Udaka, S

    1991-07-01

    An efficient heterologous protein production system was developed in Trichoderma viride, a very efficient cellulase producer. An expression vector containing the Taka-amylase A gene from Aspergillus oryzae, which was fused to the strong promoter and signal peptide sequence of the cellobiohydrolase 1 gene (cbh1) of T. viride, and the hygromycin B resistance gene was used to transform protoplasts of T. viride. Using hygromycin B resistance, a frequency of 3 transformants per microgram DNA on average was obtained. One transformant showed highly elevated alpha-amylase production, 1.0 g/l, which was shown to be under the control of the cbh1 gene promoter. Analysis of the chromosomal DNA of the transformant showed the integration of more than one copy of the vector. PMID:1368719

  2. 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

  3. In vitro sensitivity of antagonistic Trichoderma atroviride to herbicides

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    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.

  4. 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

  5. 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.

  6. Trichoderma spp. decrease Fusarium root rot in common bean Trichoderma spp. reduzem a podridão-radicular de Fusário em feijoeiro comum

    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.A eficácia de seis produtos comerciais à base de Trichoderma (PCT no controle da podridão-radicular-seca do feijoeiro (PRS foi avaliada em condições de campo. Três PCT, usados no tratamento de sementes ou aplicados no sulco de plantio, aumentaram a emergência das plântulas tanto quanto o fungicida fludioxonil. A incidência de PRS não foi afetada, mas todos os PCT e o fludioxonil reduziram a severidade em relação à testemunha. A aplicação de produtos à base de Trichoderma spp. foi tão eficaz quanto o fludioxonil no manejo da PRS.

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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. PMID:25023078

  14. БИОСИНТЕЗ КСИЛАНАЗ АБОРИГЕННЫМИ ИЗОЛЯТАМИ TRICHODERMA

    OpenAIRE

    Скворцов, Е.; Алимова, Ф.; Абузярова, Д.

    2004-01-01

    Проведены исследования, показавшие высокую активность биосинтеза ксиланаз аборигенными почвенными штаммами Trichoderma. Всего исследовано более 200 штаммов. Аборигенный штамм Trichoderma 302 показал наибольшую активность синтеза ксиланаз, 8.82±0.21 IU/ml. Еще два штамма почвенных изолята Т.303 и Т.328 показали продуктивность синтеза ксиланаз выше промышленного продуцента Trichoderma reesei, ксиланазная активность препаратов которого была 2.22±0.23 IU/ml. Ксиланазы почвенного штамма Т.302 гидр...

  15. 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.

  16. 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...

  17. 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

  18. 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.

  19. Sensibilidade de isolados de Trichoderma spp. a benomil e iprodione Sensibility of Trichoderma spp. isolates to benomyl and iprodione

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Ferreira da Silva

    1999-09-01

    Full Text Available Este trabalho teve como objetivo avaliar a sensibilidade dos isolados selvagens de Trichoderma spp.; T. viride (Tal1; T. viride (T2b; T. harzianum (Tal8; T. viride (Tligjap; T. harzianum (TW5 e T. harzianum (TMA4 às concentrações de 0; 0,25; 0,5; 1,0 e 2,5µg/ml do ingrediente ativo do fungicida benomil e 0; 2,5; 5,0; 10,0 e 100,0µg/ml do ingrediente ativo do fungicida iprodione, in vitro. A avaliação consistiu de esporulação após sete dias de incubação, mediante o preparo de suspensão de esporos e contagem em câmara de Neubauer, e do cálculo da porcentagem de redução de crescimento das colônias (RC% em meio BDA suplementado com as doses estabelecidas de cada fungicida. De modo geral, os fungicidas benomil e iprodione, presentes em meio de cultura, exerceram efeito negativo sobre o crescimento micelial e esporulação dos fungos. O isolado TW5 mostrou-se mais resistente com relação à redução de crescimento na dose de 1,0µg/ml de benomil e na dose de 10,0µg/ml de iprodione, sendo também um dos isolados que mais esporulou em presença dos fungicidas.The objective of this work was to evaluate the sensibility of wild isolates of Trichoderma spp.; T. viride (Tal1; T. viride (T2b; T. harzianum (Tal8; T. viride (Tligjap; T. harzianum (TW5 e T. harzianum (TMA4; at concentrations of 0; 0.25; 0.5; 1.0; and 2.5µg/ml of active ingredient of benomyl and at concentrations of 0; 2.5; 5.0; 10.0 e 100.0µg/ml of iprodione, in vitro. Fungitoxicity was evaluated by measuring sporulation, in Neubauer chamber, after seven days of incubation and calculation of colony growth rate. In general, benomyl and iprodione impaired micelial growth and sporulation. The isolate TW5 showed to be less affected by the fungicides. The mycelial growth was reduced at benomyl and at iprodione, and it also produced more spores in the presence of fungicides than the others isolates.

  20. 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.

  1. 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

  2. Trichoderma harzianum: a biocontrol agent against Bipolaris oryzae.

    Science.gov (United States)

    Abdel-Fattah, Gamal M; Shabana, Yasser M; Ismail, Adel E; Rashad, Younes Mohamed

    2007-08-01

    Rice brown spot, caused by Bipolaris oryzae, can be a serious disease causing a considerable yield loss. Trichoderma harzianum is an effective biocontrol agent for a number of plant fungal diseases. Thus, this research was carried out to investigate the mechanisms of action by which T. harzianum antagonizes Bipolaris oryzae in vitro, and the efficacy of spray application of a spore suspension of T. harzianum for control of rice brown spot disease under field conditions. In vitro, the antagonistic behavior of T. harzianum resulted in the overgrowth of B. oryzae by T. harzianum, while the antifungal metabolites of T. harzianum completely prevented the linear growth of B. oryzae. Light and scanning electron microscope (SEM) observations showed no evidence that mycoparasitism contributed to the aggressive nature of the tested isolate of T. harzianum against B. oryzae. Under field conditions, spraying of a spore suspension of T. harzianum at 10(8)spore ml(-1) significantly reduced the disease severity (DS) and disease incidence (DI) on the plant leaves, and also significantly increased the grain yield, total grain carbohydrate, and protein, and led to a significant increase in the total photosynthetic pigments (chlorophyll a and b and carotenoids) in rice leaves. PMID:17592758

  3. 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.

  4. Biocontrol potential of Trichoderma Sp. against plant pathogens

    Directory of Open Access Journals (Sweden)

    Anand S.

    2009-12-01

    Full Text Available Forty two strains of Trichoderma sp. were isolated from cultivated lands around Bangalore andanalyzed for their antagonistic potential against Sclerotium rolfsii and Fusarium ciceri. The potential ofbiocontrol agents ultimately lies in their capacity to control pathogens in vivo. Bioefficacy studies were henceconducted using chickpea (Cicer argentums c.v. Annigeri as an experimental plant by the roll paper towelmethod. Overall the isolates T40, T35, T30 and T25 showed better antagonistic potential in addition toenhancing plant growth. The production of chitinases to break down the mycelial cell walls of fungal plantpathogens has been implicated as a major cause of biocontrol activity (Inbar and Chet, 1995. In order tostudy the mechanism of biocontrol, ten better performing strains were plated on media, amended withcolloidal chitin and Sclerotium rolfsii cell wall extract. All the isolates showed chitinolytic activity on day threeas well as day five. Production of endochitinase and exochitinase were assayed in liquid media usingcolloidal chitin amended broth. Strains T35 and T6 displayed maximum endochitinase and exochitinaseactivity. Although all strains exhibited cellulase activity, the quantum of enzyme produced was higher in T35and T6. The results also indicate a positive correlation between enzyme production and bioefficacy.

  5. 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)

  6. Biosolubilization of raw and gamma irradiated lignite by trichoderma asperellum

    International Nuclear Information System (INIS)

    Biosolubilization is a promising technology for converting solid coal to liquid oil by addition of microorganism. Aim of this research is to compare between gamma irradiated lignite (10 kGy) with raw lignite in biosolubilization by selected fungi Trichoderma asperellum. Treatments were A (MSS + gamma irradiated lignite 5% + T. asperellum) and B (MSS + raw lignite 5% + T. asperellum) with sub-merged culture. There were two parameters observed i.e. biosolubilization product based on absorbance value at λ250nm and λ450nm and metal analysis by neutron activation analysis (NAA). The highest biosolubilization will be analyzed by FTIR and GCMS. The results showed that biosolubilization of raw lignite (B) was higher than sterilized lignite (A) based on absorbance value at λ250nm and λ450nm. The metal of lignite was decreased after incubation. FTIR analysis showed that both of treatment had similar spectra on biosolubilization products. GCMS analysis showed that both of treatment had different number of hydrocarbon, i.e. C6 - C35 (A) and C10 - C35 (B) and dominated by aromatic acids, aliphatic and phenylethers. Both of treatment product had the potency as oil substituted but its recommended to deoxygenate for higher quality. (author)

  7. 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.

  8. 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

  9. 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

  10. 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

  11. Onychomycosis caused by Scytalidium dimidiatum. Report of two cases. Review of the taxonomy of the synanamorph and anamorph forms of this coelomycete

    Directory of Open Access Journals (Sweden)

    LACAZ Carlos da Silva

    1999-01-01

    Full Text Available The authors report two cases of onychomycosis in the dystrophic form, one of them involving an HIV-positive patient, provoked by Scytalidium dimidiatum, previously called Scytalidium lignicola. The subject is reviewed from the taxonomic viewpoint, considering the anamorph Hendersonula toruloidea as a synonym of Nattrassia mangiferae, and having Scytalidium dimidiatum as the major synanamorph. According to many mycologists, Scytalidium hyalinum may be a separate species or a hyaline mutant of Scytalidium dimidiatum. Scytalidium lignicola Pesante 1957 was considered to be the type-species of the genus by ELLIS (197113 and later to be a "conidial state" of Hendersonula toruloidea by the same author, today known as Nattrassia mangiferae. The microorganism lives only on the roots of certain plants (mainly Platanus and Pinus. It produces pycnidia and is not considered to be a pathogen, although it is considered as a possible emerging agent capable of provoking opportunistic fungal lesions. The importance of this topic as one of the most outstanding in fungal taxonomy, so likely to be modified over time, as well as its interest in the field of dermatologic mycology, are emphasized.

  12. 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.

  13. 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.

  14. 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.

  15. EFFECTS OF TRICHODERMA HARZIANUM ON LETTUCE IN PROTECTED CULTIVATION

    Directory of Open Access Journals (Sweden)

    Ugur Bal

    2008-07-01

    Full Text Available Effects of Trichoderma harzianum on lettuce seedling development, plant quality characteristics at harvest and yield were investigated. Experiments were carried out in an unheated glasshouse where composted straw bales were used as the growing medium. A part of the experiment was carried out in a high tunnel where soil was the growing medium. Seeds were sown in peat and seedlings were grown in the presence of T. harzianum at dosages of 5, 10 and 15 g m-2 applied at sowing. Among the seedling characteristics studied only seedling fresh weight of cv. Yedikule was increased significantly. Effect of T. harzianum on lettuce yield was not statistically significant. In the glasshouse, the straw bales were composted with two different methods. In the method I, 134g N, 13g P, 25g K, and 76 g Ca, and in the method II, 54.3g N, 26g P, and 22.6g Ca were provided to the straw bales. The marketable yield of cv. Yedikule from the straw bales of method I was 503 g plant-1 from 15 g m-2 whereas the control remained the lowest with 425 g plant-1. For the same cultivar grown on the method II the highest marketable yield was obtained from 10 g m-2 with 303 g plant-1 compared to the control (150 g plant-1. The cv. Coolguard grown in the straw bales was tested only with the method I and the highest yield was from 15 g m-2 with 570 g plant-1 compared to 551 g plant-1 from the control. Dosage main effect regarding marketable yield in soil in the tunnel was not significant.

  16. 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.

  17. 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.

  18. 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.

  19. 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 was...

  20. 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... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Cellulase enzyme preparation derived from....1250 Cellulase enzyme preparation derived from Trichoderma longibrachiatum. (a) Cellulase...

  1. Integrated genomic and transcriptomic analysis reveals mycoparasitism as the ancestoral life style of Trichoderma

    OpenAIRE

    Kubicek, Christian P

    2011-01-01

    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.

  2. 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.

  3. 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

  4. 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...

  5. Application of solid-state fermentation for cellulase enzyme production using Trichoderma viride

    Directory of Open Access Journals (Sweden)

    Kiro Mojsov

    2010-06-01

    Full Text Available The Solid-state fermentation (SSF is alternative to submerged fermentation for production antibiotics, single cell protein, enzymes, organic acids, biofuel, etc. However, the advantages of SSF in various processes are found to be greater than in submerged fermentation. This technique not only decreases the cost of the process but also makes product cheaper for consumers. The paper describes experimental application of SSF on wheat straw for production of mycelia protein and cellulase enzymes by Trichoderma viride. This actual waste from agriculture industry was used as a nourishing base by Trichoderma viride in SSF for cellulase enzyme production. Growth and enzymes production by Trichoderma viride were evaluated on wheat straw and alkali treatment wheat straw (wet processing. The growth of the microorganism (biomass content shows maximum (123.44 mg/petri dish on alkali treatment wheat straw compared (96.36 mg/petri dish on wheat straw during of 240 hours. The results obtained demonstrate that the wheat straw waste from agriculture industry can be used as inexpensive base (carbon source for industrial production on cellulase enzymes by Trichoderma viride.

  6. 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.

  7. Parasitism of Trichoderma on Meloidogyne javanica and role of the gelatinous matrix

    Science.gov (United States)

    Trichoderma (T. asperellum-203, 44 and GH11; T. atroviride-IMI 206040 and T. harzianum-248) parasitism on Meloidogyne javanica life stages was examined in vitro. Conidium attachment and parasitism differed among the various fungi. Egg masses, their derived eggs and second-stage juveniles (J2) were p...

  8. 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

  9. 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)

    Sigler, Lynne; Hambleton, Sarah; Paré, Jean A

    2013-10-01

    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

  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. 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.

  12. 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.

  13. 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

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. Effect of variations in growth parameters on cellulase activity of Trichoderma viride NSPR006 cultured on different wood-dusts

    OpenAIRE

    O.O. Olaniyi; Fabunmi, A. O.; Akinyele, J. B.

    2013-01-01

    Aims: The biotechnology research into agro wastes has been driven by the need to screen organisms for hyper-production of novel extracellular enzymes in which cellulase plays a significant role. Therefore, the aim of the study was to pre-screen selected fungal strains and optimize cultural conditions for cellulase production by Trichoderma viride NSPR006 cultured on pretreated sawdust as lignocellulosic substrate. Methodology and results: The selected fungal isolates namely Trichoderma viride...

  19. Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease

    OpenAIRE

    Felix Carlos; De Marco Janice L

    2002-01-01

    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...

  20. 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....

  1. Trichoderma-induced plant immunity likely involves both hormonal- and camalexin-dependent mechanisms in Arabidopsis thaliana and confers resistance against necrotrophic fungus Botrytis cinerea

    OpenAIRE

    Contreras-Cornejo, Hexon Angel; Macías-Rodríguez, Lourdes; Beltrán-Peña, Elda; Herrera-Estrella, Alfredo; López-Bucio, José

    2011-01-01

    Filamentous fungi belonging to the genus Trichoderma have long been recognized as agents for the biocontrol of plant diseases. In this work, we investigated the mechanisms involved in the defense responses of Arabidopsis thaliana seedlings elicited by co-culture with Trichoderma virens and Trichoderma atroviride. Interaction of plant roots with fungal mycelium induced growth and defense responses, indicating that both processes are not inherently antagonist. Expression studies of the pathogen...

  2. 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.

  3. EVALUACIÓN DE Trichoderma asperellum COMO BIORREGULADOR DE Spongospora subterranea f. sp. subterranea EVALUATION OF Trichoderma asperellum AS BIOREGULATOR OF Spongospora subterranea f. sp. subterranea

    OpenAIRE

    Liliana María Hoyos Carvajal; Sonia Jaramillo Villegas; Sergio Orduz Peralta

    2008-01-01

    La roña de la papa causada por Spongospora subterranea, una de las principales enfermedades de la papa, es un protozoo para el cual existen limitadas estrategias de control debido a que cuenta con diversos tipos de inóculo, estructuras de resistencia y hospederos alternos. El objetivo de este trabajo fue evaluar Trichoderma asperellum T-84 y T-109 sobre S. subterranea bajo condiciones de invernadero en dos experimentos, probando en plantas hasta la novena y doceava semana, variables de peso f...

  4. EVALUACIÓN DE Trichoderma asperellum COMO BIORREGULADOR DE Spongospora subterranea f. sp. subterranea EVALUATION OF Trichoderma asperellum AS BIOREGULATOR OF Spongospora subterranea f. sp. subterranea

    Directory of Open Access Journals (Sweden)

    Liliana María Hoyos Carvajal

    2008-12-01

    Full Text Available La roña de la papa causada por Spongospora subterranea, una de las principales enfermedades de la papa, es un protozoo para el cual existen limitadas estrategias de control debido a que cuenta con diversos tipos de inóculo, estructuras de resistencia y hospederos alternos. El objetivo de este trabajo fue evaluar Trichoderma asperellum T-84 y T-109 sobre S. subterranea bajo condiciones de invernadero en dos experimentos, probando en plantas hasta la novena y doceava semana, variables de peso fresco, seco y número de nódulos producidos por el patógeno. Consistentemente, las plantas tratadas con T. asperellum aumentaron el peso fresco (Experimento 1 y peso seco (Experimento 2 y redujeron el número de nódulos de S. subterranea en raíces de papa, actuando mejor en aplicaciones solos que en mezcla. Este es un estudio preliminar que sugiere que T. asperellum puede llegar a ser a futuro un potencial agente de regulación biológica para la roña de la papa, pero que requiere estudios de la interacción papa/S. subterranea/Trichoderma para su implementación.Powdery scab caused by Spongospora subterranea, is one of the main diseases on potato crops, is a protozoo for which exist limited control strategies because it counts with diverse types of inoculum, resistance structures and alternative hosts. The objective of this work was to test Trichoderma asperellum T-84 and T-109 against S. subterranea under controlled conditions in two experiments, evaluating in plants until nine and twelve week, variables of fresh and dry weight and number of galls produced by the pathogen. Consistently the plants with T. asperellum increased fresh weight (Experiment 1 and dry weight (Experiment 2 and reduced the number of nodules of S. subterranea in potato root, better in single applications than in mixture. This is a preliminary study that suggests T. asperellum could be a potential agent of biological regulation in the future for powdery scab, but it will be

  5. 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%.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. Genome-scale investigation of phenotypically distinct but nearly clonal Trichoderma strains

    Science.gov (United States)

    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

  12. 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...

  13. High-Yield Endoglucanase Production by Trichoderma harzianum IOC-3844 Cultivated in Pretreated Sugarcane Mill Byproduct

    OpenAIRE

    Aline Machado de Castro; Marcela Costa Ferreira; Juliana Cunha da Cruz; Kelly Cristina Nascimento Rodrigues Pedro; Daniele Fernandes Carvalho; Selma Gomes Ferreira Leite; Nei Pereira

    2010-01-01

    The low-cost production of cellulolytic complexes presenting high action at mild conditions and well-balanced cellulase activities is one of the major bottlenecks for the economical viability of the production of cellulosic ethanol. In the present paper, the filamentous fungus Trichoderma harzianum IOC-3844 was used for the production of cellulases from a pretreated sugarcane bagasse (namely, cellulignin), by submerged fermentation. This fungal strain produced high contents of endoglucanase a...

  14. CONTROL OF POSTHARVEST TOMATO ROT BY SPORE SUSPENSION AND ANTIFUNGAL METABOLITES OF TRICHODERMA HARZIANUM

    OpenAIRE

    El-Katatny, Momein H.; Abeer S. Emam

    2012-01-01

    Rot of cherry tomato (Lycopersicon esculentum) fruits caused by several fungal pathogens is a detrimental disease leading to substantial yield loses worldwide. Alternaria isolates were the most common fungal species isolated from healthy or rotten fruits. Trichoderma harzianum spore suspension and culture filtrate were tested for their antagonistic activity on controlling tomato fruit rot. T. harzianum isolates suppressed or interfered with the growth of different postharvest tomato fungal pa...

  15. 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.

  16. 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.

  17. 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...

  18. 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...

  19. 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...

  20. THE ANTAGONISTIC ACTIVITY OF ACTINOMYCETES OF STREPTOMYCES GENUS IN RELATION TO TRICHODERMA KONINGII

    OpenAIRE

    Barbara Breza-Boruta; Zbigniew Paluszak

    2016-01-01

    The aim of the study was to estimate the effect of actinomycetes of genus Streptomyces on the growth of the antagonistic fungus Trichoderma koningii. 150 strains of Streptomyces spp. isolated from two potato cropping systems were used to the tests. Analyses were conducted experimentally in vitro on PDA medium with pH 6 and 7. The results obtained clearly indicate the inhibitory effect of actinomycetes on the fungus T. koningii. Of the tested population of actinomycetes only two strains did no...

  1. 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...

  2. Molecular Identification Of Trichoderma Strains Collected To Develop Plant Growth-Promoting And Biocontrol Agents

    Directory of Open Access Journals (Sweden)

    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.

  3. Biocontrol of sheath blight by Trichoderma asperellum in tropical lowland rice

    OpenAIRE

    de França, Suenny Kelly Santos; Cardoso, Aline Figueiredo; Lustosa, Denise Castro; Ramos, Edson Marcos Leal Soares; de Filippi, Marta Cristina Corsi; da Silva, Gisele Barata

    2015-01-01

    Crop damage by rice sheath blight, Rhizoctonia solani, can decrease rice yield by up to 45 %. The classical control method of rice sheath blight in the Amazon region is the application of fungicides. Therefore, we tested here the efficiency of a biocontrol agent, Trichoderma asperellum, and fungicides. Two experiments of rice cultivation were carried out with seven treatments: four isolates of T. asperellum, a mixture of the four isolates, the fungicide pencycuron, and the control. The first ...

  4. 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)] ...

  5. 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.

  6. Enantiomeric oxidation of organic sulfides by the filamentous fungi Botrytis cinerea, Eutypa lata and Trichoderma viride

    OpenAIRE

    Pinedo Rivilla, Cristina; Aleu Casatejada, Josefina; González Collado, Isidro

    2007-01-01

    The biotransformations of a series of substituted sulfides were carried out with the filamentous fungi Botrytis cinerea, Eutypa lata and Trichoderma viride. Several products underwent microbial oxidation of sulfide to sulfoxide with medium to high enantiomeric purity. With regard to sulfoxide enantioselectivity, the (R)-enantiomer was favoured in biotransformations by T. viride and E. lata while the (S)-enantiomer was favoured in those by B. cinerea. A minor amount of sulfone product...

  7. Deciphering the hormonal signalling network behind the systemic resistance induced by Trichoderma harzianum in tomato

    OpenAIRE

    Martínez-Medina, Ainhoa; Fernández, Iván; Sánchez-Guzmán, María J.; Jung, Sabine C.; Pascual, Jose A.; Pozo, María J.

    2013-01-01

    Root colonization by selected Trichoderma isolates can activate in the plant a systemic defense response that is effective against a broad-spectrum of plant pathogens. Diverse plant hormones play pivotal roles in the regulation of the defense signaling network that leads to the induction of systemic resistance triggered by beneficial organisms [induced systemic resistance (ISR)]. Among them, jasmonic acid (JA) and ethylene (ET) signaling pathways are generally essential for ISR. However, Tric...

  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. PMID:26275435

  9. Efectos del Trichoderma sp. sobre el crecimiento y desarrollo de la arveja (Pisum sativum L.

    Directory of Open Access Journals (Sweden)

    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.

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

    OpenAIRE

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

    2012-01-01

    Abstract 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 ...

  11. Characterization of a protease produced by a Trichoderma harzianum isolate which controls cocoa plant witches' broom disease

    Directory of Open Access Journals (Sweden)

    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.

  12. 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

  13. 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

  14. Partial purification and characterization of Xylanase from Trichoderma viride produced under SSF

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    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

  15. 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.

  16. 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

  17. 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

    2016-01-01

    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, 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 108 conidia/ml suspension, respectively. PMID:27168653

  18. 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

  19. Application of organic amendment and Trichoderma sp. to control basal Sclerotium rolfsii on peanut grown on partially degraded land

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    H. Suheri

    2014-04-01

    Full Text Available An experiment involving the application of organic matters, either fresh or composted, application of bioagent Trichoderma sp. was conducted to suppress basal stem rot caused by S. rolfsii on peanut. The research was conducted at dry land in Amor-Amor North Lombok District with the pump water well system. The experiments were arranged as split plot design with the main plot was pathogen inoculation and non pathogen inoculation. As sub-plot were the addition of organic matters and or addition of bioagent Trichoderma sp. plus two control treatments. Pathogen S. rolfsii applied as fragment mycelia of 1 x 10 6 fragment/ mL with the number of 10 L/plot. Inoculation of the pathogen was conducted when seedlings were about one month old. The bioagent Trichoderma sp. was applied 50 g/plot as powder in the same time of organic matters application (before planting the seeds. The results showed that application of composted organic matters reduced disease incidence 6-10%, while application as fresh organic matters reduced 1-6%. Combination between application of bioagent Trichoderma sp. with fresh and or composted organic matters suppressed disease incidence 8-10% and 8-12% respectively. Application of bioagent alone on planting medium reduced disease incidence 7.5%. Application of composted organic matters with bioagent Trichoderma sp. increased the number of fresh plant biomass 4.93-11.78 kg.

  20. 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