Trichoharzianol, a new antifungal from Trichoderma harzianum F031.

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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.

Changes induced by Trichoderma harzianum in suppressive compost controlling Fusarium wilt.

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Blaya, Josefa; López-Mondéjar, Rubén; Lloret, Eva; Pascual, Jose Antonio; Ros, Margarita

2013-09-01

The addition of species of Trichoderma to compost is a widespread technique used to control different plant diseases. The biological control activity of these species is mainly attributable to a combination of several mechanisms of action, which may affect the microbiota involved in the suppressiveness of compost. This study was therefore performed to determine the effect of inoculation of Trichoderma harzianum (T. harzianum) on compost, focusing on bacterial community structure (16S rRNA) and chitinase gene diversity. In addition, the ability of vineyard pruning waste compost, amended (GCTh) or not (GC) with T. harzianum, to suppress Fusarium wilt was evaluated. The addition of T. harzianum resulted in a high relative abundance of certain chitinolytic bacteria as well as in remarkable protection against Fusarium oxysporum comparable to that induced by compost GC. Moreover, variations in the abiotic characteristics of the media, such as pH, C, N and iron levels, were observed. Despite the lower diversity of chitinolytic bacteria found in GCTh, the high relative abundance of Streptomyces spp. may be involved in the suppressiveness of this growing media. The higher degree of compost suppressiveness achieved after the addition of T. harzianum may be due not only to its biocontrol ability, but also to changes promoted in both abiotic and biotic characteristics of the growing media. Copyright © 2013 Elsevier Inc. All rights reserved.

Enhanced root and shoot growth of wheat (Triticum aestivum L.) by Trichoderma harzianum from Turkey.

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Kucuk, Cigdem

2014-01-01

It is well known that Trichoderma species can be used as biocontrol and plant growth promote agent. In this study, Trichoderma harzianum isolates were evaluated for their growth promotion effects on wheat in greenhouse experiments. Two isolates of T. harzianum were used. The experimental design was a randomized complete block with three replications. Seeds were inoculated with conidial suspensions of each isolate. Wheat plants grown in steriled soil in pots. T. harzianum T8 and T15 isolates increased wheat length, root dry weight and shoot dry weight according to untreated control. Turkish isolates T8 and T15 did not produce damage in seeds nor in plants.

Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

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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.

Effet du compost et de Trichoderma harzianum sur la suppression ...

African Journals Online (AJOL)

SARAH

31 oct. 2013 ... induit des pourcentages de réduction qui n'ont pas dépassé 81% et 75% ... Effect of compost and trichoderma harzianum on verticillium wilt of greenhouse tomato crop ... telluriques comme Pythium, Phytophthora, Fusarium.

INTERAKSI ANTARA Trichoderma Harzianum, Penicillium SP. DAN Pseudomonas SP. SERTA KAPASITAS ANTAGONISMENYA TERHADAP Phytophthora CapsicilN VITRO*[Interaction Among Trichoderma Harzianum, Penicillium SP., Pseudomonas SP. and Antagonism Capacities Against Phy

OpenAIRE

Suharna, Nandang

2003-01-01

A preliminary study has been done to know antagonism capacities of three isolates of Trichoderma harzianum, two isolates of Penicillium sp.and one isolate of Pseudomonas sp.against Phytophthora capsici in vitro and interaction among those six antagonists.The highest antagonism capacity possessed by Penicillium sp. KN1, respectively followed by Penicillium sp.KN2,Pseudomonas sp. GH1 and the three T. harzianum isolates. Except for those three T. harzianum isolates, the two Penicillium sp.isolat...

EFIKASI TRICHODERMA HARZIANUM DENGAN BERBAGAI BAHAN ORGANIK DALAM PENGENDALIAN PENYAKIT BUSUK PANGKAL BATANG PADA LADA

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Cipta Ginting

2011-11-01

Full Text Available The objective of this research was to determine the influence of the kinds of organic matter on the efficacy of Trichoderma harzianum Rifai to control foot rot of black pepper caused by Phytophthora capsici Leonian.  Trichoderma spp. were isolated from suppressive and non-suppressive soils taken from black pepper fields with high disease incidence.  Screening of Trichoderma spp. isolates was conducted through antagonistic test with dual culture technique.  Treatments were arranged in a completely randomized design with six replications.  Treatments were rice husk, rice straw, wood dust, Arachis pintoi, mixture of the four organic matters, the mixture without T. harzianum, and without organic matter.  The test was conducted in greenhouse with media consisted of soil, organic matter, and sand (2 : 2 : 1, v/v.  After being otoclaved, the medium was infested with T. harzianum and P. capsici each with five mycelium plugs of 1-cm diameter.  Black pepper seedlings were planted 5 days after fungal infestation.  After planting the seedlings, five leaf cuts were partly inserted into the soil on each pot.  The variables observed were disease incidence on the leaf cuts inserted into the soil and disease severity on the stems and roots.  The results show that all 16 Trichoderma isolates inhibited P. capsici colonies and some isolates showed stronger inhibition than the others.  T. harzianum reduced disease severity, but there was no effect of the kinds of organic matter on the ability of T. harzianum to control foot rot.

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

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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.

The Effect of Trichoderma harzianum and Cadmium on Tolerance Index and Yield of Barley (Hordeum vulgare L.

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F. Taghavi Ghasemkheyli

2015-01-01

Full Text Available To investigate the effect of Trichoderma harzianum, as a bioabsorbant to ameliorate the harmful effects of cadmium (Cd on growth and yield of barley (Hordeum vulgare L. variety ‘Sahra’, a factorial pot experiment based on completely randomized design with three replicates was conducted. Trichoderma harzianum withtwo levels (with and without inoculation and cadmium nitrate with four levels (0, 50, 100 and 150 mg.L-1 were the treatments. Results of ANOVA revealed that there was a significant interaction between Trichoderma and cadmium nitrate in terms of biological yield, straw yield, harvest index, spike number per plant and seed number per spike. Mean comparisons showed that Trichoderma inoculation at all Cd levels significantly improved both biological and straw yields. Trichoderma at 50 and 100 mg.L-1 of Cd also increased the spike number per plant (up to 120 and 66%, respectively significantly. Increasing Cd levels decreased seed yield (19%, 1000 seed weight (18%, partitioning coefficient (57% and tolerance index (23% significantly. Inoculation of Trichoderma into growth medium had a significant effect on seed yield and tolerance index (up to 17 and 22%, respectively. In conclusion, Trichoderma harzianum inoculation at lower concentrations of Cd (50 and 100 mg.L-1 could be effective to improve growth parameters of barley plant.

Trichoderma harzianum T-78 supplementation of compost stimulates the antioxidant defence system in melon plants.

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Bernal-Vicente, Agustina; Pascual, José A; Tittarelli, Fabio; Hernández, José A; Diaz-Vivancos, Pedro

2015-08-30

Compost is emerging as an alternative plant growing medium in efforts to achieve more sustainable agriculture. The addition of specific microorganisms such as Trichoderma harzianum to plant growth substrates increases yields and reduces plant diseases, but the mechanisms of such biostimulants and the biocontrol effects are not yet fully understood. In this work we investigated how the addition of citrus and vineyard composts, either alone or in combination with T. harzianum T-78, affects the antioxidant defence system in melon plants under nursery conditions. Compost application and/or Trichoderma inoculation modulated the antioxidant defence system in melon plants. The combination of citrus compost and Trichoderma showed a biostimulant effect that correlated with an increase in ascorbate recycling enzymes (monodehydroascorbate reductase, dehydroascorbate reductase) and peroxidase. Moreover, the inoculation of both composts with Trichoderma increased the activity of antioxidant enzymes, especially those involved in ascorbate recycling. Based on the long-established relationship between ascorbic acid and plant defence responses as well as plant growth and development, it can be suggested that ascorbate recycling activities play a major role in the protection provided by Trichoderma and its biostimulant effect and that these outcomes are linked to increases in antioxidant enzymes. We can conclude that the combination of citrus compost and T. harzianum T-78 constitutes a viable, environmentally friendly strategy for improving melon plant production. © 2014 Society of Chemical Industry.

Trichoderma harzianum enhances tomato indirect defense against aphids.

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Coppola, Mariangela; Cascone, Pasquale; Chiusano, Maria Luisa; Colantuono, Chiara; Lorito, Matteo; Pennacchio, Francesco; Rao, Rosa; Woo, Sheridan Lois; Guerrieri, Emilio; Digilio, Maria Cristina

2017-12-01

Many fungal root symbionts of the genus Trichoderma are well-known for their beneficial effects on agronomic performance and protection against plant pathogens; moreover, they may enhance protection from insect pests, by triggering plant resistance mechanisms. Defense barriers against insects are induced by the activation of metabolic pathways involved in the production of defense-related plant compounds, either directly active against herbivore insects, or exerting an indirect effect, by increasing the attraction of herbivore natural enemies. In a model system composed of the tomato plant, the aphid Macrosiphum euphorbiae and the parasitoid Aphidius ervi, plant metabolic changes induced by Trichoderma harzianum and their effects on higher trophic levels have been assessed. T. harzianum T22 treatments induce a primed state that upon aphid attacks leads to an increased attraction of aphid parasitoids, mediated by the enhanced production of volatile organic compounds (VOCs) that are known to induce Aphidius ervi flight. Transcriptome sequencing of T22-treated plants infested by aphids showed a remarkable upregulation of genes involved in terpenoids biosynthesis and salicylic acid pathway, which are consistent with the observed flight response of A. ervi and the VOC bouquet profile underlying this behavioral response. © 2017 Institute of Zoology, Chinese Academy of Sciences.

Development of a multiplex Q-PCR to detect Trichoderma harzianum Rifai strain T22 in plant roots.

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Horn, Ivo R; van Rijn, Menno; Zwetsloot, Tom J J; Basmagi, Said; Dirks-Mulder, Anita; van Leeuwen, Willem B; Ravensberg, Willem J; Gravendeel, Barbara

2016-02-01

The fungal species Trichoderma harzianum is widely used as a biological agent in crop protection. To verify the continued presence of this fungus on plant roots manually inoculated with T. harzianum strain T22, a Q-PCR was designed using specific probes for this particular strain. To develop these molecular diagnostic tools, genome mining was first carried out to retrieve putative new regions by which different strains of T. harzianum could be distinguished. Subsequently, Sanger sequencing of the L-aminoacid oxidase gene (aox1) in T. harzianum was applied to determine the mutations differing between various strains isolated from the Trichoderma collection of Koppert Biological Systems. Based on the sequence information obtained, a set of hydrolysis probes was subsequently developed which discriminated T. harzianum T22 strains varying in only a single nucleotide. Probes designed for two strains uniquely recognized the respective strains in Q-PCR with a detection limit of 12,5ng DNA. Titration assays in which T. harzianum DNA from distinct strains was varied further underscored the specificity of the probes. Lastly, fungal DNA extracted from roots of greenhouse cultured tomato plants was analyzed using the probe-based assay. DNA from T. harzianum strain T22 could readily be identified on roots of greenhouse reared tomato plants inoculated with varying concentrations up to one week after treatment with a detection limit of 3e6 colony forming units of T. harzianum T22. We conclude that the Q-PCR method is a reliable and robust method for assessing the presence and quantity of T. harzianum strain T22 in manually inoculated plant material. Our method provides scope for the development of DNA based strain specific identification of additional strains of Trichoderma and other fungal biological control agents. Copyright © 2015 Elsevier B.V. All rights reserved.

Cloning, annotation and expression analysis of mycoparasitism-related genes in Trichoderma harzianum 88.

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Yao, Lin; Yang, Qian; Song, Jinzhu; Tan, Chong; Guo, Changhong; Wang, Li; Qu, Lianhai; Wang, Yun

2013-04-01

Trichoderma harzianum 88, a filamentous soil fungus, is an effective biocontrol agent against several plant pathogens. High-throughput sequencing was used here to study the mycoparasitism mechanisms of T. harzianum 88. Plate confrontation tests of T. harzianum 88 against plant pathogens were conducted, and a cDNA library was constructed from T. harzianum 88 mycelia in the presence of plant pathogen cell walls. Randomly selected transcripts from the cDNA library were compared with eukaryotic plant and fungal genomes. Of the 1,386 transcripts sequenced, the most abundant Gene Ontology (GO) classification group was "physiological process". Differential expression of 19 genes was confirmed by real-time RT-PCR at different mycoparasitism stages against plant pathogens. Gene expression analysis revealed the transcription of various genes involved in mycoparasitism of T. harzianum 88. Our study provides helpful insights into the mechanisms of T. harzianum 88-plant pathogen interactions.

Antagonism of Trichoderma harzianum ETS 323 on Botrytis cinerea mycelium in culture conditions.

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Cheng, Chi-Hua; Yang, Chia-Ann; Peng, Kou-Cheng

2012-11-01

ABSTRACT Previous studies have shown that the extracellular proteins of Trichoderma harzianum ETS 323 grown in the presence of deactivated Botrytis cinerea in culture include a putative l-amino acid oxidase and have suggested the involvement of this enzyme in the antagonistic mechanism. Here, we hypothesized that the mycoparasitic process of Trichoderma spp. against B. cinerea involves two steps; that is, an initial hyphal coiling stage and a subsequent hyphal coiling stage, with different coiling rates. The two-step antagonism of T. harzianum ETS 323 against B. cinerea during the mycoparasitic process in culture was evaluated using a biexponential equation. In addition, an l-amino acid oxidase (Th-l-AAO) was identified from T. harzianum ETS 323. The secretion of Th-l-AAO was increased when T. harzianum ETS 323 was grown with deactivated hyphae of B. cinerea. Moreover, in vitro assays indicated that Th-l-AAO effectively inhibited B. cinerea hyphal growth, caused cytosolic vacuolization in the hyphae, and led to hyphal lysis. Th-l-AAO also showed disease control against the development of B. cinerea on postharvest apple fruit and tobacco leaves. Furthermore, an apoptosis-like response, including the generation of reactive oxygen species, was observed in B. cinerea after treatment with Th-l-AAO, suggesting that Th-l-AAO triggers programmed cell death in B. cinerea. This may be associated with the two-step antagonism of T. harzianum ETS 323 against B. cinerea.

Efecto bioestimulante de Trichoderma harzianum Rifai en posturas de Leucaena, Cedro y Samán

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Leonides Castellanos González

2018-01-01

Full Text Available El presente artículo evaluó el efecto bioestimulante de Trichoderma harzianum Rifai en la producción de postura de Leucaena leucocephala (Lam de Wit., Cedrela odorata L. y Albizia saman (Jacq. Merr. Se desarrollaron tres experimentos en condiciones de vivero, uno para cada especie. Se empleó un diseño completamente aleatorio con cuatro parcelas por tratamiento. En cada experimento los tratamientos fueron: Trichoderma harzianum a razón de 20 g.L-1, 40 g.L-1 y un testigo. Se evaluó el porcentaje de germinación y las variables morfométricas diámetro y altura del tallo, así como biomasa seca en raíz y parte aérea de la planta. Los tratamientos con Trichoderma no incrementaron el porcentaje de germinación en cedro, samán y leucaena. T. harzianum incrementó la altura, el número de hojas y la biomasa seca del área foliar en las plántulas de cedro, mientras que en leucaena y samán solo provocó incrementos del diámetro basal de las plántulas.

Isolation and identification of Trichoderma harzianum from groundwater: An effective biosorbent for defluoridation of groundwater.

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Koshle, Shalini; Mahesh, S; Swamy, S Nanjunda

2016-01-01

The ability of non-viable form of Trichoderma harzianum, isolated from fluoride rich groundwater, was investigated as biosorbent for defluoridation of groundwater. Biosorption experiments were carried out at laboratory scale for removal of fluoride from groundwater. Significant effect of operational parameters on fluoride biosorption using Trichoderma harzianum as biosorbent was evaluated by varying operational parameters such as: initial fluoride concentration (2-8 mgl(-1)), biosorbent dose (0.4-1.6g/100ml), groundwater pH (6-10), temperature (30-50 degrees C) and biosorption time (30-120 min). The fluoride adsorption isotherms were modeled by Langmuir and Freundlich isotherms. Our result showed that fluoride biosorption, significantly increased with increase in groundwater pH, biosorbent dose, temperature and biosorption time, whereas increase in initial fluoride concentration reduced fluoride removal. The fluoride biosorption was rapid and maximum fluoride uptake was attained with 1.6g 100ml(-1) biosorbent within 60 min. Optimal pH 10 and temperature 50 degrees C gave maximum defluoridation efficiency. Freundlich isotherm fits well for defluoridation of groundwater using Trichoderma harzianum as biosorbent which indicated that biosorbent surface sites were heterogeneous in nature and fitted into heterogeneous site binding model.

Scanning electron microscopy observations of the interaction between Trichoderma harzianum and perithecia of Gibberella zeae.

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Inch, S; Gilbert, J

2011-01-01

Chronological events associated with the interaction between a strain of Trichoderma harzianum, T472, with known biological control activity against perithecial production of G. zeae, were studied with scanning electron microscopy to investigate the mechanisms of control. Large clusters of perithecia consisting of 5-15 perithecia formed on the autoclaved, mulched wheat straw inoculated with G. zeae alone (control) with an average of 157 perithecia per plate. Small clusters consisting of 3-6 and an average of 15 perithecia per plate perithecia formed on straw that was treated with T. harzianum. The mature perithecia from straw treated with T. harzianum produced less pigment and were lighter in color than those from the control plates. Furthermore the cells of the outer wall of these perithecia were abnormal in appearance and unevenly distributed across the surface. Immature perithecia were colonized by T. harzianum approximately 15 d after inoculation (dai) with the biocontrol agent and pathogen. Few perithecia were colonized at later stages. The affected perithecia collapsed 21 dai, compared to the perithecia in the control samples that began to collapse 28 dai. Abundant mycelium of T. harzianum was seen on the perithecia of treated samples. Perithecial structures may be resistant to penetration by the mycelium because direct penetration was not observed. Trichoderma harzianum colonized the substrate quickly and out-competed the pathogen, G. zeae.

Involvement of Trichoderma harzianum Epl-1 Protein in the Regulation of Botrytis Virulence- and Tomato Defense-Related Genes

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Gomes, Eriston V.; Ulhoa, Cirano J.; Cardoza, Rosa E.; Silva, Roberto N.; Guti?rrez, Santiago

2017-01-01

Several Trichoderma spp. are well known for their ability to: (i) act as important biocontrol agents against phytopathogenic fungi; (ii) function as biofertilizers; (iii) increase the tolerance of plants to biotic and abiotic stresses; and (iv) induce plant defense responses via the production and secretion of elicitor molecules. In this study, we analyzed the gene-regulation effects of Trichoderma harzianum Epl-1 protein during the interactions of mutant Δepl-1 or wild-type T. harzianum stra...

Variabilidade entre isolados de Trichoderma harzianum: I - Aspectos citológicos Variability among Trichoderma harzianum isolates: I - Cytological aspects

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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.

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...

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

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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

Contribución al estudio del sistema de lipasas de Trichoderma harzianum

OpenAIRE

Jorge, Maria de Lurdes Antunes

2015-01-01

[ES]Entre los hongos del género Trichoderma se han descrito algunas especies importantes como agentes de biocontrol, tales como las parejas anamorfo/teleomorfo T. harzianum/Hypocrea lixii, T. viride/H. rufa, T. atroviride/H. atroviridis y T. virens/H. virens. Trichoderma tiene una amplia distribución geográfica, está presente en casi todos los suelos y en hábitats diversos (creciendo en madera, corteza, sobre y dentro otros hongos y sustratos innumerables). Por su adaptación a distintos en...

Designing a SCAR molecular marker for monitoring Trichoderma cf. harzianum in experimental communities* #

Science.gov (United States)

Pérez, Gabriel; Verdejo, Valentina; Gondim-Porto, Clarissa; Orlando, Julieta; Carú, Margarita

2014-01-01

Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applications, such as biocontrol, where Trichoderma could be considered as an invasive species that colonizes a recipient community. The success of this invasion depends on multiple factors, which can be assayed using experimental communities as study models. Therefore, the aim of this work is to develop a species-specific sequence-characterized amplified region (SCAR) marker to monitor the colonization and growth of T. cf. harzianum when it invades experimental communities. For this study, 16 randomly amplified polymorphic DNA (RAPD) primers of 10-mer were used to generate polymorphic patterns, one of which generated a band present only in strains of T. cf. harzianum. This band was cloned, sequenced, and five primers of 20–23 mer were designed. Primer pairs 2F2/2R2 and 2F2/2R3 successfully and specifically amplified fragments of 278 and 448 bp from the T. cf. harzianum BpT10a strain DNA, respectively. Both primer pairs were also tested against the DNA from 14 strains of T. cf. harzianum and several strains of different fungal genera as specificity controls. Only the DNA from the strains of T. cf. harzianum was successfully amplified. Moreover, primer pair 2F2/2R2 was assessed by quantitative real-time polymerase chain reaction (PCR) using fungal DNA mixtures and DNA extracted from fungal experimental communities as templates. T. cf. harzianum was detectable even when as few as 100 copies of the SCAR marker were available or even when its population represented only 0.1% of the whole community. PMID:25367789

Designing a SCAR molecular marker for monitoring Trichoderma cf. harzianum in experimental communities.

Science.gov (United States)

Pérez, Gabriel; Verdejo, Valentina; Gondim-Porto, Clarissa; Orlando, Julieta; Carú, Margarita

2014-11-01

Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applications, such as biocontrol, where Trichoderma could be considered as an invasive species that colonizes a recipient community. The success of this invasion depends on multiple factors, which can be assayed using experimental communities as study models. Therefore, the aim of this work is to develop a species-specific sequence-characterized amplified region (SCAR) marker to monitor the colonization and growth of T. cf. harzianum when it invades experimental communities. For this study, 16 randomly amplified polymorphic DNA (RAPD) primers of 10-mer were used to generate polymorphic patterns, one of which generated a band present only in strains of T. cf. harzianum. This band was cloned, sequenced, and five primers of 20-23 mer were designed. Primer pairs 2F2/2R2 and 2F2/2R3 successfully and specifically amplified fragments of 278 and 448 bp from the T. cf. harzianum BpT10a strain DNA, respectively. Both primer pairs were also tested against the DNA from 14 strains of T. cf. harzianum and several strains of different fungal genera as specificity controls. Only the DNA from the strains of T. cf. harzianum was successfully amplified. Moreover, primer pair 2F2/2R2 was assessed by quantitative real-time polymerase chain reaction (PCR) using fungal DNA mixtures and DNA extracted from fungal experimental communities as templates. T. cf. harzianum was detectable even when as few as 100 copies of the SCAR marker were available or even when its population represented only 0.1% of the whole community.

Degradation of polyethylene by Trichoderma harzianum--SEM, FTIR, and NMR analyses.

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Sowmya, H V; Ramalingappa; Krishnappa, M; Thippeswamy, B

2014-10-01

Trichoderma harzianum was isolated from local dumpsites of Shivamogga District for use in the biodegradation of polyethylene. Soil sample of that dumpsite was used for isolation of T. harzianum. Degradation was carried out using autoclaved, UV-treated, and surface-sterilized polyethylene. Degradation was monitored by observing weight loss and changes in physical structure by scanning electron microscopy, Fourier transform infrared spectroscopy, and nuclear magnetic resonance spectroscopy. T. harzianum was able to degrade treated polyethylene (40%) more efficiently than autoclaved (23%) and surface-sterilized polyethylene (13%). Enzymes responsible for polyethylene degradation were screened from T. harzianum and were identified as laccase and manganese peroxidase. These enzymes were produced in large amount, and their activity was calculated using spectrophotometric method and crude extraction of enzymes was carried out. Molecular weight of laccase was determined as 88 kDa and that of manganese peroxidase was 55 kDa. The capacity of crude enzymes to degrade polyethylene was also determined. By observing these results, we can conclude that this organism may act as solution for the problem caused by polyethylene in nature.

The relation between xyr1 overexpression in Trichoderma harzianum and sugarcane bagasse saccharification performance.

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da Silva Delabona, Priscila; Rodrigues, Gisele Nunes; Zubieta, Mariane Paludetti; Ramoni, Jonas; Codima, Carla Aloia; Lima, Deise Juliana; Farinas, Cristiane Sanchez; da Cruz Pradella, José Geraldo; Seiboth, Bernhard

2017-03-20

This work investigates the influence of the positive regulator XYR1 of Trichoderma harzianum on the production of cellulolytic enzymes, using sugarcane bagasse as carbon source. Constitutive expression of xyr1 was achieved under the control of the strong Trichoderma reesei pki1 promoter. Five clones with xyr1 overexpression achieved higher xyr1 expression and greater enzymatic productivity when cultivated under submerged fermentation, hence validating the genetic construction for T. harzianum. Clone 5 presented a relative expression of xyr1 26-fold higher than the parent strain and exhibited 66, 37, and 36% higher values for filter paper activity, xylanase activity, and β-glucosidase activity, respectively, during cultivation in a stirred-tank bioreactor. The overexpression of xyr1 in T. harzianum resulted in an enzymatic complex with significantly improved performance in sugarcane bagasse saccharification, with an enhancement of 25% in the first 24h. Our results also show that constitutive overexpression of xyr1 leads to the induction of several important players in biomass degradation at early (24h) and also late (48h) timepoints of inoculation. However, we also observed that the carbon catabolite repressor CRE1 was upregulated in xyr1 overexpression mutants. These findings demonstrate the feasibility of improving cellulase production by modifying regulator expression and suggest an attractive approach for increasing total cellulase productivity in T. harzianum. Copyright © 2017 Elsevier B.V. All rights reserved.

Mycoparasitism studies of Trichoderma harzianum against Sclerotinia sclerotiorum: evaluation of antagonism and expression of cell wall-degrading enzymes genes.

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Troian, Rogério Fraga; Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Arruda, Walquiria; Ulhoa, Cirano José

2014-10-01

Trichoderma spp. are known for their biocontrol activity against several plant pathogens. A specific isolate of Trichoderma harzianum, 303/02, has the potential to inhibit the growth of Sclerotinia sclerotiorum, an important agent involved in several crop diseases. In this study, the interaction between T. harzianum 303/02 and mycelia, sclerotia and apothecia of S. sclerotiorum was studied by scanning electron microscopy. RT-qPCR was used to examine the expression of 11 genes potentially involved in biocontrol. T. harzianum 303/02 parasitizes S. sclerotiorum by forming branches that coil around the hyphae. The fungus multiplied abundantly at the sclerotia and apothecia surface, forming a dense mycelium that penetrated the inner surface of these structures. The levels of gene expression varied according to the type of structure with which T. harzianum was interacting. The data also showed the presence of synergistic action between the cell-wall degrading enzymes.

Biological Control of Meloidogyne javanica on Tomato by Trichoderma harzianum BI and Salicylic Acid

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, F. Naserinasab; , N. Sahebani; , H.R. Etebarian

2016-01-01

In this study, Trichoderma harzianum BI was evaluated for its capacity to reduce the incidence and pathogenicity of the root-knot nematode Meloidogyne javanica on tomato. Culture Şltrates of T. harzianum BI at different concentrations, (standard, 1:1, 1:10, and 1:100) were studied. In vitro studies revealed that hatching of M. javanica eggs was inhibited by the culture Şltrates and this inhibition was positively correlated with increase in the concentration of culture Şltrates. Parasitism of ...

Efficacy of soil solarization, Trichoderma harzianum, and coffee pulp amendment against Armillaria sp

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

2003-01-01

Soil solarization was evaluated singly or in combination with Trichoderma harzianum infestation or coffee pulp amendment for its effect on wood-borne inoculum of an Armillaria sp. pathogenic on tea. Solarization increased maximum soil temperatures at 10 cm depth by 9-12degreesC and reduced viability

Dose-dependent response of Trichoderma harzianum in improving drought tolerance in rice genotypes.

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Pandey, Veena; Ansari, Mohammad W; Tula, Suresh; Yadav, Sandep; Sahoo, Ranjan K; Shukla, Nandini; Bains, Gurdeep; Badal, Shail; Chandra, Subhash; Gaur, A K; Kumar, Atul; Shukla, Alok; Kumar, J; Tuteja, Narendra

2016-05-01

This study demonstrates a dose-dependent response of Trichoderma harzianum Th-56 in improving drought tolerance in rice by modulating proline, SOD, lipid peroxidation product and DHN / AQU transcript level, and the growth attributes. In the present study, the effect of colonization of different doses of T. harzianum Th-56 strain in rice genotypes were evaluated under drought stress. The rice genotypes treated with increasing dose of T. harzianum strain Th-56 showed better drought tolerance as compared with untreated control plant. There was significant change in malondialdehyde, proline, higher superoxide dismutase level, plant height, total dry matter, relative chlorophyll content, leaf rolling, leaf tip burn, and the number of scorched/senesced leaves in T. harzianum Th-56 treated rice genotypes under drought stress. This was corroborated with altered expression of aquaporin and dehydrin genes in T. harzianum Th-56 treated rice genotypes. The present findings suggest that a dose of 30 g/L was the most effective in improving drought tolerance in rice, and its potential exploitation will contribute to the advancement of rice genotypes to sustain crop productivity under drought stress. Interaction studies of T. harzianum with three aromatic rice genotypes suggested that PSD-17 was highly benefitted from T. harzianum colonization under drought stress.

Transcriptome profile of Trichoderma harzianum IOC-3844 induced by sugarcane bagasse.

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Horta, Maria Augusta Crivelente; Vicentini, Renato; Delabona, Priscila da Silva; Laborda, Prianda; Crucello, Aline; Freitas, Sindélia; Kuroshu, Reginaldo Massanobu; Polikarpov, Igor; Pradella, José Geraldo da Cruz; Souza, Anete Pereira

2014-01-01

Profiling the transcriptome that underlies biomass degradation by the fungus Trichoderma harzianum allows the identification of gene sequences with potential application in enzymatic hydrolysis processing. In the present study, the transcriptome of T. harzianum IOC-3844 was analyzed using RNA-seq technology. The sequencing generated 14.7 Gbp for downstream analyses. De novo assembly resulted in 32,396 contigs, which were submitted for identification and classified according to their identities. This analysis allowed us to define a principal set of T. harzianum genes that are involved in the degradation of cellulose and hemicellulose and the accessory genes that are involved in the depolymerization of biomass. An additional analysis of expression levels identified a set of carbohydrate-active enzymes that are upregulated under different conditions. The present study provides valuable information for future studies on biomass degradation and contributes to a better understanding of the role of the genes that are involved in this process.

A novel L-amino acid oxidase from Trichoderma harzianum ETS 323 associated with antagonism of Rhizoctonia solani.

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Yang, Chia-Ann; Cheng, Chi-Hua; Lo, Chaur-Tsuen; Liu, Shu-Ying; Lee, Jeng-Woei; Peng, Kou-Cheng

2011-05-11

Trichoderma spp. are used as biocontrol agents against phytopathogens such as Rhizoctonia solani, but their biocontrol mechanisms are poorly understood. A novel L-amino oxidase (Th-LAAO) was identified from the extracellular proteins of Trichoderma harzianum ETS 323. Here, we show a FAD-binding glycoprotein with the best substrate specificity constant for L-phenylalanine. Although the amino acid sequence of Th-LAAO revealed limited homology (16-24%) to other LAAO members, a highly conserved FAD-binding motif was identified in the N-terminus. Th-LAAO was shown to be a homodimeric protein, but the monomeric form was predominant when grown in the presence of deactivated Rhizoctonia solani. Furthermore, in vitro assays demonstrated that Th-LAAO had an antagonistic effect against Rhizoctonia solani and a stimulatory one on hyphal density and sporulation in T. harzianum ETS 323. These findings further our understanding of T. harzianum as a biocontrol agent and provide insight into the biological function of l-amino acid oxidase.

Involvement of Trichoderma harzianum Epl-1 Protein in the Regulation of Botrytis Virulence- and Tomato Defense-Related Genes.

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Gomes, Eriston V; Ulhoa, Cirano J; Cardoza, Rosa E; Silva, Roberto N; Gutiérrez, Santiago

2017-01-01

Several Trichoderma spp. are well known for their ability to: (i) act as important biocontrol agents against phytopathogenic fungi; (ii) function as biofertilizers; (iii) increase the tolerance of plants to biotic and abiotic stresses; and (iv) induce plant defense responses via the production and secretion of elicitor molecules. In this study, we analyzed the gene-regulation effects of Trichoderma harzianum Epl-1 protein during the interactions of mutant Δ epl-1 or wild-type T. harzianum strains with: (a) the phytopathogen Botrytis cinerea and (b) with tomato plants, on short (24 h hydroponic cultures) and long periods (4-weeks old plants) after Trichoderma inoculation. Our results indicate that T. harzianum Epl-1 protein affects the in vitro expression of B. cinerea virulence genes, especially those involved in the botrydial biosynthesis ( BcBOT genes), during the mycoparasitism interaction. The tomato defense-related genes were also affected, indicating that Epl-1 is involved in the elicitation of the salicylic acid pathway. Moreover, Epl-1 also regulates the priming effect in host tomato plants and contributes to enhance the interaction with the host tomato plant during the early stage of root colonization.

Involvement of Trichoderma harzianum Epl-1 Protein in the Regulation of Botrytis Virulence- and Tomato Defense-Related Genes

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Eriston V. Gomes

2017-05-01

Full Text Available Several Trichoderma spp. are well known for their ability to: (i act as important biocontrol agents against phytopathogenic fungi; (ii function as biofertilizers; (iii increase the tolerance of plants to biotic and abiotic stresses; and (iv induce plant defense responses via the production and secretion of elicitor molecules. In this study, we analyzed the gene-regulation effects of Trichoderma harzianum Epl-1 protein during the interactions of mutant Δepl-1 or wild-type T. harzianum strains with: (a the phytopathogen Botrytis cinerea and (b with tomato plants, on short (24 h hydroponic cultures and long periods (4-weeks old plants after Trichoderma inoculation. Our results indicate that T. harzianum Epl-1 protein affects the in vitro expression of B. cinerea virulence genes, especially those involved in the botrydial biosynthesis (BcBOT genes, during the mycoparasitism interaction. The tomato defense-related genes were also affected, indicating that Epl-1 is involved in the elicitation of the salicylic acid pathway. Moreover, Epl-1 also regulates the priming effect in host tomato plants and contributes to enhance the interaction with the host tomato plant during the early stage of root colonization.

Trichoderma harzianum: Inhibition of mycotoxin producing fungi and toxin biosynthesis.

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Braun, H; Woitsch, L; Hetzer, B; Geisen, R; Zange, B; Schmidt-Heydt, M

2018-04-19

A quarter of the world-wide crop is spoiled by filamentous fungi and their mycotoxins and weather extremes associated with the climate change lead to further deterioration of the situation. The ingestion of mycotoxins causes several health issues leading in the worst case to cancer in humans and animals. Common intervention strategies against mycotoxin producing fungi, such as the application of fungicides, may result in undesirable residues and in some cases to a stress induction of mycotoxin biosynthesis. Moreover, development of fungicide resistances has greatly impacted pre- and postharvest fungal diseases. Hence there is the need to develop alternative strategies to reduce fungal infestation and thus mycotoxin contamination in the food chain. Such a strategy for natural competition of important plant-pathogenic and mycotoxin producing fungi could be Trichoderma harzianum, a mycoparasitic fungus. Especially in direct comparison to certain tested fungicides, the inhibition of different tested fungal species by T. harzianum was comparable, more sustainable and in some cases more effective, too. Besides substantially reduced growth rates, a transcriptional based inhibition of mycotoxin biosynthesis in the competed Aspergillus species could be shown. Furthermore it could be clearly observed by high-resolution Scanning Electron Microscopy (SEM) that T. harzianum actively attaches to the competitor species followed by subsequent enzymatic lysis of those mycelial filaments. The analyzed isolate of T. harzianum MRI349 is not known to produce mycotoxins. In this study it could be successfully proven that T. harzianum as a biological competitor is an effective complement to the use of fungicides. Copyright © 2018 Elsevier B.V. All rights reserved.

Secretome analysis of the fungus Trichoderma harzianum grown on cellulose.

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Do Vale, Luis H F; Gómez-Mendoza, Diana P; Kim, Min-Sik; Pandey, Akhilesh; Ricart, Carlos A O; Ximenes F Filho, Edivaldo; Sousa, Marcelo V

2012-08-01

Trichoderma harzianum is a mycoparasitic filamentous fungus that produces and secretes a wide range of extracellular hydrolytic enzymes used in cell wall degradation. Due to its potential in biomass conversion, T. harzianum draws great attention from biofuel and biocontrol industries and research. Here, we report an extensive secretome analysis of T. harzianum. The fungus was grown on cellulose medium, and its secretome was analyzed by a combination of enzymology, 2DE, MALDI-MS and -MS/MS (Autoflex II), and LC-MS/MS (LTQ-Orbitrap XL). A total of 56 proteins were identified using high-resolution MS. Interestingly, although cellulases were found, the major hydrolytic enzymes secreted in the cellulose medium were chitinases and endochitinases, which may reflect the biocontrol feature of T. harzianum. The glycoside hydrolase family, including chitinases (EC 3.2.1.14), endo-N-acetylglucosaminidases (EC 3.2.1.96), hexosaminidases (EC 3.2.1.52), galactosidases (EC 3.2.1.23), xylanases (EC 3.2.1.8), exo-1,3-glucanases (EC 3.2.1.58), endoglucanases (EC 3.2.1.4), xylosidases (EC 3.2.1.37), α-L-arabinofuranosidase (EC 3.2.1.55), N-acetylhexosaminidases (EC 3.2.1.52), and other enzymes represented 51.36% of the total secretome. Few representatives were classified in the protease family (8.90%). Others (17.60%) are mostly intracellular proteins. A considerable part of the secretome was composed of hypothetical proteins (22.14%), probably because of the absence of an annotated T. harzianum genome. The T. harzianum secretome composition highlights the importance of this fungus as a rich source of hydrolytic enzymes for bioconversion and biocontrol applications. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato.

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Siddiqui, I A; Shaukat, S S

2004-01-01

To determine the influence of soil-borne fungus Trichoderma harzianum on the biocontrol performance of Pseudomonas fluorescens strain CHA0 and its 2,4-diacetylphloroglucinol (DAPG) overproducing derivative CHA0/pME3424 against Meloidogyne javanica. Amendment of the culture filtrate (CF) or methanol extract of the CF of a T. harzianum strain Th6 to P. fluorescens growth medium enhanced the production of nematicidal compound(s) by bacterial inoculants in vitro. In addition, bacteria overwhelmingly expressed phl'-'lacZ reporter gene when the medium was amended with CF of T. harzianum. Pseudomonas fluorescens and T. harzianum applied together in unsterilized sandy loam soil caused greater reduction in nematode population densities in tomato roots. Trichoderma harzianum improves root-knot nematode biocontrol by the antagonistic rhizobacterium P. fluorescens both in vitro and under glasshouse conditions. The synergistic effect of T. harzianum on the production of nematicidal compound(s) critical in biocontrol may improve the efficacy of biocontrol bacteria against plant-parasitic nematodes. Considering the inconsistent performance of the biocontrol agents under field conditions, application of a mixture of compatible T. harzianum and P. fluorescens would more closely mimic the natural situation and might broaden the spectrum of biocontrol activity with enhanced efficacy and reliability of control.

KECEPATAN PERTUMBUHAN KAPANG (Trichoderma harzianum Rifai A1300-F006 DAN AKTIVITAS SELULASE DALAM PENANGANANAN SAMPAH SELULOSA

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Pebriana Nasution

2017-04-01

Full Text Available Production of waste in urban areas reached 0.5 kg/person /day and 80% consisted of organic waste, one of many types of waste mostly generated is household waste. Generally, this household organic waste containing structural compoundsuch us long chains of cellulose.Therefore, the use of microorganism that can produce cellulase is very important to reduce the weight of garbage. Trichoderma harzianumis known as the most potential mold compared to other molds in converting cellulose. Sawdust and bran contain cellulose and hemicellulose that can be used as the main component in the mediafor its growth. The objectives of this study was: To find an effective ratio between sawdust and bran as growth mediafor Trichoderma harzianum, To know the cellulase activity of Trichoderma harzianum. This study has been conducted from April 2015 to Juli 2015 in the Laboratory of Microbiology, Department of Biology, Faculty of Mathematics and Natural Sciences, University of Andalas. The results of this study concluded that an effective ratio between sawdust and bran as a growth media for Trichoderma harzianumwhich degraded organic waste was 50:50. The highest activity of cellulase in degrading organic waste is 100%.

Production of xylan-degrading enzymes by a Trichoderma harzianum strain

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Cacais André O.Guerreiro

2001-01-01

Full Text Available Trichoderma harzianum strain 4 produced extracellular xylan-degrading enzymes, namely beta-xylanase, beta-xylosidase and alpha-arabinofuranosidase, when grown in liquid medium cultures containing oat spelt xylan as inducer. Cellulase activity was not detected. The pattern of xylan-degrading enzymes induction was influenced by the form of xylan present in the medium. They were detected in different incubation periods. Electrophoretic separation of the proteins from liquid culture filtrates by SDS-PAGE showed a variety of bands with high and low molecular weights.

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

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Felix Carlos

2002-01-01

Full Text Available Abstract Background Several Trichoderma strains have been reported to be effective in controlling plant diseases, and the action of fungal hydrolytic enzymes has been considered as the main mechanism involved in the antagonistic process. However, although Trichoderma strains were found to impair development of Crinipellis perniciosa, the causal agent of cocoa plant witches' broom disease, no fungal strain is available for effective control of this disease. We have then undertaken a program of construction of hydrolytic enzyme-overproducing Trichoderma strains aiming improvement of the fungal antagonistic capacity. The protease of an indian Trichoderma isolate showing antagonistic activity against C. perniciosa was purified to homogeneity and characterized for its kinetic properties and action on the phytopathogen cell wall. Results A protease produced by the Trichoderma harzianum isolate 1051 was purified to homogeneity by precipitation with ammonium sulfate followed by hydrophobic chromatography. The molecular mass of this protease as determined by SDS-polyacrylamide gel electrophoresis was about 18.8 kDa. Its N-terminal amino acid sequence shares no homology with any other protease. The purified enzyme substantially affected the cell wall of the phytopathogen C. perniciosa. Western-blotting analysis showed that the enzyme was present in the culture supernatant 24 h after the Trichoderma started to grow in casein-containing liquid medium. Conclusions The capacity of the Trichoderma harzianum protease to hydrolyze the cell wall of C. perniciosa indicates that this enzyme may be actually involved in the antagonistic process between the two fungi. This fact strongly suggest that hydrolytic enzyme over-producing transgenic fungi may show superior biocontrol capacity.

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

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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.

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

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Momein H. El-Katatny

2012-06-01

Full Text Available 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 pathogens albeit at different degrees. Their culture filtrate inhibited pathogen spore germination possibly due to the released extracellular diffusible metabolite(s. Besides, aberrant morphology of conidia was observed with deformation of hyphal tips. Furthermore, the resulting mycelia appeared desiccated with coagulated protoplasm leading to complete collapse of protoplasm in presence of T. harzianum culture filtrate. Application of T. harzianum spores to tomato fruits decreased disease severity significantly with the most profound effect at higher spore concentrations (108 cells per ml. Similarly, culture filtrate of T. harzianum prevented pathogen spore germination on the surface of tomato fruits leading to decreased incidence of rot symptoms at high culture filtrate concentrations. This work provides strong evidence that T. harzianum is a competent antagonist and its spore suspension and culture filtrate can be used efficiently to control postharvest tomato rot.

A novel method to prepare concentrated conidial biomass formulation of Trichoderma harzianum for seed application.

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Singh, P C; Nautiyal, C S

2012-12-01

To prepare concentrated formulation of Trichoderma harzianum MTCC-3841 (NBRI-1055) with high colony forming units (CFU), long shelf life and efficient in root colonization by a simple scrapping method. NBRI-1055 spores scrapped from potato dextrose agar plates were used to prepare a concentrated formulation after optimizing carrier material, moisture content and spore harvest time. The process provides an advantage of maintaining optimum moisture level by the addition of water rather than dehydration. The formulation had an initial 11-12 log(10) CFU g(-1). Its concentrated form reduces its application amount by 100 times (10 g 100 kg(-1) seed) and provides 3-4 log(10) CFU seed(-1). Shelf life of the product was experimentally determined at 30 and 40 °C and predicted at other temperatures following Arrhenius equation. The concentrated formulation as compared to similar products provides an extra advantage of smaller packaging for storage and transportation, cutting down product cost. Seed application of the formulation recorded significant increase in plant growth promotion. Stable and effective formulation of Trichoderma harzianum NBRI-1055 was obtained by a simple scrapping method. A new method for the production of concentrated, stable, effective and cost efficient formulation of T. harzianum has been validated for seed application. © 2012 The Society for Applied Microbiology.

qRT-PCR quantification of the biological control agent Trichoderma harzianum in peat and compost-based growing media.

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Beaulieu, Robert; López-Mondéjar, Rubén; Tittarelli, Fabio; Ros, Margarita; Pascual, José Antonio

2011-02-01

To ensure proper use of Trichoderma harzianum in agriculture, accurate data must be obtained in population monitoring. The effectiveness of qRT-PCR to quantify T. harzianum in different growing media was compared to the commonly used techniques of colony counting and qPCR. Results showed that plate counting and qPCR offered similar T. harzianum quantification patterns of an initial rapid increase in fungal population that decreased over time. However, data from qRT-PCR showed a population curve of active T. harzianum with a delayed onset of initial growth which then increased throughout the experiment. Results demonstrated that T. harzianum can successfully grow in these media and that qRT-PCR can offer a more distinct representation of active T. harzianum populations. Additionally, compost amended with T. harzianum exhibited a lower Fusarium oxysporum infection rate (67%) and lower percentage of fresh weight loss (11%) in comparison to amended peat (90% infection rate, 23% fresh weight loss). Copyright © 2010 Elsevier Ltd. All rights reserved.

Comparison of the effect of L-lysine-. cap alpha. -oxidase from Trichoderma harzianum Rifai and Trichoderma viride on nucleic acid synthesis in human tumor cells in vitro

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

1986-10-01

This paper gives comparative data on the effect of the new antitumor enzyme LO from a Soviet strain Trichoderma harzianum Rifai and from Trichoderma viride from Japan, on DNA and RNA synthesis in human ovarian carcinoma cells (CaOv strain) in culture, and also the results of the action of LO from T. harzianum Rifai on protein synthesis. Specific precursors were added before the end of the incubation time to the samples; /sup 3/H-thymidine as precursor for DNA synthesis, /sup 3/H-uridine as RNA precursor, and /sup 3/H-leucine as protein precursor. The final values of inhibition of DNA and RNA synthesis in the presence of enzyme from both sources are shown to depend in a virtually linear manner on enzyme concentration.

Influence of mineral salts upon activity of Trichoderma harzianum non-volatile metabolites on Armillaria spp. rhizomorphs

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Krystyna Przybył

2011-01-01

Full Text Available Effect of non-volatile metabolites of Trichoderma harzianum together with certain salts containing Mg++, Fe+++, Mn++, Cu++, Al+++, Ca++, K++, Na+, PO4--- and SO3--- on the production and length of rhizomorphs of Armillaria borealis, A. gallica and A. ostoyae was studied. In pure medium, T. harzianum exhibited stimulating effect on rhizomorphs of A. borealis (both number and length and A. ostoyae (only initiation. Cu++ salt totaly inhibited the initiation of rhizomorphs of Armillaria borealis, A. gallica and A. ostoyae. Effect of other compounds on the activity of T. harzianum depended on Armillaria species. The majority of chemical compounds tested supressed the activity of non-volatile metabolites of T. harzianum. Evident stimulating effect was observed under influence of sulphate salts consisting Al++ and Fe+++ on the rhizomorph number of A. borealis and A. gallica, respectively.

Liquid culture production of microsclerotia and submerged conidia by Trichoderma harzianum active against damping-off disease caused by Rhizoctonia solani.

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Kobori, Nilce N; Mascarin, Gabriel M; Jackson, Mark A; Schisler, David A

2015-04-01

Media and culturing protocols were identified that supported the formation of submerged conidia and microsclerotia (MS) by Trichoderma harzianum Rifai strain T-22 using liquid culture fermentation. Liquid media with a higher carbon concentration (36 g L(-1)) promoted MS formation at all C:N ratios tested. Hyphae aggregated to form MS after 2 d growth and after 7 d MS were fully melanized. This is the first report of MS formation by T. harzianum or any species of Trichoderma. Furthermore, submerged conidia formation was induced by liquid culture media, but yields, desiccation tolerance, and storage stability varied with C:N ratio and carbon rate. Air-dried MS granules (culture production, stabilization process, and bioefficacy of the hitherto unreported MS of T. harzianum as a potential biofungicide for use in integrated management programs against soilborne diseases. Copyright © 2014 The British Mycological Society. All rights reserved.

New insights in Trichoderma harzianum antagonism of fungal plant pathogens by secreted protein analysis.

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Monteiro, Valdirene Neves; do Nascimento Silva, Roberto; Steindorff, Andrei Stecca; Costa, Fabio Teles; Noronha, Eliane Ferreira; Ricart, Carlos André Ornelas; de Sousa, Marcelo Valle; Vainstein, Marilene Henning; Ulhoa, Cirano José

2010-10-01

Trichoderma harzianum ALL42 were capable of overgrowing and degrading Rhizoctonia solani and Macrophomina phaseolina mycelia, coiling around the hyphae with formation of apressoria and hook-like structures. Hyphae of T. harzianum ALL42 did not show any coiling around Fusarium sp. hyphae suggesting that mycoparasitism may be different among the plant pathogens. In this study, a secretome analysis was used to identify some extracellular proteins secreted by T. harzianum ALL42 after growth on cell wall of M. phaseolina, Fusarium sp., and R. solani. The secreted proteins were analyzed by two-dimensional electrophoresis and MALDI-TOF mass spectrometry. A total of 60 T. harzianum ALL42 secreted proteins excised from the gel were analyzed from the three growth conditions. While seven cell wall-induced proteins were identified, more than 53 proteins spots remain unidentified, indicating that these proteins are either novel proteins or proteins that have not yet been sequenced. Endochitinase, β-glucosidase, α-mannosidase, acid phosphatase, α-1,3-glucanase, and proteases were identified in the gel and also detected in the supernatant of culture.

Agaricus blazei as a Substrate for the Production of β-1,3-Glucanase by Trichoderma harzianum Rifai

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Roberto Da Silva

2011-01-01

Full Text Available Extracellular β-1,3-glucanase was produced by Trichoderma harzianum Rifai cultivated in the Agaricus blazei (Agaricus brasiliensis extract as a substrate in submerged fermentation. A 2²-central composite factorial design was developed using the time of culture (x1/day and Agaricus blazei extract concentration (x2/(g/L as variables, and the results were analyzed using response surface methodology (RSM. The results showed that the Agaricus blazei extract concentration was the most important variable in the production of β-1,3-glucanase, and the maximum β-1,3-glucanase activity (0.77 U/mL was obtained in one day of cultivation. The β-glucan present in the cell wall of Agaricus blazei mushroom proved to be a good substrate for inducing the production of specific β-1,3-glucanase by Trichoderma harzianum Rifai.

Salicylic acid prevents Trichoderma harzianum from entering the vascular system of roots.

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Alonso-Ramírez, Ana; Poveda, Jorge; Martín, Ignacio; Hermosa, Rosa; Monte, Enrique; Nicolás, Carlos

2014-10-01

Trichoderma is a soil-borne fungal genus that includes species with a significant impact on agriculture and industrial processes. Some Trichoderma strains exert beneficial effects in plants through root colonization, although little is known about how this interaction takes place. To better understand this process, the root colonization of wild-type Arabidopsis and the salicylic acid (SA)-impaired mutant sid2 by a green fluorescent protein (GFP)-marked Trichoderma harzianum strain was followed under confocal microscopy. Trichoderma harzianum GFP22 was able to penetrate the vascular tissue of the sid2 mutant because of the absence of callose deposition in the cell wall of root cells. In addition, a higher colonization of sid2 roots by GFP22 compared with that in Arabidopsis wild-type roots was detected by real-time polymerase chain reaction. These results, together with differences in the expression levels of plant defence genes in the roots of both interactions, support a key role for SA in Trichoderma early root colonization stages. We observed that, without the support of SA, plants were unable to prevent the arrival of the fungus in the vascular system and its spread into aerial parts, leading to later collapse. © 2014 BSPP AND JOHN WILEY & SONS LTD.

Xylanase production by Trichoderma harzianum E58

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

1989-12-01

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

Optimization of solid-state fermentation conditions for Trichoderma harzianum using an orthogonal test.

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Zhang, J D; Yang, Q

2015-03-13

The aim of this study was to develop a protocol for the production of fungal bio-pesticides with high efficiency, low cost, and non-polluting fermentation, while also increasing their survival rate under field conditions. This is the first study to develop biocontrol Trichoderma harzianum transformants TS1 that are resistant to benzimidazole fungicides. Agricultural corn stover and wheat bran waste were used as a medium and inducing carbon source for solid fermentation. Spore production was observed, and the method was optimized using single-factor tests with 4 factors at 3 levels in an orthogonal experimental design to determine the optimal culture conditions for T. harzianum TS1. In this step, we determined the best conditions for fermenting the biocontrol fungi. The optimal culture conditions for T. harzianum TS1 were cultivated for 8 days, a ratio of straw to wheat bran of 1:3, ammonium persulfate as the nitrogen source, and a water content of 30 mL. Under optimal culture conditions, the sporulation of T. harzianum TS1 reached 1.49 x 10(10) CFU/g, which was 1.46-fold higher than that achieved before optimization. Increased sporulation of T. harzianum TS1 results in better utilization of space and nutrients to achieve control of plant pathogens. This method allows for the recycling of agricultural waste straw.

Secretome analysis of the mycoparasitic fungus Trichoderma harzianum ALL 42 cultivated in different media supplemented with Fusarium solani cell wall or glucose.

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Ramada, Marcelo Henrique Soller; Steindorff, Andrei Stecca; Bloch, Carlos; Ulhoa, Cirano José

2016-02-01

Trichoderma harzianum is a fungus well known for its potential as a biocontrol agent against many fungal phytopathogens. The aim of this study was to characterize the proteins secreted by T. harzianum ALL42 when its spores were inoculated and incubated for 48 h in culture media supplemented with glucose (GLU) or with cell walls from Fusarium solani (FSCW), a phytopathogen that causes severe losses in common bean and soy crops in Brazil, as well as other crop diseases around the world. Trichoderma harzianum was able to grow in Trichoderma Liquid Enzyme Production medium (TLE) and Minimal medium (MM) supplemented with FSCW and in TLE+GLU, but was unable to grow in MM+GLU medium. Protein quantification showed that TLE+FSCW and MM+FSCW had 45- and 30- fold, respectively, higher protein concentration on supernatant when compared to TLE+GLU, and this difference was observable on 2D gel electrophoresis (2DE). A total of 94 out of 105 proteins excised from 2DE maps were identified. The only protein observed in all three conditions was epl1. In the media supplemented with FSCW, different hydrolases such as chitinases, β-1,3-glucanases, glucoamylases, α-1,3-glucanases and proteases were identified, along with other proteins with no known functions in mycoparasitism, such as npp1 and cys. Trichoderma harzianum showed a complex and diverse arsenal of proteins that are secreted in response to the presence of FSCW, with novel proteins not previously described in mycoparasitic-related studies. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The Growth of Root Rot Disease on Pepper Seed Applied by Trichoderma Harzianum Inoculum

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S. Sofian

2013-06-01

Full Text Available Normal 0 false false false IN X-NONE X-NONE MicrosoftInternetExplorer4 Root rot disease on pepper caused by Phytophthora capsici is one of the most important diseases on pepper. The using of antagonistic fungus of Trichoderma harzianum as a biological control agent of the pathogen is one of the important alternatives in controlling P. capsici without causing negative effects on the environment. The objectives of the research were to study about the ability of T. harzianum inoculum application in inhibiting the development of root-rot disease, influenced the growth of pepper seed, to studythe effective length time application of T. harzianum inoculum in inhibiting the development of root rot disease, and increased the growth of pepper seedlings. This research was arranged in a completely randomized design, with five treatments of length time application of T. harzianum inoculum i.e. control treatment without applicationtime of T. harzianum inoculum (K, application time of T. harzianum inoculum for 0 week (S0, application time of T. harzianum inoculum for 1 week (S1, application time of T. harzianum inoculum for two weeks (S2, application time of T. harzianum inoculum for three weeks (S3, and application time of T. harzianum inoculum for 4 weeks (S4 before planting. Each treatment was repeated15 times. The observed parameterswere disease percentage, the inhibition of antagonistic fungus, disease infection rate, plant height, number of leaves, wet and dry weight of plant, stem and leaves on pepper seed, and P. capsici population density. The result showed that application time of T. harzianum inoculumfor 4 weeks (S4 before planting is the most effective time in inhibiting the development of root rot disease than the other treatment sand also had significant effect on increasing the growth of pepper seed. The antagonism test showed that T. harzianum could inhibit P. capsiciin vitro. This result proves that application time of T. harzianum inoculums

Keefektivan Trichoderma harzianum sebagai Agens Pengendali Hayati Penyakit Pembuluh kayu (Vascular Streak Dieback Pada Tanaman Kakao Klon ICCRI 03 dan TSH 858

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joko susiyanto

2017-06-01

Full Text Available ABSTRAKPenyakit Pembuluh Kayu (PPK/ Vascular Streak Dieback (VSD merupakan penyakit penting yang menyerang perkebunan kakao yang disebabkan oleh patogen Oncobasidium theobromae. Penggunaan cendawan Trichoderma harzianum sebagai agens antagonis karena mempunyai kemampuan dalam menghambat pertumbuhan cendawan patogen. Pengujian keefektivan cendawan T. harzianum isolat Jember dan isolat Banyuwangi pada beberapa konsentrasi dalam mengendalikan penyakit pembuluh kayu (PPK/VSD telah dilaksanakan dikebun percobaan pusat penelitian kopi dan kakao Indonesia. Penelitian ini dilaksanakan selama 3 bulan dan pengamatan setiap minggu. Perlakuan yang digunakan dalam keefektivan yaitu: (1 T. harzianum isolat Jember dengan konsentrasi 108 spora/ml, (2 T. harzianum isolat Jember dengan konsentrasi 109 spora/ml, (3 T. harzianum isolat Jember dengan konsentrasi 1010 spora/ml, (4 T. harzianum isolat Banyuwangi konsentrasi 108 spora/ml, (5 T. harzianum isolat Banyuwangi konsentrasi 109 spora/ml, (6 T. harzianum isolat Banyuwangi konsentrasi 1010 spora/ml, dan Kontrol menggunakan air bersih. Hasil penelitian menunjukkan bahwa T. harzianum  baik isolat Jember dan Banyuwangi cukup efektif menekan perkembangan penyakit pembuluh kayu (PPK/VSD. Perlakuan T. harzianum terbaik dalam mengendalikan penyakit (PPK/VSD ditunjukkan oleh perlakuan T. harzianum  isolat Jember  109 spora/ml dengan nilai IP sebesar 0,71 % pada klon ICCRI 03, sedangkan pada klon TSH 858 isolat Banyuwangi konsentrasi 1010 spora/ml dengan nilai 7,38%. Kategori Tingkat Efikasi (TE tertinggi ditunjukkan oleh perlakuan isolat Jember konsentrasi 109 spora/ml dengan nilai sebesar 95,3% pada klon ICCRI 03, sedangkan pada klon TSH 858 isolat banyuwangi konsentrasi  1010 spora/ml dengan nilai 80,55%. Hasil uji t tanpa memperhatikan perlakuan pada Tingkat Efikasi (TE klon ICCRI 03 menunjukkan hasil berbeda nyata dengan nilai TE lebih tinggi/terbaik bila dibandingkan dengan nilai TE klon TSH 858.  �

Uji Antagonis Trichoderma harzianum Terhadap Fusarium spp. Penyebab Penyakit Layu pada Tanaman Cabai (Capsicum annum) Secara In Vitro

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Mukarlina; Khotimah, Siti; Rianti, Reny

2010-01-01

Fusarium spp., the causal agent of Fusarium wilt disease, infect sweet pepper inflicting damages on the roots, stems, leaves, flowers, and fruits. Infection of Fusarium spp. on some crops can be controlled by using Trichoderma harzianum as a biological control agent. The aims of this study were to determine: 1) the species of Fusarium infecting sweet pepper; and 2) the in vitro antagonistic potential of T. harzianum in controlling Fusarium spp. in vitro. The study was conducted fr...

Understanding the cellulolytic system of Trichoderma harzianum P49P11 and enhancing saccharification of pretreated sugarcane bagasse by supplementation with pectinase and α-L-arabinofuranosidase.

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Delabona, Priscila da Silva; Cota, Júnio; Hoffmam, Zaira Bruna; Paixão, Douglas Antonio Alvaredo; Farinas, Cristiane Sanchez; Cairo, João Paulo Lourenço Franco; Lima, Deise Juliana; Squina, Fábio Marcio; Ruller, Roberto; Pradella, José Geraldo da Cruz

2013-03-01

Supplementation of cellulase cocktails with accessory enzymes can contribute to a higher hydrolytic capacity in releasing fermentable sugars from plant biomass. This study investigated which enzymes were complementary to the enzyme set of Trichoderma harzianum in the degradation of sugarcane bagasse. Specific activities of T. harzianum extract on different substrates were compared with the extracts of Penicillium echinulatum and Trichoderma reesei, and two commercial cellulase preparations. Complementary analysis of the secretome of T. harzianum was also used to identify which enzymes were produced during growth on pretreated sugarcane bagasse. These analyses enabled the selection of the enzymes pectinase and α-L-arabinofuranosidase (AF) to be further investigated as supplements to the T. harzianum extract. The effect of enzyme supplementation on the efficiency of sugarcane bagasse saccharification was evaluated using response surface methodology. The supplementation of T. harzianum enzymatic extract with pectinase and AF increased the efficiency of hydrolysis by up to 116%. Copyright © 2012 Elsevier Ltd. All rights reserved.

Survival in soil and detection of co-transformed Trichoderma harzianum by nested PCR Sobrevivência em solo e detecção de co-transformantes de Trichoderma harzianum por PCR "nested"

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Paulo Roberto Queiroz

2004-04-01

Full Text Available The objective of this work was to evaluate the survival of two Trichoderma harzianum co-transformants, TE 10 and TE 41, carrying genes for green fluorescent protein (egfp and for resistance to benomyl, during four weeks in a contained soil microcosm. Selective culture media were used to detect viable fungal material, whose identity was confirmed by the observation of the fluorescent phenotype by direct epifluorence microscopy. PCR using two nested primer pairs specific to the egfp gene was also used to detect the transformed fungi. Although it was not possible to reliably detect the egfp gene directly from soil extracts, an enrichment step involving selective culture of soil samples in liquid medium prior to DNA extraction enabled the consistent detection of the T. harzianum co-transformants by nested PCR for the duration of the incubation period.O objetivo deste trabalho foi avaliar a sobrevivência de dois co-transformantes de Trichoderma harzianum, TE 10 e TE 41, expressando o gene da proteína de fluorescência verde (egfp e resistência a benomil, por um período de quatro semanas em microcosmo de solo, sob condições controladas. Foi utilizado um meio seletivo para detecção de material fúngico viável, o qual foi confirmado por observação quanto ao fenótipo de fluorescência em microscópio de epifluorescência direta. O fungo transformado foi detectado por PCR "nested", utilizando-se dois pares de primers específicos para o gene egfp. Foram utilizados meios líquidos enriquecidos no cultivo de amostras de solo, permitindo uma detecção consistente de co-transformantes de T. harzianum, uma vez que não foi possível a detecção do gene egfp por PCR de amostras de DNA extraídas diretamente de solo.

Efficacy of Trichoderma harzianumT22 as a biocontrol agent against root-knot nematode (Meloidogyne incognita on some soybean varieties

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T.O. Abiri

2015-01-01

Full Text Available In 2012 and 2013, a two-year field study was conducted at the University of Ilorin Teaching and Research Farm, Ilorin, the Southern Guinea Savannah Zone, Nigeria, with the aim to investigate the effect of Trichoderma harzianumT22 as a bio-control agent against a root-knot nematode (Meloidogyne incognita on some soybean varieties. The experimental field, which naturally has been known for the presence of some nematodes such as Pratylenchus, Helicitylenchus, Radopholus, Meloidogyne, Rotylenchulus, Xyphinema, was divided into two blocks, each block consisting of three plots with alleys between blocks and plots measuring 5 m and 1.5 m respectively. All treatments were replicated five times by means of a Randomized Complete Block Design. The initial soil nematode population was increased by chopping six kilograms of Meloidogyne incognita galled roots of Celosia agentea from a pure culture into all the plots. One block was treated with bio-control agent Trichoderma harzianumT22 while the second block served as a control unit. The results show that in terms of plant height, the number of branches, yield and reduction of the soil nematode population and root galls, the plants on the Trichoderma treated plots performed significantly better (P=0.05 than those in the control unit did. This therefore implies that root-knot nematodes represent a major constraint in the production of soybean while Trichoderma harzianumT22 improves the yield growth and the yield of soybean as well as better controls soil nematode populations with respect to the control trials.

Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize

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Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

2016-01-01

Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize. PMID:27830829

Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize.

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Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

2016-11-10

Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize.

POTENCIAL FARMACOINDUSTRIAL DE Trichoderma harzianum PARA FINS FARMACOTERAPÊUTICOS

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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

Cadmium induces cadmium-tolerant gene expression in the filamentous fungus Trichoderma harzianum.

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Cacciola, Santa O; Puglisi, Ivana; Faedda, Roberto; Sanzaro, Vincenzo; Pane, Antonella; Lo Piero, Angela R; Evoli, Maria; Petrone, Goffredo

2015-11-01

The filamentous fungus Trichoderma harzianum, strain IMI 393899, was able to grow in the presence of the heavy metals cadmium and mercury. The main objective of this research was to study the molecular mechanisms underlying the tolerance of the fungus T. harzianum to cadmium. The suppression subtractive hybridization (SSH) method was used for the characterization of the genes of T. harzianum implicated in cadmium tolerance compared with those expressed in the response to the stress induced by mercury. Finally, the effects of cadmium exposure were also validated by measuring the expression levels of the putative genes coding for a glucose transporter, a plasma membrane ATPase, a Cd(2+)/Zn(2+) transporter protein and a two-component system sensor histidine kinase YcbA, by real-time-PCR. By using the aforementioned SSH strategy, it was possible to identify 108 differentially expressed genes of the strain IMI 393899 of T. harzianum grown in a mineral substrate with the addition of cadmium. The expressed sequence tags identified by SSH technique were encoding different genes that may be involved in different biological processes, including those associated to primary and secondary metabolism, intracellular transport, transcription factors, cell defence, signal transduction, DNA metabolism, cell growth and protein synthesis. Finally, the results show that in the mechanism of tolerance to cadmium a possible signal transduction pathway could activate a Cd(2+)/Zn(2+) transporter protein and/or a plasma membrane ATPase that could be involved in the compartmentalization of cadmium inside the cell.

Trichoderma harzianum transformant has high extracellular alkaline proteinase expression during specific mycoparasitic interactions

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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.

Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi

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Trichothecenes are phytotoxic sesquiterpenoid compounds of fungal origin which can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in the biosynthesis of HA is the conversion of farnesyl diphosphate t...

Effect of Trichoderma isolates on tomato seedling growth response ...

African Journals Online (AJOL)

Trichoderma species are commonly used as biological control agents against phytopathogenic fungi and some isolates are able to improve plant growth. In this study, the effects of three Trichoderma isolates including Trichoderma harzianum isolate T969, T. harzianum isolate T447 and Trichoderma sp. isolate T in tomato ...

Fungal Biomass Protein Production from Trichoderma harzianum Using Rice Polishing.

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Ahmed, Sibtain; Mustafa, Ghulam; Arshad, Muhammad; Rajoka, Muhammad Ibrahim

2017-01-01

Industrially important enzymes and microbial biomass proteins have been produced from fungi for more than 50 years. High levels of crude protein as much as 45% are present in fungal biomass with balanced essential amino acids. The aim of this study was to access the potential of Trichoderma harzianum to produce fungal biomass protein from rice polishings. Maximum biomass yield was obtained at 5% (w/v) rice polishings after 72 h of incubation at 28°C at pH 4. Carbon and nitrogen ratio of 20 : 1 gave significantly higher production of fungal biomass protein. The FBP in the 75 L fermenter contained 49.50% crude protein, 32.00% true protein, 19.45% crude fiber, 9.62% ash, 11.5% cellulose content, and 0.325% RNA content. The profile of amino acids of final FBP exhibited that all essential amino acids were present in great quantities. The FBP produced by this fungus has been shown to be of good nutritional value for supplementation to poultry. The results presented in this study have practical implications in that the fungus T. harzianum could be used successfully to produce fungal biomass protein using rice polishings.

Comparative evaluation of two Trichoderma harzianum strains for major secondary metabolite production and antifungal activity.

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Ahluwalia, Vivek; Kumar, Jitendra; Rana, Virendra S; Sati, Om P; Walia, S

2015-01-01

This investigation was undertaken to identify the major secondary metabolite, produced by two Trichoderma harzianum strains (T-4 and T-5) with their antifungal activity against phytopathogenic fungi using poison food technique. The ethyl acetate extract was subjected to column chromatography using n-hexane, ethyl acetate and methanol gradually. Chromatographic separation of ethyl acetate extract of T. harzianum (T-4) resulted in the isolation and identification of palmitic acid (1), 1,8-dihydroxy-3-methylanthraquinone (2), 6-pentyl-2H-pyran-2-one (3), 2(5H)-furanone (4), stigmasterol (5) and β-sitosterol (6), while T. harzianum (T-5) gave palmitic acid (1), 1-hydroxy-3-methylanthraquinone (7), δ-decanolactone (8), 6-pentyl-2H-pyran-2-one (3), ergosterol (9), harzianopyridone (10) and 6-methyl-1,3,8-trihydroxyanthraquinone (11) as major metabolites. Among compounds screened for antifungal activity, compound 10 was found to be most active (EC50 35.9-50.2 μg mL(-1)). In conclusion, the present investigation provided significant information about antifungal activity and compounds isolated from two different strains of T. harzianum obtained from two different Himalayan locations.

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

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Samolski, Ilanit; de Luis, Alberto; Vizcaíno, Juan Antonio; Monte, Enrique; Suárez, M Belén

2009-10-13

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. 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. 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 include different stages of plant colonization, as well as

Analysis of genomic regions of Trichoderma harzianum IOC-3844 related to biomass degradation.

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Crucello, Aline; Sforça, Danilo Augusto; Horta, Maria Augusta Crivelente; dos Santos, Clelton Aparecido; Viana, Américo José Carvalho; Beloti, Lilian Luzia; de Toledo, Marcelo Augusto Szymanski; Vincentz, Michel; Kuroshu, Reginaldo Massanobu; de Souza, Anete Pereira

2015-01-01

Trichoderma harzianum IOC-3844 secretes high levels of cellulolytic-active enzymes and is therefore a promising strain for use in biotechnological applications in second-generation bioethanol production. However, the T. harzianum biomass degradation mechanism has not been well explored at the genetic level. The present work investigates six genomic regions (~150 kbp each) in this fungus that are enriched with genes related to biomass conversion. A BAC library consisting of 5,760 clones was constructed, with an average insert length of 90 kbp. The assembled BAC sequences revealed 232 predicted genes, 31.5% of which were related to catabolic pathways, including those involved in biomass degradation. An expression profile analysis based on RNA-Seq data demonstrated that putative regulatory elements, such as membrane transport proteins and transcription factors, are located in the same genomic regions as genes related to carbohydrate metabolism and exhibit similar expression profiles. Thus, we demonstrate a rapid and efficient tool that focuses on specific genomic regions by combining a BAC library with transcriptomic data. This is the first BAC-based structural genomic study of the cellulolytic fungus T. harzianum, and its findings provide new perspectives regarding the use of this species in biomass degradation processes.

Effect of Paecilomyces lilacinus, Trichoderma harzianum and Trichoderma virens fungal extracts on the hatchability of Ancylostoma eggs.

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Hofstätter, Bianca Delgado Menezes; Oliveira da Silva Fonseca, Anelise; de Souza Maia Filho, Fernando; de Souza Silveira, Julia; Persici, Beatriz Maroneze; Pötter, Luciana; Silveira, Andressa; Antoniolli, Zaida Inês; Brayer Pereira, Daniela Isabel

Ancylostoma species have demanded attention due to their zoonotic potential. The use of anthelmintics is the usual method to prevent environmental contamination by Ancylostoma eggs and larvae. Nematophagous fungi have been widely used in their biological control due to the fungus ability to capture and digest free nematode forms. The aim of this study was to evaluate the effect of four different fungal extracts of Paecilomyces lilacinus (n=2), Trichoderma harzianum (n=1) and Trichoderma virens (n=1) isolates on the hatchability of Ancylostoma eggs. Fungal extracts consisted of fungal broth culture supernatant without filtration (crude extract) and filtered broth (filtered extract), macerated mycelium (crude macerate), and macerated mycelium submitted to filtration (filtered macerate). The Ancylostoma eggs were obtained from the feces of naturally infected dogs. In vitro assays were performed in five replicates and consisted of four treatments and one control group. The activity of the fungal extracts of each evaluated fungus differed (p<0.05) from those of the control group, showing significant ovicidal activity. The hatching of the eggs suffered reduction percentages of 68.43% and 47.05% with P. lilacinus, and 56.43% with T. harzianum, when crude macerate extract was used. The reduction with the macerate extract of T. virens was slightly lower (52.25%) than that for the filtered macerate (53.64%). The results showed that all extracts were effective in reducing the hatchability of Ancylostoma eggs. The ovicidal effect observed is likely to have been caused by the action of hydrolytic enzymes secreted by the fungi. Copyright © 2016 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

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

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Ainhoa eMartinez-Medina

2013-06-01

Full Text Available Root colonization by selected Trichoderma isolates can activate in the plant a systemic defence response that is effective against a broad spectrum of plant pathogens. Diverse plant hormones play pivotal roles in the regulation of the defence signalling network that leads to the induction of systemic resistance triggered by beneficial organisms (ISR. Among them, jasmonic acid (JA and ethylene (ET signalling pathways are generally essential for ISR. However, Trichoderma ISR (TISR is believed to involve a wider variety of signalling routes, interconnected in a complex network of cross-communicating hormone pathways. Using tomato as a model, an integrative analysis of the main mechanisms involved in the systemic resistance induced by Trichoderma harzianum against the necrotrophic leaf pathogen Botrytis cinerea was performed. Root colonization by T. harzianum rendered the leaves more resistant to B. cinerea independently of major effects on plant nutrition. The analysis of disease development in shoots of tomato mutant lines impaired in the synthesis of the key defence related hormones JA, ET, salicylic acid (SA and abscisic acid (ABA and the peptide prosystemin (PS evidenced the requirement of intact JA, SA and ABA signalling pathways for a functional TISR. Expression analysis of several hormone related marker genes point to the role of priming for enhanced JA-dependent defence responses upon pathogen infection. Together, our results indicate that although TISR induced in tomato against the necrotrophs is mainly based on boosted JA-dependent responses, the pathways regulated by the plant hormones SA- and ABA are also required for successful TISR development

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

Science.gov (United States)

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, Trichoderma ISR (TISR) is believed to involve a wider variety of signaling routes, interconnected in a complex network of cross-communicating hormone pathways. Using tomato as a model, an integrative analysis of the main mechanisms involved in the systemic resistance induced by Trichoderma harzianum against the necrotrophic leaf pathogen Botrytis cinerea was performed. Root colonization by T. harzianum rendered the leaves more resistant to B. cinerea independently of major effects on plant nutrition. The analysis of disease development in shoots of tomato mutant lines impaired in the synthesis of the key defense-related hormones JA, ET, salicylic acid (SA), and abscisic acid (ABA), and the peptide prosystemin (PS) evidenced the requirement of intact JA, SA, and ABA signaling pathways for a functional TISR. Expression analysis of several hormone-related marker genes point to the role of priming for enhanced JA-dependent defense responses upon pathogen infection. Together, our results indicate that although TISR induced in tomato against necrotrophs is mainly based on boosted JA-dependent responses, the pathways regulated by the plant hormones SA- and ABA are also required for successful TISR development. PMID:23805146

Características de Trichoderma harzianum, como agente limitante en el cultivo de hongos comestibles

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Omar Romero Arenas

2009-07-01

Full Text Available El incremento dramático de incidencia y severidad de los “mohos verdes” en la producción de hongos comestibles se refleja en la aparición de formas altamente agresivas de éstos patógenos, como es el caso de los biotipos de Trichoderma harzianum (Th1, Th2., Th3 y Th4, que han sido encontrados en Europa y Norte América, donde la importancia de la patogenicidad de dicho “moho” se comprobó en 1995 con las pérdidas del 30-100% en las plantas de hongos comestibles en Chester, Pennsylvania. En México se han identificado diversos “mohos contaminantes”, entre los cuales Trichoderma spp., se encuentra frecuentemente en la producción de hongos comestibles (Agaricus bisporus, Pleurotus ostreatus y Lentinula edodes, en el 2004, un grupo de investigación detectó la presencia de cepas altamente agresivas de T. aggressivum f. aggressivum, identificadas con técnicas clásicas y moleculares, en muestras de substrato (compost contaminado, proporcionado por la principal planta de hongos de México. Actualmente se desconoce la situación sobre la distribución de Trichoderma harzianum y los problemas de contaminación en la producción de hongos comestibles, tanto de zonas rurales, como de zonas industrializadas en México, puede causar serias disminuciones en la producción de hongos comestibles y presentar pérdidas económicas para los productores de la región.

Isolation and functional analysis of Thmfs1, the first major facilitator superfamily transporter from the biocontrol fungus Trichoderma harzianum.

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Liu, Mu; Liu, Jun; Wang, Wei Min

2012-10-01

A novel major facilitator superfamily (MFS) transporter gene, Thmfs1, was isolated from Trichoderma harzianum (T. harzianum). A Thmfs1 over-expressing mutant displayed enhanced antifungal activity and fungicide tolerance, while the Thmfs1 disruption mutant showed the opposite trend. Trichodermin production in Thmfs1 disruption group (185 mg l(-1)) was decreased by less than 17 % compared to the parental strain, suggesting that Thmfs1 is not mainly responsible for trichodermin secretion. Real-time PCR showed that Thmfs1 transcript level could be induced by a certain range of trichodermin concentrations, while expression of Tri5, encoding a trichodiene synthase, was strongly inhibited under these conditions. To our knowledge, Thmfs1 is the first MFS transporter gene identified in T. harzianum.

Effect of Trichoderma harzianum on Wheat (Triticum aestivum L. Grain Yield under Different Levels of Cadmium Nitrate

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F. Taghavi Ghasemkheyli

2014-12-01

Full Text Available A pot experiment was designed to evaluate the effect of Trichoderma spp. on yield and yield components of wheat (cv. N81 under different levels of cadmium nitrate. Experiment was arranged in factorial based on completely randomized design with three replicates. Trichoderma harzianum at two levels (with and without inoculation and four levels of cadmium nitrate (0, 50, 100, 150 mg l-1 were the treatment. Results of ANOVA and mean comparisons showed that inoculation of Trichoderma increased biological yield (46% and straw yield (30% as compared to control. Cadmium pollution has led to significant decrease in harvest index, grain number per spike and partitioning coefficient up to 5, 20, 24 and 38 percent compared to control, respectively. Furthermore, cadmium and fungus interaction were significant in terms of spike number, grain weight per spike, grain yield and tolerance index. Maximum grain yield and tolerance index were recorded in Trichoderma inoculation under cadmium-free plots which nearly increased 65 and 53 percent, respectively. In conclusion, using Trichoderma under cadmium pollution could improve wheat growth, yield and tolerance index

Peptaibol, Secondary‐Metabolite, and Hydrophobin Pattern of Commercial Biocontrol Agents Formulated with Species of the Trichoderma harzianum Complex

DEFF Research Database (Denmark)

Degenkolb, Thomas; Nielsen, Kristian Fog; Dieckmann, Ralf

2015-01-01

The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics...... in plant‐protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna®, Trichosan......®, Vitalin®, Promot® WP, and TrichoMax®, formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well‐established, HPLC/MS‐based peptaibiomics approach, it could unequivocally be demonstrated that all...

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan.

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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.

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan

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Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

2015-01-01

Abstract 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

Trichoderma harzianum in combination with sheep manure amendment enhances soil suppressiveness of Fusarium wilt of tomato

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R. M. Barakat

2010-01-01

Full Text Available The effect that the biocontrol agent Trichoderma harzianum (isolate Jn14 in combination with an amendment of sheep manure has on the soil suppressiveness of Fusarium wilt of tomato was investigated over a 28-month period. A combination of T. harzianum and organic amendment at concentrations (w:w of 6 and 10% reduced tomato wilt by 21–36 % and 29–36% respectively, after 0–28 months of soil incubation. When the amendment was added at concentration of 2%, the wilt was suppressed only after 18–28 months. A combination of T. harzianum and the amendment at 6% also increased tomato plant fresh weights by 52% after 28 months, and the 10% amendment increased fresh weights by 56, 40, and 63%, after 18, 24, and 28 months respectively, compared to the experimental controls. Organic amendment at the higher concentrations further stimulated T. harzianum populations, enhanced microbial activity against Fusarium oxysporum in the soil and reduced pathogen populations. Without T. harzianum, the organic amendment at a concentration of 10% reduced disease by only 22, 24, and 23% and only after 18, 24 and 28 months of soil incubation respectively, compared with the controls. However, tomato wilt was not reduced at a 2% manure concentration in less than 12 months of incubation. Organic amendment alone at 6 and 10% reduced the pathogen population by 25% and 37% respectively after 28 months of soil incubation compared with the control. T. harzianum produced fungitoxic metabolites that reduced mycelial growth of Fusarium by 37% and conidium germination by 55% when the pathogen was grown on potato dextrose agar amended with a T. harzianum culture filtrate.

The Cerato-Platanin protein Epl-1 from Trichoderma harzianum is involved in mycoparasitism, plant resistance induction and self cell wall protection.

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Gomes, Eriston Vieira; Costa, Mariana do Nascimento; de Paula, Renato Graciano; de Azevedo, Rafael Ricci; da Silva, Francilene Lopes; Noronha, Eliane F; Ulhoa, Cirano José; Monteiro, Valdirene Neves; Cardoza, Rosa Elena; Gutiérrez, Santiago; Silva, Roberto Nascimento

2015-12-09

Trichoderma harzianum species are well known as biocontrol agents against important fungal phytopathogens. Mycoparasitism is one of the strategies used by this fungus in the biocontrol process. In this work, we analyzed the effect of Epl-1 protein, previously described as plant resistance elicitor, in expression modulation of T. harzianum genes involved in mycoparasitism process against phytopathogenic fungi; self cell wall protection and recognition; host hyphae coiling and triggering expression of defense-related genes in beans plants. The results indicated that the absence of Epl-1 protein affects the expression of all mycoparasitism genes analyzed in direct confrontation assays against phytopathogen Sclerotinia sclerotiorum as well as T. harzianum itself; the host mycoparasitic coiling process and expression modulation of plant defense genes showing different pattern compared with wild type strain. These data indicated the involvement T. harzianum Epl-1 in self and host interaction and also recognition of T. harzianum as a symbiotic fungus by the bean plants.

Elucidation of biocontrol mechanisms of Trichoderma harzianum against different plant fungal pathogens: Universal yet host specific response.

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Sharma, Vivek; Salwan, Richa; Sharma, Prem N; Kanwar, S S

2017-02-01

In the present study, different transcripts of Trichoderma harzianum ThHP-3 were evaluated for their response against four fungal pathogens Fusarium oxysporum, Colletotrichum capsici, Colletotrichum truncatum and Gloesercospora sorghi using RT-qPCR. The time course study of T. harzianum transcripts related to signal transduction, lytic enzymes, secondary metabolites and various transporters revealed variation in expression against four fungal pathogens. In a broader term, the transcripts were upregulated at various time intervals but the optimum expression of cyp3, abc, nrp, tga1, pmk, ech42 and glh20 varied with respect to host fungi. Additionally, the expression of transcripts related to transporters/cytochromes was also observed against Fusarium oxysporum after 96h whereas transcripts related to secondary metabolites and lytic enzymes showed significant difference in expression against Colletotrichum spp. from 72 to 96h. This is first study on transcriptomic response of T. harzianum against pathogenic fungi which shows their host specific response. Copyright © 2016 Elsevier B.V. All rights reserved.

MOLECULAR CHARACTERIZATION AND IDENTIFICATION OF BIOCONTROL ISOLATES OF TRICHODERMA HARZIANUM FROM EMBU DISTRICT, KENYA

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Elizabeth M Siameto

2010-10-01

Full Text Available Species in the genus Trichoderma are important as commercial source of several enzymes, biofungicides, and growth promoters. The most common biological control agents of the genus are strains of T. harzianum, T .viride and T. viriens. In this study, sixteen selected isolates of T. harzianum from different land use types in Embu, Kenya were tested for anatogonistic action against five soil borne phytopathogenic fungi (Rhizoctonia solani, Pythium sp, Fusarium graminearum, F. oxysporum f. sp phaseoli and F. oxysporum f. sp Lycopersici using dual culture assay and through production of non-volatile inhibitors. Seven isolates were further characterized using RAPD-PCR procedure to determine genetic variability. All T. harzianum isolates had considerable antagonistic effect on mycelial growth of the pathogens in dual cultures compared to the controls. Maximum inhibition occurred in Pythium sp-055E interaction (73%.The culture filtrates obtained from Czapek’s liquid medium reduced the dry weight (mg of the mycelia significantly while those from the potato dextrose broth showed minimum inhibition growth. Pythium sp was inhibited the most compared to other pathogens. Genetic similarities generated using Jaccard’s coefficient of similarity ranged from 0.231 between isolates 055E and 011E to 0.857 between isolates 010E and 015E. The technique of RAPD was efficient in demonstrating the DNA polymorphism in the isolates of T. harzianum tested showing intraspecific genetic variability. Since all T. harzianum isolates evaluated were effective in controlling colony growth of the soil borne pathogens both in dual cultures and in culture filtrates they should be tried as a broad spectrum biological control agent in the greenhouse and under field conditions.

Greenhouse evaluation of Bacillus subtilis AP-01 and Trichoderma harzianum AP-001 in controlling tobacco diseases.

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Maketon, Monchan; Apisitsantikul, Jirasak; Siriraweekul, Chatchai

2008-04-01

Two biological control agents, Bacillus subtilis AP-01 (Larminar(™)) and Trichoderma harzianum AP-001 (Trisan(™)) alone or/in combination were investigated in controlling three tobacco diseases, including bacterial wilt (Ralstonia solanacearum), damping-off (Pythium aphanidermatum), and frogeye leaf spot (Cercospora nicotiana). Tests were performed in greenhouse by soil sterilization prior to inoculation of the pathogens. Bacterial-wilt and damping off pathogens were drenched first and followed with the biological control agents and for comparison purposes, two chemical fungicides. But for frogeye leaf spot, which is an airborne fungus, a spraying procedure for every treatment including a chemical fungicide was applied instead of drenching. Results showed that neither B. subtilis AP-01 nor T harzianum AP-001 alone could control the bacterial wilt, but when combined, their controlling capabilities were as effective as a chemical treatment. These results were also similar for damping-off disease when used in combination. In addition, the combined B. subtilis AP-01 and T. harzianum AP-001 resulted in a good frogeye leaf spot control, which was not significantly different from the chemical treatment.

Production of 6-pentyl-α-pyrone by trichoderma harzianum in solid-state fermentation.

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de Souza Ramos, Aline; Fiaux, Sorele Batista; Leite, Selma Gomes Ferreira

2008-10-01

Many Trichoderma species are able to produce 6-pentyl-α-pyrone (6-PP), a lactone with coconut-like aroma. In the present work, several culture parameters were studied to enhance the production of 6-PP by Trichoderma harzianum 4040 in solid-state fermentation. Green coir powder added to a nutrient solution was used as support material for fermentation. A Plackett-Burman screening technique was applied, followed by a fractionary factorial design. The best culture conditions within the experimental domain studied were (100 g support)(-1): sucrose, 3 g; NaNO3, 0.24 g; (NH4)2SO4, 0.18 g; KH2PO4, 0.1 g; inoculum concentration, 2.2 × 10(6) spores; moisture level, 55%. The temperature established was 28°C. The fermentation under the selected conditions led to a 6-PP production six times higher (5.0 mg/g dry matter) than the initial one (0.8 mg/g dry matter) after seven days of cultivation.

Media optimization for laccase production by Trichoderma harzianum ZF-2 using response surface methodology.

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Gao, Huiju; Chu, Xiang; Wang, Yanwen; Zhou, Fei; Zhao, Kai; Mu, Zhimei; Liu, Qingxin

2013-12-01

Trichoderma harzianum ZF-2 producing laccase was isolated from decaying samples from Shandong, China, and showed dye decolorization activities. The objective of this study was to optimize its culture conditions using a statistical analysis of its laccase production. The interactions between different fermentation parameters for laccase production were characterized using a Plackett-Burman design and the response surface methodology. The different media components were initially optimized using the conventional one-factor-at-a-time method and an orthogonal test design, and a Plackett-Burman experiment was then performed to evaluate the effects on laccase production. Wheat straw powder, soybean meal, and CuSO4 were all found to have a significant influence on laccase production, and the optimal concentrations of these three factors were then sequentially investigated using the response surface methodology with a central composite design. The resulting optimal medium components for laccase production were determined as follows: wheat straw powder 7.63 g/l, soybean meal 23.07 g/l, (NH4)2SO4 1 g/l, CuSO4 0.51 g/l, Tween-20 1 g/l, MgSO4 1 g/l, and KH2PO4 0.6 g/l. Using this optimized fermentation method, the yield of laccase was increased 59.68 times to 67.258 U/ml compared with the laccase production with an unoptimized medium. This is the first report on the statistical optimization of laccase production by Trichoderma harzianum ZF-2.

Effect of trichothecene production on the plant defense response and fungal physiology: overexpression of Trichoderma arundinaceum tri4 gene in T. harzianum

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Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively,...

Truncation of a mannanase from Trichoderma harzianum improves its enzymatic properties and expression efficiency in Trichoderma reesei.

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Wang, Juan; Zeng, Desheng; Liu, Gang; Wang, Shaowen; Yu, Shaowen

2014-01-01

To obtain high expression efficiency of a mannanase gene, ThMan5A, cloned from Trichoderma harzianum MGQ2, both the full-length gene and a truncated gene (ThMan5AΔCBM) that contains only the catalytic domain, were expressed in Trichoderma reesei QM9414 using the strong constitutive promoter of the gene encoding pyruvate decarboxylase (pdc), and purified to homogeneity, respectively. We found that truncation of the gene improved its expression efficiency as well as the enzymatic properties of the encoded protein. The recombinant strain expressing ThMan5AΔCBM produced 2,460 ± 45.1 U/ml of mannanase activity in the culture supernatant; 2.3-fold higher than when expressing the full-length ThMan5A gene. In addition, the truncated mannanase had superior thermostability compared with the full-length enzyme and retained 100 % of its activity after incubation at 60 °C for 48 h. Our results clearly show that the truncated ThMan5A enzyme exhibited improved characteristics both in expression efficiency and in its thermal stability. These characteristics suggest that ThMan5AΔCBM has potential applications in the food, feed, paper, and pulp industries.

Low temperature scanning electron microscopic studies on the interaction of globodera rostochiensis woll. and trichoderma harzianum rifai

International Nuclear Information System (INIS)

Saifullah, A.; Khan, N.U.

2014-01-01

Low temperature scanning electron microscopic (LTSEM) studies revealed that Trichoderma harzianum infected mature potato cysts nematode eggs by penetrating directly the cyst wall or via natural opening of mouth. Mycelial penetration on cyst wall or egg surface has been seen. The penetration of cyst wall or egg surface was either chemical or mechanical (directly or with appresorium) or both. Freeze fractionation showed the presence of mycelia inside the eggs. (author)

Evaluation of seed coating formulations of Trichoderma harzianum on cucumber seeds against pre- and post-emergence damping-off caused by Pythium ultimum

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Seed coating formulations of Trichoderma harzianum were evaluated on cucumber seeds to control pre- and post-emergence damping-off caused by Pythium ultimum in greenhouse studies. Results showed that coating formulation H reduced the disease incidence significantly, and had the potential for commer...

Quantification and role of organic acids in cucumber root exudates in Trichoderma harzianum T-E5 colonization.

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Zhang, Fengge; Meng, Xiaohui; Yang, Xingming; Ran, Wei; Shen, Qirong

2014-10-01

The ability to colonize on plant roots is recognized as one of the most important characteristics of the beneficial fungi Trichoderma spp. The aim of this study is to prove that the utilization of organic acids is a major trait of Trichoderma harzianum T-E5 for colonization of cucumber roots. A series experiments in split-root hydroponic system and in vitro were designed to demonstrate the association between the utilization of organic acids and T-E5 colonization on cucumber roots. In the split-root hydroponic system, inoculation with T-E5 (T) significantly increased the biomass of cucumber plants compared with CK (non-inoculation with T-E5). The T-E5 hyphae densely covering the cucumber root surface were observed by scanning electron microscopy (SEM). Three organic acids (oxalic acid, malic acid and citric acid) were identified from both the CK and T treatments by HPLC and LC/ESI-MS procedures. The amounts of oxalic acid and malic acid in T were significantly higher than those in CK. All the organic acids exhibited different and significant stimulation effects on the mycelial growth and conidial germination of T-E5 in vitro. An additional hydroponic experiment demonstrated the positive effects of organic acids on the T-E5 colonization of cucumber roots. In conclusion, the present study revealed that certain organic acids could be used as nutritional sources for Trichoderma harzianum T-E5 to reinforce its population on cucumber roots. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Molecular characterization of intergeneric hybrid between Aspergillus oryzae and Trichoderma harzianum by protoplast fusion.

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Patil, N S; Patil, S M; Govindwar, S P; Jadhav, J P

2015-02-01

Protoplast fusion between Aspergillus oryzae and Trichoderma harzianum and application of fusant in degradation of shellfish waste. The filamentous chitinolytic fungal strains A. oryzae NCIM 1272 and T. harzianum NCIM 1185 were selected as parents for protoplast fusion. Viable protoplasts were released from fungal mycelium using enzyme cocktail containing 5 mg ml(-1) lysing enzymes from T. harzianum, 0.06 mg ml(-1) β-glucuronidase from Helix pomatia and 1 mg ml(-1) purified Penicillium ochrochloron chitinase in 0.8 mol l(-1) sorbitol as an osmotic stabilizer. Intergeneric protoplast fusion was carried out using 60% polyethylene glycol as a fusogen. At optimum conditions, the regeneration frequency of the fused protoplasts on colloidal chitin medium and fusion frequency were calculated. Fusant showed higher rate of growth pattern, chitinase activity and protein content than parents. Fusant formation was confirmed by morphological markers, viz. colony morphology and spore size and denaturation gradient gel electrophoresis (DGGE). This study revealed protoplast fusion between A. oryzae and T. harzianum significantly enhanced chitinase activity which ultimately provides potential strain for degradation of shellfish waste. Consistency in the molecular characterization results using DGGE is the major outcome of this study which can be emerged as a fundamental step in fusant identification. Now it is need to provide attention over effective chitin degradation to manage shrimp processing issues. In this aspect, ability of fusant to degrade shellfish waste efficiently in short incubation time revealed discovery of potential strain in the reclamation of seafood processing crustacean bio-waste. © 2014 The Society for Applied Microbiology.

The expression of extracellular fungal cell wall hydrolytic enzymes in different Trichoderma harzianum isolates correlates with their ability to control Pyrenochaeta lycopersici

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LUZ MARÍA PÉREZ

2002-01-01

Full Text Available Four isolates of Trichoderma harzianum (ThN3, Th11, Th12 and Th16 were selected for their ability to control the in vitro development of the tomato root pathogen Pyrenochaeta lycopersici. Analysis of the mechanisms involved in biocontrol showed that the formation of non-volatile metabolites appears to be one of those involved in biocontrol of P. lycopersici by all T. harzianum isolates tested. Nevertheless, the higher secretion of chitinases, both in number of isoenzymes and activity by the Th11 strain, correlated well with its higher ability to control this agent in laboratory and greenhouse experiments as compared to the other T. harzianum isolates tested. The secretion of ß -1,3-endoglucanases and/or proteases appeared to have less significance than endochitinases in the biological control of P. lycopersici

Involvement of G proteins and cAMP in the production of chitinolytic enzymes by Trichoderma harzianum Envolvimento de proteínas G e cAMP na produção de enzimas quitinolíticas por Trichoderma harzianum

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Alexandre A.P. Firmino

2002-06-01

Full Text Available The effect of G protein modulators and cyclic AMP (cAMP on N-acetylglucosaminidase (NAGase production was investigated during 84 h of growth of a Trichoderma harzianum strain in chitin-containing medium. Caffeine (5 mM, N6--2'-O-dibutyryladenosine 3'5'-cyclic monophosphate sodium salt (dBcAMP (1 mM and 3-isobutyl-1-methylxanthine (IBMX (2 mM decreased extracellular NAGase activity by 80%, 77% and 37%, respectively. AlCl3/KF (100 µM/10 mM and 200 µM/ 20 mM decreased the activity by 85% and 95%, respectively. Cholera (10 µ/mL and pertussis (20 µ/mL toxins also affected NAGase activity, causing a decrease of approximately 75%. Upon all treatments, protein bands of approximately 73 kDa, 68 kDa and 45 kDa had their signals diminished whilst a 50 kDa band was enhanced only by treatment with cholera and pertussis toxins. N-terminal sequencing analysis identified the 73 kDa and 68 kDa proteins as being T. harzianum NAGase in two different truncated forms whereas the 45 kDa band comprised a T. harzianum endochitinase. The 50 kDa protein showed sequence similarity to Coriolus vesicolor cellobiohydrolase. The above results suggest that a signaling pathway comprising G-proteins, adenylate cyclase and cAMP may be involved in the synthesis of T. harzianum chitinases.O efeito de cAMP e de moduladores de proteínas G sobre a produção de N-acetilglicosaminidase (NAGase foi investigado durante o crescimento de Trichoderma harzianum em meio contendo quitina. Cafeína (5 mM, dBcAMP (1mM e IBMX (2 mM provocaram diminuições na atividade extracelular de NAGase em 80%, 77% e 37%, respectivamente. Por outro lado, a presença de AlCl3/KF nas concentrações de 100 µM/10 mM e 200 µM/ 20 mM causou decréscimo na atividade em 85% e 95%, respectivamente. A toxina do cólera (10 µ/mL e a toxina pertussis (20 µ/mL também afetaram a atividade de NAGase, causando um decréscimo de aproximadamente 75%. Análises eletroforéticas mostraram que todos os tratamentos

Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme.

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Pellegrini, Vanessa O A; Serpa, Viviane Isabel; Godoy, Andre S; Camilo, Cesar M; Bernardes, Amanda; Rezende, Camila A; Junior, Nei Pereira; Franco Cairo, João Paulo L; Squina, Fabio M; Polikarpov, Igor

2015-11-01

Trichoderma filamentous fungi have been investigated due to their ability to secrete cellulases which find various biotechnological applications such as biomass hydrolysis and cellulosic ethanol production. Previous studies demonstrated that Trichoderma harzianum IOC-3844 has a high degree of cellulolytic activity and potential for biomass hydrolysis. However, enzymatic, biochemical, and structural studies of cellulases from T. harzianum are scarce. This work reports biochemical characterization of the recombinant endoglucanase I from T. harzianum, ThCel7B, and its catalytic core domain. The constructs display optimum activity at 55 °C and a surprisingly acidic pH optimum of 3.0. The full-length enzyme is able to hydrolyze a variety of substrates, with high specific activity: 75 U/mg for β-glucan, 46 U/mg toward xyloglucan, 39 U/mg for lichenan, 26 U/mg for carboxymethyl cellulose, 18 U/mg for 4-nitrophenyl β-D-cellobioside, 16 U/mg for rye arabinoxylan, and 12 U/mg toward xylan. The enzyme also hydrolyzed filter paper, phosphoric acid swollen cellulose, Sigmacell 20, Avicel PH-101, and cellulose, albeit with lower efficiency. The ThCel7B catalytic domain displays similar substrate diversity. Fluorescence-based thermal shift assays showed that thermal stability is highest at pH 5.0. We determined kinetic parameters and analyzed a pattern of oligosaccharide substrates hydrolysis, revealing cellobiose as a final product of C6 degradation. Finally, we visualized effects of ThCel7B on oat spelt using scanning electron microscopy, demonstrating the morphological changes of the substrate during the hydrolysis. The acidic behavior of ThCel7B and its considerable thermostability hold a promise of its industrial applications and other biotechnological uses under extremely acidic conditions.

ETHANOL PRECIPITATION OF GLYCOSYL HYDROLASES PRODUCED BY Trichoderma harzianum P49P11

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M. A. Mariño

2015-06-01

Full Text Available AbstractThis study aimed to concentrate glycosyl hydrolases produced by Trichoderma harzianum P49P11 by ethanol precipitation. The variables tested besides ethanol concentration were temperature and pH. The precipitation with 90% (v/v ethanol at pH 5.0 recovered more than 98% of the xylanase activity, regard less of the temperature (5.0, 15.0, or 25.0 °C. The maximum recovery of cellulase activity as FPase was 77% by precipitation carried out at this same pH and ethanol concentration but at 5.0 °C. Therefore, ethanol precipitation can be considered to be an efficient technique for xylanase concentration and, to a certain extent, also for the cellulase complex.

Oxidative damage induced by heat stress could be relieved by nitric oxide in Trichoderma harzianum LTR-2.

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Yu, Yang; Yang, Zijun; Guo, Kai; Li, Zhe; Zhou, Hongzi; Wei, Yanli; Li, Jishun; Zhang, Xinjian; Harvey, Paul; Yang, Hetong

2015-04-01

Trichoderma harzianum is an important commercial biocontrol fungal agent. The temperature has been shown to be an important parameter and strain-specific to the mycelia growth of fungi, but less report makes the known of the mechanisms in T. harzianum. In our study, a 6-h treatment of heat increased the thiobarbituric acid reactive substances (TBARS) and nitric oxide (NO) concentration in mycelia to 212 and 230 % the level of the control, respectively. The exogenous NO donor sodium nitroprusside (150 μM) reduced the TBARS concentration to 53 % of that under heat stress (HS). At the same time, the NO-specific scavenger at 250 μM, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-1-oxyl-3-oxide, prevented the exogenous NO-relieved TBARS accumulation under HS. The increased NO concentration under HS was reduced 41 % by the NO synthase (NOS) inhibitor L-N(G)-nitroarginine methyl ester, but not the nitrate reductase (NR) inhibitor tungstate. Our study exhibited that NO can protect the mycelia of T. harzianum from HS and reduce the oxidative damage by enhancing the activity of NOS and NR.

Novel mutations in β-tubulin gene in Trichoderma harzianum mutants resistant to methyl benzimidazol-2-yl carbamate.

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Li, M; Zhang, H Y; Liang, B

2013-01-01

Twelve-low resistant (LR) mutants of Trichoderma harzianum with the capability of grow fast at 0.8 μg/mL methyl benzimidazol-2-yl carbamate (MBC) were obtained using UV mutagenesis. MR and HR mutants which could grow fast at 10 and 100 μg/mL MBC, respectively, were isolated by step-up selection protocols in which UV-treated mutants were induced and mycelial sector screening was made in plates with growth medium. Subsequently, β-tubulin genes of 14 mutants were cloned to describe-the molecular lesion likely to be responsible-for MBC resistance. Comparison of the β-tubulin sequences of the mutant and sensitive strains of T. harzianum revealed 2 new MBC-binding sites differed from those in other plant pathogens. A single mutation at-amino acid 168, having Phe (TTC) instead of Ser (TCC)', was demonstrated for the HR mutant; a double mutation in amino acid 13 resulting in the substitution of Gly (GGC) by Val (GTG) was observed in β-tubulin gene of MR mutant. On the other hand, no substitutions were identified in the β-tubulin gene and its 5'-flanking regions in 12 LR mutants of T. harzianum.

Carbohydrate-active enzymes in Trichoderma harzianum: a bioinformatic analysis bioprospecting for key enzymes for the biofuels industry.

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Ferreira Filho, Jaire Alves; Horta, Maria Augusta Crivelente; Beloti, Lilian Luzia; Dos Santos, Clelton Aparecido; de Souza, Anete Pereira

2017-10-12

Trichoderma harzianum is used in biotechnology applications due to its ability to produce powerful enzymes for the conversion of lignocellulosic substrates into soluble sugars. Active enzymes involved in carbohydrate metabolism are defined as carbohydrate-active enzymes (CAZymes), and the most abundant family in the CAZy database is the glycoside hydrolases. The enzymes of this family play a fundamental role in the decomposition of plant biomass. In this study, the CAZymes of T. harzianum were identified and classified using bioinformatic approaches after which the expression profiles of all annotated CAZymes were assessed via RNA-Seq, and a phylogenetic analysis was performed. A total of 430 CAZymes (3.7% of the total proteins for this organism) were annotated in T. harzianum, including 259 glycoside hydrolases (GHs), 101 glycosyl transferases (GTs), 6 polysaccharide lyases (PLs), 22 carbohydrate esterases (CEs), 42 auxiliary activities (AAs) and 46 carbohydrate-binding modules (CBMs). Among the identified T. harzianum CAZymes, 47% were predicted to harbor a signal peptide sequence and were therefore classified as secreted proteins. The GH families were the CAZyme class with the greatest number of expressed genes, including GH18 (23 genes), GH3 (17 genes), GH16 (16 genes), GH2 (13 genes) and GH5 (12 genes). A phylogenetic analysis of the proteins in the AA9/GH61, CE5 and GH55 families showed high functional variation among the proteins. Identifying the main proteins used by T. harzianum for biomass degradation can ensure new advances in the biofuel production field. Herein, we annotated and characterized the expression levels of all of the CAZymes from T. harzianum, which may contribute to future studies focusing on the functional and structural characterization of the identified proteins.

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

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

2012-03-01

The on-site production of cellulases is an important strategy for the development of sustainable second-generation ethanol production processes. This study concerns the use of a specific cellulolytic enzyme complex for hydrolysis of pretreated sugar cane bagasse. Glycosyl hydrolases (FPase, xylanase, and β-glucosidase) were produced using a new strain of Trichoderma harzianum, isolated from the Amazon rainforest and cultivated under different conditions. The influence of the carbon source was first investigated using shake-flask cultures. Selected carbon sources were then further studied under different pH conditions using a stirred tank bioreactor. Enzymatic activities up to 121 FPU/g, 8000 IU/g, and 1730 IU/g of delignified steam-exploded bagasse+sucrose were achieved for cellulase, xylanase and β-glucosidase, respectively. This enzymatic complex was used to hydrolyze pretreated sugar cane bagasse. A comparative evaluation, using an enzymatic extract from Trichoderma reesei RUTC30, indicated similar performance of the T. harzianum enzyme complex, being a potential candidate for on-site production of enzymes. Copyright © 2011 Elsevier Ltd. All rights reserved.

Inoculation of Trichoderma harzianum on Zea mays its effect on the addition of nitrogen fertilizer at 50%

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Tavera-Zavala Dulce Daniela

2017-08-01

Full Text Available The crop Zea mays (maize requires nitrogen fertilizer (NF usually as NH4NO3 (ammonium nitrate, which applied in excess causes loss of productivity in soil. An alternative to reduce and optimize the dose of NF in crop Z. mays is to inoculate it with Trichoderma harzianum. The main objective was to analyze the effect of three doses of T. harzianum in Z. mays at 50% of NF. This experiment was performed in a greenhouse under an experimental design of random blocks, with 5 treatment and 5 replicates, the re-sponse variables used were: phenology: seedling height (SH and root length (RL, and biomass: aerial and radical fresh/dry weight (AFW/ADW/(RFW/RDW at seedlings and flowering stage, the experimental data were analyzed by Tukey 0.05%. The results showed a positive effect of the specific density of all viable structures of T. harzianum in Z. mays since was observed 92% of seed germination, numerical value statistical difference to the 81% in Z. mays without inoculum and NF at 100% or relative control (RC. At seedling Z. mays with T. harzianum 40 g/100 g seeds registered an ADW of 0.32 g and a RDW of 0.25 g, these values were statistical different to the 0.21 g of ADW, and 0.19 g of RDW in Z. mays without inoculum and fed with NF at 100% or RC. The above mentioned suggests that T. harzi-anum transform seed and root exudates in plant growth promoting substances (PGPS, optimizing the use of NH4NO3 and allowing its reduction until 50% without causing a nutritional deficit on normal Z. mays growth.

Expression profiles of defence related cDNAs in oil palm (Elaeis guineensis Jacq.) inoculated with mycorrhizae and Trichoderma harzianum Rifai T32.

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Tan, Yung-Chie; Wong, Mui-Yun; Ho, Chai-Ling

2015-11-01

Basal stem rot is one of the major diseases of oil palm (Elaies guineensis Jacq.) caused by pathogenic Ganoderma species. Trichoderma and mycorrhizae were proposed to be able to reduce the disease severity. However, their roles in improving oil palm defence system by possibly inducing defence-related genes in the host are not well characterized. To better understand that, transcript profiles of eleven putative defence-related cDNAs in the roots of oil palm inoculated with Trichoderma harzianum T32 and mycorrhizae at different time points were studied. Transcripts encoding putative Bowman-Birk protease inhibitor (EgBBI2) and defensin (EgDFS) increased more than 2 fold in mycorrhizae-treated roots at 6 weeks post inoculation (wpi) compared to those in controls. Transcripts encoding putative dehydrin (EgDHN), glycine-rich RNA binding protein (EgGRRBP), isoflavone reductase (EgIFR), type 2 ribosome inactivating protein (EgT2RIP), and EgDFS increased in the oil palm roots treated with T. harzianum at 6 and/or 12 wpi compared to those in the controls. Some of these genes were also expressed in oil palm roots treated with Ganoderma boninense. This study provides an insight of some defence-related genes induced by Trichoderma and mycorrhizae, and their roles as potential agents to boost the plant defence system. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Production of 6-pentyl-α-pyrone by trichoderma harzianum in solid-state fermentation Produção de 6-pentil-α-pirona por Trichoderma harzianum em fermentação em estado sólido

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Aline de Souza Ramos

2008-12-01

Full Text Available Many Trichoderma species are able to produce 6-pentyl-α-pyrone (6-PP, a lactone with coconut-like aroma. In the present work, several culture parameters were studied to enhance the production of 6-PP by Trichoderma harzianum 4040 in solid-state fermentation. Green coir powder added to a nutrient solution was used as support material for fermentation. A Plackett-Burman screening technique was applied, followed by a fractionary factorial design. The best culture conditions within the experimental domain studied were (100 g support-1: sucrose, 3 g; NaNO3, 0.24 g; (NH42SO4, 0.18 g; KH2PO4, 0.1 g; inoculum concentration, 2.2 x 10(6 spores; moisture level, 55%. The temperature established was 28ºC. The fermentation under the selected conditions led to a 6-PP production six times higher (5.0 mg/g dry matter than the initial one (0.8 mg/g dry matter after seven days of cultivation.Muitas espécies do gênero Trichoderma são capazes de produzir a substância 6-pentil-α-pirona (6-PP, uma lactona com aroma característico de coco. No presente trabalho, vários parâmetros de cultura foram estudados para aumentar a produção de 6-PP por Trichoderma harzianum 4040 em fermentação em estado sólido. Pó da casca de coco verde adicionado à uma solução nutriente foi usado como material de suporte para a fermentação. Um planejamento experimental de varredura segundo a técnica de Plackett-Burman foi aplicado, seguido de um planejamento fatorial fracionário. No domínio experimental estudado, as melhores condições de cultura foram (100 g suporte-1: sacarose, 3 g; NaNO3, 0,24 g; (NH42SO4, 0,18 g; KH2PO4, 0,1 g; produção do inóculo, 2,2 x 10(6 esporos; umidade, 55%. A temperatura estabelecida foi de 28ºC. Esse estudo conduziu à concentração de 6-PP seis vezes maior (5,0 mg/g de matéria seca do que a inicial (0,8 mg/g de matéria seca após sete dias de cultivo.

Involvement of the Transcriptional Coactivator ThMBF1 in the Biocontrol Activity of Trichoderma harzianum.

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Rubio, M Belén; Pardal, Alonso J; Cardoza, Rosa E; Gutiérrez, Santiago; Monte, Enrique; Hermosa, Rosa

2017-01-01

Trichoderma harzianum is a filamentous fungus well adapted to different ecological niches. Owing to its ability to antagonize a wide range of plant pathogens, it is used as a biological control agent in agriculture. Selected strains of T. harzianum are also able to increase the tolerance of plants to biotic and abiotic stresses. However, little is known about the regulatory elements of the T. harzianum transcriptional machinery and their role in the biocontrol by this species. We had previously reported the involvement of the transcription factor THCTF1 in the T. harzianum production of the secondary metabolite 6-pentyl-pyrone, an important volatile compound related to interspecies cross-talk. Here, we performed a subtractive hybridization to explore the genes regulated by THCTF1, allowing us to identify a multiprotein bridging factor 1 ( mbf1 ) homolog. The gene from T. harzianum T34 was isolated and characterized, and the generated Thmbf1 overexpressing transformants were used to investigate the role of this gene in the biocontrol abilities of the fungus against two plant pathogens. The transformants showed a reduced antifungal activity against Fusarium oxysporum f. sp. lycopersici race 2 (FO) and Botrytis cinerea (BC) in confrontation assays on discontinuous medium, indicating that the Thmbf1 gene could affect T. harzianum production of volatile organic compounds (VOC) with antifungal activity. Moreover, cellophane and dialysis membrane assays indicated that Thmbf1 overexpression affected the production of low molecular weight secreted compounds with antifungal activity against FO. Intriguingly, no correlation in the expression profiles, either in rich or minimal medium, was observed between Thmbf1 and the master regulator gene cross-pathway control ( cpc1 ). Greenhouse assays allowed us to evaluate the biocontrol potential of T. harzianum strains against BC and FO on susceptible tomato plants. The wild type strain T34 significantly reduced the necrotic leaf

Involvement of the Transcriptional Coactivator ThMBF1 in the Biocontrol Activity of Trichoderma harzianum

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M. Belén Rubio

2017-11-01

Full Text Available Trichoderma harzianum is a filamentous fungus well adapted to different ecological niches. Owing to its ability to antagonize a wide range of plant pathogens, it is used as a biological control agent in agriculture. Selected strains of T. harzianum are also able to increase the tolerance of plants to biotic and abiotic stresses. However, little is known about the regulatory elements of the T. harzianum transcriptional machinery and their role in the biocontrol by this species. We had previously reported the involvement of the transcription factor THCTF1 in the T. harzianum production of the secondary metabolite 6-pentyl-pyrone, an important volatile compound related to interspecies cross-talk. Here, we performed a subtractive hybridization to explore the genes regulated by THCTF1, allowing us to identify a multiprotein bridging factor 1 (mbf1 homolog. The gene from T. harzianum T34 was isolated and characterized, and the generated Thmbf1 overexpressing transformants were used to investigate the role of this gene in the biocontrol abilities of the fungus against two plant pathogens. The transformants showed a reduced antifungal activity against Fusarium oxysporum f. sp. lycopersici race 2 (FO and Botrytis cinerea (BC in confrontation assays on discontinuous medium, indicating that the Thmbf1 gene could affect T. harzianum production of volatile organic compounds (VOC with antifungal activity. Moreover, cellophane and dialysis membrane assays indicated that Thmbf1 overexpression affected the production of low molecular weight secreted compounds with antifungal activity against FO. Intriguingly, no correlation in the expression profiles, either in rich or minimal medium, was observed between Thmbf1 and the master regulator gene cross-pathway control (cpc1. Greenhouse assays allowed us to evaluate the biocontrol potential of T. harzianum strains against BC and FO on susceptible tomato plants. The wild type strain T34 significantly reduced the

The Variations of Glycolysis and TCA Cycle Intermediate Levels Grown in Iron and Copper Mediums of Trichoderma harzianum.

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Tavsan, Zehra; Ayar Kayali, Hulya

2015-05-01

The efficiency of optimal metabolic function by microorganism depends on various parameters, especially essential metal supplementation. In the present study, the effects of iron and copper metals on metabolism were investigated by determination of glycolysis and tricarboxylic acid (TCA) cycle metabolites' levels with respect to the metal concentrations and incubation period in Trichoderma harzianum. The pyruvate and citrate levels of T. harzianum increased up to 15 mg/L of copper via redirection of carbon flux though glycolysis by suppression of pentose phosphate pathway (PPP). However, the α-ketoglutarate levels decreased at concentration higher than 5 mg/L of copper to overcome damage of oxidative stress. The fumarate levels correlated with the α-ketoglutarate levels because of substrate limitation. Besides, in T. harzianum cells grown in various concentrations of iron-containing medium, the intracellular pyruvate, citrate, and α-ketoglutarate levels showed positive correlation with iron concentration due to modifying of expression of glycolysis and TCA cycle enzymes via a mechanism involving cofactor or allosteric regulation. However, as a result of consuming of prior substrates required for fumarate production, its levels rose up to 10 mg/L.

Quantification of the biocontrol agent Trichoderma harzianum with real-time TaqMan PCR and its potential extrapolation to the hyphal biomass.

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López-Mondéjar, Rubén; Antón, Anabel; Raidl, Stefan; Ros, Margarita; Pascual, José Antonio

2010-04-01

The species of the genus Trichoderma are used successfully as biocontrol agents against a wide range of phytopathogenic fungi. Among them, Trichoderma harzianum is especially effective. However, to develop more effective fungal biocontrol strategies in organic substrates and soil, tools for monitoring the control agents are required. Real-time PCR is potentially an effective tool for the quantification of fungi in environmental samples. The aim of this study consisted of the development and application of a real-time PCR-based method to the quantification of T. harzianum, and the extrapolation of these data to fungal biomass values. A set of primers and a TaqMan probe for the ITS region of the fungal genome were designed and tested, and amplification was correlated to biomass measurements obtained with optical microscopy and image analysis, of the hyphal length of the mycelium of the colony. A correlation of 0.76 between ITS copies and biomass was obtained. The extrapolation of the quantity of ITS copies, calculated based on real-time PCR data, into quantities of fungal biomass provides potentially a more accurate value of the quantity of soil fungi. Copyright 2009 Elsevier Ltd. All rights reserved.

Cellulase production by Trichoderma harzianum in static and mixed solid-state fermentation reactors under nonaseptic conditions

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Deschamps, F.; Giuliano, C.; Asther, M.; Huet, M.C.; Roussos, S.

1985-09-01

Cellulase production from lignocellulosic materials was studied in solid-state cultivation by both static and mixed techniques under nonaseptic conditions. The effects of fermentation conditions, such as moisture content, pH, temperature, and aeration, on cellulase production by Trichoderma harzianum using a mixture of wheat straw (80%) and bran (20%) were investigated. With a moisture content of 74% and a pH of 5.8, 18 IU filter paper activity and 198 IU endoglucanase activity/g initial substrate content were obtained in 66 hours. The extension from static column cultivation to stirred tank reactor of 65 l capacity gave similar yields of cellulase.

Potency of six isolates of biocontrol agents endophytic Trichoderma against fusarium wilt on banana

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J Taribuka

2017-01-01

Full Text Available Fusarium wilt caused by F. oxysporum f.sp. cubense is one of very damaging banana plant diseases which can cause plant death. Disease control using intensive chemical fungicides will have negative impacts on the environment and humans. Endophytic Trichoderma is one of the biological control agents which can reduce the amount of inoculum of pathogens, so it can reduce disease intensity. The objectives of this study was to assess the ability of endophytic Trichoderma in inducing plant resistance against fusarium wilt. Endophytic Trichoderma was obtained from healthy roots of banana from three regencies in Yogyakarta, namely Trichoderma harzianum.swn-1, T. harzianum.swn-2, T. harzianum.psr-1, T. asperrellum, T. gamsii, and T. koningiopsis. Research on induced resintance was conducted in the greenhouse with polybag using Completely Randomized Design with 14 treatments and 3 replications. The results showed that the ability of Trichoderma gamsii antagonism against F. oxysporum f.sp. cubense was 60.61%. T. asperellum and T. harzianum.swn-2 could suppress this disease resulted in disease intensity of 8.33% which categorize as resistant. Trichoderma harzianum.psr-1 was significantly different in stimulating plant vegetative growth. Induced resistance by using endophytic Trichoderma spp. against  F. oxysporum f.sp. cubense showed increase in total phenolic compounds on the third and fourth weeks as well as peroxidase activity on the third, fourth and fifth weeks.  Observation of lignification on  the fifth week  showed that lignification occurred in root xylem

The platelet-activating factor acetylhydrolase gene derived from Trichoderma harzianum induces maize resistance to Curvularia lunata through the jasmonic acid signaling pathway.

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Yu, Chuanjin; Fan, Lili; Gao, Jinxin; Wang, Meng; Wu, Qiong; Tang, Jun; Li, Yaqian; Chen, Jie

2015-01-01

Platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum was upregulated by the interaction of T. harzianum with maize roots or the foliar pathogen Curvularia lunata. PAF-AH was associated with chitinase and cellulase expressions, but especially with chitinase, because its activity in the KO40 transformant (PAF-AH disruption transformant) was lower, compared with the wild-type strain T28. The result demonstrated that the colonization of maize roots by T. harzianum induced systemic protection of leaves inoculated with C. lunata. Such protection was associated with the expression of inducible jasmonic acid pathway-related genes. Moreover, the data from liquid chromatography-mass spectrometry confirmed that the concentration of jasmonic acid in maize leaves was associated with the expression level of defense-related genes, suggesting that PAF-AH induced resistance to the foliar pathogen. Our findings showed that PAF-AH had an important function in inducing systemic resistance to maize leaf spot pathogen.

Effect of endophytic pseudomonas aeruinosa and trichoderma harzianum on soil-borne diseases, mycorrhizae and induction of systemic resistance in okra grown in soil amended with vernonia anthelmintica (L.) seeds powder

International Nuclear Information System (INIS)

Shafique, H. A.; Noreen, R.; Haque, S. E. U.; Sultana, V.; Ara, J.

2015-01-01

Biostimulants are used in agricultural practices for plant growth improvement. These fertilizers improve microbial activity and cause a negative impact on soil-borne pathogens. In recent years, stimulating plant natural defense is considered as most promising alternative strategy for crop productivity. The present study was carried out to examine the effect of endophytic Pseudomonas aeruginosa and Trichoderma harzianum in soil amendment with Vernonia anthelmintica seeds powder, on root rotting fungi, plant growth, mycorrhizal population around roots, phosphorous uptake and stimulation of plant defense markers like poylphenol and antioxidant status in okra. Combine application of Vernonia with Pseudomonas aeruginosa and Trichoderma harzianum significantly (p<0.05) suppressed Rhizoctonia solani and Fusarium oxysporum with complete reduction of Macrophomina phaseolina and Fusarium solani. Pseudomonas aeruginosa and T. harzianum alone or in Vernonia amended soil significantly reduced nematode galls on roots. Organic amendment also improved plant resistance against root diseases as evident from enhanced DPPH radical scavenging capacity and polyphenol content in treated plants as compare to control. VA Mycorrhizal spores were found significantly (p<0.05) higher in number around roots received Pseudomonas aeruginosa or T. harzianum alone or in Vernonia amended soil. Whereas, higher concentrations of phosphorus in okra shoots were found in plants received biocontrol agents in amended soil. Mixed application of PGPR and T. harzianum in amended soil produced tallest plants than other treatments. Soil amendment with Vernonia seed powder alone or with biocontrol agents offer a non-chemical means of plant disease control. (author)

Overexpression of erg1 gene in Trichoderma harzianum CECT 2413: effect on the induction of tomato defence-related genes.

Science.gov (United States)

Cardoza, R E; Malmierca, M G; Gutiérrez, S

2014-09-01

To investigate the effect of the overexpression of erg1 gene of Trichoderma harzianum CECT 2413 (T34) on the Trichoderma-plant interactions and in the biocontrol ability of this fungus. Transformants of T34 strain overexpressing erg1 gene did not show effect on the ergosterol level, although a drastic decrease in the squalene level was observed in the transformants at 96 h of growth. During interaction with plants, the erg1 overexpression resulted in a reduction of the priming ability of several tomato defence-related genes belonging to the salicylate pathway, and also of the TomLoxA gene, which is related to the jasmonate pathway. Interestingly, other jasmonate-related genes, such as PINI and PINII, were slightly induced. The erg1 overexpressed transformants also showed a reduced ability to colonize tomato roots. The ergosterol biosynthetic pathway might play an important role in regulating Trichoderma-plant interactions, although this role does not seem to be restricted to the final product; instead, other intermediates such as squalene, whose role in the Trichoderma-plant interaction has not been characterized, would also play an important role. The functional analysis of genes involved in the synthesis of ergosterol could provide additional strategies to improve the ability of biocontrol of the Trichoderma strains and their interaction with plants. © 2014 The Society for Applied Microbiology.

Xylanase Production from Trichoderma harzianum 1073 D3 with Alternative Carbon and Nitrogen Sources

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Isil Seyis

2005-01-01

Full Text Available The effect of some natural wastes (orange pomace, orange peel, lemon pomace, lemon peel, apple pomace, pear peel, banana peel, melon peel and hazelnut shell on the production of xylanase from Trichoderma harzianum 1073 D3 has been studied and maximum activity has been observed on melon peel (26.5 U/mg of protein followed by apple pomace and hazelnut shell. Also, molasses could be used as an additional carbon source as it decreased the production time approximately by 50 %. Finally, potential alternatives of organic nitrogen source (cotton leaf and soybean residue wastes were analyzed and it was concluded that peptone could be replaced with these residues especially when economics of the process is the major objective.

Biocontrol potential of Trichoderma harzianum in controlling wilt disease of pistachio caused by Verticillium dahliae

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Fotoohiyan Zeinab

2017-06-01

Full Text Available Verticillium wilt caused by Verticillium dahliae, is one of the most devastating diseases in pistachio orchards in the world including Iran. In search for an effective non-chemical strategy for the management of this disease, we evaluated the biocontrol potential of Trichoderma harzianum isolates obtained from the rhizosphere of healthy pistachio trees in different locations of the Kerman province of Iran against V. dahliae under laboratory and greenhouse conditions. Dual culture tests in the laboratory were conducted in a completely randomized design using 72 T. harzianum isolates. Twenty isolates showed the highest in vitro antagonistic activity. The results indicated that all 20 isolates were capable of inhibiting the mycelial growth of V. dahliae significantly. Among them, isolates Tr8 and Tr19 were the most effective by 88.89% and 85.12% inhibition, respectively. Extracted cell free metabolites of all effective isolates also inhibited the growth of V. dahliae in the culture medium significantly. According to the results, isolates Tr4 and Tr6 inhibited fungal pathogen growth by 94.94% and 88.15% respectively, through production of non-volatile metabolites. In the evaluation of volatile metabolites, isolates Tr5 and Tr4 were the most effective by 26.27% and 24.49% growth inhibition, respectively. Based on the results of the in vitro experiments, the five most effective isolates were selected for evaluation under greenhouse conditions for their biocontrol potential in controlling Verticillium wilt of pistachio. Results of the greenhouse, (in vivo experiments were positive and indicated that the occurrence of wilt disease in plants treated with the antagonists alone or in combination with pathogenic fungus was lower than in plants inoculated with pathogen alone. The overall results of this study suggest that Trichoderma fungal antagonist may be an effective biocontrol agent for the control of Verticillium wilt of pistachio.

Evaluation of the effect of efficient microorganisms and Trichoderma harzianum application on the production of onion plantlets (Allium cepa L.

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Ramón Liriano González

2015-02-01

Full Text Available The research was done in the intensive-farming areas of the Cooperative of Credits and Services “Ramón Ruiz del Sol”, in Los Arabos, Matanzas. The objective was to evaluate the application effects of effective microorganism (EM and Trichoderma harzianum on the main growth indexes of onion seedlings (Allium cepa L. during nursery stage. Six treatments were studied ( control, organic matter at 24 kg.m-2, T. harzianum at 30 g.m-2, Effective Microorganisms at 15 mL.m-2 at the moment of sowing and 15 and 30 days after the germination of the seeds, organic matter at 24 kg.m-2 + T. harzianum 30 g.m-2 + Efficient Microorganisms at 15 mL.m-2 at 15 and 30 days after germination of the seeds, and T. harzianum 30 g.m-2 + Efficient Microorganisms at 15 mL.m-2 at 15 and 30 days after the germination of the seeds. The experiment was arranged in a randomized block design. 55 days after seed germination, the height of the seedlings, number of leaves per seedlings, diameter of the false stem, root length, as well as the fresh and dry weight of the radical system and the foliate area were evaluated. The results proved that the Effective Microorganisms (EM and T. harzianum application improved the production of quality onion seedlings. The treatment 5 (organic matter 24 kg.m-2 + T. harzianum 30 g.m-2 + Efficient Microorganisms at 15 mL.m-2 at 15 and 30 days after germination had better results in the evaluated variables.

Evaluation in vitro de l'activité antagoniste d'isolats de Trichoderma harzianum Pers. contre trois espèces fongiques pathogènes de l'oignon au Burkina Faso

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Dabire, TG.

2016-01-01

Full Text Available In vitro Evaluation of Antagonistic Activity of Trichoderma harzianum Pers. Isolates against Three Fungal Species Pathogens of Onion in Burkina Faso. The antagonistic effect of five Trichoderma harzianum Pers. isolates of which four from Burkina Faso and one from South Africa was evaluated in vitro against Aspergillus niger van Tieghem, Fusarium oxysporum f.sp. cepae Schlecht. Emend. Snyd & Hans and Fusarium solani (Mart. Sacc. regarded as the main onion fungal pathogens in Burkina Faso. The fungal pathogens species and T. harzianum isolates were directly confronted in Petri dishes for seven days. The pathogen radial growth in presence or absence of antagonist was measured. The two isolates with the highest antagonism rates were grown with each pathogen in confrontation at distance. The culture took place in two Petri dishes sealed with parafilm for seven days. The radial growth of each pathogen was measured. All isolates of T. harzianum inhibited the fungal pathogens growth with antagonism ratio ranging from 0.61 to 0.93 and 0.01 to 0.24 respectively by direct and remote confrontation. The isolates ThTab from Burkina Faso and ThAfs from South Africa showed the best antagonism effect and a myco-parasitic action on all tested pathogens. The antagonistic action modes of these isolates and their possible biological control use are discussed.

Solubilisation of Phosphate and Micronutrients by Trichoderma harzianum and Its Relationship with the Promotion of Tomato Plant Growth.

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Li, Rui-Xia; Cai, Feng; Pang, Guan; Shen, Qi-Rong; Li, Rong; Chen, Wei

2015-01-01

Trichoderma harzianum strain SQR-T037 is a biocontrol agent that has been shown to enhance the uptake of nutrients (macro- and microelements) by plants in fields. The objective of this study was to investigate the contribution of SQR-T037 to P and microelement (Fe, Mn, Cu and Zn) nutrition in tomato plants grown in soil and in hydroponic conditions. Inoculation with SQR-T037 significantly improved the biomass and nutrient uptake of tomato seedlings grown in a nutrient-limiting soil. So we investigated the capability of SQR-T037 to solubilise sparingly soluble minerals in vitro via four known mechanisms: acidification by organic acids, chelation by siderophores, redox by ferric reductase and hydrolysis by phytase. SQR-T037 was able to solubilise phytate, Fe2O3, CuO, and metallic Zn but not Ca3(PO4)2 or MnO2. Organic acids, including lactic acid, citric acid, tartaric acid and succinic acid, were detected by HPLC and LC/MS in two Trichoderma cultures. Additionally, we inoculated tomato seedlings with SQR-T037 using a hydroponic system with specific nutrient deficiencies (i.e., nutrient solutions deficient in P, Fe, Cu or Zn and supplemented with their corresponding solid minerals) to better study the effects of Trichoderma inoculation on plant growth and nutrition. Inoculated seedlings grown in Cu-deficient hydroponic conditions exhibited increases in dry plant biomass (92%) and Cu uptake (42%) relative to control plants. However, we did not observe a significant effect on seedling biomass in plants grown in the Fe- and Zn-deficient hydroponic conditions; by contrast, the biomass decreased by 82% in the P-deficient hydroponic condition. Thus, we demonstrated that Trichoderma SQR-T037 competed for P (phytate) and Zn with tomato seedlings by suppressing root development, releasing phytase and/or chelating minerals. The results of this study suggest that the induction of increased or suppressed plant growth occurs through the direct effect of T. harzianum on root

Harzianolide, a novel plant growth regulator and systemic resistance elicitor from Trichoderma harzianum.

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Cai, Feng; Yu, Guanghui; Wang, Ping; Wei, Zhong; Fu, Lin; Shen, Qirong; Chen, Wei

2013-12-01

A detailed understanding of the effect of natural products on plant growth and protection will underpin new product development for plant production. The isolation and characterization of a known secondary metabolite named harzianolide from Trichoderma harzianum strain SQR-T037 were described, and the bioactivity of the purified compound as well as the crude metabolite extract in plant growth promotion and systemic resistance induction was investigated in this study. The results showed that harzianolide significantly promoted tomato seedling growth by up to 2.5-fold (dry weight) at a concentration of 0.1 ppm compared with the control. The result of root scan suggested that Trichoderma secondary metabolites may influence the early stages of plant growth through better root development for the enhancement of root length and tips. Both of the purified harzianolide and crude metabolite extract increased the activity of some defense-related enzymes to response to oxidative stress. Examination of six defense-related gene expression by real-time reverse transcription-PCR analysis revealed that harzianolide induces the expression of genes involved in the salicylic acid (PR1 and GLU) and jasmonate/ethylene (JERF3) signaling pathways while crude metabolite extract inhibited some gene expression (CHI-II and PGIP) related to basal defense in tomato plants. Further experiment showed that a subsequent challenge of harzianolide-pretreated plants with the pathogen Sclerotinia sclerotiorum resulted in higher systemic resistance by the reduction of lesion size. These results indicate that secondary metabolites of Trichoderma spp., like harzianolide, may play a novel role in both plant growth regulation and plant defense responses. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Functional Analysis of the Trichoderma harzianum nox1 Gene, Encoding an NADPH Oxidase, Relates Production of Reactive Oxygen Species to Specific Biocontrol Activity against Pythium ultimum▿†

Science.gov (United States)

Montero-Barrientos, M.; Hermosa, R.; Cardoza, R. E.; Gutiérrez, S.; Monte, E.

2011-01-01

The synthesis of reactive oxygen species (ROS) is one of the first events following pathogenic interactions in eukaryotic cells, and NADPH oxidases are involved in the formation of such ROS. The nox1 gene of Trichoderma harzianum was cloned, and its role in antagonism against phytopathogens was analyzed in nox1-overexpressed transformants. The increased levels of nox1 expression in these transformants were accompanied by an increase in ROS production during their direct confrontation with Pythium ultimum. The transformants displayed an increased hydrolytic pattern, as determined by comparing protease, cellulase, and chitinase activities with those for the wild type. In confrontation assays against P. ultimum the nox1-overexpressed transformants were more effective than the wild type, but not in assays against Botrytis cinerea or Rhizoctonia solani. A transcriptomic analysis using a Trichoderma high-density oligonucleotide (HDO) microarray also showed that, compared to gene expression for the interaction of wild-type T. harzianum and P. ultimum, genes related to protease, cellulase, and chitinase activities were differentially upregulated in the interaction of a nox1-overexpressed transformant with this pathogen. Our results show that nox1 is involved in T. harzianum ROS production and antagonism against P. ultimum. PMID:21421791

EFIKASI ISOLAT TRICHODERMA TERPILIH DENGAN BAHAN ORGANIK UNTUK MENGENDALIKAN PENYAKIT BUSUK PANGKAL BATANG PADA LADA DI LAPANGAN

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Cipta Ginting

2017-05-01

Full Text Available Efficacy of selected Trichoderma isolate and organic matter to control foot rot of black pepper in the field. The objective of this experiment was to determine the efficacy of selected Trichoderma isolatand organic matter to control the disease. Dual culture method was used to select a Trichoderma isolate. The experiment to evaluate the efficacy of selected isolate consisted of control (no application of T. harzianum or organic matter, T. harzianum and rice straw, and T. harzianum and coffee husk. As starter, T. harzianum was grown in broken rice. Two liters of organic matter was infested with suspension of 10 g starter in 100 ml steril water and incubated for 2 weeks. The mixture was applied around the base of black pepper stem. The results showed that all plants treated with T. harzianum and organic matter did not show disease symptom. Ten percent of the control plants showed symptoms. However, statistical analysis showed that the occurrence of the disease was not significantly different between treatments. Applications ofT. harzianum and rice straw increased the density of Trichoderma for 1 and 2 months after application. T. harzianum and coffee husk increased the density of the fungus 1 month after application.

The Evidence of Non n-glycan Linked Mannose in Exochitinase 42kDa, from Trichoderma harzianum BIO10671 Glycosylation

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Muskhazli, M.

2006-01-01

Full Text Available Chitinase 42 kDa produced by Trichoderma harzianum has been proven as a prime compound to be excreted onto the hyphae of the pathogen causing localised cell wall lysis at the point of interaction. Later it will initiate the process of the host cell becomes empty of cytoplasm, disintegrates and shows a rapid collapse. This study investigates the existence of N-glycan linked mannose in chitinase 42 kDa produced by the Malaysian T. harzianum strain BIO10671. The chitinase 42 kDa from T. harzianum BIO10671 was initially purified using anion exchange chromatography prior to a series of experiments such as immunoblotting against the chitinase 42 kDa antibody, lectin staining for detecting any terminal linked mannose, and galactofuranose detection to determine the presence of galatofuranase components in glycoproteins. The enzyme purification harvested about 12-fold of chitinase 42 kDa from T. harzianum BIO10671 with strong indication of the chitinase 42 kDa presence on SDS-Page. This was confirmed by immunoblotting with a strong response around 42 kDa after overnight incubation in chitinase 42 kDa antibody suggesting that the gene for chitinase 42 kDa was greatly expressed in this strain. There are no intervation of galatofuranose on any of the terminal mannose in chitinase 42 kDa as shown by negative results on samples treated with or without endoglycosidase-H and lectin staining. Therefore, it can be concludeed that glycosylation occurred in the chitinase 42 kDa from T. harzianum 42 kDa was not in the form of N-glycan linked mannose as expected.

Compatibilidad de Trichoderma harzianum Rifai con fungicidas del arroz y su efecto sobre tres fitopatógenos fúngicos

OpenAIRE

Manuel Francisco Rodríguez Saldaña; Pausides Milanés Virelles; Ernesto Juniors Pérez Torres; Yurisandra Sierra Reyes

2016-01-01

Resumen La investigación se realizó en el Laboratorio Provincial de Sanidad Vegetal en Camagüey, en la etapa comprendida de septiembre de 2013 a septiembre de 2015, en condiciones in vitro donde se determinó la compatibilidad y capacidad antagónica frente a pesticidas usados en el arroz, de Trichoderma harzianum Rifai cepa A-34 sobre los patógenos del arroz (Bipolaris oryzae Breda de Haan, Saracladium oryzae (Sawada) w. Gams & D.Hawksworth y Magnaporthe grisea (Hebert) Barr). Las ev...

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation*

Science.gov (United States)

Li, Ya-qian; Song, Kai; Li, Ya-chai; Chen, Jie

2016-01-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×108 spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×108 spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents. PMID:27487807

Statistical culture-based strategies to enhance chlamydospore production by Trichoderma harzianum SH2303 in liquid fermentation.

Science.gov (United States)

Li, Ya-Qian; Song, Kai; Li, Ya-Chai; Chen, Jie

2016-08-01

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10(8) spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10(8) spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.

Cellulose hydrolysis by fungi. 2. Cellulase production by Trichoderma harzianum in liquid medium fermentation

Energy Technology Data Exchange (ETDEWEB)

Roussos, S.; Raimbault, M. (Laboratoire de Microbiologie ORSTOM, Centre de Recherche IRCHA, 91 - Vert-le-Petit (France))

Microcrystalline cellulose (cellulose Avicel, Merck) supported growth of Trichoderma harzianum and induced production of cellulases in liquid cultures. After 50 h growth, the total cellulasic activities present in both the supernatant and the mycelium were 3,000 IU/l of carboxymethyl cellulose, 400 IU/l of filter paper activity, and 4 IU/l of cotton activity corresponding to 1.7 g/l of proteins. Cellulase production could be increased by a preliminary treatment of cellulose, and pH regulation during growth. The influence of inoculum concentration was studied and an optimum of 3 X 10/sup 7/ conidia/g dry weight of substrate was demonstrated. Using a synthetic culture medium, a soluble factor of germination was demonstrated which could be leached out by 3 successive washings of conidia.

Biological control of white mold by Trichoderma harzianum in common bean under field conditions

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Daniel Diego Costa Carvalho

2015-12-01

Full Text Available Abstract: The objective of this work was to evaluate Trichoderma harzianum isolates for biological control of white mold in common bean (Phaseolus vulgaris. Five isolates were evaluated for biocontrol of white mold in 'Perola' common bean under field conditions, in the 2009 and 2010 crop seasons. A commercial isolate (1306 and a control treatment were included. Foliar applications at 2x109 conidia mL-1 were performed at 42 and 52 days after sowing (DAS, in 2009, and at 52 DAS in 2010. The CEN287, CEN316, and 1306 isolates decreased the number of Sclerotinia sclerotiorum apothecia per square meter in comparison to the control, in both crop seasons. CEN287, CEN316, and 1306 decreased white mold severity during the experimental period, when compared to the control.

Biocontrol with Trichoderma species for the management of postharvest crown rot of banana

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G. Sangeetha

2009-09-01

Full Text Available Lasiodiplodia theobromae and Colletotrichum musae cause the postharvest crown rot disease complex of banana. In vitro experiments evaluated the effect of twelve isolates of Trichoderma spp. from the soil of organic banana orchards (‘native isolates’ and eight isolates of Trichoderma spp. from culture collections (‘introduced isolates’ on the two pathogens. The native and introduced Trichoderma spp. had varied antagonistic effects against the two pathogens. Eight Trichoderma spp. isolates effective in the in vitro assays were evaluated singly on fruits both at room temperature and in cold storage. Single antagonists did not satisfactorily control crown rot on the fruits as compared with the fungicide carbendazim. However, two isolates of T. viride, one of T. harzianum and one of T. koningii performed well when applied singly, and these were selected for evaluation in isolate mixtures. There was very little antagonism between these isolates. Of 11 two-way, three-way and four-way mixtures of these isolates, the four-way and a three-way mixtures reduced crown rot incidence, both at room temperature and in cold storage, giving better control than carbendazim. The study identified consortia of compatible Trichoderma antagonists with superior biocontrol potential for the management of the postharvest crown rot complex of banana.

Induction of SA-signaling pathway and ethylene biosynthesis in Trichoderma harzianum-treated tomato plants after infection of the root-knot nematode Meloidogyne incognita.

Science.gov (United States)

Leonetti, Paola; Zonno, Maria Chiara; Molinari, Sergio; Altomare, Claudio

2017-04-01

Salicylic acid-signaling pathway and ethylene biosynthesis were induced in tomato treated with Trichoderma harzianum when infected by root-knot nematodes and limited the infection by activation of SAR and ethylene production. Soil pre-treatment with Trichoderma harzianum (Th) strains ITEM 908 (T908) and T908-5 decreased susceptibility of tomato to Meloidogyne incognita, as assessed by restriction in nematode reproduction and development. The effect of T. harzianum treatments on plant defense was detected by monitoring the expression of the genes PR-1/PR-5 and JERF3/ACO, markers of the SA- and JA/ET-dependent signaling pathways, respectively. The compatible nematode-plant interaction in absence of fungi caused a marked suppression of PR-1, PR-5, and ACO gene expressions, either locally or systemically, whilst expression of JERF3 gene resulted unaffected. Conversely, when plants were pre-treated with Th-strains, over-expression of PR-1, PR-5, and ACO genes was observed in roots 5 days after nematode inoculation. JERF3 gene expression did not change in Th-colonized plants challenged with nematodes. In the absence of nematodes, Trichoderma-root interaction was characterized by the inhibition of both SA-dependent signaling pathway and ET biosynthesis, and, in the case of PR-1 and ACO genes, this inhibition was systemic. JERF3 gene expression was systemically restricted only at the very early stages of plant-fungi interaction. Data presented indicate that Th-colonization primed roots for Systemic Acquired Resistance (SAR) against root-knot nematodes and reacted to nematode infection more efficiently than untreated plants. Such a response probably involves also activation of ET production, through an augmented transcription of the ACO gene, which encodes for the enzyme catalyzing the last step of ET biosynthesis. JA signaling and Induced Systemic Resistance (ISR) do not seem to be involved in the biocontrol action of the tested Th-strains against RKNs.

Structural basis of transport function in major facilitator superfamily protein from Trichoderma harzianum.

Science.gov (United States)

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

2017-02-01

Trichothecenes are the sesquiterpenes secreted by Trichoderma spp. residing in the rhizosphere. These compounds have been reported to act as plant growth promoters and bio-control agents. The structural knowledge for the transporter proteins of their efflux remained limited. In this study, three-dimensional structure of Thmfs1 protein, a trichothecene transporter from Trichoderma harzianum, was homology modelled and further Molecular Dynamics (MD) simulations were used to decipher its mechanism. Fourteen transmembrane helices of Thmfs1 protein are observed contributing to an inward-open conformation. The transport channel and ligand binding sites in Thmfs1 are identified based on heuristic, iterative algorithm and structural alignment with homologous proteins. MD simulations were performed to reveal the differential structural behaviour occurring in the ligand free and ligand bound forms. We found that two discrete trichothecene binding sites are located on either side of the central transport tunnel running from the cytoplasmic side to the extracellular side across the Thmfs1 protein. Detailed analysis of the MD trajectories showed an alternative access mechanism between N and C-terminal domains contributing to its function. These results also demonstrate that the transport of trichodermin occurs via hopping mechanism in which the substrate molecule jumps from one binding site to another lining the transport tunnel. Copyright © 2016 Elsevier B.V. All rights reserved.

Differential Response of Extracellular Proteases of Trichoderma Harzianum Against Fungal Phytopathogens.

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Sharma, Vivek; Salwan, Richa; Sharma, Prem N

2016-09-01

In the present study, production of extracellular proteases by Trichoderma harzianum was evaluated based on the relative gene expression and spectrophotometric assay. The fungal isolates were grown in Czapek Dox Broth medium supplemented with deactivated mycelium of plant fungal pathogens such as Fusarium oxysporum, Colletotrichum capsici, Gloeocercospora sorghi, and Colletotrichum truncatum. The maximum protease activity was detected after 48 h of incubation against Colletotrichum spp. Similarly in qRT-PCR, the relative gene expression of four proteases varied from 48 to 96 h against host pathogens in a time-independent manner. Among proteases, statistically significant upregulation of asp, asp, and srp was observed against Colletotrichum spp., followed by F. oxysporum. But in the case of pepM22, maximum upregulation was observed against F. oxysporum. The variation in enzyme assay and qRT-PCR of proteases at different time intervals against various fungal phytopathogens could be due to the limitation of using casein as a substrate for all types of proteases or protease-encoding transcripts selected for qRT-PCR, which may not be true representative of total protease activity.

Seed biopriming with novel strain of Trichoderma harzianum for the control of toxigenic Fusarium verticillioides and fumonisins in maize

DEFF Research Database (Denmark)

Nayaka, S.Chandra; Niranjana, S.R.; Shankar, A.C. Uday

2010-01-01

Fusarium verticillioides is one of the most important fungal pathogens in maize causing both pre- and post-harvest losses and also capable of producing Fumonisins. In the present study attempts have been made for screening potential T. harzianum from native rhizosphere and to study its effect...... on Fusarium ear rot disease, fumonisin accumulation in different maize cultivars grown in India. Eight isolates of T. harzianum were isolated and T. harzianum isolate Th-8 exhibited better antifungal activity than carbendizim. Th-8 was formulated in different solid substrates like wheat bran, paddy husk...... of 10 g/kg. Treated seeds were subjected to evaluate F. verticillioides incidence, seed germination, seedling vigour and field emergence, yield, thousand seed weight and fumonisin production. It was found that the pure culture of T. harzianum was more effective in reducing the F. verticillioides...

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

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Strakowska, Judyta; Błaszczyk, Lidia; Chełkowski, Jerzy

2014-07-01

The degradation of native cellulose to glucose monomers is a complex process, which requires the synergistic action of the extracellular enzymes produced by cellulolytic microorganisms. Among fungi, the enzymatic systems that can degrade native cellulose have been extensively studied for species belonging to the genera of Trichoderma. The majority of the cellulolytic enzymes described so far have been examples of Trichoderma reesei, extremely specialized in the efficient degradation of plant cell wall cellulose. Other Trichoderma species, such as T. harzianum, T. koningii, T. longibrachiatum, and T. viride, known for their capacity to produce cellulolytic enzymes, have been isolated from various ecological niches, where they have proved successful in various heterotrophic interactions. As saprotrophs, these species are considered to make a contribution to the degradation of lignocellulosic plant material. Their cellulolytic potential is also used in interactions with plants, especially in plant root colonization. However, the role of cellulolytic enzymes in species forming endophytic associations with plants or in those existing in the substratum for mushroom cultivation remains unknown. The present review discusses the current state of knowledge about cellulolytic enzymes production by Trichoderma species and the encoding genes, as well as the involvement of these proteins in the lifestyle of Trichoderma. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

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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

Purification and characterization of a beta-Glucanase produced by Trichoderma harzianum showing biocontrol potential

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Janice Lisboa de Marco

2007-01-01

Full Text Available A beta-1,3-glucanase was produced by Trichoderma harzianum in cultures containing chitin as the sole substrate. Two proteins showing beta-1,3-glucanase activity were purified to apparent homogeneity by hydrophobic chromatography. The molecular masses of these proteins were 29 and 36 kDa. The 36 kDa protein was further characterized. It was active on a broad pH range, and maximal activity was detected at pH 5.0. The optimum temperature of the 36 kDa beta-1,3-glucanase was 50ºC, but the purified enzyme was very sensitive to temperature. It lost about 60% or more of the activity after incubation for 30 min at 45, 50 and 60ºC. The apparent K M and Vmax for hydrolysis of laminarin at pH 5.0 and 37ºC, were 0.099 mg of reducing sugar/mL and 0.3 mg of reducing sugar/min.mL, respectively. The enzyme was insensitive to organic compound and metal ions, except for the ferric ion which inhibited about 100% of the original activity at the concentration of 1 mM. In contrast to other hydrolytic enzymes (a chitinase and a protease produced by the same T. harzianum isolate (1051, the beta-1,3-glucanase showed no effect on the cell wall of the phytopathogenic fungus Crinipellis perniciosa.Uma beta-1,3-glucanase foi produzida por Trichoderma harzianum em cultura contendo quitina como fonte de carbono. Duas proteínas com atividade de beta-1,3-glucanase foram purificadas através de cromatografia de interação hidrofóbica. As massas moleculares destas proteínas foram de 29 kDa e 36 kDa. A proteína de 36 kDa foi caracterizada quanto à influência das condições de pH e temperatura. A atividade máxima foi encontrada em pH 5,0 e temperatura de 50ºC. A proteína purificada mostrou-se muito sensível à temperatura. Aproximadamente 60% da atividade original foi perdida por incubação da proteína a 45ºC, 50ºC e 60ºC, por 30 min. O K M aparente e a Vmax para hidrólise de laminarina em pH 5,0 à 37ºC, foram de 0,099 mg de açúcar redutor/mL e 0,3 mg de a

An assessment of adaptive and antagonistic properties of Trichoderma sp. strains in vegetable waste composts

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Wolna-Maruwka Agnieszka

2017-12-01

Full Text Available The experiment consisted in monitoring the count of moulds and three selected Trichoderma sp. isolates (T1 - Trichoderma atroviride, T2 - Trichoderma harzianum, T3 - Trichoderma harzianum in vegetable (onion and tomato waste composted with additives (straw, pig manure. Additionally, the aim of the study was to determine the type of interaction occurring between autochthonous fungi isolated from composts after the end of the thermophilic phase and Trichoderma sp. strains applied in the experiment. Number of microorganisms was determined by the plate method, next the identification was confirmed. The rating scale developed by Mańka was used to determine the type of interactions occurring between microorganisms. The greatest count of moulds in onion waste composts was noted in the object which had simultaneously been inoculated with two strains T1 - T. atroviride and T3 - T. harzianum. The greatest count of moulds was noted in the tomato waste composts inoculated with T2 - T. harzianum strain. Microscope identification revealed that Penicillum sp., Rhizopus sp., Alternaria sp. and Mucor sp. strains were predominant in onion waste composts. In tomato waste composts Penicillium was the predominant genus, followed by Rhizopus. The test of antagonism revealed the inhibitory effect of Trichoderma isolates on most autochthonous strains of moulds. Tomato waste composts proved to be better substrates for the growth and development of Trichoderma sp. isolates. The results of the study show that vegetable waste can be used in agriculture as carriers of antagonistic microorganisms.

Production of a recombinant swollenin from Trichoderma harzianum in Escherichia coli and its potential synergistic role in biomass degradation.

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Santos, Clelton A; Ferreira-Filho, Jaire A; O'Donovan, Anthonia; Gupta, Vijai K; Tuohy, Maria G; Souza, Anete P

2017-05-16

Fungal swollenins (SWOs) constitute a class of accessory proteins that are homologous to canonical plant expansins. Expansins and expansin-related proteins are well known for acting in the deagglomeration of cellulose structure by loosening macrofibrils. Consequently, SWOs can increase the accessibility and efficiency of the other enzymes involved in the saccharification of cellulosic substrates. Thus, SWOs are promising targets for improving the hydrolysis of plant biomass and for use as an additive to enhance the efficiency of an enzyme cocktail designed for the production of biofuels. Here, we report the initial characterization of an SWO from Trichoderma harzianum (ThSwo) that was successfully produced using Escherichia coli as a host. Initially, transcriptome and secretome data were used to compare swo gene expression and the amount of secreted ThSwo. The results from structural modeling and phylogenetic analysis of the ThSwo protein showed that ThSwo does preserve some structural features of the plant expansins and family-45 glycosyl hydrolase enzymes, but it evolutionarily diverges from both of these protein classes. Recombinant ThSwo was purified at a high yield and with high purity and showed secondary folding similar to that of a native fungal SWO. Bioactivity assays revealed that the purified recombinant ThSwo created a rough and amorphous surface on Avicel and displayed a high synergistic effect with a commercial xylanase from T. viride, enhancing its hydrolytic performance up to 147 ± 7%. Many aspects of the structure and mechanism of action of fungal SWOs remain unknown. In the present study, we produced a recombinant, active SWO from T. harzianum using a prokaryotic host and confirmed its potential synergistic role in biomass degradation. Our work paves the way for further studies evaluating the structure and function of this protein, especially regarding its use in biotechnology.

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

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

2009-09-01

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

Characterization of the cellulolytic secretome of Trichoderma harzianum during growth on sugarcane bagasse and analysis of the activity boosting effects of swollenin.

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A L Rocha, Vanessa; N Maeda, Roberto; Pereira, Nei; F Kern, Marcelo; Elias, Luisa; Simister, Rachael; Steele-King, Clare; Gómez, Leonardo D; McQueen-Mason, Simon J

2016-03-01

This study demonstrates the production of an active enzyme cocktail produced by growing Trichoderma harzianum on sugarcane bagasse. The component enzymes were identified by LCMS-MS. Glycosyl hydrolases were the most abundant class of proteins, representing 67% of total secreted protein. Other carbohydrate active enzymes involved in cell wall deconstruction included lytic polysaccharide mono-oxygenases (AA9), carbohydrate-binding modules, carbohydrate esterases and swollenin, all present at levels of 1%. In total, proteases and lipases represented 5 and 1% of the total secretome, respectively, with the rest of the secretome being made up of proteins of unknown or putative function. This enzyme cocktail was efficient in catalysing the hydrolysis of sugarcane bagasse cellulolignin to fermentable sugars for potential use in ethanol production. Apart from mapping the secretome of T. harzianum, which is a very important tool to understand the catalytic performance of enzyme cocktails, the gene coding for T. harzianum swollenin was expressed in Aspergillus niger. This novel aspect in this work, allowed increasing the swollenin concentration by 95 fold. This is the first report about the heterologous expression of swollenin from T. harzianum, and the findings are of interest in enriching enzyme cocktail with this important accessory protein which takes part in the cellulose amorphogenesis. Despite lacking detectable glycoside activity, the addition of swollenin of T. harzianum increased by two-fold the hydrolysis efficiency of a commercial cellulase cocktail. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:327-336, 2016. © 2016 American Institute of Chemical Engineers.

Peptaibol, secondary-metabolite, and hydrophobin pattern of commercial biocontrol agents formulated with species of the Trichoderma harzianum complex.

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Degenkolb, Thomas; Fog Nielsen, Kristian; Dieckmann, Ralf; Branco-Rocha, Fabiano; Chaverri, Priscila; Samuels, Gary J; Thrane, Ulf; von Döhren, Hans; Vilcinskas, Andreas; Brückner, Hans

2015-04-01

The production of bioactive polypeptides (peptaibiotics) in vivo is a sophisticated adaptation strategy of both mycoparasitic and saprotrophic Trichoderma species for colonizing and defending their natural habitats. This feature is of major practical importance, as the detection of peptaibiotics in plant-protective Trichoderma species, which are successfully used against economically relevant bacterial and fungal plant pathogens, certainly contributes to a better understanding of these complex antagonistic interactions. We analyzed five commercial biocontrol agents (BCAs), namely Canna(®) , Trichosan(®) , Vitalin(®) , Promot(®) WP, and TrichoMax(®) , formulated with recently described species of the Trichoderma harzianum complex, viz. T. afroharzianum, T. simmonsii, and T. guizhouense. By using the well-established, HPLC/MS-based peptaibiomics approach, it could unequivocally be demonstrated that all of these formulations contained new and recurrent peptaibols, i.e., peptaibiotics carrying an acetylated N-terminus, the C-terminus of which is reduced to a 1,2-amino alcohol. Their chain lengths, including the amino alcohol, were 11, 14, and 18 residues, respectively. Peptaibols were also to be the dominating secondary metabolites in plate cultures of the four strains obtained from four of the Trichoderma- based BCAs, contributing 95% of the UHPLC-UV/VIS peak areas and 99% of the total ion count MS peak area from solid media. Furthermore, species-specific hydrophobins, as well as non-peptaibiotic secondary metabolites, were detected, the latter being known for their antifungal, siderophore, or plant-growth-promoting activities. Notably, none of the isolates produced low-molecular weight mycotoxins. Copyright © 2015 Verlag Helvetica Chimica Acta AG, Zürich.

Production of trichodiene by Trichoderma harzianum alters the perception of this biocontrol strain by plants and antagonized fungi.

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Malmierca, Mónica G; McCormick, Susan P; Cardoza, Rosa E; Alexander, Nancy J; Monte, Enrique; Gutiérrez, Santiago

2015-08-01

Trichothecenes are phytotoxic sesquiterpenic mycotoxins that can act as virulence factors in plant diseases. Harzianum A (HA) is a non-phytotoxic trichothecene produced by Trichoderma arundinaceum. The first step in HA biosynthesis is the conversion of farnesyl diphosphate to trichodiene (TD), a volatile organic compound (VOC), catalysed by a sesquiterpene synthase encoded by the tri5 gene. Expression of tri5 in the biocontrol strain Trichoderma harzianum CECT 2413 resulted in production of TD in parallel with a reduction of ergosterol biosynthesis and an unexpected increase in the level of squalene. Transformants expressing tri5 displayed low chitinase activity and induced expression of Botrytis cinerea BOT genes, although their total antagonistic potential against phytopathogenic fungi was not reduced. VOCs released by the tri5-transformant induced expression of tomato defence genes related to salicylic acid (SA), and TD itself strongly induced the expression of SA-responsive genes and reduced the development of lateral roots. Together, these results suggest that TD acts as a signalling VOC in the interactions of Trichoderma with plants and other microorganisms by modulating the perception of this fungus to a given environment. Moreover, the TD ability to induce systemic defences indicates that complex trichothecene structures may not be necessary for inducing such responses. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

Fusarium oxysporum induces the production of proteins and volatile organic compounds by Trichoderma harzianum T-E5.

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Zhang, Fengge; Yang, Xingming; Ran, Wei; Shen, Qirong

2014-10-01

Trichoderma species have been used widely as biocontrol agents for the suppression of soil-borne pathogens. However, some antagonistic mechanisms of Trichoderma are not well characterized. In this study, a series of laboratory experiments were designed to characterize the importance of mycoparasitism, exoenzymes, and volatile organic compounds (VOCs) by Trichoderma harzianum T-E5 for the control of Fusarium oxysporum f. sp. cucumerinum (FOC). We further tested whether these mechanisms were inducible and upregulated in presence of FOC. The results were as follows: T-E5 heavily parasitized FOC by coiling and twisting the entire mycelium of the pathogen in dual cultures. T-E5 growing medium conditioned with deactivated FOC (T2) showed more proteins and higher cell wall-degrading enzyme activities than T1, suggesting that FOC could induce the upregulation of exoenzymes. The presence of deactivated FOC (T2') also resulted in the upregulation of VOCs that five and eight different types T-E5-derived VOCs were identified from T1' and T2', respectively. Further, the excreted VOCs in T2' showed significantly higher antifungal activities against FOC than T1'. In conclusion, mycoparasitism of T-E5 against FOC involved mycelium contact and the production of complex extracellular substances. Together, these data provide clues to help further clarify the interactions between these fungi. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Enzyme Production and Nitrogen Fixation by Free, Immobilized and Coimmobilized Inoculants of Trichoderma harzianum and Azospirillum brasilense and Their Possible Role in Growth Promotion of Tomato

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Momein H. El-Katatny

2010-01-01

Full Text Available A plant growth-promoting rhizobacterium (Azospirillum brasilense strain Az and a biocontrol fungus (Trichoderma harzianum strain T24 have been evaluated for their individual and combined production of hydrolytic enzymes, nitrogen fixation and their possible role in growth promotion of tomato seedlings. The studied organisms were inoculated as free or calcium alginate-encapsulated cells. All freshly prepared macrobeads showed high encapsulation capacity (EC/% of inocula compared with dry macrobeads. Results of enzyme production did not exhibit consistent pattern of the effect of encapsulation process on enzyme production. Beads entrapping bacterial and/or fungal cells were used successfully in 3 repeated cycles in the presence of fresh sterile culture medium in each growth cycle. Enzyme production by immobilized bacterial and/or fungal cells increased as the growth cycles were repeated. Co-culturing of A. brasilense with T. harzianum (free or immobilized in semisolid nitrogen deficient medium (N-free medium enabled A. brasilense to fix nitrogen on pectin, chitin and carboxymethyl cellulose. The activity of nitrogen fixation by A. brasilense in the case of single and combined cultures with Trichoderma (using dry encapsulated beads into the sterile soil increased with the addition of carbon source. Most of inoculations with free or alginate macrobead formulations of T. harzianum and/or A. brasilense showed significant increase in the growth parameters of tomato seedlings. The root system grew more profusely in the case of all seeds treated with A. brasilense. The growth parameters of Az/T24-treated seeds using dry coimmobilized macrobeads were higher than those of the untreated control. Moreover, the effect was improved significantly in soil enriched with different C sources. Enhanced tomato seedling growth after the co-inoculation could be due to the synergistic effect of both Trichoderma and Azospirillum. Finally, co-inoculation with Azospirillum

Experimental mixture design as a tool to enhance glycosyl hydrolases production by a new Trichoderma harzianum P49P11 strain cultivated under controlled bioreactor submerged fermentation.

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Delabona, Priscila da Silva; Farinas, Cristiane Sanchez; Lima, Deise Juliana da Silva; Pradella, José Geraldo da Cruz

2013-03-01

This work investigates the glycosyl hydrolase (GH) profile of a new Trichoderma harzianum strain cultivated under controlled bioreactor submerged fermentation. The influence of different medium components (delignified steam-exploded sugarcane bagasse, sucrose, and soybean flour) on GH biosynthesis was assessed using experimental mixture design (EMD). Additionally, the effect of increased component concentrations in culture media selected from the EMD was studied. It was found that that a mixed culture medium could significantly maximize GH biosynthesis rate, especially for xylanase enzymes which achieved a 2-fold increment. Overall, it was demonstrated that T. harzianumP49P11 enzymes have a great potential to be used in the deconstruction of biomass. Copyright © 2012 Elsevier Ltd. All rights reserved.

Trichoderma harzianum MTCC 5179 impacts the population and functional dynamics of microbial community in the rhizosphere of black pepper (Piper nigrum L.).

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Umadevi, Palaniyandi; Anandaraj, Muthuswamy; Srivastav, Vivek; Benjamin, Sailas

2017-11-29

Employing Illumina Hiseq whole genome metagenome sequencing approach, we studied the impact of Trichoderma harzianum on altering the microbial community and its functional dynamics in the rhizhosphere soil of black pepper (Piper nigrum L.). The metagenomic datasets from the rhizosphere with (treatment) and without (control) T. harzianum inoculation were annotated using dual approach, i.e., stand alone and MG-RAST. The probiotic application of T. harzianum in the rhizhosphere soil of black pepper impacted the population dynamics of rhizosphere bacteria, archae, eukaryote as reflected through the selective recruitment of bacteria [Acidobacteriaceae bacterium (p=1.24e-12), Candidatus koribacter versatilis (p=2.66e-10)] and fungi [(Fusarium oxysporum (p=0.013), Talaromyces stipitatus (p=0.219) and Pestalotiopsis fici (p=0.443)] in terms of abundance in population and bacterial chemotaxis (p=0.012), iron metabolism (p=2.97e-5) with the reduction in abundance for pathogenicity islands (p=7.30e-3), phages and prophages (p=7.30e-3) with regard to functional abundance. Interestingly, it was found that the enriched functional metagenomic signatures on phytoremediation such as benzoate transport and degradation (p=2.34e-4), and degradation of heterocyclic aromatic compounds (p=3.59e-13) in the treatment influenced the rhizosphere micro ecosystem favoring growth and health of pepper plant. The population dynamics and functional richness of rhizosphere ecosystem in black pepper influenced by the treatment with T. harzianum provides the ecological importance of T. harzianum in the cultivation of black pepper. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Synergistic effects of plant defense elicitors and Trichoderma harzianum on enhanced induction of antioxidant defense system in tomato against Fusarium wilt disease.

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Zehra, Andleeb; Meena, Mukesh; Dubey, Manish Kumar; Aamir, Mohd; Upadhyay, R S

2017-11-02

Plant defense against their pathogens can be induced by a complex network of different inducers. The present study investigates the synergistic effect of Trichoderma harzianum, exogenous salicylic acid (SA) and methyl jasmonate (MeJA) over the response and regulation of the antioxidant defense mechanisms and lipid peroxidation in tomato plants against Fusarium wilt disease. In the present work, tomato plants were infected by Fusarium oxysporum f. sp. lycopersici 3 days after inoculated with T. harzianum and/or sprayed daily for 3 days with chemical inducers (SA and MeJA). Plants were analysed at 0, 24, 48, 72 and 96 h after inoculation with Fusarium oxysporum f. sp. lycopersici. Infection of tomato plants by pathogen led to strong reduction in the dry weight of roots and shoots with the enhanced concentration of H 2 O 2 and varying degree of lipid peroxidation. Concurrently, exogenous SA, when applied with pathogen greatly enhanced H 2 O 2 content as well as activities of antioxidant enzymes except catalase (CAT) and ascorbate peroxidase (APx). The pathogen challenged plants pretreated with T. harzianum and MeJA together exhibited less lipid peroxidation and as well as the elevated level of ascorbic acid and enhanced activities of antioxidant enzymes. All applied treatments protected tomato seedlings against Fusarium wilt disease but the percentage of protection was found higher in plants pretreated with the combination of T. harzianum and chemical inducers.

Biological role of Trichoderma harzianum-derived platelet-activating factor acetylhydrolase (PAF-AH) on stress response and antagonism.

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Yu, Chuanjin; Fan, Lili; Wu, Qiong; Fu, Kehe; Gao, Shigang; Wang, Meng; Gao, Jinxin; Li, Yaqian; Chen, Jie

2014-01-01

We investigated the properties of platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum. The enzyme, comprised of 572 amino acids, shares high homology with PAF-AH proteins from T. koningii and other microbial species. The optimum enzymatic activity of PAF-AH occurred at pH 6 in the absence of Ca2+ and it localized in the cytoplasm, and we observed the upregulation of PAF-AH expression in response to carbon starvation and strong heat shock. Furthermore, PAF-AH knockout transformant growth occurred more slowly than wild type cells and over-expression strains grown in SM medium at 37°C and 42°C. In addition, PAF-AH expression significantly increased under a series of maize root induction assay. Eicosanoic acid and ergosterol levels decreased in the PAF-AH knockouts compared to wild type cells, as revealed by GC/MS analysis. We also determined stress responses mediated by PAF-AH were related to proteins HEX1, Cu/Zn superoxide dismutase, and cytochrome c. Finally, PAF-AH exhibited antagonistic activity against Rhizoctonia solani in plate confrontation assays. Our results indicate PAF-AH may play an important role in T. harzianum stress response and antagonism under diverse environmental conditions.

Metabolites of Trichoderma species isolated from damp building materials.

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McMullin, David R; Renaud, Justin B; Barasubiye, Tharcisse; Sumarah, Mark W; Miller, J David

2017-07-01

Buildings that have been flooded often have high concentrations of Trichoderma spores in the air while drying. Inhaled spores and spore and mycelial fragments contain large amounts of fungal glucan and natural products that contribute to the symptoms associated with indoor mould exposures. In this study, we considered both small molecules and peptaibol profiles of T. atroviride, T. koningiopsis, T. citrinoviride, and T. harzianum strains obtained from damp buildings in eastern Canada. Twenty-residue peptaibols and sorbicillin-derived metabolites (1-6) including a new structure, (R)-vertinolide (1), were characterized from T. citrinoviride. Trichoderma koningiopsis produced several koninginins (7-10), trikoningin KA V, and the 11-residue lipopeptaibols trikoningin KB I and trikoningin KB II. Trichoderma atroviride biosynthesized a mixture of 19-residue trichorzianine-like peptaibols, whereas T. harzianum produced 18-residue trichokindin-like peptaibols and the 11-residue harzianin HB I that was subsequently identified from the studied T. citrinoviride strain. Two α-pyrones, 6-pentyl-pyran-2-one (11) and an oxidized analog (12), were produced by both T. atroviride and T. harzianum. Aside from exposure to low molecular weight natural products, inhalation of Trichoderma spores and mycelial fragments may result in exposure to membrane-disrupting peptaibols. This investigation contributes to a more comprehensive understanding of the biologically active natural products produced by fungi commonly found in damp buildings.

Differential display of abundantly expressed genes of Trichoderma harzianum during colonization of tomato-germinating seeds and roots.

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Mehrabi-Koushki, Mehdi; Rouhani, Hamid; Mahdikhani-Moghaddam, Esmat

2012-11-01

The identification of Trichoderma genes whose expression is altered during early stages of interaction with developing roots of germinated seeds is an important step toward understanding the rhizosphere competency of Trichoderma spp. The potential of 13 Trichoderma strains to colonize tomato root and promote plant growth has been evaluated. All used strains successfully propagated in spermosphere and continued their growth in rhizoplane simultaneously root enlargement while the strains T6 and T7 were the most abundant in the apical segment of roots. Root colonization in most strains associated with promoting the roots and shoots growth while they significantly increased up to 43 and 40 % roots and shoots dry weights, respectively. Differential display reverse transcriptase-PCR (DDRT-PCR) has been developed to detect differentially expressed genes in the previously selected strain, Trichoderma harzianum T7, during colonization stages of tomato-germinating seeds and roots. Amplified DDRT-PCR products were analyzed on gel agarose and 62 differential bands excised, purified, cloned, and sequenced. Obtained ESTs were submit-queried to NCBI database by BLASTx search and gene ontology hierarchy. Most of transcripts (29 EST) corresponds to known and hypothetical proteins such as secretion-related small GTPase, 40S ribosomal protein S3a, 3-hydroxybutyryl-CoA dehydrogenase, DNA repair protein rad50, lipid phosphate phosphatase-related protein type 3, nuclear essential protein, phospholipase A2, fatty acid desaturase, nuclear pore complex subunit Nup133, ubiquitin-activating enzyme, and 60S ribosomal protein L40. Also, 13 of these sequences showed no homology (E > 0.05) with public databases and considered as novel genes. Some of these ESTs corresponded to genes encodes enzymes potentially involved in nutritional support of microorganisms which have obvious importance in the establishment of Trichoderma in spermosphere and rhizosphere, via potentially functioning in

Biogenic silver nanoparticles based on trichoderma harzianum: synthesis, characterization, toxicity evaluation and biological activity

Science.gov (United States)

Guilger, Mariana; Pasquoto-Stigliani, Tatiane; Bilesky-Jose, Natália; Grillo, Renato; Abhilash, P. C.; Fraceto, Leonardo Fernandes; Lima, Renata De

2017-03-01

White mold is an agricultural disease caused by the fungus Sclerotinia sclerotiorum, which affects important crops. There are different ways of controlling this organism, but none provides inhibition of its resistance structures (sclerotia). Nanotechnology offers promising applications in agricultural area. Here, silver nanoparticles were biogenically synthesized using the fungus Trichoderma harzianum and characterized. Cytotoxicity and genotoxicity were evaluated, and the nanoparticles were initially tested against white mold sclerotia. Their effects on soybean were also investigated with no effects observed. The nanoparticles showed potential against S. sclerotiorum, inhibiting sclerotia germination and mycelial growth. Nanoparticle characterization data indicated spherical morphology, satisfactory polydispersity and size distribution. Cytotoxicity and genotoxicity assays showed that the nanoparticles caused both the effects, although, the most toxic concentrations were above those applied for white mold control. Given the potential of the nanoparticles against S. sclerotiorum, we conclude that this study presents a first step for a new alternative in white mold control.

Characterization of Malaysian Trichoderma isolates using random amplified microsatellites (RAMS).

Science.gov (United States)

Siddiquee, Shafiquzzaman; Tan, Soon Guan; Yusuf, Umi Kalsom; Fatihah, Nur Hasan Nudin; Hasan, Md Mainul

2012-01-01

Trichoderma species are commercially applied as biocontrol agents against numerous plant pathogenic fungi due to their production of antifungal metabolites, competition for nutrients and space, and mycoparasitism. However, currently the identification of Trichoderma species from throughout the world based on micro-morphological descriptions is tedious and prone to error. The correct identification of Trichoderma species is important as several traits are species-specific. The Random Amplified Microsatellites (RAMS) analysis done using five primers in this study showed different degrees of the genetic similarity among 42 isolates of this genus. The genetic similarity values were found to be in the range of 12.50-85.11% based on a total of 76 bands scored in the Trichoderma isolates. Of these 76 bands, 96.05% were polymorphic, 3.95% were monomorphic and 16% were exclusive bands. Two bands (250 bp and 200 bp) produced by primer LR-5 and one band (250 bp) by primer P1A were present in all the Trichoderma isolates collected from healthy and infected oil palm plantation soils. Cluster analysis based on UPGMA of the RAMS marker data showed that T. harzianum, T. virens and T. longibrachiatum isolates were grouped into different clades and lineages. In this study we found that although T. aureoviride isolates were morphologically different when compared to T. harzianum isolates, the UPGMA cluster analysis showed that the majority isolates of T. aureoviride (seven from nine) were closely related to the isolates of T. harzianum.

Application of Trichoderma harziunum T22 as a biofertilizer ...

African Journals Online (AJOL)

This study was conducted to investigate the effect of Trichoderma harzianum strain (T22) on the growth and development of maize (Zea mays) plants. T. harzianum was applied to the grains in two different treatments. The first one was performed by inoculating the soil with the air dried growth of the tested fungus in three ...

Saccharification of rice straw by cellulase from a local Trichoderma harzianum SNRS3 for biobutanol production.

Science.gov (United States)

Rahnama, Nooshin; Foo, Hooi Ling; Abdul Rahman, Nor Aini; Ariff, Arbakariya; Md Shah, Umi Kalsom

2014-12-12

Rice straw has shown to be a promising agricultural by-product in the bioconversion of biomass to value-added products. Hydrolysis of cellulose, a main constituent of lignocellulosic biomass, is a requirement for fermentable sugar production and its subsequent bioconversion to biofuels such as biobutanol. The high cost of commercial enzymes is a major impediment to the industrial application of cellulases. Therefore, the use of local microbial enzymes has been suggested. Trichoderma harzianum strains are potential CMCase and β-glucosidase producers. However, few researches have been reported on cellulase production by T. harzianum and the subsequent use of the crude cellulase for cellulose enzymatic hydrolysis. For cellulose hydrolysis to be efficiently performed, the presence of the whole set of cellulase components including exoglucanase, endoglucanase, and β-glucosidase at a considerable concentration is required. Biomass recalcitrance is also a bottleneck in the bioconversion of agricultural residues to value-added products. An effective pretreatment could be of central significance in the bioconversion of biomass to biofuels. Rice straw pretreated using various concentrations of NaOH was subjected to enzymatic hydrolysis. The saccharification of rice straw pretreated with 2% (w/v) NaOH using crude cellulase from local T. harzianum SNRS3 resulted in the production of 29.87 g/L reducing sugar and a yield of 0.6 g/g substrate. The use of rice straw hydrolysate as carbon source for biobutanol fermentation by Clostridium acetobutylicum ATCC 824 resulted in an ABE yield, ABE productivity, and biobutanol yield of 0.27 g/g glucose, 0.04 g/L/h and 0.16 g/g glucose, respectively. As a potential β-glucosidase producer, T. harzianum SNRS3 used in this study was able to produce β-glucosidase at the activity of 173.71 U/g substrate. However, for cellulose hydrolysis to be efficient, Filter Paper Activity at a considerable concentration is also required to initiate the

Expression of the β-1,3-glucanase gene bgn13.1 from Trichoderma harzianum in strawberry increases tolerance to crown rot diseases but interferes with plant growth.

Science.gov (United States)

Mercado, José A; Barceló, Marta; Pliego, Clara; Rey, Manuel; Caballero, José L; Muñoz-Blanco, Juan; Ruano-Rosa, David; López-Herrera, Carlos; de Los Santos, Berta; Romero-Muñoz, Fernando; Pliego-Alfaro, Fernando

2015-12-01

The expression of antifungal genes from Trichoderma harzianum, mainly chitinases, has been used to confer plant resistance to fungal diseases. However, the biotechnological potential of glucanase genes from Trichoderma has been scarcely assessed. In this research, transgenic strawberry plants expressing the β-1,3-glucanase gene bgn13.1 from T. harzianum, under the control of the CaMV35S promoter, have been generated. After acclimatization, five out of 12 independent lines analysed showed a stunted phenotype when growing in the greenhouse. Moreover, most of the lines displayed a reduced yield due to both a reduction in the number of fruit per plant and a lower fruit size. Several transgenic lines showing higher glucanase activity in leaves than control plants were selected for pathogenicity tests. When inoculated with Colletotrichum acutatum, one of the most important strawberry pathogens, transgenic lines showed lower anthracnose symptoms in leaf and crown than control. In the three lines selected, the percentage of plants showing anthracnose symptoms in crown decreased from 61 % to a mean value of 16.5 %, in control and transgenic lines, respectively. Some transgenic lines also showed an enhanced resistance to Rosellinia necatrix, a soil-borne pathogen causing root and crown rot in strawberry. These results indicate that bgn13.1 from T. harzianum can be used to increase strawberry tolerance to crown rot diseases, although its constitutive expression affects plant growth and fruit yield. Alternative strategies such as the use of tissue specific promoters might avoid the negative effects of bgn13.1 expression in plant performance.

Combined enzymatic hydrolysis and fermentation of aspenwood using enzymes derived from Trichoderma harzianum E58

Energy Technology Data Exchange (ETDEWEB)

1990-05-01

Energy, Mines and Resources Canada supported a project with Forintek Canada Corp. directed toward the conversion of aspenwood to ethanol. This conversion is carried out through three sequential steps, steam explosion/extraction, hydrolysis and fermentation. This investigation involved study of the factors which governed the rate and extent of cellulose hydrolysis. The physical and chemical state of the material to be hydrolysed, enzyme concentation and adsorption onto residue, end-product characterization and inhibition, recycling of enzymes and cellulose, and growth media for the fungus were among the variables examined. The research demonstrated the interdependency between pretreatment, cellulose hydrolysis, hemicellulose fermentation and enzyme production. It was also determined that because of the amount of cellulose required for enzyme production and the difficulties encountered in recovering/recycling the celluloses, further work is required in order to commercialize an enzymatic hydrolysis process based on Trichoderma harzianum E58.

Integrated control of wood destroying basidiomycetes combining Cu-based wood preservatives and Trichoderma spp.

Science.gov (United States)

Ribera, Javier; Fink, Siegfried; Bas, Maria Del Carmen; Schwarze, Francis W M R

2017-01-01

The production of new generation of wood preservatives (without addition of a co-biocide) in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720) was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium). T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55-65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0-25%). Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens previously exposed to T

Integrated control of wood destroying basidiomycetes combining Cu-based wood preservatives and Trichoderma spp.

Directory of Open Access Journals (Sweden)

Javier Ribera

Full Text Available The production of new generation of wood preservatives (without addition of a co-biocide in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720 was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium. T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55-65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0-25%. Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens

Antagonistic studies and hyphal interactions of the new antagonist Aspergillus piperis against some phytopathogenic fungi in vitro in comparison with Trichoderma harzianum.

Science.gov (United States)

El-Debaiky, Samah A

2017-12-01

The present study represents, for the first time, the detailed studies about the hyphal interactions of Aspergillus piperis, as a new antagonist, against some isolated plant pathogenic fungi (Alternaria alternata, Alternaria solani, Botrytis cinerea, Sclerotium cepivorum and Sclerotinia sclerotiorum) in vitro. The bio-controlling capability of A. piperis against the tested phytopathogens was tested using the dual culture method. This experiment revealed that A. piperis had antagonistic activity and reduced the growth of the tested phytopathogens and grew over their mycelia in the paired plates. Also, several antagonistic mechanisms were recorded, in this study, between A. piperis and the tested phytopathogens using the microscopic examination. The bio-controlling activity and the antagonistic mechanisms exhibited by the new antagonist, A. piperis were compared with those obtained by the common antagonist, Trichoderma harzianum against the same phytopathogens. The obtained results showed that, A. piperis was more effective than T. harzianum in inhibiting all the tested species in the dual culture plates. The best result was 81.85% inhibition percentage against S. sclerotiorum by A. piperis while, T. harzianum exhibits only 45.18%. Moreover, several antagonistic mechanisms and hyphal interactions were investigated among the hyphae of both A.piperis and T. harzianum and the hyphae of the tested phytopathogens. These mechanisms were summarized as; mycoparasitism (coiling and penetration of the hyphae) and antibiosis in the form of lysis of the hyphal cells and spores, denaturation and breaking of the hyphae. The indirect interaction (antibiosis) and the direct mycoparasitism were observed by A. piperis against all the tested phytopathogens, but it attacked the hyphae and conidiophores of A. alternata by only the antibiosis interaction. The microscopic examination revealed also that T. harzianum attacked the tested phytopathogens by both antibiosis and mycoparasitism

Differential expression of oil palm pathology genes during interactions with Ganoderma boninense and Trichoderma harzianum.

Science.gov (United States)

Alizadeh, Fahimeh; Abdullah, Siti Nor Akmar; Khodavandi, Alireza; Abdullah, Faridah; Yusuf, Umi Kalsom; Chong, Pei Pei

2011-07-01

The expression profiles of Δ9 stearoyl-acyl carrier protein desaturase (SAD1 and SAD2) and type 3 metallothionein (MT3-A and MT3-B) were investigated in seedlings of oil palm (Elaeis guineensis) artificially inoculated with the pathogenic fungus Ganoderma boninense and the symbiotic fungus Trichoderma harzianum. Expression of SAD1 and MT3-A in roots and SAD2 in leaves were significantly up-regulated in G. boninense inoculated seedlings at 21 d after treatment when physical symptoms had not yet appeared and thereafter decreased to basal levels when symptoms became visible. Our finding demonstrated that the SAD1 expression in leaves was significantly down-regulated to negligible levels at 42 and 63 d after treatment. The transcripts of MT3 genes were synthesized in G. boninense inoculated leaves at 42 d after treatment, and the analyses did not show detectable expression of these genes before 42 d after treatment. In T. harzianum inoculated seedlings, the expression levels of SAD1 and SAD2 increased gradually and were stronger in roots than leaves, while for MT3-A and MT3-B, the expression levels were induced in leaves at 3d after treatment and subsequently maintained at same levels until 63d after treatment. The MT3-A expression was significantly up-regulated in roots at 3d after treatment and thereafter were maintained at this level. Both SAD and MT3 expression were maintained at maximum levels or at levels higher than basal. This study demonstrates that oil palm was able to distinguish between pathogenic and symbiotic fungal interactions, thus resulting in different transcriptional activation profiles of SAD and MT3 genes. Increases in expression levels of SAD and MT3 would lead to enhanced resistance against G. boninense and down-regulation of genes confer potential for invasive growth of the pathogen. Differences in expression profiles of SAD and MT3 relate to plant resistance mechanisms while supporting growth enhancing effects of symbiotic T. harzianum

Application of Trichoderma harzianum in the control of basal stem rot of oil palms

Institute of Scientific and Technical Information of China (English)

Abdullah F; Ilias G N M

2004-01-01

@@ The palm, Elaeis guineensis, has its origins in Africa but is planted on a commercial basis in several countries Statistics for 2002 showed that in the lead for land mass under oil palm cultivation is Indonesia, at 3,769,000 ha, followed by Malaysia at 3,376,000 ha; however, the world' s leading producer of palm oil is still Malaysia, since the 1970's. Both countries are predicted to produce 82.4%of the world's palm oil production by the year 2005. However, the palm is susceptible to basal stem rot, a devastating disease which results in direct loss of field stands and to which no effective chemical control is yet available. Caused by Ganoderma boninense, infected palms appear symptomless, at the first sign of disease, at least 50 % of the internal trunk tissue stem would have actually rotted. This study investigated the efficacy of Trichoderma harzianum (isolate FA 1132) as a biological control agent, using 6-month old oil palm seedlings as models and the experiment performed in a greenhouse at 29-30 ℃ ambient conditions. The plants were artificially infected with G. boninense and a conidial suspension of 1 × 109-9 × 109 spores/mL was applied as a soil drench at 1L/plant every 2 weeks for 20weeks. The parameters examined were efficacy of the biocontrol agent and the effect of Trichodermaincorporated mulch in addition to the soil drench. Efficacy was assessed in terms disease severity index (DSI) where a higher percentage indicates a higher severity. Results showed that infection first sets in on untreated plants at week 12 and got worse progressively. The completely untreated plants were all infected and the DSI at 20 weeks after infection (wa. i.) was 92. 5%. Plants given only a Trichoderma -infused food base supplement without conidial suspension gave a DSI of 70% whereas those given a conidial soil drench without supplemental food base gave a DSI of 85% at 20 w.a.i.Infected plants given a conidial treatment together with a food base supplement gave a DSI

Biocontrol potential of Trichoderma harzianum isolate T-aloe against Sclerotinia sclerotiorum in soybean.

Science.gov (United States)

Zhang, Fuli; Ge, Honglian; Zhang, Fan; Guo, Ning; Wang, Yucheng; Chen, Long; Ji, Xiue; Li, Chengwei

2016-03-01

Sclerotinia stem rot, caused by Sclerotinia sclerotiorum (Lib.) de Bary is a major disease of soybean (Glycine max (L.) Merr.). At present, we revealed the three-way interaction between Trichoderma harzianum T-aloe, pathogen S. sclerotiorum and soybean plants in order to demonstrate biocontrol mechanism and evaluate biocontrol potential of T-aloe against S. sclerotiorum in soybean. In our experiments, T-aloe inhibited the growth of S. sclerotiorum with an efficiency of 56.3% in dual culture tests. T-aloe hyphae grew in parallel or intertwined with S. sclerotiorum hyphae and produced hooked contact branches, indicating mycoparasitism. Plate tests showed that T-aloe culture filtrate inhibited S. sclerotiorum growth with an inhibition efficiency of 51.2% and sclerotia production. T-aloe pretreatment showed growth-promoting effect on soybean plants. The activities of peroxidase, superoxide dismutase, and catalase increased, and the hydrogen peroxide (H2O2) as well as the superoxide radical (O2(-)) content in soybean leaves decreased after T-aloe pretreatment in response to S. sclerotiorum pathogen challenge. T-aloe treatment diminished damage caused by pathogen stress on soybean leaf cell membrane, and increased chlorophyll as well as total phenol contents. The defense-related genes PR1, PR2, and PR3 were expressed in the leaves of T-aloe-treated plants. In summary, T-aloe displayed biocontrol potential against S. sclerotiorum. This is the first report of unraveling biocontrol potential of Trichoderma Spp. to soybean sclerotinia stem rot from the three-way interaction between the biocontrol agent, pathogen S. sclerotiorum and soybean plants. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Daniele Franco Martins Machado

2015-02-01

Full Text Available Neste trabalho, o objetivo foi avaliar o efeito de isolados de Trichoderma spp. na emergência de plântulas e no crescimento de mudas de cambará (Gochnatia polymorpha. Utilizou-se, em casa de vegetação, substrato esterilizado e não esterilizado, sendo avaliados os efeitos de quatro isolados de trichoderma: TSM1 e TSM2 de Trichoderma viride, 2B2 e 2B22 de Trichoderma harzianum mais um mix preparado com a mistura desses quatro isolados, além de dois produtos comerciais à base de trichoderma. A análise dos dados permitiu concluir que os isolados de trichoderma testados não interferem na emergência das plântulas, mas os isolados 2B2 e 2B22 de T. harzianum apresentam potenciais como promotores de crescimento de mudas de cambará.

BIOACTIVE METABOLITES FROM TRICHODERMA HARZIANUM ...

African Journals Online (AJOL)

a

ABSTRACT. The tea plant, Camellia sinensis (L.) O. Kuntze is an important crop in the agriculturally based economy of Kenya. Many diseases affect the tea plant but the most prevalent is armillaria root rot caused by the fungus Armillaria mellea. Compounds from the fermentation of. Trichoderma species in different media ...

Screening of antagonistic bacteria against the green mold disease (Trichoderma harzianum Rifai of Grey Oyster Mushroom (Pleurotus pulmonarius (Fr. Quel.

Directory of Open Access Journals (Sweden)

Nualsri, C.

2005-01-01

Full Text Available A total of 174 strains of bacteria antagonistic against the green mold (Trichoderma harzianum, isolated from cultivating bags and fruiting bodies of the mushrooms, were screened for effects on mushroom mycelia and ability to control the green mold disease. Twenty-eight of them promoted the primodia formation of the Pleurotus pulmonarius mycelia on agar plates. Twenty-two isolates were selected and further tested in a mushroom house. Cell suspension of each isolate was prepared and sprayed onto the spawn surface of P. pulmonarius. Fifteen isolates shortened the times required from watering to 2nd and 3rd flushing and increased yield of the basidiocarps by 1.1-34.3% over 30 days. Six isolates of bacteria which showed an inhibitory effect against T. harzianum, enhanced primordia formation and increased yield of P. pulmonarius were selected and used for control testing in a cultivation house. The suspension of each isolate was sprayed onto the spawn surface immediately after exposure to the air in the mushroom house, followed by spore suspension of T. harzianum two days later. The number of infected bags was counted at 30 days after inoculation and the cumulative yield was compared after 60 days. The results showed that bacteria isolate B012-022 was highly effective in suppressing the green mold disease.Only 6.7% of the cultivating bags were found to be infected by T. harzianum when bacteria isolate B012-022 was applied. Cumulative yield obtained from 900 g of 94% sawdust + 5% rice bran + 1% Ca(OH2 was 300.0 g/bag after 60 days, 71.1% higher than the bags infected by the green mold and without bacterial spraying. Identification of the six bacterial isolates showed all to be Bacillus spp.

Synthesis of a nanoporous molecularly imprinted polymers for dibutyl Phthalate extracted from Trichoderma Harzianum

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Maede Shahiri Tabarestani

2016-07-01

Full Text Available In this study, molecularly imprinted polymers were synthesized for dibutyl phthalate as a bioactive chemical compound with antifungal activity which produced by Trichoderma Harzianum (JX1738521. The molecularly imprinted polymers were synthesized via precipitation polymerization method from methacrylic acid, dibutyl phthalate and trimetylolpropantrimethacrylate as a functional monomer, template and cross-linker, respectively. After removal of the template by the eluent from the MIPs, the leached nanoparticles of the MIPs had a good binding capacity as equal 830 mg/g. The polymer particles have been evaluated by field emission scan electron microscopy and Brunauer–Emmett–Teller  techniques. The excellent specific surface area in the molecularly imprinted polymers as equal to 690.301 m2/g comparatively to non-imprinted polymers (ca. 89.894 m2/g, confirms that the nanoporous MIPs were synthesized, successfully. The results indicated that the nanoporous MIPs can be used in solid phase extraction. This is a novel method for separation of the bioactive compounds from fungi secondary metabolites in biological control.

Compatibilidad de Trichoderma harzianum Rifai con fungicidas del arroz y su efecto sobre tres fitopatógenos fúngicos

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Manuel Francisco Rodríguez Saldaña

2016-04-01

Full Text Available Resumen La investigación se realizó en el Laboratorio Provincial de Sanidad Vegetal en Camagüey, en la etapa comprendida de septiembre de 2013 a septiembre de 2015, en condiciones in vitro donde se determinó la compatibilidad y capacidad antagónica frente a pesticidas usados en el arroz, de Trichoderma harzianum Rifai cepa A-34 sobre los patógenos del arroz (Bipolaris oryzae Breda de Haan, Saracladium oryzae (Sawada w. Gams & D.Hawksworth y Magnaporthe grisea (Hebert Barr. Las evaluaciones mediante métodos tradicionales de aislamiento microbiológico del crecimiento micelial, la esporulación y la germinación conidial del antagonista para determinar si ejerce los mecanismos de acción de antibiosis, competencia, y parasitismo ante los patógenos fúngicos se efectuaron a partir desde las 24 hasta las 216 horas empleando. Para el análisis estadístico se empleó un diseño bifactorial en el cultivo dual, además de las escalas para la determinación de la capacidad antagónica de los microorganismos. Los ingredientes activos tebuconazol + procloraz, trifloxistrobin+ ciproconazol, y epoxiconazol + kresoxim- metil, afectaron el crecimiento micelial del antagonista, por otra parte el antagonista frente a los ingredientes activos carbendazim, oxicloruro de cobre, azoxistrobina y tebuconazol + triadimenol tuvo crecimiento micelial, esporulación e interacción con los patógenos afectándose el crecimiento de los mismos a través de los mecanismos de acción de enrollamiento, penetración, granulación , y lisis celular desde las 96 hasta las 216 horas. Trichoderma harzianum Rifai compatibility with fungicides and its effect on three fungal plant rice pathogens. ABSTRACT The research was conducted at the Provincial Laboratory of Plant Protection in Camagüey, in the fall period September 2013 to September 2015, under in vitro conditions where compatibility and antagonistic capacity was determined against used in rice pesticides, Trichoderma

Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum.

Science.gov (United States)

Borisova, Anna S; Eneyskaya, Elena V; Jana, Suvamay; Badino, Silke F; Kari, Jeppe; Amore, Antonella; Karlsson, Magnus; Hansson, Henrik; Sandgren, Mats; Himmel, Michael E; Westh, Peter; Payne, Christina M; Kulminskaya, Anna A; Ståhlberg, Jerry

2018-01-01

The ascomycete fungus Trichoderma reesei is the predominant source of enzymes for industrial conversion of lignocellulose. Its glycoside hydrolase family 7 cellobiohydrolase (GH7 CBH) Tre Cel7A constitutes nearly half of the enzyme cocktail by weight and is the major workhorse in the cellulose hydrolysis process. The orthologs from Trichoderma atroviride ( Tat Cel7A) and Trichoderma harzianum ( Tha Cel7A) show high sequence identity with Tre Cel7A, ~ 80%, and represent naturally evolved combinations of cellulose-binding tunnel-enclosing loop motifs, which have been suggested to influence intrinsic cellobiohydrolase properties, such as endo-initiation, processivity, and off-rate. The Tat Cel7A, Tha Cel7A, and Tre Cel7A enzymes were characterized for comparison of function. The catalytic domain of Tat Cel7A was crystallized, and two structures were determined: without ligand and with thio-cellotriose in the active site. Initial hydrolysis of bacterial cellulose was faster with Tat Cel7A than either Tha Cel7A or Tre Cel7A. In synergistic saccharification of pretreated corn stover, both Tat Cel7A and Tha Cel7A were more efficient than Tre Cel7A, although Tat Cel7A was more sensitive to thermal inactivation. Structural analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We hypothesize that the combination of loop motifs is the main determinant for the observed differences in Cel7A activity on cellulosic substrates. Fine-tuning of the loop flexibility appears to be an important evolutionary target in Trichoderma spp., a conclusion supported by the RCA data. Our results indicate that, for industrial use, it would be beneficial to combine loop motifs from Tat Cel7A with the thermostability features of Tre Cel7A. Furthermore

Combined enzymatic hydrolysis and fermentation of aspenwood using enzymes derived from Trichoderma harzianum E58

Energy Technology Data Exchange (ETDEWEB)

1989-05-01

A project was initiated to study the conversion of aspenwood to ethanol, butanol or butanediol. The conversion method consisted of steam explosion pretreatment, followed by the enzymatic hydrolysis of the carbohydrate polymers, cellulose and hemicellulose. The enzyme was derived from a wild strain of the fungus Trichoderma harzianum E58, chosen because it produces a cellulose system that can degrade crystalline cellulose to glucose. The aspenwood was steamed at 240{degree}C for 80 seconds and then water and alkali extracted. The insoluble residue was 84% cellulose and was used for both enzyme production and the production of glucose, which was fermented to ethanol. Before fermentation of the water-soluble fraction was possible, the acetylxylan had to be hydrolyzed and the inhibitors (glucose, galactose, acetic and uronic acids, and lignin- and sugar-degradation products) removed. Enzymatic hydrolysis was found to generate less fermentation inhibitors than sulfuric acid hydrolysis. Due to market factors, fermentation research centred on the production of ethanol from hemicellulose, using the yeast Pichia stipitis. Although lignin had no effect on hydrolysis, it increased the bulk to be handled, in combination with small amounts of cellulose was found to strongly adsorb the cellulose enzymes, and broke down to produce inhibitors of the cellulose complex of T. harzanium and the enzyme production phase. Thus, it was advantageous to remove the lignin prior to enzyme production and cellular hydrolysis. None of the strategies were successful in decreasing the amount of cellulose required for enzyme production. It was concluded that T. harzianum E58 is unsuitable for use in a commercial bioconversion project. 59 refs., 31 figs., 31 tabs.

[Effect of Glomus versiforme and Trichoderma harzianum on growth and quality of Salvia miltiorrhiza].

Science.gov (United States)

Wang, Xue; Chen, Mei-Lan; Yang, Guang; Li, Xiao-Ming; Li, Peng-Ying; Chen, Min

2014-05-01

The present study aimed to investigate the effect of Glomus versiforme and Trichodema harzianum on the growth and quality of Salvia miltiorrhiza continuous cropping under field conditions. The field plot experiment was conducted, these active components in the plant were analyzed by HPLC, the root diseases incidence rate of S. miltiorrhiza determined by observation and counting, and relative parameters were measured. The data was statistically processed. The result showed that inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly decreased the root diseases incidence rate of S. miltiorrhiza, and combined inoculation of G. versiforme with T. harzianum was better than other treatments. All treatments improved accumulation of active ingredients in root. Inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly increased the content of salvianolic acid B and cryptotanshinone of root (P harzianum and combined inoculation of G. versiforme with T. harzianum significantly enhanced the content of tanshinone I and tanshinone II(A) of the root (P harzianum can effectively reduce the root diseases incidence of continuous cropping S. miltiorrhiza, and improve the quality of S. miltiorrhiza.

ANTAGONISMO IN VITRO DE Trichoderma harzianum Rifai SOBRE Fusarium oxysporum Schlecht f. sp passiflorae EN MARACUYÁ (Passiflora edulis Sims var. Flavicarpa DEL MUNICIPIO ZONA BANANERA COLOMBIANA ANTAGONISM IN VITRO OF Trichoderma harzianum Rifai AGAINST Fusarium oxysporum Schlecht f. sp passiflorae IN PASSION FRUIT (Passiflora edulis Sims var. Flavicarpa FROM COLOMBIAN BANANERA ZONE MUNICIPALITY

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Reinel José Fernández Barbosa

2009-06-01

determining the antagonistic capacity of 6 strains of Trichoderma harzianum against the pathogen. 3 commercial strains were evaluated (TCC-001, TCC-005 and TCC-006 and 3 isolated from soil cultivated with palm oil in the Research Center Caribia of Corpoica (TCN-009, TCN-010, TCN-014. These were evaluated under conditions in vitro using the technique of dual cultivation in Petri plates with Agar Sabouraud. Competition for nutritious and space, micoparasitism and Percentage of Radial Growth Inhibition (PRGI was evaluated, incubating them for 10 days at 28 ºC. A totally randomized design settled down, with 13 treatments and 3 repetitions. All the isolates of T. harzianum exceeded in growth to F. oxysporum with a pathogen growth radiu averages (PGR of 7.42 cm in dual culture. While the pathogen showed average radiu of 1.99 cm. TCN-009 and TCC-006 expressed the best results in competition for nutrients and space to grow 4 times faster than F. oxysporum and to reduce 3 times less the PRGI compared with the control, without significant differences between treatments (P=0.0001; also, produced the largest PRGI to 10 days with values of 64.61 and 65.91% respectively. There was no significant difference when comparing commercial and native isolates, but for the indigenous nature of the isolation, TCN-009 is, an in vitro level, more promising in the search for an antagonist in the control F. oxysporum in similar agroclimatic regions to the Bananera Zone Colombian.

Evaluación de Trichoderma harzianum como propuesta alternativa al uso de sustancias químicas sintéticas para el control de Botrytis sp. en el cultivo de rosas (Rosa sp.) variedad aubade en la Finca Florícola Valle Verde Cayambe

OpenAIRE

Jara Guaña, Andrea Margarita

2014-01-01

The principal objective in the present research was "to evaluate the efficacy of Trichoderma harzianum as an alternative to the use of synthetic chemicals for the control of Botrytis sp, in the roses cultivation (Rosa sp). Aubade range, to reduce environmental impact through an experimental process, which was performed in the flower farm located in Valle Verde in Ayora Region Cayambe, Pichincha province. The specific objectives were: to compare the effects of control harzianum on Botrytis ...

Role of Bradyrhizobium japonicum and Trichoderma spp. in the control of root rot disease of soybean

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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.

Trichoderma amazonicum, a new endophytic species on Hevea brasiliensis and H. guianensis from the Amazon basin

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A new species of Trichoderma (teleomorph Hypocrea, Ascomycota, Sordariomycetes, Hypocreales, Hypocreaceae), T. amazonicum, endophytic on the living sapwood and leaves of Hevea spp. trees is described. Trichoderma amazonicum is distinguished from closely related species in the Harzianum clade (e.g. ...

Biological control of Meloidogyne incognita by Trichoderma ...

African Journals Online (AJOL)

Biological control against the root-knot nematode, Meloidogyne incognita was proven to occur in tomato, Solanum lycopersicom, soil-drenched with different isolates of Trichoderma harzianum and a commercial suspension of Serratia marcescens (Nemaless). The potential of such biocontrol agents to trigger plant defense ...

Cloning and sequencing of a cellobiohydrolase gene from Trichoderma harzianum FP108

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Patrick Guilfoile; Ron Burns; Zu-Yi Gu; Matt Amundson; Fu-Hsian Chang

1999-01-01

A cbbl cellobiohydrolase gene was cloned and sequenced from the fungus Trichoderrna harzianum FP108. The cloning was performed by PCR amplification of T. harzianum genomic DNA, using PCR primers whose sequence was based on the cbbl gene from Tricboderma reesei. The 3' end of the gene was isolated by inverse...

Comparative growth of trichoderma strains in different nutritional sources, using bioscreen c automated system Crescimento de linhagens de Trichoderma em diferentes fontes nutricionais, empregando o sistema automatizado Bioscreen C.

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Bianca Caroline Rossi-Rodrigues

2009-06-01

Full Text Available Trichoderma is one of the fungi genera that produce important metabolites for industry. The growth of these organisms is a consequence of the nutritional sources used as also of the physical conditions employed to cultivate them. In this work, the automated Bioscreen C system was used to evaluate the influence of different nutritional sources on the growth of Trichoderma strains (T. hamatum, T. harzianum, T. viride, and T. longibrachiatum isolated from the soil in the Juréia-Itatins Ecological Station (JIES, São Paulo State - Brazil.The cultures were grown in liquid culture media containing different carbon- (2%; w/v and nitrogen (1%; w/v sources at 28ºC, pH 6.5, and agitated at 150 rpm for 72 h. The results showed, as expected, that glucose is superior to sucrose as a growth-stimulating carbon source in the Trichoderma strains studied, while yeast extract and tryptone were good growth-stimulating nitrogen sources in the cultivation of T. hamatum and T. harzianum.Trichoderma é um dos gêneros de fungos produtores de metabólitos de interesse industrial. O crescimento destes organismos é conseqüência das fontes nutricionais utilizadas, juntamente com as condições físicas de cultivo. Neste trabalho, o sistema automatizado Bioscreen C foi utilizado para avaliar a influência de diferentes fontes nutricionais sobre o crescimento de linhagens de Trichoderma (T. hamatum, T. harzianum, T. viride e T. longibrachiatum isoladas do solo da Estação Ecológica da Juréia-Itatins (JIES, São Paulo - Brasil. Os cultivos foram feitos em meios líquidos de cultura contendo diferentes fontes de carbono (2%; w / v e nitrogênio (1%; w / v a 28ºC, pH 6,5 e agitados a 150 rpm durante 72 h. Os resultados mostraram, conforme esperado, que a glicose é melhor do que a sacarose como fonte de carbono indutora de crescimento das linhagens de Trichoderma testadas, enquanto que, o extrato de leveduras e a triptona foram boas fontes de nitrogênio indutoras de

Nitrogen Metabolism and Growth Enhancement in Tomato Plants Challenged with Trichoderma harzianum Expressing the Aspergillus nidulans Acetamidase amdS Gene

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Domínguez, Sara; Rubio, M. Belén; Cardoza, Rosa E.; Gutiérrez, Santiago; Nicolás, Carlos; Bettiol, Wagner; Hermosa, Rosa; Monte, Enrique

2016-01-01

Trichoderma is a fungal genus that includes species that are currently being used as biological control agents and/or as biofertilizers. In addition to the direct application of Trichoderma spp. as biocontrol agents in plant protection, recent studies have focused on the beneficial responses exerted on plants, stimulating the growth, activating the defenses, and/or improving nutrient uptake. The amdS gene, encoding an acetamidase of Aspergillus, has been used as a selectable marker for the transformation of filamentous fungi, including Trichoderma spp., but the physiological effects of the introduction of this gene into the genome of these microorganisms still remains unexplored. No evidence of amdS orthologous genes has been detected within the Trichoderma spp. genomes and the amdS heterologous expression in Trichoderma harzianum T34 did not affect the growth of this fungus in media lacking acetamide. However, it did confer the ability for the fungus to use this amide as a nitrogen source. Although a similar antagonistic behavior was observed for T34 and amdS transformants in dual cultures against Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum, a significantly higher antifungal activity was detected in amdS transformants against F. oxysporum, compared to that of T34, in membrane assays on media lacking acetamide. In Trichoderma-tomato interaction assays, amdS transformants were able to promote plant growth to a greater extent than the wild-type T34, although compared with this strain the transformants showed similar capability to colonize tomato roots. Gene expression patterns from aerial parts of 3-week-old tomato plants treated with T34 and the amdS transformants have also been investigated using GeneChip Tomato Genome Arrays. The downregulation of defense genes and the upregulation of carbon and nitrogen metabolism genes observed in the microarrays were accompanied by (i) enhanced growth, (ii) increased carbon and nitrogen levels, and (iii) a

Nitrogen Metabolism and Growth Enhancement in Tomato Plants Challenged with Trichoderma harzianum Expressing the Aspergillus nidulans Acetamidase amdS Gene.

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Domínguez, Sara; Rubio, M Belén; Cardoza, Rosa E; Gutiérrez, Santiago; Nicolás, Carlos; Bettiol, Wagner; Hermosa, Rosa; Monte, Enrique

2016-01-01

Trichoderma is a fungal genus that includes species that are currently being used as biological control agents and/or as biofertilizers. In addition to the direct application of Trichoderma spp. as biocontrol agents in plant protection, recent studies have focused on the beneficial responses exerted on plants, stimulating the growth, activating the defenses, and/or improving nutrient uptake. The amdS gene, encoding an acetamidase of Aspergillus, has been used as a selectable marker for the transformation of filamentous fungi, including Trichoderma spp., but the physiological effects of the introduction of this gene into the genome of these microorganisms still remains unexplored. No evidence of amdS orthologous genes has been detected within the Trichoderma spp. genomes and the amdS heterologous expression in Trichoderma harzianum T34 did not affect the growth of this fungus in media lacking acetamide. However, it did confer the ability for the fungus to use this amide as a nitrogen source. Although a similar antagonistic behavior was observed for T34 and amdS transformants in dual cultures against Rhizoctonia solani, Botrytis cinerea, and Fusarium oxysporum, a significantly higher antifungal activity was detected in amdS transformants against F. oxysporum, compared to that of T34, in membrane assays on media lacking acetamide. In Trichoderma-tomato interaction assays, amdS transformants were able to promote plant growth to a greater extent than the wild-type T34, although compared with this strain the transformants showed similar capability to colonize tomato roots. Gene expression patterns from aerial parts of 3-week-old tomato plants treated with T34 and the amdS transformants have also been investigated using GeneChip Tomato Genome Arrays. The downregulation of defense genes and the upregulation of carbon and nitrogen metabolism genes observed in the microarrays were accompanied by (i) enhanced growth, (ii) increased carbon and nitrogen levels, and (iii) a

Trichoderma harzianum T-22 induces systemic resistance in tomato infected by Cucumber mosaic virus

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Antonella Vitti

2016-10-01

Full Text Available Understanding the induction of plant defenses against viruses using biocontrol agents is essential for developing new strategies against these pathogens, given the ineffectiveness of chemical treatments. The ability of Trichoderma harzianum, strain T-22 (T22 to control Cucumber mosaic virus (CMV in Solanum lycopersicum var. cerasiforme plants and the changes in the physiology of tomato treated/infected with T22/CMV were examined. Plant growth-promoting effects, photosynthetic performance, reactive oxygen species (ROS scavenging enzymes, and phytohormones were investigated. T22 improved tomato growth in terms of plant height and improved photosynthesis, total chlorophyll content and plant gas exchange. In contrast, CMV induced a negative effect on dry matter accumulation and inhibited the photosynthetic capacity. The analysis of plant hormones demonstrated that treating with T22 before or simultaneously to CMV infection, led to a systemic resistance by jasmonic acid/ethylene and salicylic acid signaling pathways. Conversely, systemic resistance was abscissic acid-dependent when T22 treatment was administered after the CMV infection. In conclusion, the data reported here indicate that the T22-based strategy may be the most effective measure against CMV.

Impact of salicylic acid- and jasmonic acid-regulated defences on root colonization by Trichoderma harzianum T-78.

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Martínez-Medina, Ainhoa; Appels, Freek V W; van Wees, Saskia C M

2017-08-03

We recently found that the beneficial fungus Trichoderma harzianum T-78 primes tomato plants for salicylic acid (SA)- and jasmonic acid (JA)-regulated defenses, resulting in enhanced resistance against the root knot nematode Meloidogyne incognita. By using SA- and JA-impaired mutant lines and exogenous hormonal application, here we investigated whether the SA- and JA-pathways also have a role in T-78 root colonization of Arabidopsis thaliana. Endophytic colonization by T-78 was faster in the SA-impaired mutant sid2 than in the wild type. Moreover, elicitation of SA-dependent defenses by SA application reduced T-78 colonization, indicating that the SA-pathway affects T-78 endophytism. In contrast, elicitation of the JA-pathway, which antagonized SA-dependent defenses, resulted in enhanced endophytic colonization by T-78. These findings are in line with our previous observation that SA-dependent defenses are repressed by T-78, which likely aids colonization by the endophytic fungus.

Construction and functional analysis of Trichoderma harzianum mutants that modulate maize resistance to the pathogen Curvularia lunata.

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Fan, Lili; Fu, Kehe; Yu, Chuanjin; Ma, Jia; Li, Yaqian; Chen, Jie

2014-01-01

Agrobacterium tumefaciens-mediated transformation (ATMT) was used to generate an insertional mutant library of the mycelial fungus Trichoderma harzianum. From a total of 450 mutants, six mutants that showed significant influence on maize resistance to C. lunata were analyzed in detail. Maize coated with these mutants was more susceptible to C. lunata compared with those coated with a wild-type (WT) strain. Similar to other fungal ATMT libraries, all six mutants were single copy integrations, which occurred preferentially in noncoding regions (except two mutants) and were frequently accompanied by the loss of border sequences. Two mutants (T66 and T312) that were linked to resistance were characterized further. Maize seeds coated with T66 and T312 were more susceptible to C. lunata than those treated with WT. Moreover, the mutants affected the resistance of maize to C. lunata by enhancing jasmonate-responsive gene expression. T66 and T312 induced maize resistance to C. lunata infection through a jasmonic acid-dependent pathway.

Mathematical modeling of enzyme production using Trichoderma harzianum P49P11 and sugarcane bagasse as carbon source.

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Gelain, Lucas; da Cruz Pradella, José Geraldo; da Costa, Aline Carvalho

2015-12-01

A mathematical model to describe the kinetics of enzyme production by the filamentous fungus Trichoderma harzianum P49P11 was developed using a low cost substrate as main carbon source (pretreated sugarcane bagasse). The model describes the cell growth, variation of substrate concentration and production of three kinds of enzymes (cellulases, beta-glucosidase and xylanase) in different sugarcane bagasse concentrations (5; 10; 20; 30; 40 gL(-1)). The 10 gL(-1) concentration was used to validate the model and the other to parameter estimation. The model for enzyme production has terms implicitly representing induction and repression. Substrate variation was represented by a simple degradation rate. The models seem to represent well the kinetics with a good fit for the majority of the assays. Validation results indicate that the models are adequate to represent the kinetics for a biotechnological process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effects of Trichoderma harzianum Rifai over Plasmodiophora brassicae Woronin in broccoli, in Escagüey, municipality of Rangel, Mérida state

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Mirna Labrador Morales

2014-04-01

Full Text Available The effectiveness of Trichoderma harzianum in suppressing clubroot of brassicas, which is caused by Plasmodiophora brassicae, was tested on broccoli (Brassica oleracea var. italica Plenck, in field conditions, in Escagüey, municipality of Rangel, Merida State, Venezuela. The experiment showed that the applications of a biopesticide based on this antagonist had a significant effect on the percentage of health plants and the crop yields, in dependence of the dose used. In addition, the relation benefits/cost was also favorable. These results showed that, for these particular conditions, P. brassicae is an adequate biological alternative to control the clubroot of brassicas, no aggressive to environment and human beings, useful for the transition phase toward a sustainable agriculture, without chemical pesticides.

Effects of Trichoderma harzianum Rifai over Plasmodiophora brassicae Woronin in broccoli, in Escagüey, municipality of Rangel, Mérida State

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Mirna Labrador Morales

2016-03-01

Full Text Available The effectiveness of Trichoderma harzianum in suppressing clubroot of brassicas, which is caused by Plasmodiophora brassicae, was tested on broccoli (Brassica oleracea var. italica Plenck, in field conditions, in Escagüey, municipality of Rangel, Merida State, Venezuela. The experiment showed that the applications of a biopesticide based on this antagonist had a significant effect on the percentage of health plants and the crop yields, in dependence of the dose used. In addition, the relation benefits/cost was also favorable. These results showed that, for these particular conditions, P. brassicae is an adequate biological alternative to control the clubroot of brassicas, no aggressive to environment and human beings, useful for the transition phase toward a sustainable agriculture, without chemical pesticides.

Antibiosis of Trichoderma spp strains native to northeastern Mexico against the pathogenic fungus Macrophomina phaseolina

Science.gov (United States)

Mendoza, José Luis Hernández; Pérez, María Isabel Sánchez; Prieto, Juan Manuel González; Velásquez, Jesús DiCarlo Quiroz; Olivares, Jesús Gerardo García; Langarica, Homar Rene Gill

2015-01-01

Abstract Sampling of agricultural soils from the Mexican northeastern region was performed to detect Trichoderma spp., genetically characterize it, and assess its potential use as a biologic control agent against Macrophomina phaseolina. M. phaseolina is a phytopathogen that attacks over 500 species of cultivated plants and causes heavy losses in the regional sorghum crop. Sampling was performed immediately after sorghum or corn harvest in an area that was approximately 170 km from the Mexico-USA border. Sixteen isolates were obtained in total. Using colony morphology and sequencing the internal transcribed spacers (ITS) 1 and 4 of 18S rDNA, 14 strains were identified as Trichoderma harzianum, T. koningiopsis and T. virens. Subsequently, their antagonistic activity against M. phaseolina was evaluated in vitro, and 11 isolates showed antagonism by competition and stopped M. phaseolina growth. In 4 of these isolates, the antibiosis phenomenon was observed through the formation of an intermediate band without growth between colonies. One strain, HTE808, was identified as Trichoderma koningiopsis and grew rapidly; when it came into contact with the M. phaseolina colony, it continued to grow and sporulated until it covered the entire petri dish. Microscopic examination confirmed that it has a high level of hyperparasitism and is thus considered to have high potential for use in the control of this phytopathogen. PMID:26691467

Insights into the structure and function of fungal β-mannosidases from glycoside hydrolase family 2 based on multiple crystal structures of the Trichoderma harzianum enzyme.

Science.gov (United States)

Nascimento, Alessandro S; Muniz, Joao Renato C; Aparício, Ricardo; Golubev, Alexander M; Polikarpov, Igor

2014-09-01

Hemicellulose is an important part of the plant cell wall biomass, and is relevant to cellulosic ethanol technologies. β-Mannosidases are enzymes capable of cleaving nonreducing residues of β-d-mannose from β-d-mannosides and hemicellulose mannose-containing polysaccharides, such as mannans and galactomannans. β-Mannosidases are distributed between glycoside hydrolase (GH) families 1, 2, and 5, and only a handful of the enzymes have been structurally characterized to date. The only published X-ray structure of a GH family 2 mannosidase is that of the bacterial Bacteroides thetaiotaomicron enzyme. No structures of eukaryotic mannosidases of this family are currently available. To fill this gap, we set out to solve the structure of Trichoderma harzianum GH family 2 β-mannosidase and to refine it to 1.9-Å resolution. Structural comparisons of the T. harzianum GH2 β-mannosidase highlight similarities in its structural architecture with other members of GH family 2, reveal the molecular mechanism of β-mannoside binding and recognition, and shed light on its putative galactomannan-binding site. Coordinates and observed structure factor amplitudes have been deposited with the Protein Data Bank (4CVU and 4UOJ). The T. harzianum β-mannosidase 2A nucleotide sequence has GenBank accession number BankIt1712036 GeneMark.hmm KJ624918. © 2014 FEBS.

Butanediol production from cellulose and hemicellulose by Klebsiella pneumoniae grown in sequential coculture with Trichoderma harzianum

Energy Technology Data Exchange (ETDEWEB)

Yu, E.K.C.; Deschatelets, L.; Louis-Seize, G.; Saddler, J.N.

1985-10-01

The bioconverison of cellulose and hemicellulose substrates to 2,3-butanediol by a sequential coculture approach was investigated with the cellulolytic fungus Trichoderma harzianum E58 and the fermentative bacterium Klebsiella pneumoniae. Vogel medium optimal for the production of the cellulolytic and xylanolytic enzymes of the fungus was found to be inhibitory to butanediol fermentation. This inhibition appeared to be due to a synergistic effect of various ingredients, particularly the salts, present in the fungal medium. The removal or replacement of such ingredients from Vogel medium led to the relief of fermentation inhibition, but the treatments also resulted in a significant decrease in fungal enzyme production. Resting cells of K. pneumoniae could be used for butanediol production in the fungal medium, indicating that the inhibitory effect on solvent production under such conditions was due to the indirect result of growing inhibition of the bacterial cells. The resting-cell approach could be combined with a fed-batch system for the direct conversion of 8 to 10% (wt/vol) of Solka-Floc or aspenwood xylan to butanediol at over 30% of the theoretical conversion efficiencies.

Compatibility study of Trichoderma harzianum Rifai and rice fungicides, and effects on three fungal plant pathogens

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Manuel Francisco Rodríguez Saldaña

2017-04-01

Full Text Available This research took place at the Provincial Plant Sanitation Laboratory, in Camaguey, Cuba, between September 2013 and September 2015. The in vitro compatibility and antagonistic capacity of Trichoderma harzianum Rifai (strain A-34 on rice pathogens (Bipolaris oryzae Breda de Haan, Sarocladium oryzae (Sawada w., Gams and D. Hawksworth and Magnaporthe grisea (Hebert Barr, was determined against pesticides used on rice. Assessment using traditional methods of microbiological isolation of mycelial growth, sporulation and conidial germination of the antagonist, to determine if the action mechanisms (antibiosis, competence, parasitism against fungal pathogens, was made between 24 and 216 hours of application. A bifactorial design in dual culture was used for statistical analysis, along with scales for determination of microbial antagonistic capacity. Active ingredients tebuconazol + procloraz, trifloxistrobin+ ciproconazole, and epoxiconazole + kresoxim-methyl, affected mycelial growth of the antagonist. Moreover, the antagonist against active ingredients carbendazim, copper oxychloride, azoxystrobin and tebuconazo + triadimenol showed mycelial growth, sporulation and pathogen interaction, affecting their growth by means of coiling, penetration, granulation, and cell lysis, between 96 and 216 hours.

Sequence analysis and gene expression of putative oil palm chitinase and chitinase-like proteins in response to colonization of Ganoderma boninense and Trichoderma harzianum.

Science.gov (United States)

Yeoh, K-A; Othman, A; Meon, S; Abdullah, F; Ho, C-L

2013-01-01

Chitinases are glycosyl hydrolases that cleave the β-1,4-glycosidic linkages between N-acetylglucosamine residues in chitin which is a major component of fungal cell wall. Plant chitinases hydrolyze fungal chitin to chitin oligosaccharides that serve as elicitors of plant defense system against fungal pathogens. However, plants synthesize many chitinase isozymes and some of them are not pathogenesis-related. In this study, three full-length cDNA sequences encoding a putative chitinase (EgChit3-1) and two chitinase-like proteins (EgChit1-1 and EgChit5-1) have been cloned from oil palm (Elaeis guineensis) by polymerase chain reaction (PCR). The abundance of these transcripts in the roots and leaves of oil palm seedlings treated with Ganoderma boninense (a fungal pathogen) or Trichoderma harzianum (an avirulent symbiont), and a combination of both fungi at 3, 6 and 12 weeks post infection were profiled by real time quantitative reverse-transcription (qRT)-PCR. Our findings showed that the gene expression of EgChit3-1 increased significantly in the roots of oil palm seedlings treated with either G. boninense or T. harzianum and a combination of both; whereas the gene expression of EgChit1-1 in the treated roots of oil palm seedlings was not significantly higher compared to those of the untreated oil palm roots. The gene expression of EgChit5-1 was only higher in the roots of oil palm seedlings treated with T. harzianum compared to those of the untreated oil palm roots. In addition, the gene expression of EgChit1-1 and EgChit3-1 showed a significantly higher gene expression in the leaf samples of oil palm seedlings treated with either G. boninense or T. harzianum.

Inhibición in vitro de aislamientos nativos de Trichoderma en presencia de la cepa comercial T22

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Gabriela Hernández – Ramírez

2013-01-01

Full Text Available Título en ingles: In vitro inhibition from native isolates of Trichoderma against commercial strain T22.  Título corto: Inhibición in vitro de aislamientos nativos Resumen El objetivo de este trabajo fue evaluar 14 aislamientos nativos del género Trichoderma (Tn obtenidos de la rizósfera del cultivo de caña de azúcar de acuerdo con su efecto de inhibición In vitro en cultivos duales contra la cepa comercial de Trichoderma harzianum (T22. Los aislamientos nativos con mayor índice de inhibición fueron los denominados Palma (T12 y Hubero (T11, con un valor de 2, mientras que T01, T04, T05 y T08 presentaron un valor de 1. Además, el mayor porcentaje de inhibición fue causado por el aislamiento T03, con una media de 92.32 ± 2.64%, sin diferencias estadísticas con los nativos T07, con un valor inhibición de 88.99 ± 1.46%, T01 87.93 ± 2.28%, y T12; 87.71 ± 3.16. En este caso, los aislamientos T07 y T12 mostraron capacidad inhibitoria con mayores atributos para su selección como agentes de control biológico. Al evaluar la tolerancia In vitro de los aislamientos nativos de Trichoderma a la actividad del agua (aw, se observó efecto significativo sobre el desarrollo micelial de los aislamientos de Trichoderma nativo evaluados. Los aislamientos T03 (7919.3 ± 932.53 y T12 (6388.2 ± 623.40 presentaron el mejor desarrollo radial. Estos resultados sugieren que los aislamientos T03 y T12 podrían desarrollarse mejor bajo condiciones de estrés hídrico. Por lo anterior, concluimos que los aislamientos T07 y T12, son los mejores candidatos como agentes inhibidores y se sugiere evaluarlos In vitro o invernadero contra el agente causal de la enfermedad del muermo rojo en caña de azúcar (Colletotrichum falcatum. Palabras clave: Control Biológico; tolerancia;  ompetencia; estrés hídrico; actividad de agua. Abstract The objective of this work was to evaluate 14 native e Trichoderma genus isolates (Tn obtained from the rhyzosphere of

Nitrogen metabolism and growth enhancement in tomato plants challenged with Trichoderma harzianum expressing the Aspergillus nidulans acetamidase amdS gene

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Sara Domínguez

2016-08-01

Full Text Available Trichoderma is a fungal genus that includes species that are currently being used as biological control agents and/or as biofertilizers. In addition to the direct application of Trichoderma spp. as biocontrol agents in plant protection, recent studies have focused on the beneficial responses exerted on plants, stimulating the growth, activating the defenses, and/or improving nutrient uptake. The amdS gene, encoding an acetamidase of Aspergillus, has been used as a selectable marker for the transformation of filamentous fungi, including Trichoderma spp., but the physiological effects of the introduction of this gene into the genome of these microorganisms still remains unexplored. No evidence of amdS orthologous genes has been detected within the Trichoderma spp. genomes and the amdS heterologous expression in T. harzianum T34 did not affect the growth of this fungus in media lacking acetamide. However, it did confer the ability for the fungus to use this amide as a nitrogen source. Although a similar antagonistic behavior was observed for T34 and amdS transformants in dual cultures against Rhizoctonia solani, Botrytis cinerea and Fusarium oxysporum, a significantly higher antifungal activity was detected in amdS transformants against F. oxysporum, compared to that of T34, in membrane assays on media lacking acetamide. In Trichoderma-tomato interaction assays, amdS transformants were able to promote plant growth to a greater extent than the wild-type T34, although compared with this strain the transformants showed similar capability to colonize tomato roots. Gene expression patterns from aerial parts of 3-week-old tomato plants treated with T34 and the amdS transformants have also been investigated using GeneChip Tomato Genome Arrays. The downregulation of defense genes and the upregulation of carbon and nitrogen metabolism genes observed in the microarrays were accompanied by i enhanced growth, ii increased carbon and nitrogen levels and iii a

Effects of Trichothecene Production on the Plant Defense Response and Fungal Physiology: Overexpression of the Trichoderma arundinaceum tri4 Gene in T. harzianum.

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Cardoza, R E; McCormick, S P; Malmierca, M G; Olivera, E R; Alexander, N J; Monte, E; Gutiérrez, S

2015-09-01

Trichothecenes are fungal sesquiterpenoid compounds, the majority of which have phytotoxic activity. They contaminate food and feed stocks, resulting in potential harm to animals and human beings. Trichoderma brevicompactum and T. arundinaceum produce trichodermin and harzianum A (HA), respectively, two trichothecenes that show different bioactive properties. Both compounds have remarkable antibiotic and cytotoxic activities, but in addition, trichodermin is highly phytotoxic, while HA lacks this activity when analyzed in vivo. Analysis of Fusarium trichothecene intermediates led to the conclusion that most of them, with the exception of the hydrocarbon precursor trichodiene (TD), have a detectable phytotoxic activity which is not directly related to the structural complexity of the intermediate. In the present work, the HA intermediate 12,13-epoxytrichothec-9-ene (EPT) was produced by expression of the T. arundinaceum tri4 gene in a transgenic T. harzianum strain that already produces TD after transformation with the T. arundinaceum tri5 gene. Purified EPT did not show antifungal or phytotoxic activity, while purified HA showed both antifungal and phytotoxic activities. However, the use of the transgenic T. harzianum tri4 strain induced a downregulation of defense-related genes in tomato plants and also downregulated plant genes involved in fungal root colonization. The production of EPT by the transgenic tri4 strain raised levels of erg1 expression and reduced squalene accumulation while not affecting levels of ergosterol. Together, these results indicate the complex interactions among trichothecene intermediates, fungal antagonists, and host plants. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Trichoderma harzianum Produces a New Thermally Stable Acid Phosphatase, with Potential for Biotechnological Application.

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Souza, Amanda Araújo; Leitão, Vanessa Oliveira; Ramada, Marcelo Henrique; Mehdad, Azadeh; Georg, Raphaela de Castro; Ulhôa, Cirano José; de Freitas, Sonia Maria

2016-01-01

Acid phosphatases (ACPases) are produced by a variety of fungi and have gained attention due their biotechnological potential in industrial, diagnosis and bioremediation processes. These enzymes play a specific role in scavenging, mobilization and acquisition of phosphate, enhancing soil fertility and plant growth. In this study, a new ACPase from Trichoderma harzianum, named ACPase II, was purified and characterized as a glycoprotein belonging to the acid phosphatase family. ACPase II presents an optimum pH and temperature of 3.8 and 65 °C, respectively, and is stable at 55 °C for 120 min, retaining 60% of its activity. The enzyme did not require metal divalent ions, but was inhibited by inorganic phosphate and tungstate. Affinity for several phosphate substrates was observed, including phytate, which is the major component of phosphorus in plant foods. The inhibition of ACPase II by tungstate and phosphate at different pH values is consistent with the inability of the substrate to occupy its active site due to electrostatic contacts that promote conformational changes, as indicated by fluorescence spectroscopy. A higher affinity for tungstate rather than phosphate at pH 4.0 was observed, in accordance with its highest inhibitory effect. Results indicate considerable biotechnological potential of the ACPase II in soil environments.

The potential application of fungus Trichoderma harzianum Rifai in biodegradation of detergent and industry

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Jakovljević Violeta D.

2015-01-01

Full Text Available The potential application of fungus Trichoderma harzianum Rifai in biodegradation of commercial detergent (MERIX, Henkel, Serbia was in the focus of this study. The fungus was isolated from wastewater samples of the Rasina River, downstream where the industrial wastewaters of factory Henkel (Krusevac, Serbia discharge into river. The fungus was cultivated in liquid growth medium by Czapek with addition of detergent at a concentration of 0.3% during 16 days. Analysis of fermentation broth evaluated the chemical and biochemical changes of pH, redox potential, activity of alkaline and acid invertase as well as activity of alkaline protease. In addition, the influence of detergent on fungal growth and total dry weight biomass was determined. At the same time, detergent disappearance in terms of methylene blue active substances in the medium was measured. The detergent at a concentration of 0.3% influenced significant decrease of pH value and increase of redox potential. The detergent showed inhibitory effect on acid invertase activity and stimulatory effect on alkaline invertase and protease activity. The fungus decomposed about 74.24% of tested detergent during 16 days, but total dry weight biomass reduced about 20% in relation to control. [Projekat Ministarstva nauke Republike Srbije, br. III 43004

Biocontrol potential of Pasteuria penetrans, Pochonia chlamydosporia, Paecilomyces lilacinus and Trichoderma harzianum against Meloidogyne incognita in

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Tariq MUKHTAR

2013-05-01

Full Text Available The root-knot nematode, Meloidogyne incognita, is a sedentary endoparasitic plant pathogen with a very wide host range, which causes annual crop losses amounting to millions of dollars. The small number of available nematicides and restrictions on the use of non-fumigant nematicides due to high toxicity to humans and non-target organisms hinder effective nematode control. A possible alternative to chemical nematicides is the use of biological control agents for the management of this nematode. In the present study, the efficacy of four biocontrol agents was tested against M. incognita at different doses. The biocontrol agents Pasteuria penetrans, Pochonia chlamydosporia, Paecilomyces lilacinus and Trichoderma harzianum were mass produced and mixed with the formalin sterilized soil at the rates of 2 × 103, 4 × 103, 6 × 103, 8 × 103, and 1 × 104 endospores/chlamydospores/cfu per g of soil. Okra seeds (cv. Sabz Pari were sown in pots of soil amended with the different agents, and 10 d after emergence, the plants were inoculated with 2000 freshly hatched second stage juveniles of M. incognita. Data on plant growth parameters and nematode infestations were recorded 7 weeks after inoculation. The antagonists varied significantly in enhancing various growth parameters and reducing nematode infestations in a dose-responsive manner. Both P. penetrans and P. lilacinus were equally effective and caused maximum reductions in number of galls, egg masses, nematode fecundity and build up as compared with T. harzianum and P. chlamydosporia. Reductions in these parameters at the concentration of 8 × 103 were statistically similar with those caused at the concentration of 1 × 104 chlamydospores/ endospores/cfu. Our results indicate that application of antagonists can suppress galling and reproduction of M. incognita resulting in enhancement of plant growth.

Antagonismo in vitro de Trichoderma harzianum Rifai sobre Fusarium solani (Mart. Sacc., asociado a la marchitez en maracuyá

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Carol Libeth Suárez Meza

2008-07-01

Full Text Available Cultivos de maracuyá de la región Caribe colombiana presentan problemas de marchitez y el hongo Fusarium solani (Mart. Sacc., está asociado a esta patología. Plantas de maracuyá de la Estación Experimental (E.E Caribia de Corpoica poseen este problema y por tal razón se determinó el antagonismo in vitro de aislamientos de Trichoderma harzianum Rifai, frente a F. solani, aislado de plantas enfermas de maracuyá (Passiflora edulis. Utilizando la técnica de cultivo dual en platos Petri con Agar Sabouraud, se evaluaron competencia por nutrientes y espacio, micoparasitismo y porcentaje de inhibición del crecimiento radial (PICR, empleando un diseño estadístico factorial 2x6x1 con arreglo completamente aleatorio. Se obtuvieron tres aislamientos nativos de T. harzianum (TCN-009, TCN-010, TCN-014 de suelo de Palma de Aceite (Elaeis guineensis de la E.E Caribia, y se compararon con tres aislamientos comerciales (TCC-001, TCC-005, TCC-006. TCC-001 y TCN-014, reportaron ser más competentes por nutrientes y espacio, con el mayor radio de crecimiento de 7,50 y 7,32 cm el día 10, comparado a FSM-011 en el cual solo fue de 2,30 cm. Aunque, TCN-014 mostró micoparasitismo grado 4 con ambos aislamientos de F. solani y TCC-005 únicamente con FSM-012, el cual fue más susceptible a ser micoparasitado. En cuanto al PICR, los tratamientos con mejores porcentajes de inhibición fueron TA-9, TA-12 y TA-6 con valores de 70,56, 68,52 y 65,32% respectivamente. El aislamiento del patógeno mayormente inhibido fue FSM-011. Todos estos resultados demuestran que hubo antagonismo in vitro al utilizar los aislamientos nativos y comerciales de T. harzianum sobre F. solani.  Palabras clave: Passiflora, competencia, micoparasitismo, inhibición, antagonismo.

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

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

1985-06-01

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

Analysis of Phaseolus vulgaris response to its association with Trichoderma harzianum (ALL-42) in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani.

Science.gov (United States)

Pereira, Jackeline L; Queiroz, Rayner M L; Charneau, Sébastien O; Felix, Carlos R; Ricart, Carlos A O; da Silva, Francilene Lopes; Steindorff, Andrei Stecca; Ulhoa, Cirano J; Noronha, Eliane F

2014-01-01

The present study was carried out to evaluate the ability of Trichoderma harzianum (ALL 42-isolated from Brazilian Cerrado soil) to promote common bean growth and to modulate its metabolism and defense response in the presence or absence of the phytopathogenic fungi Rhizoctonia solani and Fusarium solani using a proteomic approach. T. harzianum was able to promote common bean plants growth as shown by the increase in root/foliar areas and by size in comparison to plants grown in its absence. The interaction was shown to modulate the expression of defense-related genes (Glu1, pod3 and lox1) in roots of P. vulgaris. Proteomic maps constructed using roots and leaves of plants challenged or unchallenged by T. harzianum and phytopathogenic fungi showed differences. Reference gels presented differences in spot distribution (absence/presence) and relative volumes of common spots (up or down-regulation). Differential spots were identified by peptide fingerprinting MALDI-TOF mass spectrometry. A total of 48 identified spots (19 for leaves and 29 for roots) were grouped into protein functional classes. For leaves, 33%, 22% and 11% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively. For roots, 17.2%, 24.1% and 10.3% of the identified proteins were categorized as pertaining to the groups: metabolism, defense response and oxidative stress response, respectively.

Comparative studies on the induction of Trichoderma harzianum mutanase by α-(1→3)-glucan-rich fruiting bodies and mycelia of Laetiporus sulphureus.

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Wiater, Adrian; Pleszczyńska, Małgorzata; Szczodrak, Janusz; Janusz, Grzegorz

2012-01-01

Mutanase (α-(1→3)-glucanase) is a little-known inductive enzyme that is potentially useful in dentistry. Here, it was shown that the cell wall preparation (CWP) obtained from the fruiting body or vegetative mycelium of polypore fungus Laetiporus sulphureus is rich in α-(1→3)-glucan and can be successfully used for mutanase induction in Trichoderma harzianum. The content of this biopolymer in the CWP depended on the age of fruiting bodies and increased along with their maturation. In the case of CWP prepared from vegetative mycelia, the amount of α-(1→3)-glucan depended on the mycelium age and also on the kind of medium used for its cultivation. All CWPs prepared from the individually harvested fruiting body specimens induced high mutanase activity (0.53-0.82 U/mL) in T. harzianum after 3 days of cultivation. As for the CWPs obtained from the hyphal mycelia of L. sulpureus, the maximal enzyme productivity (0.34 U/mL after 3 days of incubation) was recorded for CWP prepared from the 3 week-old mycelium cultivated in Sabouraud medium. Statistically, a high positive correlation was found between the total percentage content of α-(1→3)-glucan in the CWP and the mutanase activity.

Inoculation of tomato seedlings with Trichoderma Harzianum and Arbuscular Mycorrhizal Fungi and their effect on growth and control of wilt in tomato seedlings

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Margaret W. Mwangi

2011-06-01

Full Text Available A green house study was conducted to investigate the ability of an isolate of Trichoderma harzianum (P52 and arbuscular mycorrhizal fungi (AMF in enhancing growth and control of a wilt pathogen caused by Fusarium oxysporum f. sp. lycopersici in tomato seedlings. The plants were grown in plastic pots filled with sterilized soils. There were four treatments applied as follows; P52, AMF, AMF + P52 and a control. A completely randomized design was used and growth measurements and disease assessment taken after 3, 6 and 9 weeks. Treatments that significantly (P < 0.05 enhanced heights and root dry weights were P52, AMF and a treatment with a combination of both P52 and AMF when compared the control. The treatment with both P52 and AMF significantly (P < 0.05 enhanced all growth parameters (heights; shoot and root dry weight investigated compared to the control. Disease severity was generally lower in tomato plants grown with isolate P52 and AMF fungi either individually or when combined together, though the effect was not statistically significant (P0.05. A treatment combination of P52 + AMF had less trend of severity as compared to each individual fungus. T. harzianum and AMF can be used to enhance growth in tomato seedlings.

Quantification and characterisation of Trichoderma spp. from different ecosystems.

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Sariah, M; Choo, C W; Zakaria, H; Norihan, M S

2005-01-01

Basal stem rot of oil palm caused by Ganoderma boninense is of major economic importance. Observations of the low incidence of disease due to Ganoderma species in natural stands, suggest that the disease is kept under control by some biological means. Trichoderma spp. are saprophytic fungi with high antagonistic activities against soil-borne pathogens. However, their abundance and distribution are soil and crop specific. Trichoderma species have been found to be concentrated in the A1 (0-30 cm) and Be soil horizons (30-60 cm), although the abundance of Trichoderma was not significantly different between the oil palm and non-oil palm ecosystems. Characterisation of Trichoderma isolates based on cultural, morphological and DNA polymorphism showed that T. harzianum, T. virens, T. koningii and T. longibrachiatum made up 72, 14, 10 and 4% of the total Trichoderma isolates isolated. As Trichoderma species are present in the oil palm ecosystem, but at lower numbers and in locations different from those desired, soil augmentation with antagonistic Trichoderma spp. can be developed as a strategy towards integrated management of basal stem rot of oil palm.

In vitro antagonism of cotton seedlings fungi and characterization of chitinase isozyme activities in Trichoderma harzianum.

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Asran-Amal, A; Moustafa-Mahmoud, S M; Sabet, K K; El Banna, O H

2010-04-01

The antagonistic fungus Trichoderma harzianum is widely recognized as a potential biocontrol agent against several soil-borne plant pathogens. T. harzinum is rich source of chitinoltic enzymes. In vitro screening of 5 isolates of T. harzinum, one isolate of Chaetomium globosum and one isolate of Conetherium mentance, revealed that all of them had reduced growth area of Macrophomina phaseolina, Fusarium solani and Rhizoctonia solani on PDA medium, significantly. The inhibition percentage ranged from 77.9 % to 55.9% for M. phaseolina and 59.2% to 40.4% for R. solani by T. harzinum and C. mentance, respectively. Inhibition for F. solani ranged from 76.5% to 55.7% by T. harzinum and C. globosum, respectively. Isozyme gel electrophoresis was used to assess chitinase activity secreted by selected isolates of T. harzinum under different pH degrees and temperatures. Obtained results indicated that activity of chitinase isozyme produced at 30 °C was higher than 15-20 °C for all tested isolates and activity of chitinase produced by isolates No. 4 and 5 of T. harzinum at pH (7-7.5) was higher than at pH 6, respectively.

Exposing of Trichoderma spp. to gamma radiation for stimulating its pesticide biodegradation activity

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Afify, A.E.M.R.; Ibrahim, G.M.; Abo El Seoud, M.A.; Helal, I.M.M.; Kassem, B.W.

2012-01-01

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.

HFB7 - A novel orphan hydrophobin of the Harzianum and Virens clades of Trichoderma, is involved in response to biotic and abiotic stresses.

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Przylucka, Agnes; Akcapinar, Gunseli Bayram; Chenthamara, Komal; Cai, Feng; Grujic, Marica; Karpenko, Juriy; Livoi, Miriam; Shen, Qirong; Kubicek, Christian P; Druzhinina, Irina S

2017-05-01

Hydrophobins are small secreted cysteine-rich proteins exclusively found in fungi. They are able to self-assemble in single molecular layers at hydrophobic-hydrophilic interfaces and can therefore be directly involved in establishment of fungi in their habitat. The genomes of filamentous mycotrophic fungi Trichoderma encode a rich diversity of hydrophobins, which are divided in several groups based on their structure and evolution. Here we describe a new member of class II hydrophobins, HFB7, that has a taxonomically restricted occurrence in Harzianum and Virens clades of Trichoderma. Evolutionary analysis reveals that HFB7 proteins form a separate clade distinct from other Trichoderma class II hydrophobins and that genes encoding them evolve under positive selection pressure. Homology modelling of HFB7 structure in comparison to T. reesei HFB2 reveals that the two large hydrophobic patches on the surface of the protein are remarkably conserved between the two hydrophobins despite significant difference in their primary structures. Expression of hfb7 gene in T. virens increases at interactions with other fungi and a plant and in response to a diversity of abiotic stress conditions, and is also upregulated during formation of aerial mycelium in a standing liquid culture. This upregulation significantly exceeds that of expression of hfb7 under a strong constitutive promoter, and T. virens strains overexpressing hfb7 thus display only changes in traits characterized by low hfb7 expression, i.e. faster growth in submerged liquid culture. The hfb7 gene is not expressed in conidia. Our data allow to conclude that this protein is involved in defence of Trichoderma against a diversity of stress factors related to the oxidative stress. Moreover, HFB7 likely helps in the establishment of the fungus in wetlands or other conditions related to high humidity. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Trichodiene Production in a Trichoderma harzianum erg1-Silenced Strain Provides Evidence of the Importance of the Sterol Biosynthetic Pathway in Inducing Plant Defense-Related Gene Expression.

Science.gov (United States)

Malmierca, M G; McCormick, S P; Cardoza, R E; Monte, E; Alexander, N J; Gutiérrez, S

2015-11-01

Trichoderma species are often used as biocontrol agents against plant-pathogenic fungi. A complex molecular interaction occurs among the biocontrol agent, the antagonistic fungus, and the plant. Terpenes and sterols produced by the biocontrol fungus have been found to affect gene expression in both the antagonistic fungus and the plant. The terpene trichodiene (TD) elicits the expression of genes related to tomato defense and to Botrytis virulence. We show here that TD itself is able to induce the expression of Botrytis genes involved in the synthesis of botrydial (BOT) and also induces terpene gene expression in Trichoderma spp. The terpene ergosterol, in addition to its role as a structural component of the fungal cell membranes, acts as an elicitor of defense response in plants. In the present work, using a transformant of T. harzianum, which is silenced in the erg1 gene and accumulates high levels of squalene, we show that this ergosterol precursor also acts as an important elicitor molecule of tomato defense-related genes and induces Botrytis genes involved in BOT biosynthesis, in both cases, in a concentration-dependent manner. Our data emphasize the importance of a balance of squalene and ergosterol in fungal interactions as well as in the biocontrol activity of Trichoderma spp.

Abiotic stresses affect Trichoderma harzianum T39-induced resistance to downy mildew in grapevine.

Science.gov (United States)

Roatti, Benedetta; Perazzolli, Michele; Gessler, Cesare; Pertot, Ilaria

2013-12-01

Enhancement of plant defense through the application of resistance inducers seems a promising alternative to chemical fungicides for controlling crop diseases but the efficacy can be affected by abiotic factors in the field. Plants respond to abiotic stresses with hormonal signals that may interfere with the mechanisms of induced systemic resistance (ISR) to pathogens. In this study, we exposed grapevines to heat, drought, or both to investigate the effects of abiotic stresses on grapevine resistance induced by Trichoderma harzianum T39 (T39) to downy mildew. Whereas the efficacy of T39-induced resistance was not affected by exposure to heat or drought, it was significantly reduced by combined abiotic stresses. Decrease of leaf water potential and upregulation of heat-stress markers confirmed that plants reacted to abiotic stresses. Basal expression of defense-related genes and their upregulation during T39-induced resistance were attenuated by abiotic stresses, in agreement with the reduced efficacy of T39. The evidence reported here suggests that exposure of crops to abiotic stress should be carefully considered to optimize the use of resistance inducers, especially in view of future global climate changes. Expression analysis of ISR marker genes could be helpful to identify when plants are responding to abiotic stresses, in order to optimize treatments with resistance inducers in field.

Fungicidal compounds from a marine Ascidian-associated fungus Trichoderma harzianum

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PrabhaDevi; Ciavatta, M.L.; Wahidullah, S.; Vuppala, S.; DeSouza, L.

. Of the five isolated secondary metabolites, compounds 2 and 5 are being reported here for the first time from T. harzianum. Compounds 1 and 4 inhibited the growth of Sclerotium rolfsii causing sclerotium wilt or rot disease in tropical plants. Compound 2 and 5...

Trichoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani.

Science.gov (United States)

Singh, B N; Singh, A; Singh, B R; Singh, H B

2014-03-01

To investigate the efficacy of Trichoderma harzianum NBRI-1055 (denoted as 'T-1055') in suppression of seedling blight of sunflower caused by Rhizoctonia solani Kühn and their impact on host defence responses. T-1055 was applied as seed treatment, soil application and combined application (seed treatment + soil application). Higher protection afforded by combined application of T-1055 was associated with the marked induction of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), peroxidase (PO) and cinnamyl alcohol dehydrogenase (CAD) activities. The activities of PAL and PPO reached maximum at 10 days after sowing (DAS), while PO and CAD levels reached maximum at 12 DAS. This was further supported by the accumulation of total phenolic content that showed an increase up to threefold at 14 DAS. In addition, HPLC analysis revealed that the contents of ferulic and p-coumaric acids increased by 6·3 and 4·6 times, respectively, at 14 DAS. Amount of gallic acid was also little more than double. Lignin deposition in sunflower root increased by 2·7, 3·4 and 3·7 times through combined application of T-1055 at 16, 18 and 20 DAS, respectively. Combined application also increased the accumulation of PR-2 and PR-3 proteins by 3·3 and 3·9 times, respectively, at 12 DAS in followed by seed treatment alone. The combined application of T-1055 triggered defence responses in an enhanced level in sunflower than the soil and seed alone and provided better protection against Rhizoctonia seedling blight. Rhizospheric fungal bioagent 'T-1055' can enhance protection in sunflower against the R. solani pathogen through augmented elicitation of host defence responses. © 2013 The Society for Applied Microbiology.

Biocontrol Effectiveness of Indigenous Trichoderma Species against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici on Tomato

OpenAIRE

Hajji Lobna; Chattaoui Mayssa; Regaieg Hajer; M'Hamdi-Boughalleb Naima; Rhouma Ali; Horrigue-Raouani Najet

2016-01-01

In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rat...

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

OpenAIRE

Marbun, Harif Nepen

2016-01-01

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

Identification and Characterization of Trichoderma Species Damaging Shiitake Mushroom Bed-Logs Infested by Camptomyia Pest.

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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.

mRNA expression of EgCHI1, EgCHI2, and EgCHI3 in oil palm leaves (Elaeis guineesis Jacq.) after treatment with Ganoderma boninense pat. and Trichoderma harzianum Rifai.

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Naher, Laila; Tan, Soon Guan; Ho, Chai Ling; Yusuf, Umi Kalsom; Ahmad, Siti Hazar; Abdullah, Faridah

2012-01-01

Basal stem rot (BSR) disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment.

Relevance of trichothecenes in fungal physiology: Disruption of tri5 in Trichoderma arundinaceum

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Trichothecenes are sesquiterpenoid mycotoxins produced mainly by Fusarium species. Harzianum A (HA), a non-phytotoxic trichothecene produced by Trichoderma arundinaceum, has recently been found to have antagonistic activity against fungal plant pathogens and to induce plant genes involved in defense...

Phytohormone profiles induced by trichoderma isolates correspond with their biocontrol and plant growth-promoting activity on melon plants.

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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.

Biodiversity of Trichoderma Community in the Tidal Flats and Wetland of Southeastern China.

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Saravanakumar, Kandasamy; Yu, Chuanjin; Dou, Kai; Wang, Meng; Li, Yaqian; Chen, Jie

2016-01-01

To investigate the biodiversity of Trichoderma (Hypocreaceae) and their relation to sediment physical and chemical properties, we collected a total of 491 sediment samples from coastal wetlands (tidal flat and wetland) in Southeast China. Further, we applied two types of molecular approaches such as culture dependent and independent methods for identification of Trichoderma spp. A total of 254 isolates were obtained and identified to 13 species such as T. aureoviride, T. asperellum, T. harzianum, T. atroviride, T. koningiopsis, T. longibrachiatum, T. koningii. T. tawa, T. viridescens, T. virens, T. hamatum, T. viride, and T. velutinum by the culture-dependent (CD) method of these, T. tawa was newly described in China. Subsequently, the culture indepented method of 454 pyrosequencing analysis revealed a total of six species such as T. citrinoviride, T. virens, T. polysporum, T. harzianum/Hypocrea lixii and two unknown species. Notably, T. citrinoviride and T. polysporum were not found by the CD method. Therefore, this work revealed that the combination of these two methods could show the higher biodiversity of Trichoderma spp., than either of this method alone. Among the sampling sites, Hangzhou, Zhejiang Province, exhibited rich biodiversity and low in Fengxian. Correlation and Redundancy discriminant analysis (RDA) revealed that sediment properties of temperature, redox potential (Eh) and pH significantly influenced the biodiversity of Trichoderma spp.

Biodiversity of Trichoderma Community in the Tidal Flats and Wetland of Southeastern China

Science.gov (United States)

Saravanakumar, Kandasamy; Yu, Chuanjin; Dou, Kai; Wang, Meng; Li, Yaqian; Chen, Jie

2016-01-01

To investigate the biodiversity of Trichoderma (Hypocreaceae) and their relation to sediment physical and chemical properties, we collected a total of 491 sediment samples from coastal wetlands (tidal flat and wetland) in Southeast China. Further, we applied two types of molecular approaches such as culture dependent and independent methods for identification of Trichoderma spp. A total of 254 isolates were obtained and identified to 13 species such as T. aureoviride, T. asperellum, T. harzianum, T. atroviride, T. koningiopsis, T. longibrachiatum, T. koningii. T. tawa, T. viridescens, T. virens, T. hamatum, T. viride, and T. velutinum by the culture-dependent (CD) method of these, T. tawa was newly described in China. Subsequently, the culture indepented method of 454 pyrosequencing analysis revealed a total of six species such as T. citrinoviride, T. virens, T. polysporum, T. harzianum/Hypocrea lixii and two unknown species. Notably, T. citrinoviride and T. polysporum were not found by the CD method. Therefore, this work revealed that the combination of these two methods could show the higher biodiversity of Trichoderma spp., than either of this method alone. Among the sampling sites, Hangzhou, Zhejiang Province, exhibited rich biodiversity and low in Fengxian. Correlation and Redundancy discriminant analysis (RDA) revealed that sediment properties of temperature, redox potential (Eh) and pH significantly influenced the biodiversity of Trichoderma spp. PMID:28002436

Biodiversity of Trichoderma Community in the Tidal Flats and Wetland of Southeastern China.

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Kandasamy Saravanakumar

Full Text Available To investigate the biodiversity of Trichoderma (Hypocreaceae and their relation to sediment physical and chemical properties, we collected a total of 491 sediment samples from coastal wetlands (tidal flat and wetland in Southeast China. Further, we applied two types of molecular approaches such as culture dependent and independent methods for identification of Trichoderma spp. A total of 254 isolates were obtained and identified to 13 species such as T. aureoviride, T. asperellum, T. harzianum, T. atroviride, T. koningiopsis, T. longibrachiatum, T. koningii. T. tawa, T. viridescens, T. virens, T. hamatum, T. viride, and T. velutinum by the culture-dependent (CD method of these, T. tawa was newly described in China. Subsequently, the culture indepented method of 454 pyrosequencing analysis revealed a total of six species such as T. citrinoviride, T. virens, T. polysporum, T. harzianum/Hypocrea lixii and two unknown species. Notably, T. citrinoviride and T. polysporum were not found by the CD method. Therefore, this work revealed that the combination of these two methods could show the higher biodiversity of Trichoderma spp., than either of this method alone. Among the sampling sites, Hangzhou, Zhejiang Province, exhibited rich biodiversity and low in Fengxian. Correlation and Redundancy discriminant analysis (RDA revealed that sediment properties of temperature, redox potential (Eh and pH significantly influenced the biodiversity of Trichoderma spp.

Suppressive Effect of Trichoderma spp. on toxigenic Fusarium species.

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Błaszczyk, Lidia; Basińska-Barczak, Aneta; Ćwiek-Kupczyńska, Hanna; Gromadzka, Karolina; Popiel, Delfina; Stępień, Łukasz

2017-03-30

The aim of the present study was to examine the abilities of twenty-four isolates belonging to ten different Trichoderma species (i.e., Trichoderma atroviride, Trichoderma citrinoviride, Trichoderma cremeum, Trichoderma hamatum, Trichoderma harzianum, Trichoderma koningiopsis, Trichoderma longibrachiatum, Trichoderma longipile, Trichoderma viride and Trichoderma viridescens) to inhibit the mycelial growth and mycotoxin production by five Fusarium strains (i.e., Fusarium avenaceum, Fusarium cerealis, Fusarium culmorum, Fusarium graminearum and Fusarium temperatum). Dual-culture bioassay on potato dextrose agar (PDA) medium clearly documented that all of the Trichoderma strains used in the study were capable of influencing the mycelial growth of at least four of all five Fusarium species on the fourth day after co-inoculation, when there was the first apparent physical contact between antagonist and pathogen. The qualitative evaluation of the interaction between the colonies after 14 days of co-culturing on PDA medium showed that ten Trichoderma strains completely overgrew and sporulated on the colony at least one of the tested Fusarium species. Whereas, the microscopic assay provided evidence that only T. atroviride AN240 and T. viride AN255 formed dense coils around the hyphae of the pathogen from where penetration took place. Of all screened Trichoderma strains, T. atroviride AN240 was also found to be the most efficient (69-100% toxin reduction) suppressors of mycotoxins (deoxynivalenol, 3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, nivalenol, zearalenone, beauvericin, moniliformin) production by all five Fusarium species on solid substrates. This research suggests that T. atroviride AN240 can be a promising candidate for the biological control of toxigenic Fusarium species.

Trichoderma harzianum como promotor del crecimiento vegetal del maracuyá (Passiflora edulis var. flavicarpa Degener

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Cubillos-Hinojosa Juan

2009-04-01

Full Text Available

Se realizó un experimento en condiciones de laboratorio e invernadero, con el propósito de evaluar el efecto de la cepa nativa TCN-014 y la cepa comercial TCC-005 de Trichoderma harzianum sobre la germinación y el crecimiento temprano del maracuyá. Se adecuaron inóculos de 104, 106 y 108 conidias/mL para cada cepa y se aplicaron a semillas de maracuyá; se evaluó el número de semillas germinadas durante 15 días; se calculó el porcentaje de germinación, el índice de velocidad de germinación y el tiempo medio de germinación. Posteriormente las semillas germinadas se llevaron a condiciones de invernadero y transcurridos dos meses se midió la altura de las plántulas, el grosor del tallo, el número de hojas, la longitud de la raíz y el peso seco total. Todos los tratamientos estimularon la germinación de las semillas y el desarrollo de las plántulas; sin embargo la cepa nativa en concentraciones 106 y 108 conidias/mL mostró resultados superiores frente a la cepa comercial. Los resultados sugieren una acción efectiva de T. harzianum como promotor de crecimiento vegetal, mostrando que tiene potencial para la elaboración de un bioproducto útil para el manejo ecológico del cultivo de maracuyá.

mRNA Expression of EgCHI1, EgCHI2, and EgCHI3 in Oil Palm Leaves (Elaeis guineesis Jacq. after Treatment with Ganoderma boninense Pat. and Trichoderma harzianum Rifai

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Laila Naher

2012-01-01

Full Text Available Background. Basal stem rot (BSR disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq. chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both. Methods. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR. Results. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants. Conclusion. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment.

mRNA Expression of EgCHI1, EgCHI2, and EgCHI3 in Oil Palm Leaves (Elaeis guineesis Jacq.) after Treatment with Ganoderma boninense Pat. and Trichoderma harzianum Rifai

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Naher, Laila; Tan, Soon Guan; Ho, Chai Ling; Yusuf, Umi Kalsom; Ahmad, Siti Hazar; Abdullah, Faridah

2012-01-01

Background. Basal stem rot (BSR) disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both. Methods. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR. Results. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants. Conclusion. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment. PMID:22919345

Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species.

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Jiang, Yuan; Wang, Jin-Liang; Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

2016-01-01

We surveyed the Trichoderma (Hypocreales, Ascomycota) biodiversity in agricultural fields in four major agricultural provinces of East China. Trichoderma strains were identified based on molecular approaches and morphological characteristics. In three sampled seasons (spring, summer and autumn), 2078 strains were isolated and identified to 17 known species: T. harzianum (429 isolates), T. asperellum (425), T. hamatum (397), T. virens (340), T. koningiopsis (248), T. brevicompactum (73), T. atroviride (73), T. fertile (26), T. longibrachiatum (22), T. pleuroticola (16), T. erinaceum (16), T. oblongisporum (2), T. polysporum (2), T. spirale (2), T. capillare (2), T. velutinum (2), and T. saturnisporum (1). T. harzianum, T. asperellum, T. hamatum, and T. virens were identified as the dominant species with dominance (Y) values of 0.057, 0.052, 0.048, and 0.039, respectively. The species amount, isolate numbers and the dominant species of Trichoderma varied between provinces. Zhejiang Province has shown the highest diversity, which was reflected in the highest species amount (14) and the highest Shannon-Wiener diversity index of Trichoderma haplotypes (1.46). We observed that relative frequencies of T. hamatum and T. koningiopsis under rice soil were higher than those under wheat and maize soil, indicating the preference of Trichoderma to different crops. Remarkable seasonal variation was shown, with summer exhibiting the highest biodiversity of the studied seasons. These results show that Trichoderma biodiversity in agricultural fields varies by region, crop, and season. Zhejiang Province (the southernmost province in the investigated area) had more T. hamatum than Shandong Province (the northernmost province), not only in isolate amounts but also in haplotype amounts. Furthermore, at haplotype level, only T. hamatum showed a gradient distribution from south to north in correspondence analysis among the four dominant species. The above results would contribute to the

Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species

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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

Trichoderma Biodiversity of Agricultural Fields in East China Reveals a Gradient Distribution of Species.

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Yuan Jiang

Full Text Available 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

Response of growth characters and yield of wheat (Triticum aestivum L. to co-inoculation of farmyard manure, Trichoderma spp. and Psudomunas spp.

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A. Shahsavari

2016-04-01

Full Text Available In order to investigate the effects of cattle manure, benefit fungi of Trichoderma species and Psudomunas spp. bacteria on seedling emergence parameters, growth and yield of wheat (Triticum aestivum L. a pot experiment was carried out in factorial (23 arrangement based on a randomized complete design with three replications. The treatments were three levels of cattle manure (10, 20 and 30 t.ha-1, three Trichoderma species (T. viridae, T. harzianum, T. hamatum and either application or non- application of Psudomunas bacteria. Analysis of data showed that control plants and 10 t.ha-1 cattle manure treatments had significantly effect on emergence percentage and field emergence rate compared to 20 and 30 t.ha-1 cattle manure. In the current experiment, the maximum grain yield was observed in 20 t.ha-1 inoculated with both T. viridae and T. harzianum. Application of 10 and 20 t.ha-1cattle manure markedly increased harvest index and biomass by 39.72 and 19.47%, respectively compared to control treatment (no manure application. Also, T. viridae fugues improved plant biomass compared to T. harzianum. The fungus of T. harzianum enhanced harvest index rather than T. viridae and T. hamatum. Application of Psudomunas bacteria significantly increased plant biomass and harvest index compared to pots without bacteria application. Results showed that colony counts of three Trichoderma species in the soil rhizosphere enhanced when rates of cattle manure application increased. The most of soil microbial population was observed in 30 t.ha-1 level of cattle manure inoculated with T. harzianum (74.68 × 108cfu mg-1 dry soil.

Trichoderma species mediated differential tolerance against biotic stress of phytopathogens in Cicer arietinum L.

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Saxena, Amrita; Raghuwanshi, Richa; Singh, Harikesh Bahadur

2015-02-01

Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, β-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chitinase Expression Due to Reduction in Fusaric Acid Level in an Antagonistic Trichoderma harzianum S17TH.

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Sharma, Vivek; Bhandari, Pamita; Singh, Bikram; Bhatacharya, Amita; Shanmugam, Veerubommu

2013-06-01

To study the effect of reduction in phytotoxin level on fungal chitinases, antagonistic Trichoderma spp. were screened for their ability to reduce the level of fusaric acid (FA), the phytotoxin produced by Fusarium spp. A T. harzianum isolate S17TH was able to tolerate high levels of FA (up to 500 ppm) but was unable to reduce the toxin to a significant level (non-toxic) added to minimal synthetic broth (MSB). However, the isolate was able to reduce 400 ppm FA in the liquid medium after 7 days to a non-toxic level and displayed similar level of antagonism over the control (without FA). In studies of the effect of the reduction in FA (400 ppm) level on chitinase gene expression in PCR assays, nag1 was significantly repressed but ech42 expression was only slightly repressed. Chitinase activity was either reduced or absent in the extracellular proteins of MSB supplemented with 400 ppm FA, which could be attributed to the effect of residual FA or its breakdown products through unknown mechanisms. Selection of S17TH as a toxin insensitive isolate that could commensurate the negative effect on chitinase activity makes it a potential antagonist against Fusarium spp.

Mycoparasitism studies of Trichoderma species against three phytopathogenic fungi: evaluation of antagonism and hydrolytic enzyme production.

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Qualhato, Thiago Fernandes; Lopes, Fabyano Alvares Cardoso; Steindorff, Andrei Stecca; Brandão, Renata Silva; Jesuino, Rosália Santos Amorim; Ulhoa, Cirano José

2013-09-01

Trichoderma spp. are used for biocontrol of several plant pathogens. However, their efficient interaction with the host needs to be accompanied by production of secondary metabolites and cell wall-degrading enzymes. Three parameters were evaluated after interaction between four Trichoderma species and plant-pathogenic fungi: Fusarium solani, Rhizoctonia solani and Sclerotinia sclerotiorum. Trichoderma harzianum and T. asperellum were the most effective antagonists against the pathogens. Most of the Trichoderma species produced toxic volatile metabolites, having significant effects on growth and development of the plant pathogens. When these species were grown in liquid cultures with cell walls from these plant pathogens, they produced and secreted β-1,3-glucanase, NAGAse, chitinase, acid phosphatase, acid proteases and alginate lyase.

Evaluación de Trichoderma sobre hongos contaminantes de semillas de palma híbrida interespecífica OxG (Elaeis oleifera x Elaeis guineensis

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Oscar Eduardo Ladino Rey

2017-01-01

Full Text Available The germination process of hybrid palm interspecific OxG (E. oleifera x E guineensis. seeds in the company Indupalma ltda has filed a decrease in production, which is reflected in economic losses due to the existence of filamentous fungi that affect seeds. Previous studies have shown the potential controller Trichoderma sp. on these fungi. Commercial and native strains of T. harzianum and T. viride were evaluated against prevalent fungal contaminants of interspecific hybrid palm seeds. They were isolated and identified eleven strains of pathogenic fungi from interspecific hybrid palm seeds that were removed from the different stages of germination. Dual plate tests were conducted at prevalent fungi and the percent inhibition of radial growth (PICO and mycoparasitism were determined. These fungi were inhibited in growth by the antagonistic effect of T. harzianum and T viride. Similarly, T.viride showed a great potential as a biocontrol agent to inhibit over 60% seven out of eleven strains of contaminating fungi.

Enhanced cellulase production by Trichoderma harzianum by cultivation on glycerol followed by induction on cellulosic substrates.

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Delabona, Priscila da Silva; Lima, Deise Juliana; Robl, Diogo; Rabelo, Sarita Cândida; Farinas, Cristiane Sanchez; Pradella, José Geraldo da Cruz

2016-05-01

The use of glycerol obtained as an intermediate of the biodiesel manufacturing process as carbon source for microbial growth is a potential alternative strategy for the production of enzymes and other high-value bioproducts. This work evaluates the production of cellulase enzymes using glycerol for high cell density growth of Trichoderma harzianum followed by induction with a cellulosic material. Firstly, the influence of the carbon source used in the pre-culture step was investigated in terms of total protein secretion and fungal morphology. Enzymatic productivity was then determined for cultivation strategies using different types and concentrations of carbon source, as well as different feeding procedures (batch and fed-batch). The best strategy for cellulase production was then further studied on a larger scale using a stirred tank bioreactor. The proposed strategy for cellulase production, using glycerol to achieve high cell density growth followed by induction with pretreated sugarcane bagasse, achieved enzymatic activities up to 2.27 ± 0.37 FPU/mL, 106.40 ± 8.87 IU/mL, and 9.04 ± 0.39 IU/mL of cellulase, xylanase, and β-glucosidase, respectively. These values were 2 times higher when compared to the control experiments using glucose instead of glycerol. This novel strategy proved to be a promising approach for improving cellulolytic enzymes production, and could potentially contribute to adding value to biomass within the biofuels sector.

Thc6 protein, isolated from Trichoderma harzianum, can induce maize defense response against Curvularia lunata.

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Fan, Lili; Fu, Kehe; Yu, Chuanjin; Li, Yingying; Li, Yaqian; Chen, Jie

2015-05-01

Mutant T66 was isolated from 450 mutants (constructed with Agrobacterium tumefaciens-mediated transformation method) of Trichoderma harzianum. Maize seeds coated with T66 were more susceptible to Curvularia lunata when compared with those coated with wild-type (WT) strain. The disease index of maize treated with T66 and WT were 62.5 and 42.1%, respectively. Further research showed T-DNA has inserted into the ORF of one gene, which resulted in the functional difference between WT and T66. The gene was cloned and named Thc6, which encodes a novel 327 amino acid protein. To investigate its function, we obtained knockout, complementation, and overexpression mutants of Thc6. Challenge inoculation studies suggested that the Thc6 overexpression mutant can reduce the disease index of maize inbred line Huangzao 4 against the leaf spot pathogen (C. lunata). Meanwhile, The Thc6 mutants were found to affect the resistance of maize inbred line Huangzao 4 against C. lunata by enhancing the activation of jasmonate-responsive genes expression. Liquid chromatography-mass spectrometry (LC-MS) data further confirmed that the concentration of jasmonate in the induced maize exhibits a parallel change tendency with the expression level of defense-related genes. Hence, the Thc6 gene could be participated in the induced resistance of maize inbred line Huangzao 4 against C. lunata infection through a jasmonic acid-dependent pathway. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fungal Screening on Olive Oil for Extracellular Triacylglycerol Lipases: Selection of a Trichoderma harzianum Strain and Genome Wide Search for the Genes

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Canseco-Pérez, Miguel Angel; Castillo-Avila, Genny Margarita; Islas-Flores, Ignacio; Apolinar-Hernández, Max M.; Rivera-Muñoz, Gerardo; Gamboa-Angulo, Marcela; Couoh-Uicab, Yeny

2018-01-01

A lipolytic screening with fungal strains isolated from lignocellulosic waste collected in banana plantation dumps was carried out. A Trichoderma harzianum strain (B13-1) showed good extracellular lipolytic activity (205 UmL−1). Subsequently, functional screening of the lipolytic activity on Rhodamine B enriched with olive oil as the only carbon source was performed. The successful growth of the strain allows us to suggest that a true lipase is responsible for the lipolytic activity in the B13-1 strain. In order to identify the gene(s) encoding the protein responsible for the lipolytic activity, in silico identification and characterization of triacylglycerol lipases from T. harzianum is reported for the first time. A survey in the genome of this fungus retrieved 50 lipases; however, bioinformatic analyses and putative functional descriptions in different databases allowed us to choose seven lipases as candidates. Suitability of the bioinformatic screening to select the candidates was confirmed by reverse transcription polymerase chain reaction (RT-PCR). The gene codifying 526309 was expressed when the fungus grew in a medium with olive oil as carbon source. This protein shares homology with commercial lipases, making it a candidate for further applications. The success in identifying a lipase gene inducible with olive oil and the suitability of the functional screening and bioinformatic survey carried out herein, support the premise that the strategy can be used in other microorganisms with sequenced genomes to search for true lipases, or other enzymes belonging to large protein families. PMID:29370083

Eficiência de Trichoderma harzianum e Gliocladium viride na redução da incidência de Botrytis cinerea em tomateiro cultivado sob ambiente protegido Efficiency of Trichoderma harzianum and Gliocladium viride in decreasing the incidence of Botrytis cinerea in tomato cultivated in protected environment

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Bruno Brito Lisboa

2007-10-01

Full Text Available A produção de tomates no Estado do RS ocupa um importante papel sócio-econômico, que pode ser constatado pelo crescimento do cultivo dessa hortaliça em ambiente protegido. Essa técnica permite a produção de tomates em período de entressafra; no entanto, ocorrem também condições favoráveis para o desenvolvimento de doenças fúngicas como o mofo cinzento provocado por Botrytis cinerea. O surgimento de raças de patógenos resistentes a fungicidas químicos vem fazendo com que o controle biológico torne-se uma alternativa necessária. Neste trabalho foi realizada seleção in vitro de 24 isolados do fungo Trichoderma harzianum e 12 de Gliocladium viride que inibiram o desenvolvimento do patógeno B. cinerea. Foram selecionados dois isolados (TRIC-30 e GLIO-10 para serem testados em experimentos em condições de campo com tomates cultivados sob ambiente protegido, nos quais a pulverização foliar semanal com uma suspensão com 2x10(7 conídios mL-1 reduziu significativamente a incidência do mofo cinzento, enquanto a aplicação dos antagonistas nas sementes, no substrato e na cova, no momento do plantio, não reduziu a incidência do patógeno.The production of tomato in the State of Rio Grande do Sul performs an important economical and social role that can be evidenced by the increase in cultivation of this vegetable in protected environment. This practice allows the production of tomato during the off-season periods. However, it can also promote favorable conditions to the development of gray mold caused by Botrytis cinerea, and the arising of pathogen races resistant to fungicides is turning biological control into a necessary alternative. In the present work, an in vitro selection among 24 isolates of the fungus Trichoderma harzianum and 12 of Gliocladium viride that inhibited the development of the pathogen B. cinerea was carried out. Two isolates (TRIC-30 e GLIO-10 were selected to be tested in an experiment in field

Secretomic survey of Trichoderma harzianum grown on plant biomass substrates.

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Gómez-Mendoza, Diana Paola; Junqueira, Magno; do Vale, Luis Henrique Ferreira; Domont, Gilberto Barbosa; Ferreira Filho, Edivaldo Ximenes; Sousa, Marcelo Valle de; Ricart, Carlos André Ornelas

2014-04-04

The present work aims at characterizing T. harzianum secretome when the fungus is grown in synthetic medium supplemented with one of the four substrates: glucose, cellulose, xylan, and sugarcane bagasse (SB). The characterization was done by enzymatic assays and proteomic analysis using 2-DE/MALDI-TOF and gel-free shotgun LC-MS/MS. The results showed that SB induced the highest cellulolytic and xylanolytic activities when compared with the other substrates, while remarkable differences in terms of number and distribution of protein spots in 2-DE gels were also observed among the samples. Additionally, treatment of the secretomes with PNGase F revealed that most spot trails in 2-DE gels corresponded to N-glycosylated proteoforms. The LC-MS/MS analysis of the samples identified 626 different protein groups, including carbohydrate-active enzymes and accessory, noncatalytic, and cell-wall-associated proteins. Although the SB-induced secretome displayed the highest cellulolytic and xylanolytic activities, it did not correspond to a higher proteome complexity because CM-cellulose-induced secretome was significantly more diverse. Among the identified proteins, 73% were exclusive to one condition, while only 5% were present in all samples. Therefore, this study disclosed the variation of T. harzianum secretome in response to different substrates and revealed the diversity of the fungus enzymatic toolbox.

Systemic Resistance to Powdery Mildew in Brassica napus (AACC) and Raphanus alboglabra (RRCC) by Trichoderma harzianum TH12.

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Alkooranee, Jawadayn Talib; Yin, Yongtai; Aledan, Tamarah Raad; Jiang, Yingfen; Lu, Guangyuan; Wu, Jiangsheng; Li, Maoteng

2015-01-01

Trichoderma harzianum TH12 is a microbial pesticide for certain rapeseed diseases. The mechanism of systemic resistance induced by TH12 or its cell-free culture filtrate (CF) in Brassica napus (AACC) and Raphanus alboglabra (RRCC) to powdery mildew disease caused by ascomycete Erysiphe cruciferarum was investigated. In this study, we conducted the first large-scale global study on the cellular and molecular aspects of B. napus and R. alboglabra infected with E. cruciferarum. The histological study showed the resistance of R. alboglabra to powdery mildew disease. The growth of fungal colonies was not observed on R. alboglabra leaves at 1, 2, 4, 6, 8, and 10 days post-inoculation (dpi), whereas this was clearly observed on B. napus leaves after 6 dpi. In addition, the gene expression of six plant defense-related genes, namely, PR-1, PR-2 (a marker for SA signaling), PR-3, PDF 1.2 (a marker for JA/ET signaling), CHI620, and CHI570, for both genotypes were analyzed in the leaves of B. napus and R. alboglabra after treatment with TH12 or CF and compared with the non-treated ones. The qRT-PCR results showed that the PR-1 and PR-2 expression levels increased in E. cruciferarum-infected leaves, but decreased in the TH12-treated leaves compared with leaves treated with CF. The expression levels of PR-3 and PDF1.2 decreased in plants infected by E. cruciferarum. However, expression levels increased when the leaves were treated with TH12. For the first time, we disclosed the nature of gene expression in B. napus and R. alboglabra to explore the resistance pathways in the leaves of both genotypes infected and non-infected by powdery mildew and inoculated or non-inoculated with elicitor factors. Results suggested that R. alboglabra exhibited resistance to powdery mildew disease, and the application of T. harzianum and its CF are a useful tool to facilitate new protection methods for resist or susceptible plants.

Biocontrol efficacy of different isolates of Trichoderma against soil borne pathogen Rhizoctonia solani.

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Asad, Saeed Ahmad; Ali, Naeem; Hameed, Abdul; Khan, Sabaz Ali; Ahmad, Rafiq; Bilal, Muhammad; Shahzad, Muhammad; Tabassum, Ayesha

2014-01-01

In this study, the biocontrol abilities of water-soluble and volatile metabolites of three different isolates of Trichoderma (T. asperellum, T. harzianum and Trichoderma spp.) against soil borne plant pathogen Rhizoctonia solani were investigated both in vitro and in vivo. The results showed for the first time that mycelial growth inhibition of the pathogen was 74.4-67.8% with water-soluble metabolites as compared to 15.3-10.6% with volatile metabolites in vitro. In vivo antagonistic activity of Trichoderma isolates against R. solani was evaluated on bean plants under laboratory and greenhouse conditions. We observed that T. asperellum was more effective and consistent, lowering disease incidence up to 19.3% in laboratory and 30.5% in green house conditions. These results showed that three isolates of Trichoderma could be used as effective biocontrol agents against R. solani.

Some ecological aspects of the fungi trichoderma spp and bliocladium spp in Palma de Vino soils farm, la Dorada, Caldas, Colombia

International Nuclear Information System (INIS)

Viteri, Silvio E; Zarta, D; Salgado, N

2000-01-01

Trichoderma spp and Gliocladium spp have been reported as control agents of some of the fungi that cause root rot diseases in various crops. Despite this potential the there is no information on their ecology in tropical soils. This study was conducted at the Palma de Vino farm, La Dorada, Caldas with the aim of contributing to the understanding of their ecology in their natural habitat in the tropics. Representative soil and rhizosphere samples were collected and analyzed for some physical, chemical, and biological properties. The results showed clearly that: 1) both genera, form part of the native microflora of those soils. 2) The Trichoderma spp population fluctuated between 5.7x10 2 and 6.4x10 3 and between 7.4x10 2 and 2x10 4 UFC g - 1 in the soil and rhizoplane, respectively; the Gliocladium spp population could not be estimated, probably due to their low numbers. 3) Within the genus Trichoderma the species T. Hamatum, T. Harzianum, and T. Koningu could be identified. 4) The Trichoderma spp populations, especially those from de rhizoplane, showed a clear relation with de soil pH. As far as we know, these results represent the first source of information on the ecology of these two agronomic important fungi, under tropical conditions

Involvement of Trichoderma trichothecenes in the biocontrol activity and in the induction of plant defense related genes

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Trichoderma species produce trichothecenes, most notably trichodermin and harzianum A (HA), by a biosynthetic pathway in which several of the involved proteins have significant differences in functionality, compared to their Fusarium orthologues. In addition, the genes encoding these proteins show a...

Phylogeny of the Clinically Relevant Species of the Emerging Fungus Trichoderma and Their Antifungal Susceptibilities

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

2014-01-01

A set of 73 isolates of the emerging fungus Trichoderma isolated from human and animal clinical specimens were characterized morphologically and molecularly using a multilocus sequence analysis that included the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and fragments of the translation elongation factor 1 alpha (Tef1), endochitinase CHI18-5 (Chi18-5), and actin 1 (Act1) genes. The most frequent species was Trichoderma longibrachiatum (26%), followed by Trichoderma citrinoviride (18%), the Hypocrea lixii/Trichoderma harzianum species complex (15%), the newly described species Trichoderma bissettii (12%), and Trichoderma orientale (11%). The most common anatomical sites of isolation in human clinical specimens were the respiratory tract (40%), followed by deep tissue (30%) and superficial tissues (26%), while all the animal-associated isolates were obtained from superficial tissue samples. Susceptibilities of the isolates to eight antifungal drugs in vitro showed mostly high MICs, except for voriconazole and the echinocandins. PMID:24719448

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

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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.

Potential of Trichoderma species on Helminthosporium causing leaf spot on cane palm, Chrysalidocarpus lutescens.

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Jegathambigai, V; Karunaratne, M D S D; Svinningen, A; Mikunthan, G

2008-01-01

The cane palm, Chrysalidocarpus lutescens is one among the plant material of the export industries in Sri Lanka. The export quality of C. lutescens was declined due to the repeated occurrence of a leaf spot caused by Helminthosporium. Widespread occurrence of the leaf spot affected the cane palm production and succumb it to a huge setback in the floriculture industry in Sri Lanka. Being an export industry eco-friendly means of disease control was the prime focus for a better management of such vulnerable disease. Trichoderma is a potential bio agent, which has definite role in suppressing the inoculum of Helminthosporium sp. This study aims to evaluate the efficacy of Trichoderma species to control naturally established leaf spot in cane palm under field conditions. Three isolates of T. viride and two isolates of T. harzianum were evaluated. All the Trichoderma species performed significantly in reducing the disease incidence. T. viride + T. harzianum combination (1 x 10(10) cfu/ml) was the best compared to chemical in decreasing the mean disease severity index and improving the frequency of healthy plants. The colour of the leaves regained due to the application of Trichoderma sp. The results revealed that leaf spot incidence was lowered significantly in cane palms treated with Trichoderma species followed by treatment with combination of Trichoderma sp. and fungicides. The fungicide mixture (hexaconozole 50 g/l + Isoprothiolane 400 g/l) failed to lower the disease incidence and had no effect in suppressing the inocula of Helminthosporium, although recommended. Mixing of Trichoderma species with fungicide did not exhibit any additive effect. The combination of different species of Trichoderma would target species of Helminthosporium that exist as a complex group under field conditions. The results also proved that the existence of heterogeneity in Helminthosporium that could be tackled and effectively controlled by a combination of different species of the bio

Trichoderma ROLE IN AGROFORESTRY-CACAOTAL SYSTEMS AS AN ANTAGONAL AGENT

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Úrsula del Carmen López-Ferrer

2017-05-01

Full Text Available Agricultural and cocoa agroforestry systems are important for food production and biodiversity conservation. Among this diversity there is a group of fungi of the genus Trichoderma that present antagonistic effects against phytopathogens and this action can be used as a form of biological control of plant pathogens. In the agroforestry-cacao system the diseases with the highest frequency and with the greatest impact on cocoa production (Theobroma cacao are black rot (Phytophthora spp., Broom broom (Moniliophthora perniciosa and moniliasis (Moniliophthora roreri. The objective of this work was to perform an analysis of the main theoretical and practical aspects about the genus Trichoderma and its role in agriculture as an antagonistic agent. One of the microscopic features in the delimitation of the genus, especially by the presence of structures called phalid. The antagonistic mechanisms used by Trichoderma spp. Are described as competition, antibiosis and mycoparasitism. Mycoparasitism is having a relevance on the implications of extracellular enzymes such as chitinases, cellulases, β-1-3-glucanases and proteases that lyse or digest the walls of fungi, Moniliophthora roreri disease. This fungus can inhibit the growth of other fungi and bacteria by producing several volatile and non-volatile secondary metabolites. On the other hand, it participates in the production of regulators of growth and stimulation of the division, differentiation and cellular growth in the plant by the elicitor agent. Trichoderma species that are commercialized for biological control, growth promoter and biofertilizer are T. viride, T. polysporum and T. harzianum. The T. virens and T. harzianum species are most used for the antagonistic control of M. roreri, Phytophthora spp., and M. perniciosa in agroforestry-cacao systems (Theobroma cacao L. with optimal results to the inhibitory effect for these diseases.

Evaluation of Trichoderma spp. for biocontrol of tomato sudden caused by Pythium aphanidermatum following flooding in tropical hot season.

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Le, H T T; Black, L L; Sikora, R A

2003-01-01

Tomato sudden death is a major problem in tomato production in tropical lowland areas. The plant wilts and dies following artificial or natural flooding for 48-72 hrs in the summer season. Occurrence of this disease is related to aggressiveness of Pythium aphanidermatum on tomato at high soil temperature (>30 degrees C). Several methods such as using biological control agents, fungicides and other cultural practices were applied in attempts to control tomato sudden death. Three Trichoderma harzianum and two Trichoderma virens isolates were evaluated for biocontrol of the disease in the greenhouse and in the field T. harzianum and T. virens isolates were separately used to treat the seed, potting medium and also incorporated into the soil before transplanting. Field soil was naturally infested with P. aphanidermatum, while greenhouse soil was inoculated with the pathogen 10 days after transplanting. All treatments were flooded for 48 hrs at 32 degrees C soil temperature. Results from this study show that most tested T. harzianum and T. virens isolates have little promise for control of tomato sudden death following flooding. The percentage of tomato plants that wilted after growing in soil treated with either T. harzianum or T. virens and P. aphanidermatum was not significantly different when compared to the soil treated with P. aphanidermatum alone. No wilted plants were observed in the control (non treated soil).

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

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

1991-12-31

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

EFFECT OF Trichoderma harzianum AND ARBUSCULAR MYCORRHIZAL FUNGI ON GROWTH IN TOMATO (Lycopersicum esculentum MILL SEEDLINGS, NAPIER (Pennisetum purpureum L AND TEA (Camellia sinensis L CUTTINGS

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Margaret Mwangi

2010-09-01

Full Text Available A green-house study was conducted to investigate the ability of an isolate of Trichoderma harzianum (P52 and arbuscular mycorrhizal fungi (AMF in enhancing growth in tomato seedlings, tea and napier grass cuttings. The effect of these bio-inoculants on growth was compared with the influence of Diammonium phosphate (DAP fertilizer and the interactions of these three factors (P52, AMF and DAP. The plants were grown in plastic pots filled with sterilized soils. A completely randomized design was used and growth measurements taken on height shoot and root dry weights.  It was observed that isolate P52 and DAP fertilizer individually enhanced growth in tomatoes, tea and napier while AMF only enhanced growth in tomatoes. Combinations of P52 and DAP; P52, AMF and DAP enhanced growth significantly (P

Trichoderma strains- Silybum marianum hairy root cultures interactions

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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

Tolerance to chitosan by Trichoderma species is associated with low membrane fluidity.

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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. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Botrydial and botcinins produced by Botrytis cinerea regulate expression of Trichoderma arundinaceum genes involved in trichothecene biosynthesis

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Trichoderma arundinaceum (Ta37) and Botrytis cinerea produce the sesquiterpenes harzianum A (HA) and botrydial (BOT), respectively, and also the polyketides aspinolides (Asp) and botcinines (Botc), respectively. In the present work, we analyzed the role of BOT and Botcs in the T. arundinaceum-B. cin...

Assessment of the rhizosphere competency and pentachlorophenol-metabolizing activity of a pesticide-degrading strain of Trichoderma harzianum introduced into the root zone of corn seedlings.

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Rigot, Jerĵme; Matsumura, Fumio

2002-05-01

To develop a dependable approach to introduce laboratory selected, pesticide-degrading microorganisms into soil environments for the purpose of in situ bioremediation, we tested the possibility of utilizing plant rhizospheres as the vehicle. We first established the rhizosphere competency of a strain of the soil fungus Trichoderma harzianum, a biocontrol species well studied by plant pathologists to colonize plant rhizosphere in many parts of the world. The strain we chose, T.h.2023 is resistant to many fungicides, and it has been shown to metabolize several pesticides. Second, we found that it readily metabolized pentachlorophenol (PCP), which is quickly and stochiometrically converted to pentachloroanisole (PCA) in liquid culture. Taking advantage of this specific feature, we have developed a sensitive metabolic marker approach that allowed us to monitor for the continuous presence and activity of this fungal strain in the corn rhizosphere soil in situ over time.

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

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Oskiera Michał

2015-06-01

Full Text Available Trichoderma strains that are beneficial to both the growth and health of plants can be used as plant growth-promoting fungi (PGPF or biological control agents (BCA in agricultural and horticultural practices. In order to select PGPF or BCA strains, their biological properties and taxonomy must be carefully studied. In this study, 104 strains of Trichoderma collected at geographically different locations in Poland for selection as PGPF or BCA were identified by DNA barcoding, based on the sequences of internal transcribed spacers 1 and 2 (ITS1 and 2 of the ribosomal RNA gene cluster and on the sequences of translation elongation factor 1 alpha (tef1, chitinase 18-5 (chi18-5, and RNA polymerase II subunit (rpb2 gene fragments. Most of the strains were classified as: T. atroviride (38%, T. harzianum (21%, T. lentiforme (9%, T. virens (9%, and T. simmonsii (6%. Single strains belonging to T. atrobrunneum, T. citrinoviride, T. crassum, T. gamsii, T. hamatum, T. spirale, T. tomentosum, and T. viridescens were identified. Three strains that are potentially pathogenic to cultivated mushrooms belonging to T. pleuroticola and T. aggressivum f. europaeum were also identified. Four strains: TRS4, TRS29, TRS33, and TRS73 were classified to Trichoderma spp. and molecular identification was inconclusive at the species level. Phylogeny analysis showed that three of these strains TRS4, TRS29, and TRS33 belong to Trichoderma species that is not yet taxonomically established and strain TRS73 belongs to the T. harzianum complex, however, the species could not be identified with certainty.

Influence of Rhizoctonia solani and Trichoderma spp. in growth of bean (Phaseolus vulgaris L.) and in the induction of plant defense-related genes

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Mayo, Sara; Gutiérrez, Santiago; Malmierca, Monica G.; Lorenzana, Alicia; Campelo, M. Piedad; Hermosa, Rosa; Casquero, Pedro A.

2015-01-01

Many Trichoderma species are well-known for their ability to promote plant growth and defense. We study how the interaction of bean plants with R. solani and/or Trichoderma affect the plants growth and the level of expression of defense-related genes. Trichoderma isolates were evaluated in vitro for their potential to antagonize R. solani. Bioassays were performed in climatic chambers and development of the plants was evaluated. The effect of Trichoderma treatment and/or R. solani infection on the expression of bean defense-related genes was analyzed by real-time PCR and the production of ergosterol and squalene was quantified. In vitro growth inhibition of R. solani was between 86 and 58%. In in vivo assays, the bean plants treated with Trichoderma harzianum T019 always had an increased size respect to control and the plants treated with this isolate did not decrease their size in presence of R. solani. The interaction of plants with R. solani and/or Trichoderma affects the level of expression of seven defense-related genes. Squalene and ergosterol production differences were found among the Trichoderma isolates, T019 showing the highest values for both compounds. T. harzianum T019 shows a positive effect on the level of resistance of bean plants to R. solani. This strain induces the expression of plant defense-related genes and produces a higher level of ergosterol, indicating its ability to grow at a higher rate in the soil, which would explain its positive effects on plant growth and defense in the presence of the pathogen. PMID:26442006

Heavy Metal Tolerance and Removal Capacity of Trichoderma species Isolated from Mine Tailings in Itogon, Benguet

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Myra Tansengco

2017-11-01

Full Text Available Waste from mining industries contains various heavy metals that can pollute the environment. Bioremediation using efficient fungi can help in eliminating these heavy metal contaminants. This study focused on the isolation, identification, and characterization of heavy metal-resistant fungi from mine tailings in Itogon, Benguet. Isolation of fungi was done by serial dilution and spread plate techniques on potato dextrose agar (PDA with an individual heavy metal, i.e. chromium (Cr, copper (Cu, lead (Pb, zinc (Zn, and nickel (Ni. Of the 29 fungal isolates, four species were selected and molecularly identified as Trichoderma virens, T. harzianum, T. saturnisporum, and T. gamsii. Growth tolerance on PDA with increasing concentrations (200-1000 ppm of an individual heavy metal indicated the following trend: T. virens > T. harzianum > T. gamsii > T. saturnisporum. Growth test indicates that all Trichoderma isolates can tolerate high levels of Cr and Pb, however tolerance to Cu, Zn, and Ni was species specific. Shakeflask culture using T. virens showed high lead removal (91-96% over broad pH range while and at neutral pH, T. virens had 70% and 63% reductions for Cu and Cr, respectively. Results of this study highlights the potential of Trichoderma isolates for biological wastewater treatment in mining industries.

Biological control of Rhizoctonia solani on potato by using indigenous Trichoderma spp.

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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.

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

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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.

Estudio del antagonismo de algunas especies de Trichoderma sobre Fusarium Oxysporum y Rhizoctonia Solani Antagonism studies of Trichoderma sp.p.. with Fusarium oxysporum and Rhizoctonia solani

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Elias Ricardo

1989-12-01

Full Text Available En este trabajo se estudió el antagonismo de algunos aislamientos del hongo Trichoderma obtenidos de suelos colornbianos en el control de Fusarium oxysporum y Rhizoctonia solani. En los ensayos "in vitre" se observó un marcado antagonismo entre las colonias de los aislamientos de Trichoderma sobre R. sotsni, con una reducción apreciable
del tamaño de la colonia y un antaqonismo menor sobre F. oxysporum. En los ensayos de parasitismo a nivel microscópico, se observó una gran interacción entre alqunos
de los aislamientos de T. harzianum y T. hamatum y el patógéno R. solani rnanifestado por el enrollamiento, penetración, fragmentación y lisis de las hifas del patoqeno.
Los aislamientos de Trichoderma causaron un retraso en la aparición de los síntomas, una reducción en la severidad de la
enfermedad. y un menor número de plantas enfermas ocasionadas por F. oxysporum f. sp, cucumerinum en pepino cohombro, y su efecto fue superior en todos los casos a la
aplicación del fungicida benomil. Los aislamientos del antagonista aumentaron la qerminación de las semillas, la emergencia y el tamaño de las plántulas y redujeron la severidad de la enfermedad ocasionada por R. solani en fríjol.Several experiments were conducted to study the antagonism of 17 isolates of Trichoderma from Colombian soils with Fusarium oxysporum and Rhizoctonia solani. In "in vitro" tests, a high antagonism between colonies was found being greater the antagonism of Trichoderma with R. solani. At the microscopic level it was observed a great interaction between T. harzianum and T. hamatum with R. solani in such a way that the hyphae of the pathogen showed coiling, penetration, fragmentation and lysis. The Trichoderma isolates caused reduction in the disease severity, in the incubation period and a lower number of diseased cucumber plants when they were inoeulated with F. oxysporum f. sp, cucumerinum and these effects were better than Benomyl

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

NARCIS (Netherlands)

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

2003-01-01

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

  The fungus Trichoderma spp. on vegetables

DEFF Research Database (Denmark)

Winding, Anne

are important for risk assessment of MPCA and are the objectives of this project. Trichoderma spp. on tomatoes and cucumbers grown in greenhouses and on broccoli and celery leaf grown outdoors were quantified during a growth season. A MPCA with T. harzianum was applied in a greenhouse growing tomatoes...

Combinatorial efficacy of Trichoderma spp. and Pseudomonas fluorescens to enhance suppression of cell wall degrading enzymes produced by Fusarium wilt of Arachis hypogaea.L

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P Rajeswari

2017-12-01

Full Text Available Fusarium oxysporum, the soil borne pathogen causes vascular wilt, on majority of crop plants. It has been demonstrated that two different species of Trichoderma and Pseudomonas fluorescens suppress disease by different mechanisms. Therefore, application of a mixture of these biocontrol agents, and thus of several suppressive mechanisms, may represent a viable control strategy. A necessity for biocontrol by combinations of biocontrol agents can be the compatibility of the co-inoculated micro-organisms. Hence, compatibility between Trichoderma spp. and Pseudomonas fluorescens that have the ability to suppress Fusarium oxysporum in vitro on the activity of pectinolytic enzymes of Fusarium oxysporum. The activity of pectinolytic enzymes, i.e. pectin methyl esterase, endo and exo polymethylgalacturonases and exo and endo pectin trans eliminases produced by Fusarium oxysporum (Control was higher. Maximum inhibition of pectin methylesterase, exo and endo polymethylgalacturonase and exo and endopectin trans eliminase was shown by culture filtrate of Trichoderma viride + Pseudomonas fluorescens (Tv+Pf (1+2%, followed by Trichoderma harzianum + Pseudomonas fluorescens, (Th +Pf (1.5+2% and Trichoderma viride + Trichoderma harzianum (Tv+Th (1+1.5%. However, pathogenecity suppression of Fusarium oxysporum, a causative of Arachis hypogaea. L by the compatible combination of Trichodema viride + Pseudomonas fluorescens (1+2% was significantly better as compared to the single bio-agent. This indicates that specific interactions between biocontrol agents influence suppression of pathogenicity factors directly by combinations of these compatible bio-agents.

Identification of mycoparasitism-related genes against the phytopathogen Sclerotinia sclerotiorum through transcriptome and expression profile analysis in Trichoderma harzianum.

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Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Coelho, Alexandre Siqueira Guedes; Miller, Robert Neil Gerard; Pappas, Georgios Joannis; Ulhoa, Cirano José; Noronha, Eliane Ferreira

2014-03-18

The species of T. harzianum are well known for their biocontrol activity against plant pathogens. However, few studies have been conducted to further our understanding of its role as a biological control agent against S. sclerotiorum, a pathogen involved in several crop diseases around the world. In this study, we have used RNA-seq and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum gene expression during growth on cell wall of S. sclerotiorum (SSCW) or glucose. RT-qPCR was also used to examine genes potentially involved in biocontrol, during confrontation between T. harzianum and S. sclerotiorum. Data obtained from six RNA-seq libraries were aligned onto the T. harzianum CBS 226.95 reference genome and compared after annotation using the Blast2GO suite. A total of 297 differentially expressed genes were found in mycelia grown for 12, 24 and 36 h under the two different conditions: supplemented with glucose or SSCW. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on SSCW or glucose. We identified various genes of biotechnological value encoding proteins with functions such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. To validate the expression profile, RT-qPCR was performed using 20 randomly chosen genes. RT-qPCR expression profiles were in complete agreement with the RNA-Seq data for 17 of the genes evaluated. The other three showed differences at one or two growth times. During the confrontation assay, some genes were up-regulated during and after contact, as shown in the presence of SSCW which is commonly used as a model to mimic this interaction. The present study is the first initiative to use RNA-seq for identification of differentially expressed genes in T. harzianum strain TR274, in response to the phytopathogenic fungus S. sclerotiorum. It provides insights into the mechanisms of

The Combination of Trichoderma harzianum and Chemical Fertilization Leads to the Deregulation of Phytohormone Networking, Preventing the Adaptive Responses of Tomato Plants to Salt Stress.

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Rubio, M B; Hermosa, Rosa; Vicente, Rubén; Gómez-Acosta, Fabio A; Morcuende, Rosa; Monte, Enrique; Bettiol, Wagner

2017-01-01

Plants have evolved effective mechanisms to avoid or reduce the potential damage caused by abiotic stresses. In addition to biocontrol abilities, Trichoderma genus fungi promote growth and alleviate the adverse effects caused by saline stress in plants. Morphological, physiological, and molecular changes were analyzed in salt-stressed tomato plants grown under greenhouse conditions in order to investigate the effects of chemical and biological fertilizations. The application of Trichoderma harzianum T34 to tomato seeds had very positive effects on plant growth, independently of chemical fertilization. The application of salt stress significantly changed the parameters related to growth and gas-exchange rates in tomato plants subject to chemical fertilization. However, the gas-exchange parameters were not affected in unfertilized plants under the same moderate saline stress. The combined application of T34 and salt significantly reduced the fresh and dry weights of NPK-fertilized plants, while the opposite effects were detected when no chemical fertilization was applied. Decaying symptoms were observed in salt-stressed and chemically fertilized plants previously treated with T34. This damaged phenotype was linked to significantly higher intercellular CO 2 and slight increases in stomatal conductance and transpiration, and to the deregulation of phytohormone networking in terms of significantly lower expression levels of the salt overlay sensitivity 1 ( SOS1 ) gene, and the genes involved in signaling abscisic acid-, ethylene-, and salicylic acid-dependent pathways and ROS production, in comparison with those observed in salt-challenged NPK-fertilized plants.

Effect of mycosynthesized silver nanoparticles from filtrate of Trichoderma harzianum against larvae and pupa of dengue vector Aedes aegypti L.

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Sundaravadivelan, Chandran; Padmanabhan, Madanagopal Nalini

2014-03-01

Mosquitoes transmit dreadful diseases, causing millions of deaths every year. Therefore, screening for larvicidal and pupicidal activity of microbial extracts attributes could lead to development of new and improved mosquito control methods that are economical and safe for nontarget organisms and are ecofriendly. Synthetic chemical insecticides occupy predominant position in control strategies. These hazardous chemicals exert unwarranted toxicity and lethal effects on nontarget organisms, develop physiological resistance in target, and cause adverse environmental effect. For vector control, fungal-mediated natural products have been a priority in this area at present. In the current study, effective larvicidal and pupicidal effect of mycosynthesized silver nanoparticles (Ag NPs) using an entomopathogenic fungi Trichoderma harzianum against developmental stages of the dengue vector Aedes aegypti was investigated. An attractive possibility of green nanotechnology is to use microorganisms in the synthesis of nanosilver especially Ag NPs. The mycosynthesized Ag NPs were characterized to find their unique properties through UV-visible spectrophotometer, X-ray diffraction analysis, Fourier transform infrared, and surface characteristics by scanning electron microscopy. To analyze the bioefficacy, different test concentrations for extracellular filtrate (0.2, 0.4, 0.6, 0.8, and 1.0 %) and Ag NPs (0.05, 0.10, 0.15, 0.20, and 0.25 %) were prepared to a final volume of 200 mL using deionized water; 20 larvae of each instars (I-IV) and pupa were exposed to each test concentration separately which included a set of control (distilled water) group with five replicates. Characterization of the synthesized Ag NPs were about 10-20 nm without aggregation. Susceptibility of larval instars to synthesized Ag NPs was higher than the extracellular filtrate of T. harzianum alone after 24-h exposure, where the highest mortality was recorded as 92 and 96 % for first and second instars and

Novel aspinolide production by Trichoderma arundinaceum with a potential role in Botrytis cinerea antagonistic activity and plant defense priming

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Harzianum A (HA), a trichothecene produced by Trichoderma arundinaceum, has recently been described to have antagonistic activity against fungal plant pathogens and to induce plant defence genes. In the present work, we have shown that a tri5 genedisrupted mutant that lacks HA production overproduce...

Purification and Biochemical Characterization of a Neutral Serine Protease from Trichoderma harzianum. Use in Antibacterial Peptide Production from a Fish By-Product Hydrolysate.

Science.gov (United States)

Aissaoui, Neyssene; Chobert, Jean-Marc; Haertlé, Thomas; Marzouki, M Nejib; Abidi, Ferid

2017-06-01

This study reports the purification and biochemical characterization of an extracellular neutral protease from the fungus Trichoderma harzianum. The protease (Th-Protease) was purified from the culture supernatant to homogeneity by a three-step procedure with 14.2% recovery and 9.06-fold increase in specific activity. The purified enzyme appeared as a single protein band after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular mass of about 20 kDa. The optimum pH and temperature for the proteolytic activity were pH 7.0 and 40 °C, respectively. The enzyme was then investigated for its potential application in the production of antibacterial peptides. Interestingly, Scorpaena notata viscera protein hydrolysate prepared using the purified serine protease (Th-Protease) showed remarkable in vitro antibacterial activities. A peptide with a high antibacterial activity was further purified by a three-step procedure, and its sequence was identified as FPIGMGHGSRPA. The result of this study offers a promising alternative to produce natural antibacterial peptides from fish protein hydrolysate.

Control biológico del Tizón Tardío Phytophthora infestans en papa Solanum tuberosum a través de consorcios microbianos formados por hongos nativos del género Trichoderma sp.

OpenAIRE

Bustamante Gavilanes, Adriana Elizabeth

2015-01-01

Esta investigación evalúo la eficiencia de consorcios microbianos formados por hongos nativos del género Trichoderma sp. como biocontroladores del tizón tardío Phytophthora infestans en papa Solanum tuberosum. En las pruebas de antagonismo las cepas del género Trichoderma sp. inhibieron en promedio entre 83% y 87% al patógeno. En base a los resultados obtenidos en el ensayo de campo mostró que el tratamiento T3: Trichoderma harzianum+ Trichoderma sp. + Trichoderma atroviride es más efectivo ...

Assessment of metabolic capacity of Trichoderma inhamatum Bol12 QD biocontrol on native strains of Phytophthora infestans in vitro

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Puño Ramon

2011-08-01

Full Text Available Plant pathogen Phytophthora infestans is a cause of decreased crop yield of tomato, to control these losses, farmers use chemicals. This has consequences for the environment, human health and beneficial organisms in the ecosystem. The objective was to obtain and identify native isolates of Trichoderma spp. In soil planted with tomato Tlayacapan, Morelos (Mexico, Alternaria solani problems and Phytophthora infestans, also determine their antagonistic capacity in vitro. Trichoderma was isolated directly from soil by dilution in culture medium plate with potato dextrose agar (PDA. On the other side plate dilutions of yeast T. QD Bol12 inhamatum crops produced in batch for 30 days to compare the effectiveness of biocontrol. The filtered yeast inhibited mycelial growth kinetic of the agent in laboratory with the 1:2 dilution growth was 32.5% for the 1:4 dilution mycelial growth was 69.1% and finally to the dilution of 1:8 of the yeast biocontrol mycelium grew to 95.2%. To demonstrate the inhibitory activity on the pathogen in field crops, there were 3 L batch for four months. The application of three doses (undiluted, diluted 1:2 and 1:4 plus a control dilution water only was performed in a complete block design with four replications randomly with the tomato crop, belonging to the variety Santa Cruz Kada Gigante in the plots of the Academic Rural United Campesina Carmen Pampa. Statistical analysis by Duncan's test showed that the pure leaven reduce infection by Phytophthora infestans significantly in tomato. Appeared another tomato plant pathogen, Septoria lycopersici, in the course of fieldwork. We also evaluated the effect of the dose of yeast to this disease, and also noticed a significant reduction with all doses of yeast. These experiments demonstrated that the seeds of T. QD Bol12 inhamatum have biocontrol effect on the tomato crop. The antagonistic capacity was assessed using the cellophane and the kind of antagonism with the dual culture

Combination of Trichoderma harzianum endochitinase and a membrane-affecting fungicide on control of Alternaria leaf spot in transgenic broccoli plants.

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Mora, A; Earle, E D

2001-04-01

Progeny from transgenic broccoli (cv. Green Comet) expressing a Trichoderma harzianum endochitinase gene were used to assess the interaction between endochitinase and the fungicide Bayleton in the control of Alternaria brassicicola. In vitro assays have shown synergistic effects of endochitinase and fungicides on fungal pathogens. Our study examined the in planta effects of endochitinase and Bayleton, individually and in combination. Two month old transgenic and non-transgenic plants were sprayed with ED50 levels of Bayleton and/or inoculated with an A. brassicicola spore suspension. Disease levels in non-sprayed transgenic plants were not statistically different from sprayed transgenic plants nor from sprayed non-transgenic controls. Thus endochitinase-transgenic plants alone provided a significant reduction of disease severity, comparable to the protection by fungicide on non-transgenic plants. Comparison of the expected additive and observed effects revealed no synergism between endochitinase and Bayleton (at ED50 level), and usually less than an additive effect. Some transgenic lines sprayed with fungicide at doses higher than ED50 showed resistance similar to the non-sprayed transgenic lines, again suggesting no synergistic effect. Lack of synergism may be due to incomplete digestion of the cell wall by endochitinase, so that the effect of Bayleton at the cell membrane is not enhanced.

Antagonistic activity Trichoderma harzianum Rifai on the causal agent of rice blast (Pyricularia grisea Sacc.

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Ernesto Juniors Pérez Torres

2017-10-01

Full Text Available With the objective to evaluate the antagonistic activity of T. harzianum (strain A-34 on the causal agent of rice blast (P. grisea, were developed several in vitro experiments. It was evaluated the biocontrol mechanisms such as competition through mounted the percent inhibition of radial growth of hyphae of P. grisea from 24 to 240 hours and the antagonistic capacity. In addition, was evaluated micoparasitism to inclination the observation of events Microscopy winding, penetration, vacuolization, lysis, and antibiosis by observing 24 hours a confrontation between the hyphae of the phytopathogenic fungus and biological control agent. It was obtained at 120 hours 100 % inhibition of micelial growth of causal agent, what corresponded with the degree 1 of antagonistic capacity (scale and is recorded as a hyperparasitic action on P. grisea. It was evidenced an antibiotic effect of metabolites produced by T. harzianum (strain A-34 to 24 hours of confrontation, where there was time interaction between the hyphae of microorganisms with 14,3 % inhibition, also was evidence the micoparasitism events by penetration, vacuolization and lysis in the cells of phytopathogenic fungus. These results demonstrated the ability of T. harzianum (strain A-34 on causal agent of rice blast (P. grisea.

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

Components of a rice-oilseed rape production system augmented with trichoderma sp. Tri-1 control sclerotinia sclerotiorum on oilseed rape

Science.gov (United States)

Sclerotinia sclerotiorum causes serious yield losses on many crops throughout the world. In two field trials conducted at the same location in consecutive years, a treatment containing formulated Trichoderma harzianum-1 (Tri-1) resulted in oilseed rape seed yield that was significantly greater than...

Detoxification of toxic phorbol esters from Malaysian Jatropha curcas Linn. kernel by Trichoderma spp. and endophytic fungi.

Science.gov (United States)

Najjar, Azhar; Abdullah, Norhani; Saad, Wan Zuhainis; Ahmad, Syahida; Oskoueian, Ehsan; Abas, Faridah; Gherbawy, Youssuf

2014-02-05

The presence of phorbol esters (PEs) with toxic properties limits the use of Jatropha curcas kernel in the animal feed industry. Therefore, suitable methods to detoxify PEs have to be developed to render the material safe as a feed ingredient. In the present study, the biological treatment of the extracted PEs-rich fraction with non-pathogenic fungi (Trichoderma harzianum JQ350879.1, T. harzianum JQ517493.1, Paecilomyces sinensis JQ350881.1, Cladosporium cladosporioides JQ517491.1, Fusarium chlamydosporum JQ350882.1, F. chlamydosporum JQ517492.1 and F. chlamydosporum JQ350880.1) was conducted by fermentation in broth cultures. The PEs were detected by liquid chromatography-diode array detector-electrospray ionization mass spectrometry (LC-DAD-ESIMS) and quantitatively monitored by HPLC using phorbol-12-myristate 13-acetate as the standard. At day 30 of incubation, two T. harzianum spp., P. sinensis and C. cladosporioides significantly (p rich fraction for growth. In the cytotoxicity assay, cell viabilities of Chang liver and NIH 3T3 fibroblast cell lines were less than 1% with the untreated PEs-rich fraction, but 84.3%-96.5% with the fungal treated PEs-rich fraction. There was no inhibition on cell viability for normal fungal growth supernatants. To conclude, Trichoderma spp., Paecilomyces sp. and Cladosporium sp. are potential microbes for the detoxification of PEs.

Trichothecenes and aspinolides produced by Trichoderma arundinaceum regulate expression of Botrytis cinerea genes involved in virulence and growth

Science.gov (United States)

Trichoderma arundinaceum (Ta37) and Botrytis cinerea (B05.10) produce the sesquiterpenoids harzianum A (HA) and botrydial (BOT), respectively. Ta'Tri5, an HA non-producer mutant, produces high levels of the polyketide compounds aspinolides (Asp) B and C. We analyzed the role of HA and Asp in the B. ...

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.

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

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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.

Diversity and effect of Trichoderma spp. associated with green mold disease on Lentinula edodes in China.

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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. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Species specific substrates and products choices of 4-O-acetyltransferase from Trichoderma brevicompactum.

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Sharma, Shikha; Kumari, Indu; Hussain, Razak; Ahmed, Mushtaq; Akhter, Yusuf

2017-09-01

Antagonistic species of Trichoderma such as T. harzianum, T. viride, T. virens and T. koningii are well-known biocontrol agents that have been reported to suppress pathogenic soil microbes and enhance the growth of crop plants. Secondary metabolites (SMs) including trichothecenes are responsible for its biocontrol activities. The trichothecenes, trichodermin and harzianum A (HA) are produced in species dependent manner respectively, by Trichoderma brevicompactum (TB) and Trichoderma arundinaceum (TA). The last step in the pathway involves the conversion of trichodermol into trichodermin or HA alternatively, which is catalyzed by 4-O-acetyltransferase (encoded by tri3 gene). Comparative sequence analysis of acetyltransferase enzyme of TB with other chloramphenicol acetyltransferase (CAT) family proteins revealed the conserved motif involved in the catalysis. Multiple substrate binding studies were carried out to explore the mechanism behind the two different outcomes. His188 was found to have a role in initial substrate binding. In the case of trichodermin synthesis, represented by ternary complex 1, the trichodermol and acetic anhydride (AAn), the two substrates come very close to each other during molecular simulation analysis so that interactions become possible between them and acetyl group may get transferred from AAn to trichodermol, and Tyr476 residue mediates this phenomenon resulting in the formation of trichodermin. However, in case of the HA biosynthesis using the TB version of enzyme, represented by ternary complex 2, the two substrates, trichodermol and octa-2Z,4E,6E-trienedioic acid (OCTA) did not show any such interactions. Copyright © 2017 Elsevier Inc. All rights reserved.

Identification, characterization and phylogenetic analysis of antifungal Trichoderma from tomato rhizosphere.

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

2016-01-01

The use of Trichoderma isolates with efficient antagonistic activity represents a potentially effective and alternative disease management strategy to replace health hazardous chemical control. In this context, twenty isolates were obtained from tomato rhizosphere and evaluated by their antagonistic activity against four fungal pathogens ( Fusarium oxysporum f. sp. lycopersici , Alternaria alternata , Colletotrichum gloeosporoides and Rhizoctonia solani ). The production of extracellular cell wall degrading enzymes of tested isolates was also measured. All the isolates significantly reduced the mycelial growth of tested pathogens but the amount of growth reduction varied significantly as well. There was a positive correlation between the antagonistic capacity of Trichoderma isolates towards fungal pathogens and their lytic enzyme production. The Trichoderma isolates were initially sorted according to morphology and based on the translation elongation factor 1-α gene sequence similarity, the isolates were designated as Trichoderma harzianum , T. koningii , T. asperellum , T. virens and T. viride . PCA analysis explained 31.53, 61.95, 62.22 and 60.25% genetic variation among Trichoderma isolates based on RAPD, REP-, ERIC- and BOX element analysis, respectively. ERG - 1 gene, encoding a squalene epoxidase has been used for the first time for diversity analysis of antagonistic Trichoderma from tomato rhizosphere. Phylogenetic analysis of ERG -1 gene sequences revealed close relatedness of ERG -1sequences with earlier reported sequences of Hypocrea lixii , T. arundinaceum and T. reesei. However, ERG -1 gene also showed heterogeneity among some antagonistic isolates and indicated the possibility of occurrence of squalene epoxidase driven triterpene biosynthesis as an alternative biocontrol mechanism in Trichoderma species.

Trichoderma-Based Biostimulants Modulate Rhizosphere Microbial Populations and Improve N Uptake Efficiency, Yield, and Nutritional Quality of Leafy Vegetables

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Nunzio Fiorentino

2018-06-01

Full Text Available Microbial inoculants such as Trichoderma-based products are receiving great interest among researchers and agricultural producers for their potential to improve crop productivity, nutritional quality as well as resistance to plant pathogens/pests and numerous environmental stresses. Two greenhouse experiments were conducted to assess the effects of Trichoderma-based biostimulants under suboptimal, optimal and supraoptimal levels of nitrogen (N fertilization in two leafy vegetables: Iceberg lettuce (Lactuca sativa L. and rocket (Eruca sativa Mill.. The yield, nutritional characteristics, N uptake and mineral composition were analyzed for each vegetable crop after inoculation with Trichoderma strains T. virens (GV41 or T. harzianum (T22, and results were compared to non-inoculated plants. In addition, the effect of the Trichoderma-based biostimulants on microbes associated with the rhizosphere in terms of prokaryotic and eukaryotic composition and concentration using DGGE was also evaluated. Trichoderma-based biostimulants, in particular GV41, positively increased lettuce and rocket yield in the unfertilized plots. The highest marketable lettuce fresh yield was recorded with either of the biostimulant inoculations when plants were supplied with optimal levels of N. The inoculation of rocket with GV41, and to a lesser degree with T22, elicited an increase in total ascorbic acid under both optimal and high N conditions. T. virens GV41 increased N-use efficiency of lettuce, and favored the uptake of native N present in the soil of both lettuce and rocket. The positive effect of biostimulants on nutrient uptake and crop growth was species-dependent, being more marked with lettuce. The best biostimulation effects from the Trichoderma treatments were observed in both crops when grown under low N availability. The Trichoderma inoculation strongly influenced the composition of eukaryotic populations in the rhizosphere, in particularly exerting different

The Anti-Fasciolasis Properties of Silver Nanoparticles Produced by Trichoderma harzianum and Their Improvement of the Anti-Fasciolasis Drug Triclabendazole

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AbdelElah A. Banaja

2013-11-01

Full Text Available Recently, new strains of Fasciola demonstrated drug resistance, which increased the need for new drugs or improvement of the present drugs. Nanotechnology is expected to open some new opportunities to fight and prevent diseases using an atomic scale tailoring of materials. The ability to uncover the structure and function of biosystems at the nanoscale, stimulates research leading to improvement in biology, biotechnology, medicine and healthcare. The size of nanomaterials is similar to that of most biological molecules and structures; therefore, nanomaterials can be useful for both in vivo and in vitro biomedical research and applications. Therefore, this work aimed to isolate fungal strains from Taif soil samples, which have the ability to synthesize silver nanoparticles. The fungus Trichoderma harzianum, when challenged with silver nitrate solution, accumulated silver nanoparticles (AgNBs on the surface of its cell wall in 72 h. These nanoparticles, dislodged by ultrasonication, showed an absorption peak at 420 nm in a UV-visible spectrum, corresponding to the plasmon resonance of silver nanoparticles. The transmission electron micrographs of dislodged nanoparticles in aqueous solution showed the production of reasonably monodisperse silver nanoparticles (average particle size: 4.66 nm by the fungus. The percentage of non hatching eggs treated with the Triclabendazole drug was 69.67%, while this percentage increased to 89.67% in combination with drug and AgNPs.

Production of hydrolytic enzymes by Trichoderma isolates with antagonistic activity against Crinipellis perniciosa, the causal agent of witches' broom of cocoa

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Marco Janice Lisboa De

2003-01-01

Full Text Available Two isolates of Trichoderma, which reduce the incidence of witches'broom disease caused in cocoa by Crinipellis perniciosa, were evaluated for their potential to produce hydrolases in liquid medium. Very low or no hydrolytic activity was produced in the absence of any substrate. The activities of chitinase, N-acetylglucosaminidase, beta-1,3-glucanase, total cellulase, endoglucanase, aryl- beta-glucosidase, beta-glucosidase, protease and amylase increased dramatically within 72-120 h of growth in the presence of specific substrates. Except for N-acetylglucosaminidase and beta-glucosidase Trichoderma harzianum isolate 1051 produced the largest amounts of hydrolases. The possible involvement of these enzymes in the antagonistic interaction between Trichoderma and C. perniciosa is discussed.

Proteomic analysis of grapevine resistance induced by Trichoderma harzianum T39 reveals specific defence pathways activated against downy mildew

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Perazzolli, Michele

2012-01-01

Downy mildew is caused by the oomycete Plasmopara viticola and is one of the most serious diseases of grapevine. The beneficial microorganism Trichoderma harzianum T39 (T39) has previously been shown to induce plant-mediated resistance and to reduce the severity of downy mildew in susceptible grapevines. In order to better understand the cellular processes associated with T39-induced resistance, the proteomic and histochemical changes activated by T39 in grapevine were investigated before and 1 day after P. viticola inoculation. A comprehensive proteomic analysis of T39-induced resistance in grapevine was performed using an eight-plex iTRAQ protocol, resulting in the identification and quantification of a total of 800 proteins. Most of the proteins directly affected by T39 were found to be involved in signal transduction, indicating activation of a complete microbial recognition machinery. Moreover, T39-induced resistance was associated with rapid accumulation of reactive oxygen species and callose at infection sites, as well as changes in abundance of proteins involved in response to stress and redox balance, indicating an active defence response to downy mildew. On the other hand, proteins affected by P. viticola in control plants mainly decreased in abundance, possibly reflecting the establishment of a compatible interaction. Finally, the high-throughput iTRAQ protocol allowed de novo peptide sequencing, which will be used to improve annotation of the Vitis vinifera cv. Pinot Noir proteome. PMID:23105132

Identification of differentially expressed genes from Trichoderma harzianum during growth on cell wall of Fusarium solani as a tool for biotechnological application.

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Vieira, Pabline Marinho; Coelho, Alexandre Siqueira Guedes; Steindorff, Andrei Stecca; de Siqueira, Saulo José Linhares; Silva, Roberto do Nascimento; Ulhoa, Cirano José

2013-03-15

The species of T. harzianum are well known for their biocontrol activity against many plant pathogens. However, there is a lack of studies concerning its use as a biological control agent against F. solani, a pathogen involved in several crop diseases. In this study, we have used subtractive library hybridization (SSH) and quantitative real-time PCR (RT-qPCR) techniques in order to explore changes in T. harzianum genes expression during growth on cell wall of F. solani (FSCW) or glucose. RT-qPCR was also used to examine the regulation of 18 genes, potentially involved in biocontrol, during confrontation between T. harzianum and F. solani. Data obtained from two subtractive libraries were compared after annotation using the Blast2GO suite. A total of 417 and 78 readable EST sequence were annotated in the FSCW and glucose libraries, respectively. Functional annotation of these genes identified diverse biological processes and molecular functions required during T. harzianum growth on FSCW or glucose. We identified various genes of biotechnological value encoding to proteins which function such as transporters, hydrolytic activity, adherence, appressorium development and pathogenesis. Fifteen genes were up-regulated and sixteen were down-regulated at least at one-time point during growth of T. harzianum in FSCW. During the confrontation assay most of the genes were up-regulated, mainly after contact, when the interaction has been established. This study demonstrates that T. harzianum expressed different genes when grown on FSCW compared to glucose. It provides insights into the mechanisms of gene expression involved in mycoparasitism of T. harzianum against F. solani. The identification and evaluation of these genes may contribute to the development of an efficient biological control agent.

Changes in hyphal morphology and activity of phenoloxidases during interactions between selected ectomycorrhizal fungi and two species of Trichoderma.

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Mucha, Joanna

2011-06-01

Patterns of phenoloxidase activity can be used to characterize fungi of different life styles, and changes in phenoloxidase synthesis were suspected to play a role in the interaction between ectomycorrhizal and two species of Trichoderma. Confrontation between the ectomycorrhizal fungi Amanita muscaria and Laccaria laccata with species of Trichoderma resulted in induction of laccase synthesis, and the laccase enzyme was bound to mycelia of ectomycorrhizal fungi. Tyrosinase release was noted only during interaction of L. laccata strains with Trichoderma harzianum and T. virens. Ectomycorrhizal fungi, especially strains of Suillus bovinus and S. luteus, inhibited growth of Trichoderma species and caused morphological changes in its colonies in the zone of interaction. In contrast, hyphal changes occurred less often in the ectomycorrhizal fungi tested. Species of Suillus are suggested to present a different mechanism in their interaction with other fungi than A. muscaria and L. laccata.

Trichoderma harzianum containing 1-aminocyclopropane-1-carboxylate deaminase and chitinase improved growth and diminished adverse effect caused by Fusarium oxysporum in soybean.

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Zhang, Fuli; Chen, Can; Zhang, Fan; Gao, Lidong; Liu, Jidong; Chen, Long; Fan, Xiaoning; Liu, Chang; Zhang, Ke; He, Yuting; Chen, Chen; Ji, Xiue

2017-03-01

An isolate, named Trichoderma harzianum T-soybean, showed growth-promoting for soybean seedlings and induced resistance to Fusarium oxysporum under greenhouse. Compared to control soybean seedlings, fresh weight, dry weight, lateral root number, chlorophyll content, root activity and soluble protein of plants pretreated with T-soybean increased, but initial pod height reduced. Furthermore, we found that T-soybean inhibited the growth of F. oxysporum by parasitic function. In addition, plate test results showed that culture filtrates of T-soybean also inhibited significantly F. oxysporum growth. Meanwhile, T-soybean treatment obviously reduced disease severity and induced quickly the H 2 O 2 and O 2 - burst as well as pathogenesis related protein gene (PR3) expression after F. oxysporum inoculation, and subsequently diminished the cell damage in soybean caused by the pathogen challenge. Reactive oxygen species (ROS) scavenging enzymes activity analysis showed that the activities of peroxidase (POD), polyphenol oxidase (PPO) and superoxide dismutase (SOD) increased significantly in T-soybean pretreated plants. These results suggested that T-soybean treatment induced resistance in soybean seedlings to F. oxysporum by companying the production of ROS and the increasing of ROS scavenging enzymes activity as well as PR3 expression. Copyright © 2016 Elsevier GmbH. All rights reserved.

Potential of Trichoderma spp. strains for the bioremediation of soils contaminated with petroleum

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Marcia Pesántez

2016-10-01

Full Text Available Fungi species can degrade xenobiotic compounds contaminating the soil, including hydrocarbons. The objective of this work was to determine the potential of three strains of Trichoderma, isolated from soil contaminated with petroleum, for bioremediation. Trichoderma harzianum CCECH-Te1, Trichoderma viride CCECH-Te2 and Trichoderma psedokoningii CCECH-Te3 were included in one assay with each independent strain. The inoculum was adjusted to a concentration of 1x1010 conidia ml-1 which was applied to soil contaminated by an oil spill. After 96 days of inoculation, soil samples were taken at 10 and 15 cm depth. The content of total hydrocarbons, polycyclic aromatic hydrocarbons and heavy metals such as cadmium, nickel and lead were determined. With the data, it was calculated the percentage of removal of the analyzed compounds by each strain. At 10 cm and 15 cm depth, it was observed the removal of the compounds in percentages that reached between 47 and 69.1% in the hydrocarbons and up to 53.72% in the heavy metals. It which denoted the potential of the three strains for bioremediation in contaminated soils.   Keywords: heavy metals, polycyclic aromatic hydrocarbons, xenobiotic compounds

A mini-bag technique for evaluation of fungicide effects on Trichoderma spp in mushroom compost.

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Abosriwil, Salem O; Clancy, Kevin J

2004-04-01

An in vivo technique was developed to observe colonisation of mushroom compost by Trichoderma spp. Isolates of T. harzianum (Th2), T. harzianum (Th1), T. koningii (Tk) and T. viride (Tv) were artificially introduced into compost using a mini-bag system. Wheat grains, colonised by Trichoderma spp, were placed centrally on a layer of compost at the bottom of 1-litre polythene bags which were then filled with 350 g of spawned or un-spawned compost, and partially sealed. After 14 and 21 days incubation at 27 degrees C, the bags were assessed for recovery of Trichoderma from middle and top zones using a needle stab re-isolation technique and a visual colonisation scoring system. Visible green mould contamination, similar to that observed in practice, developed within 21 days. The visual colonisation scoring was reliably related to the re-isolation success. In this evaluation, Trichoderma spp showed considerable differences in their relative abilities to colonise spawned and un-spawned compost, with Th2 isolates being consistently superior to the other isolates of Th1, Tk and Tv in colonising spawned compost. This technique was employed to evaluate the effects of fungicides on the colonisation of mushroom compost by three Trichoderma spp: Th2, Th1 and Tk, using 1-litre and 5-litre mini-bag systems. Aqueous suspensions of benomyl, carbendazim, thiabendazole, prochloraz and prochloraz+carbendazim incorporated into the compost at 50 mg litre(-1), or applied to spawn at 50 mg kg(-1), reduced the colonisation by Trichoderma spp. Prochloraz and prochloraz+carbendazim were superior to benomyl, carbendazim or thiabendazole in reducing compost colonisation by Th2, Th1 and Tk, with Th2 being the most persistent type, capable of colonising treated compost in the presence of all five fungicides. The prochloraz+carbendazim mixture, not normally used in mushroom production, was equal to or better than prochloraz alone. The incidence of green mould colonisation by Th2 was as extensive

Antagonistic Activity of Trichoderma ISolates against Sclerotium rolfsii : Screening of Efficient Isolates from Morocco Soils for Biological Control

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N. Khattabi

2004-12-01

Full Text Available Seventy Trichoderma spp. isolates collected from different regions of Morocco were tested for their capacity to inhibit in vitro mycelial growth of Sclerotium rolfsii, and for their effect on the viability of S. rolfsii sclerotia in the soil. The Trichoderma spp. isolates inhibited mycelial growth of S. rolfsii to various degrees, with 52% of isolates expressing an average inhibition, varying between 45 and 55%. The effect on the viability of sclerotia in the soil also varied between isolates of Trichoderma, with the majority (84% having a slight effect. A group of twenty isolates identified as Trichoderma harzianum when tested in sterilized soil, significantly reduced sclerotial viability though not in natural soil. Four of these isolates (Nz, Kb2, Kb3 and Kf1 showed good antagonistic activity against S. rolfsii and were also highly competitive in natural soil. These isolates would therefore be candidates for development in biological control program.

Cepa de Trichoderma útil para el tratamiento y/o prevención de infecciones provocadas por hongos pertenecientes al género Verticillium

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Barroso-Albarracín, Juan Bautista; Chaki, Mounira; Mercado-Blanco, Jesús; Valverde-Corredor, Antonio; López García, Antonio Alejandro

2011-01-01

La invención se refiere al uso de la cepa de hongos Trichoderma harzianum CECT 2413 para la prevención y/o tratamiento de infecciones de plantas y/o suelos provocadas por hongos fitopatógenos pertenecientes al género Verticíllium, preferiblemente por los patotipos defoliante o no defoliante de Verticillium dahliae, agente causante de la Verticilosis del olivo. Además, la invención propone el uso de esta cepa de hongos en combinación con otros hongos del género Trichoderma.

Detoxification of Toxic Phorbol Esters from Malaysian Jatropha curcas Linn. Kernel by Trichoderma spp. and Endophytic Fungi

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Azhar Najjar

2014-02-01

Full Text Available The presence of phorbol esters (PEs with toxic properties limits the use of Jatropha curcas kernel in the animal feed industry. Therefore, suitable methods to detoxify PEs have to be developed to render the material safe as a feed ingredient. In the present study, the biological treatment of the extracted PEs-rich fraction with non-pathogenic fungi (Trichoderma harzianum JQ350879.1, T. harzianum JQ517493.1, Paecilomyces sinensis JQ350881.1, Cladosporium cladosporioides JQ517491.1, Fusarium chlamydosporum JQ350882.1, F. chlamydosporum JQ517492.1 and F. chlamydosporum JQ350880.1 was conducted by fermentation in broth cultures. The PEs were detected by liquid chromatography-diode array detector-electrospray ionization mass spectrometry (LC-DAD-ESIMS and quantitatively monitored by HPLC using phorbol-12-myristate 13-acetate as the standard. At day 30 of incubation, two T. harzianum spp., P. sinensis and C. cladosporioides significantly (p < 0.05 removed PEs with percentage losses of 96.9%–99.7%, while F. chlamydosporum strains showed percentage losses of 88.9%–92.2%. All fungal strains could utilize the PEs-rich fraction for growth. In the cytotoxicity assay, cell viabilities of Chang liver and NIH 3T3 fibroblast cell lines were less than 1% with the untreated PEs-rich fraction, but 84.3%–96.5% with the fungal treated PEs-rich fraction. There was no inhibition on cell viability for normal fungal growth supernatants. To conclude, Trichoderma spp., Paecilomyces sp. and Cladosporium sp. are potential microbes for the detoxification of PEs.

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

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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.

Biological control of Fusarium solani f. sp. phaseoli the causal agent of root rot of bean using Bacillus subtilis CA32 and Trichoderma harzianum RU01

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Saman Abeysinghe

2007-09-01

Full Text Available Root rot, caused by Fusarium solani f. sp. phaseoli, is one of the main root diseases impacting production of common bean in Sri Lanka. Rhizobacteria were screened in dual Petri plate assays to select antagonistic strains against F. solani f. sp. phaseoli. B. subtilis CA32 effectively antagonized the pathogen. T. harzianum RU01 also showed the antagonistic activity. The efficacy of the B. subtilis CA32 and the T. harzianum RU01 were tested in greenhouse pot experiments against F. solani f. sp. phaseoli. Seed bacterization with B. subtilis CA32 and T. harzianum RU01 significantly protected bean seedlings from F. solani f. sp. phaseoli compared to the untreated control plants. Plant protection was more pronounced in T. harzianum RU01 treated plants than bacterized plants. Enhanced root growth was observed only T. harzianum RU01 treated plants, suggesting that the biotic modifications of the mycorrhizosphere as a result of colonization with T. harzianum RU01.

Phylogeny and taxonomical investigation of Trichoderma spp. from Indian region of Indo-Burma Biodiversity hot spot region with special reference to Manipur.

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Kamala, Th; Devi, S Indira; Sharma, K Chandradev; Kennedy, K

2015-01-01

Towards assessing the genetic diversity and occurrence of Trichoderma species from the Indian region of Indo-Burma Biodiversity hotspot, a total of 193 Trichoderma strains were isolated from cultivated soils of nine different districts of Manipur comprising 4 different agroclimatic zones. The isolates were grouped based on the morphological characteristics. ITS-RFLP of the rDNA region using three restriction digestion enzymes: Mob1, Taq1, and Hinf1, showed interspecific variations among 65 isolates of Trichoderma. Based on ITS sequence data, a total of 22 different types of representative Trichoderma species were reported and phylogenetic analysis showed 4 well-separated main clades in which T. harzianum was found to be the most prevalent spp. among all the Trichoderma spp. Combined molecular and phenotypic data leads to the development of a taxonomy of all the 22 different Trichoderma spp., which was reported for the first time from this unique region. All these species were found to produce different extrolites and enzymes responsible for the biocontrol activities against the harmful fungal phytopathogens that hamper in food production. This potential indigenous Trichoderma spp. can be targeted for the development of suitable bioformulation against soil and seedborne pathogens in sustainable agricultural practice.

Antagonism of Trichoderma species on Cladosporium herbarum and their enzimatic characterization Antagonismo de espécies de Trichoderma sobre Cladosporium herbarum e suas caracterização enzimática

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Maria Angélica G. Barbosa

2001-06-01

Full Text Available The verrucose caused by Cladosporium herbarum reduces production and quality of Passion fruit (Passiflora edulis Sims., a largely consumed tropical fruit. This work aimed to investigate the antagonism of Trichoderma species (T. polysporum, T. koningii, T. viride and T. harzianum against Cladosporium herbarum, and to study the production of extracellular hydrolytic enzymes by the pathogen and the antagonists. The results showed considerable antagonistic potential for the biocontrol of C. herbarum isolates by all Trichoderma species, except T. koningii. The most distinguished effect was observed for T. polysporum. In relation to the pattern of esterase obtained by electrophoresis in poliacrylamyde gel, the major activity presented by the isolates was observed after five days of incubation. The C. herbarum isolates produced extracellular enzymes, lipase, pectinase, cellulase, and protease, all possibly related to the infection process. Amylase excretion, not known to be associated with phytopathogens, was detected in Trichoderma species, but not in C. herbarum. In addition to amylase, all Trichoderma species tested produced also extracellular cellulase and pectinase, except T. koningii in relation to the latter enzyme. The demonstration of various esterase isoenzymes in zymograms of the Trichoderma species and C. herbarum isolates was markedly improved by washing the mycelia with detergents or EDTA. This fact suggested that a major fraction of extracelular enzymes may remain attached to outside fungal cell wall after being excreted.O maracujá (Passiflora edulis Sims., um fruto tropical amplamente consumido, tem sua produção e a qualidade dos seus frutos reduzidos pela verrugose causada por Cladosporium herbarum. Este trabalho objetivou investigar o antagonismo de espécies de Trichoderma (T. polysporum, T.koningii, T. viride e T. harzianum contra C. herbarum, e estudar a produção de enzimas hidrolíticas extracelulares pelo fitopatógeno e

Evaluation of Trichoderma Isolates for Biological Control of Rhizoctonia Root Rot of Bean in Zanjan

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M. Khodae

2016-03-01

measured by dividing the difference between the radial growth of control and antagonized cultures of the pathogen by the radial growth of the control and multiplied by 100. All experiments were replicated three times. The third test was conducted with growing of Trichoderma and Rhizoctonia on microscopic slides covered with PDA and the interaction of hypha was studied using a microscope. For assessment of in vivo antagonistic activity, a greenhouse experiment was designed in 11 treatments with 3 replicates. Inoculum of R. solani AG-4 was prepared by inoculation of sterilized wheat grains with the fungus and the soil was inoculated with infected wheat grains. Bean seeds (red bean, cultivar Naz were treated with conidial suspension of Trichoderma (107 ml-1 conidia and planted in pots. Disease severity was recorded using a scale (1 to 5 according to Nelson and coworkers (1996 30 days after sowing. Shoot length, root length, shoot weight and root weight also were measured. For all data the analysis of variance (ANOVA was performed using SPSS Version 18. Table 1- Trichoderma isolates used for their antagonistic activity against Rhizoctonia solani isolates\tspecies T36\tT.viride T125\tT.viride T131\tT.viride T93\tT.virens T89\tT.parseomosum T25\tT.harzianum T12-0\tT.virens T12-N\tT.harzianum T19\tT.sp. T6\tT.sp. T95\tT.harzianum Results and Discussion: The results indicated that the mycoparasitism was the main mechanism accounting for antagonistic activity of Trichoderma isolates. In a competitive saprophytic ability test in a dual culture on PDA, T25, T93, T12-N and T12-0 performed more effective than the other isolates, as T25 covered the whole colony surface of R. solani, grew and sproulated on the pathogen mycelia after five days. Volatile metabolites of all Trichoderma isolates inhibited mycelial growth, whereas T12-N with % 32/73 at the same time culture and T6 with %56/08 at 72 h early culture of Trichoderma isolates had the highest effect on mycelial growth of R. solani

Production of spores of Trichoderma harzianum on sugar cane molasses and bagasse pith in solid state fermentation for biocontrol

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Jose A. Rodríguez-León

1999-01-01

Full Text Available Solid state fermentation was carried out for the production of spores from Trichoderma harzianum No 53 using sugar cane bagasse pith as solid matrix and sugar cane molasses as carbon and energy source. Different nitrogen sources such as urea, (NH42SO4 , NH4H2PO4 and (NH42HPO4 were added in the media to test their effect on spores production. Among these, urea was found most useful that resulted high no of spores (1x10(9/gDM. The influence of temperature and initial moisture of the substrate was studied through a 2² experimental plan design. No statistical differences were found within the range of 30-35ºC and 60-70% for temperature and moisture respectively. The biotechnological parameters of the process were derived from the Oxygen Uptake Rate (OUR pattern, which corresponded to the order of 10(9spores/g moist material. The specific growth rate, maintenance coefficient and the yield based on O2 consumption were 0.108 h-1, 0.001 g.O2/g.biomass.h and 2.7 g biomass/g O2 consumed, respectively.Esporos de Tricoderma harzianum Nº 53 foram produzidos por fermentação no estado sólido (FES utilizando bagaço de cana como suporte e melaço de cana como fonte de carbono. Diferentes fontes de nitrogênio foram testadas (uréia, (NH42 SO4 , NH4H2PO4 e (NH42HPO4 na produção de esporos. As mais elevadas concentrações de esporos (10(9 esporos/g de suporte úmido foram obtidas utilizando a uréia como fonte de nitrogênio. O efeito da temperatura e umidade inicial foram estudadas através da utilização da planificação experimental utilizando um modelo 2². Não foi encontrada diferença estatística na produção de esporos na faixa de temperatura compreendida entre 30-35 ° C e umidade inicial de 60-70%. Os parâmetros biotecnológicos foram determinados através da taxa de oxigênio consumido (OUR correspondente a uma produção de 10(9 esporos/g de suporte úmido. A taxa de crescimento especifico, coeficiente de manutenção e rendimento foram

Phylogeny and Taxonomical Investigation of Trichoderma spp. from Indian Region of Indo-Burma Biodiversity Hot Spot Region with Special Reference to Manipur

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Kamala, Th.; Devi, S. Indira; Sharma, K. Chandradev; Kennedy, K.

2015-01-01

Towards assessing the genetic diversity and occurrence of Trichoderma species from the Indian region of Indo-Burma Biodiversity hotspot, a total of 193 Trichoderma strains were isolated from cultivated soils of nine different districts of Manipur comprising 4 different agroclimatic zones. The isolates were grouped based on the morphological characteristics. ITS-RFLP of the rDNA region using three restriction digestion enzymes: Mob1, Taq1, and Hinf1, showed interspecific variations among 65 isolates of Trichoderma. Based on ITS sequence data, a total of 22 different types of representative Trichoderma species were reported and phylogenetic analysis showed 4 well-separated main clades in which T. harzianum was found to be the most prevalent spp. among all the Trichoderma spp. Combined molecular and phenotypic data leads to the development of a taxonomy of all the 22 different Trichoderma spp., which was reported for the first time from this unique region. All these species were found to produce different extrolites and enzymes responsible for the biocontrol activities against the harmful fungal phytopathogens that hamper in food production. This potential indigenous Trichoderma spp. can be targeted for the development of suitable bioformulation against soil and seedborne pathogens in sustainable agricultural practice. PMID:25699268

The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens.

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Rashmi, Singh; Maurya, Sudarshan; Upadhyay, Ram Sanmukh

2016-01-01

The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2%) and Trichoderma atroviride (40.3%) were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69U/mL) and β-1,3-glucanase activity (61.9U/mL). Th-m1 also showed high competitive saprophytic ability (CSA) among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%). Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

The improvement of competitive saprophytic capabilities of Trichoderma species through the use of chemical mutagens

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Rashmi Singh

2016-03-01

Full Text Available Abstract The antagonistic potential of Trichoderma strains was assayed by studying the effect of their culture filtrate on the radial growth of Sclerotium rolfsii, the causal agent of chickpea collar rot. Trichoderma harzianum-1432 (42.2% and Trichoderma atroviride (40.3% were found to be strong antagonists. To enhance their antagonistic potential, mutagenesis of these two selected strains was performed. Two mutants, Th-m1 and T. atroviride m1, were found to be more effective than their parent strains. The enzymatic activities of the selected parent and mutant strains were assayed, and although both mutants were found to have enhanced enzymatic activities compared to their respective parent strains, Th-m1 possessed the maximum cellulase (5.69 U/mL and β-1,3-glucanase activity (61.9 U/mL. Th-m1 also showed high competitive saprophytic ability (CSA among all of the selected parent and mutant strains, and during field experiments, Th-m1 was found to successfully possess enhanced disease control (82.9%.

Structural and Phylogenetic Analysis of Laccases from Trichoderma: A Bioinformatic Approach

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Cázares-García, Saila Viridiana; Vázquez-Garcidueñas, Ma. Soledad; Vázquez-Marrufo, Gerardo

2013-01-01

The genus Trichoderma includes species of great biotechnological value, both for their mycoparasitic activities and for their ability to produce extracellular hydrolytic enzymes. Although activity of extracellular laccase has previously been reported in Trichoderma spp., the possible number of isoenzymes is still unknown, as are the structural and functional characteristics of both the genes and the putative proteins. In this study, the system of laccases sensu stricto in the Trichoderma species, the genomes of which are publicly available, were analyzed using bioinformatic tools. The intron/exon structure of the genes and the identification of specific motifs in the sequence of amino acids of the proteins generated in silico allow for clear differentiation between extracellular and intracellular enzymes. Phylogenetic analysis suggests that the common ancestor of the genus possessed a functional gene for each one of these enzymes, which is a characteristic preserved in T. atroviride and T. virens. This analysis also reveals that T. harzianum and T. reesei only retained the intracellular activity, whereas T. asperellum added an extracellular isoenzyme acquired through horizontal gene transfer during the mycoparasitic process. The evolutionary analysis shows that in general, extracellular laccases are subjected to purifying selection, and intracellular laccases show neutral evolution. The data provided by the present study will enable the generation of experimental approximations to better understand the physiological role of laccases in the genus Trichoderma and to increase their biotechnological potential. PMID:23383142

Influence of protoplast fusion between two Trichoderma spp. on extracellular enzymes production and antagonistic activity.

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Hassan, Mohamed M

2014-11-02

Biological control plays a crucial role in grapevine pathogens disease management. The cell-wall degrading enzymes chitinase, cellulase and β-glucanase have been suggested to be essential for the mycoparasitism activity of Trichoderma species against grapevine fungal pathogens. In order to develop a useful strain as a single source of these vital enzymes, it was intended to incorporate the characteristics of two parental fungicides tolerant mutants of Trichoderma belonging to the high chitinase producing species T. harzianum and the high cellulase producing species T. viride , by fusing their protoplasts. The phylogeny of the parental strains was carried out using a sequence of the 5.8S-ITS region. The BLAST of the obtained sequence identified these isolates as T. harzianum and T. viride . Protoplasts were isolated using lysing enzymes and were fused using polyethylene glycol. The fused protoplasts have been regenerated on protoplast regeneration minimal medium supplemented with two selective fungicides. Among the 40 fast growing fusants, 17 fusants were selected based on their enhanced growth on selective media for further studies. The fusant strains were growing 60%-70% faster than the parents up to third generation. All the 17 selected fusants exhibited morphological variations. Some fusant strains displayed threefold increased chitinase enzyme activity and twofold increase in β-glucanase enzyme activity compared to the parent strains. Most fusants showed powerful antagonistic activity against Macrophomin aphaseolina , Pythium ultimum and Sclerotium rolfsii pathogens. Fusant number 15 showed the highest inhibition percentage (92.8%) against M. phaseolina and P. ultimum, while fusant number 9 showed the highest inhibition percentage (98.2%) against the growth of S. rolfsii. A hyphal intertwining and degradation phenomenon was observed by scanning electron microscope. The Trichoderma antagonistic effect against pathogenic fungal mycelia was due to the

Promoção do crescimento do feijoeiro e controle da antracnose por Trichoderma spp

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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.

Effect of Trichoderma harzianum biomass and Bradyrhizobium sp. strain NC 92 to control leaf blight disease of bambara groundnut (Vigna subterranea caused by Rhizoctonia solani in the field

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Mana Kanjanamaneesathian

2007-01-01

Full Text Available Four hundred and sixty two strains of Trichoderma spp. were isolated from 23 soil samples in which groundnut (Arachis hypogaea L. and bambara groundnut (Vigna subterranea L. had been planted in Songkhla, Phattalung, Nakhon Si Thammarat, Narathiwat and Yala provinces. These fungi were tested against Rhizoctonia solani, a causal agent of leaf blight of bambara groundnut, using dual culture technique on PDA medium. Among 462 isolates tested, 226 isolates had an ability to overgrow R. solani completely. Further testing found 13 isolates having the ability to parasitize mycelia of R. solani. Among these isolates, ThB-1-54 produced a cellulolytic enzyme on congo-red agar. This isolate was later identified as T. harzianum Rifai. In the field test, applying biomass of the isolate ThB-1-54 cultured on ground mesocarp fiber of oil palm, the combination of the isolate ThB-1-54 on ground mesocarp fiber of oil palm and Bradyrhizobium sp. (strain NC 92, or fungicide (iprodione had no effect on disease severity, yield, or the amount of total nitrogen content in stems or seeds of bambara groundnut plant.

Detecting the Hormonal Pathways in Oilseed Rape behind Induced Systemic Resistance by Trichoderma harzianum TH12 to Sclerotinia sclerotiorum.

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Alkooranee, Jawadayn Talib; Aledan, Tamarah Raad; Ali, Ali Kadhim; Lu, Guangyuan; Zhang, Xuekun; Wu, Jiangsheng; Fu, Chunhua; Li, Maoteng

2017-01-01

Plants have the ability to resist pathogen attack after infection or treatment with biotic and abiotic elicitors. In oilseed rape plant Brassica napus AACC and in the artificially synthesized Raphanus alboglabra RRCC, the root-colonizing Trichoderma harzianum TH12 fungus triggers induced systemic resistance (ISR), and its culture filtrate (CF) triggers a systemic acquired resistance (SAR) response against infection by the Sclerotinia sclerotiorum. Salicylic acid (SA) and jasmonic acid/ethylene (JA/ET) are plant hormone signals that play important roles in the regulation of ISR and SAR. In this study, at six different time points (1, 2, 4, 6, 8 and 10 days post-infection [dpi]), six resistance genes were used as markers of signaling pathways: JA/ET signaling used AOC3, PDF1.2 and ERF2 genes, while PR-1, TGA5 and TGA6 genes were used as markers of SA signaling. The results of quantitative real-time polymerase chain reaction (qRT-PCR) showed that AOC3, PDF1.2 and ERF2 expression levels in infected leaves of AACC and RRCC increase at 1 and 2 dpi with S. sclerotiorum or inoculation with TH12. PR-1, TGA5 and TGA6 expression levels increased at 8 and 10 dpi in infected leaves. PR-1, TGA5 and TGA6 expression levels increased early in plants treated with CF in both of the healthy genotypes. Furthermore, induction of SA- and JA/ET-dependent defense decreased disease symptoms in infected leaves at different times. The results suggest that the RRCC genotype exhibits resistance to disease and that the ability of TH12 and its CF to induce systemic resistance in susceptible and resistant oilseed rape genotypes exists. In addition, the results indicate for the first time that in RRCC the SA signaling pathway is involved in resistance to necrotrophic pathogens.

Trichoderma species from the cacao agroecosystem with biocontrol potential of Moniliophthora roreri

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Omar Reyes-Figueroa

2016-01-01

Full Text Available La moniliasis del cacao ( Moniliophthora roreri es la principal limitante parasítica de la producción de cacao ( Theobroma cacao en México. Una alternativa sostenible para el control de la enfermedad es el uso del hongo Trichoderma . El objetivo del presente estudio fue seleccionar aislamientos nativos de Trichoderma con las mejores características antagónicas y fisiológicas in vitro, para el control de M. roreri . Para ello, se caracterizaron 50 aislamientos de Trichoderma , obtenidos del agroecosistema cacao. El crecimiento micelial y la producción de conidios a 25, 30 y 35 °C se consideraron variables fisiológicas. El micoparasitismo, antibiosis y antagonismo potencial fueron las variables antagónicas. Se encontraron diferencias significativas ( P = 0.0001 en todas las variables evaluadas . El intervalo de temperatura óptima para el crecimiento micelial y producción de conidios fue de 25 a 30 °C. El micoparasitismo varió de 0 a 100 % y solo los aislamientos de seis especies mostraron esta característica. La antibiosis osciló entre 6.8 y 55.5 %, y el antagonismo potencial varió de 3.4 a 69 %. Trichoderma virens (TTC017 y T. harzianum (TTC090, TTC039, TTC073 mostraron el mejor biocontrol potencial in vitro , por lo que son cepas prometedoras para futuras investigaciones sobre control biológico de la moniliasis del cacao.

Airborne signals from Trichoderma fungi stimulate iron uptake responses in roots resulting in priming of jasmonic acid-dependent defences in shoots of Arabidopsis thaliana and Solanum lycopersicum.

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Martínez-Medina, Ainhoa; Van Wees, Saskia C M; Pieterse, Corné M J

2017-11-01

Root colonization by Trichoderma fungi can trigger induced systemic resistance (ISR). In Arabidopsis, Trichoderma-ISR relies on the transcription factor MYB72, which plays a dual role in the onset of ISR and the activation of Fe uptake responses. Volatile compounds (VCs) from rhizobacteria are important elicitors of MYB72 in Arabidopsis roots. Here, we investigated the mode of action of VCs from Trichoderma fungi in the onset of ISR and Fe uptake responses. VCs from Trichoderma asperellum and Trichoderma harzianum were applied in an in vitro split-plate system with Arabidopsis or tomato seedlings. Locally, Trichoderma-VCs triggered MYB72 expression and molecular, physiological and morphological Fe uptake mechanisms in Arabidopsis roots. In leaves, Trichoderma-VCs primed jasmonic acid-dependent defences, leading to an enhanced resistance against Botrytis cinerea. By using Arabidopsis micrografts of VCs-exposed rootstocks and non-exposed scions, we demonstrated that perception of Trichoderma-VCs by the roots leads to a systemic signal that primes shoots for enhanced defences. Trichoderma-VCs also elicited Fe deficiency responses and shoot immunity in tomato, suggesting that this phenomenon is expressed in different plant species. Our results indicate that Trichoderma-VCs trigger locally a readjustment of Fe homeostasis in roots, which links to systemic elicitation of ISR by priming of jasmonic acid-dependent defences. © 2017 John Wiley & Sons Ltd.

Genetic analysis reveals diversity and genetic relationship among Trichoderma isolates from potting media, cultivated soil and uncultivated soil.

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Al-Sadi, Abdullah M; Al-Oweisi, Fatma A; Edwards, Simon G; Al-Nadabi, Hamed; Al-Fahdi, Ahmed M

2015-07-28

Trichoderma is one of the most common fungi in soil. However, little information is available concerning the diversity of Trichoderma in soil with no previous history of cultivation. This study was conducted to investigate the most common species and the level of genetic relatedness of Trichoderma species from uncultivated soil in relation to cultivated soil and potting media. A total of 24, 15 and 13 Trichoderma isolates were recovered from 84 potting media samples, 45 cultivated soil samples and 65 uncultivated soil samples, respectively. Analysis based on the internal transcribed spacer region of the ribosomal RNA (rRNA) and the translation elongation factor gene (EF1) indicated the presence of 9 Trichoderma species: T. harzianum (16 isolates), T. asperellum (13), T. citrinoviride (9), T. orientalis (3), T. ghanense (3), T. hamatum (3), T. longibrachiatum (2), T. atroviride (2), and T. viride (1). All species were found to occur in potting media samples, while five Trichoderma species were recovered from the cultivated soils and four from the uncultivated soils. AFLP analysis of the 52 Trichoderma isolates produced 52 genotypes and 993 polymorphic loci. Low to moderate levels of genetic diversity were found within populations of Trichoderma species (H = 0.0780 to 0.2208). Analysis of Molecular Variance indicated the presence of very low levels of genetic differentiation (Fst = 0.0002 to 0.0139) among populations of the same Trichoderma species obtained from the potting media, cultivated soil and uncultivated soil. The study provides evidence for occurrence of Trichoderma isolates in soil with no previous history of cultivation. The lack of genetic differentiation among Trichoderma populations from potting media, cultivated soil and uncultivated soil suggests that some factors could have been responsible for moving Trichoderma propagules among the three substrates. The study reports for the first time the presence of 4 Trichoderma species in Oman: T

Antipathy of Trichoderma against Sclerotium rolfsii Sacc.: Evaluation of Cell Wall-Degrading Enzymatic Activities and Molecular Diversity Analysis of Antagonists.

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Hirpara, Darshna G; Gajera, Harsukh P; Hirpara, Hitesh Z; Golakiya, Balubhai A

2017-01-01

The fungus Trichoderma is a teleomorph of the Hypocrea genus and associated with biological control of plant diseases. The microscopic, biochemical, and molecular characterization of Trichoderma was carried out and evaluated for in vitro antagonistic activity against the fungal pathogen Sclerotium rolfsii causing stem rot disease in groundnut. In total, 11 isolates of Trichoderma were examined for antagonism at 6 and 12 days after inoculation (DAI). Out of 11, T. virens NBAII Tvs12 evidenced the highest (87.91%) growth inhibition of the test pathogen followed by T. koningii MTCC 796 (67.03%), T. viride NBAII Tv23 (63.74%), and T. harzianum NBAII Th1 (60.44%). Strong mycoparasitism was observed in the best antagonist Tvs12 strain during 6-12 DAI. The specific activity of cell wall-degrading enzymes - chitinase and β-1,3-glucanase - was positively correlated with growth inhibition of the test pathogen. In total, 18 simple sequence repeat (SSR) polymorphisms were reported to amplify 202 alleles across 11 Trichoderma isolates. The average polymorphism information content for SSR markers was found to be 0.80. The best antagonist Tvs 12 was identified with 7 unique SSR alleles amplified by 5 SSR markers. Clustering patterns of 11 Trichoderma strains showed the best antagonist T. virens NBAII Tvs 12 outgrouped with a minimum 3% similarity from the rest of Trichoderma. © 2017 S. Karger AG, Basel.

Structural and mechanistic analysis of engineered trichodiene synthase enzymes from Trichoderma harzianum: towards higher catalytic activities empowering sustainable agriculture.

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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.

Fungal and plant gene expression during the colonization of cacao seedlings by endophytic isolates of four Trichoderma species.

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Bailey, B A; Bae, H; Strem, M D; Roberts, D P; Thomas, S E; Crozier, J; Samuels, G J; Choi, Ik-Young; Holmes, K A

2006-11-01

Endophytic isolates of Trichoderma species are being considered as biocontrol agents for diseases of Theobroma cacao (cacao). Gene expression was studied during the interaction between cacao seedlings and four endophytic Trichoderma isolates, T. ovalisporum-DIS 70a, T. hamatum-DIS 219b, T. harzianum-DIS 219f, and Trichoderma sp.-DIS 172ai. Isolates DIS 70a, DIS 219b, and DIS 219f were mycoparasitic on the pathogen Moniliophthora roreri, and DIS 172ai produced metabolites that inhibited growth of M. roreri in culture. ESTs (116) responsive to endophytic colonization of cacao were identified using differential display and their expression analyzed using macroarrays. Nineteen cacao ESTs and 17 Trichoderma ESTs were chosen for real-time quantitative PCR analysis. Seven cacao ESTs were induced during colonization by the Trichoderma isolates. These included putative genes for ornithine decarboxylase (P1), GST-like proteins (P4), zinc finger protein (P13), wound-induced protein (P26), EF-calcium-binding protein (P29), carbohydrate oxidase (P59), and an unknown protein (U4). Two plant ESTs, extensin-like protein (P12) and major intrinsic protein (P31), were repressed due to colonization. The plant gene expression profile was dependent on the Trichoderma isolate colonizing the cacao seedling. The fungal ESTs induced in colonized cacao seedlings also varied with the Trichoderma isolate used. The most highly induced fungal ESTs were putative glucosyl hydrolase family 2 (F3), glucosyl hydrolase family 7 (F7), serine protease (F11), and alcohol oxidase (F19). The pattern of altered gene expression suggests a complex system of genetic cross talk occurs between the cacao tree and Trichoderma isolates during the establishment of the endophytic association.

Assisted phytoextraction of heavy metals: compost and Trichoderma effects on giant reed (Arundo donax L. uptake and soil N-cycle microflora

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Nunzio Fiorentino

2013-12-01

Full Text Available Little information is available as to the real effectiveness of the phytoextraction remediation technique, since laboratory experiments are still the most common way in which this is measured. Given this, an experiment on a cadmium-polluted soil was carried out in open field conditions in Southern Italy with the aim of assessing the growth and the phytoextraction potential of giant reed (Arundo donax L. Compost fertilisation and Trichoderma harzianum A6 inoculations were used to verify the possibility of increasing the metal uptake of the crop. Biomass yield of giant reed in the first growth season (average 12.8 Mg ha–1 was not affected by the Cd concentration in the soil and this increased significantly with compost fertilisation (13.8 Mg ha–1. Both compost fertilisation and T. harzianum inoculation increased cadmium uptake and translocation in leaves. Nitrifying bacteria was shown to be a useful tool to biomonitor soil quality. These results proved the suitability of the giant reed for assisted-phytoremedation with the use of compost fertilisation and T. harzianum.

Potential of genes and gene products from Trichoderma sp. and Gliocladium sp. for the development of biological pesticides.

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Lorito, M; Hayes, C K; Zoina, A; Scala, F; Del Sorbo, G; Woo, S L; Harman, G E

1994-12-01

Fungal cell wall degrading enzymes produced by the biocontrol fungi Trichoderma harzianum and Gliocladium virens are strong inhibitors of spore germination and hyphal elongation of a number of phytopathogenic fungi. The purified enzymes include chitinolytic enzymes with different modes of action or different substrate specificity and glucanolytic enzymes with exo-activity. A variety of synergistic interactions were found when different enzymes were combined or associated with biotic or abiotic antifungal agents. The levels of inhibition obtained by using enzyme combinations were, in some cases, comparable with commercial fungicides. Moreover, the antifungal interaction between enzymes and common fungicides allowed the reduction of the chemical doses up to 200-fold. Chitinolytic and glucanolytic enzymes from T. harzianum were able to improve substantially the antifungal ability of a biocontrol strain of Enterobacter cloacae. DNA fragments containing genes encoding for different chitinolytic enzymes were isolated from a cDNA library of T. harzianum and cloned for mechanistic studies and biocontrol purposes. Our results provide additional information on the role of lytic enzymes in processes of biocontrol and strongly suggest the use of lytic enzymes and their genes for biological control of plant diseases.

SELEKSI DAN UJI ANTAGONISME TRICHODERMA SPP. ISOLAT TAHAN FUNGISIDA NABATI TERHADAP PERTUMBUHAN PHYTOPHTHORA CAPSICI

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Joko Prasetyo, Efri & Radix Suharjo .

2011-11-01

Full Text Available Screening and testing of isolate of Trichoderma spp. resistant to botanical fungicides on the growth of P. capsici. The research was conducted at the Plant Disease laboratory, Plant Protection Department, Faculty of Agriculture, Lampung University from March – November 2007. The aim of the research was to find isolate of Trichoderma spp. resistant to botanical fungicide  which still have its capability to inhibit the growth of P. capsici. Completely randomized design with three replicates was used in the research. Data collected in this research were isolates of Trichoderma spp. resistant to botanical fungicides and inhibition percentage of isolates of Trichoderma spp. resistant to botanical fungicide on the growth of P. capsici. The data were analysed with anova and continued with LSD test at 5% of significant level. As much as 9 isolates of Trichoderma spp. resistant to botanical fungicide were found. Three isolates of T. viride could survive in the 100% of tumeric powder (C, east indian galangale powder (A, and clove leaf powder (B. Two isolates of T. harzianum could survive in the 100% of tumeric powder (A and clove leaf powder (A. One isolate survived in the 90% of east indian galangale powder (D. Two isolates of T. koningii also could survive in the 100% of tumeric powder (A  and clove leaf powder (A. One isolate survived in the 90% of east indian galangale powder (C. All of the isolates  inhibited  P. capsici. All of the isolates found statistically had the same capability to inhibit the growth of P. capsici.

Potential antagonism of some Trichoderma strains isolated from Moroccan soil against three phytopathogenic fungi of great economic importance

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Wafaa MOKHTARI

2017-09-01

Full Text Available In this study, 17 Trichoderma strains were isolated from different soils (crop fields and Argan forests in Morocco. Purified monospore cultures were identified using molecular methods and tested for their potential antagonism against three phytopathogenic fungi (Fusarium oxyxporum, verticillium dahlia and rhizoctonia solani. After DNA extraction, translation elongation factor (tef1 was amplified in extracts of 17 strains, sequenced and compared with their ex-types. As a result, three species were identified among the strains, which clustered in two different subclades of Trichoderma: the species T. afroharzianum, and T. guizhouense belong to the Harzianum clade, while T. longibrachiatum belongs to the Longibrachiatum clade. Investigation of potential antagonistic effects of these strains against the soil-borne phytopathogens F. oxysporum, R. solani and V. dahliae was conducted in a dual culture plate assay, using 17 promising Trichoderma strains that have been selected based on a polymerase chain reaction (PCR screening approach. In vitro, Trichoderma isolates showed effective antagonistic performance by decreasing soil borne pathogens mycelium radial growth. Trichoderma afroharzianum showed the highest Percentage of Radial Inhibition Growth (PRIG %. The highest PRIG% = 98% was for 8A2.3 isolate against R. solani and the lowest PRIG%= 67% for T9i10 against F. oxysporum. On the other hand, T9i12, which is T. reesei species, led to a high radial inhibition of pathogens’ mycelium.

Identification of Trichoderma Species Using Partial Sequencing of nrRNA and tef1α Genes with Report of Trichoderma capillare in Iran Mycoflore

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mehdi Mehrabi-Koushki

2018-01-01

reverse primers by Macrogen Company. The Sequences were edited and assembled using BioEdit v. 7.0.9.0 (10 and DNA Baser Sequence Assembeler v4 programs (2013, Heracle BioSoft, www.DnaBaser.com, respectively. These sequences were submit-queried against the NCBI non-redundant database and related to known DNA sequences by BLASTn algorithm to assign putative identity. They also were subjected to the TrichO Key (5 and TrichoBLAST (15 for more characterization. The phylogenetic tree was constructed through maximum likelihood analysis based on tef1α sequence under K2+G model. The tree was rooted to close species of N. macroconidialis. Result and Discussion: Approximately 550 and 850 bases of the ITS and tef1α regions were sequenced from the isolates studied and then deposited in the GenBank (Table 2. The annotation of indexed sequences showed which multiple insertion-type frame shifts have interestingly occurred into the reading frame of tef1α gene belonging to isolate of T. capillare Isf-7 (Fig. 3. To identify isolates of Trichoderma, ITS and tef1α sequences were subjected to the TrichO Key (5, TrichoBLAST (15 and BLASTn Search. The analysis of ITS and tef1α sequences (Table 2, Fig. 2, in combination with morphology (Table 1, showed which the isolates place in seven species as follow: T. harzianum Rifai, T. virens (J.H. Mill., Giddens & A.A. Foster Arx, T. pleuroticola Yu & Park, T. asperellum Samuels, Lieckf & Nirenberg, T. koningiopsis Oudem., T. brevicompactum Kraus, Kubicek & Gams and T. capillare Samuels & Kubicek. In BLASTn search, ITS and tef1α regions separately provided unambiguous identification for isolates of T. virens, T. koningiopsis and T. brevicompactum while ITS region provided ambiguous identification for Isolates of Trichoderma harzianum, T. capillare, T. pleuroticola and T. asperellum. Here, tef1α region could provide more accurate identification as good DNA barcoding (Table 2. The isolates showed the sequence identity ranging from 96 to 100% for tef

Distribution and Genetic Variability of Fusarium oxysporum Associated with Tomato Diseases in Algeria and a Biocontrol Strategy with Indigenous Trichoderma spp.

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Debbi, Ali; Boureghda, Houda; Monte, Enrique; Hermosa, Rosa

2018-01-01

Fifty fungal isolates were sampled from diseased tomato plants as result of a survey conducted in seven tomato crop areas in Algeria from 2012 to 2015. Morphological criteria and PCR-based identification, using the primers PF02 and PF03, assigned 29 out of 50 isolates to Fusarium oxysporum ( Fo ). The banding patterns amplified for genes SIX1, SIX3 and SIX4 served to identify races 2 and 3 of Fo f. sp. lycopersici (FOL), and Fo f. sp. radicis lycopersici (FORL) among the Algerian isolates. All FOL isolates showed pathogenicity on the susceptible tomato cv. "Super Marmande," while nine of out 10 Algerian FORL isolates were pathogenic on tomato cv. "Rio Grande." Inter simple sequence repeat (ISSR) fingerprints showed high genetic diversity among Algerian Fo isolates. Seventeen Algerian Trichoderma isolates were also obtained and assigned to the species T. asperellum (12 isolates), T. harzianum (four isolates) and T. ghanense (one isolate) based on ITS and tef1 α gene sequences. Different in vitro tests identified the antagonistic potential of native Trichoderma isolates against FORL and FOL. Greenhouse biocontrol assays performed on "SM" tomato plants with T. ghanense T8 and T. asperellum T9 and T17, and three Fo isolates showed that isolate T8 performed well against FORL and FOL. This finding was based on an incidence reduction of crown and root rot and Fusarium wilt diseases by 53.1 and 48.3%, respectively.

Distribution and Genetic Variability of Fusarium oxysporum Associated with Tomato Diseases in Algeria and a Biocontrol Strategy with Indigenous Trichoderma spp.

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Ali Debbi

2018-02-01

Full Text Available Fifty fungal isolates were sampled from diseased tomato plants as result of a survey conducted in seven tomato crop areas in Algeria from 2012 to 2015. Morphological criteria and PCR-based identification, using the primers PF02 and PF03, assigned 29 out of 50 isolates to Fusarium oxysporum (Fo. The banding patterns amplified for genes SIX1, SIX3 and SIX4 served to identify races 2 and 3 of Fo f. sp. lycopersici (FOL, and Fo f. sp. radicis lycopersici (FORL among the Algerian isolates. All FOL isolates showed pathogenicity on the susceptible tomato cv. “Super Marmande,” while nine of out 10 Algerian FORL isolates were pathogenic on tomato cv. “Rio Grande.” Inter simple sequence repeat (ISSR fingerprints showed high genetic diversity among Algerian Fo isolates. Seventeen Algerian Trichoderma isolates were also obtained and assigned to the species T. asperellum (12 isolates, T. harzianum (four isolates and T. ghanense (one isolate based on ITS and tef1α gene sequences. Different in vitro tests identified the antagonistic potential of native Trichoderma isolates against FORL and FOL. Greenhouse biocontrol assays performed on “SM” tomato plants with T. ghanense T8 and T. asperellum T9 and T17, and three Fo isolates showed that isolate T8 performed well against FORL and FOL. This finding was based on an incidence reduction of crown and root rot and Fusarium wilt diseases by 53.1 and 48.3%, respectively.

Биологическая и биохимическая стабильность метаболитов почвенного сапрофита Trichoderma harzianum Rifai

OpenAIRE

Пакина, Е.; Шнейдер, Ю.; Смирнова, И.

2009-01-01

Исследована стабильность метаболитов почвенного сапрофита Trichoderma harzianum Rifai в процессе длительного хранения. Показано, что в процессе длительного хранения культуры почвенный сапрофит Trichoderma harzianum Rifai сохраняет свою ферментативную и биологическую активность.

Adaptation of Trichoderma Species to Pesticide Confidor and Evaluation of their Growth Ability in the Media Containing Confidor

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Farnaz Ershadfath

2015-12-01

Full Text Available Introduction: Contamination caused by pesticides is considered as one of the environmental problems. Bioremediation is exploiting the ability of microorganisms to remove pollutants. Trichoderma species are free-living fungi that exist naturally in the environment. These fungi have the ability to uptake some contaminants biologically. The aim of this study is to evaluate the effect of Confidor, as an environmental contaminant, on the growth ability of Trichoderma sp. as a contaminant absorber. Materials and methods: Five species of Trichoderma fungi were cultured in PDA media. Then the fungi were adapted with 3 different concentrations of Confidor gradually (5, 10 and 20 mg/l. The diameter of the fungal colonies growing in different concentrations of the toxin, were measured after 24 hr and were compared with the control samples (medium without toxin. Results: Results showed that in all species of fungi the colony diameters increased significantly with increasing toxin concentrations. The largest colony diameter was related to T.tomentosum, T.asperellum and T.harzianum (88.88, 87.5 and 86.95%, respectively at the concentration of 20 mg of toxic. Also, in all studied fungal species, in the medium containing 20 (mg/ l of toxic, the aerial hyphae expanded much thicker and faster than other concentrations. Discussion and conclusion: The results indicate a significant increase in the growth ability of Trichoderma strains with increasing Confidor concentration. Therefore it could be concluded that Trichoderma fungi have a high potentiality for biodegradation of Confidor.

IDENTIFICAÇÃO E UTILIZAÇÃO DE Trichoderma spp. ARMAZENADOS E NATIVOS NO BIOCONTROLE DE Sclerotinia sclerotiorum

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GERARDA BEATRIZ PINTO DA SILVA

2015-01-01

Full Text Available The fungus Sclerotinia sclerotiorum , handles significant losses in lettuce production. Being a soil borne fungus, its management is difficult, and an alternative is the use of biological control using species of the Trichoderma genus. Thus, the objectives of this study were to identify native species of Trichoderma spp. presents in the soil with (CP and without white mold (SP, evaluate the growth rate and in vitro antagonism of Trichoderma spp. against S. sclerotiorum and to verify the biocontrol potential of Trichoderma spp. microbi- olized lettuce seeds, growing in substrate infested with S. sclerotiorum . Trichoderma spp. isolates were obtained from areas with and without history of white mold or stored in water. Mycelial growth rate and sporu- lation of the Trichoderma spp. isolates and control of Trichoderma spp. versus S. sclerotiorum in the in vitro essays. For the in vivo essay, lettuce seeds were microbiolized with Trichoderma spp. and the substrate was infested with S. sclerotiorum . The native isolates of Trichoderma identified belong to T. koningiopsis and T. asperellum species. The CP isolates had higher mycelial growth rates when compared to the SP isolates and stored while the stored isolates showed better responses in confrontation. The application of Trichoderma spp. promoted higher seedlings quality compared to control, as well as good seedlings development in the presence of the pathogen.

Downy mildew resistance induced by Trichoderma harzianum T39 in susceptible grapevines partially mimics transcriptional changes of resistant genotypes

Science.gov (United States)

2012-01-01

Background Downy mildew, caused by Plasmopara viticola, is one of the most severe diseases of grapevine and is commonly controlled by fungicide treatments. The beneficial microorganism Trichoderma harzianum T39 (T39) can induce resistance to downy mildew, although the molecular events associated with this process have not yet been elucidated in grapevine. A next generation RNA sequencing (RNA-Seq) approach was used to study global transcriptional changes associated with resistance induced by T39 in Vitis vinifera Pinot Noir leaves. The long-term aim was to develop strategies to optimize the use of this agent for downy mildew control. Results More than 14.8 million paired-end reads were obtained for each biological replicate of T39-treated and control leaf samples collected before and 24 h after P. viticola inoculation. RNA-Seq analysis resulted in the identification of 7,024 differentially expressed genes, highlighting the complex transcriptional reprogramming of grapevine leaves during resistance induction and in response to pathogen inoculation. Our data show that T39 has a dual effect: it directly modulates genes related to the microbial recognition machinery, and it enhances the expression of defence-related processes after pathogen inoculation. Whereas several genes were commonly affected by P. viticola in control and T39-treated plants, opposing modulation of genes related to responses to stress and protein metabolism was found. T39-induced resistance partially inhibited some disease-related processes and specifically activated defence responses after P. viticola inoculation, causing a significant reduction of downy mildew symptoms. Conclusions The global transcriptional analysis revealed that defence processes known to be implicated in the reaction of resistant genotypes to downy mildew were partially activated by T39-induced resistance in susceptible grapevines. Genes identified in this work are an important source of markers for selecting novel

Identification of melatonin in Trichoderma spp. and detection of melatonin content under controlled-stress growth conditions from T. asperellum.

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Liu, Tong; Zhao, Fengzhou; Liu, Zhen; Zuo, Yuhu; Hou, Jumei; Wang, Yanjie

2016-07-01

T. koningii, T. harzianum, T. asperellum, T. longibrachiatum, and T. viride were analyzed using liquid chromatography-tandem mass spectrometry to determine whether melatonin is present. Results showed that there were abundant amounts of endogenous melatonin in five Trichoderma species, but no melatonin was found in any of the culture filtrates. T. asperellum had the highest amount of melatonin (27.588 ± 0.326 μg g(-1) dry mass), followed by T. koningii, T. harzianum, T. longibrachiatum, and T. viride. The endogenous melatonin content of T. asperellum in controlled-stress growth conditions was also detected. The data showed that chemical stressors (CdCl2 , CuSO4 , and H2 O2 ) provoked an increase in endogenous melatonin levels. CdCl2 had the highest stimulatory effect on melatonin production, as the product reached reaching up to three times the melatonin content of the control. NaCl stimulated a decrease of melatonin. Acidic conditions (pH 3 and pH 5) as well as slightly alkaline conditions (pH 9) resulted in an increase in the melatonin content, whereas pH11 resulted in a significant decrease in the melatonin content, only 12.276 ± 0.205 μg g(-1) dry mass. The current study is first to report melatonin content and the change of melatonin content under different stress situations in Trichoderma spp. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Electrochemical DNA biosensor for the detection of Trichoderma harzianum based on a gold electrode modified with a composite membrane made from an ionic liquid, ZnO nanoparticles and chitosan, and by using acridine orange as a redox indicator

International Nuclear Information System (INIS)

Siddiquee, S.; Yusof, N.A.; Salleh, A.B.; Tan, S.G.; Bakar, F.A.

2011-01-01

An electrochemical DNA biosensor was developed that is based on a gold electrode modified with a nanocomposite membrane made from an ionic liquid, ZnO nanoparticles and chitosan. A single-stranded DNA probe was immobilized on this electrode. Acridine orange was used as the hybridization probe for monitoring the hybridization of the target DNA. The biosensor was capable of detecting target DNA in the concentration range from 1.0 x 10 -14 to 1.8 x 10 -4 mol L -1 , with a detection limit of 1.0 x 10 -15 mol L -1 . The approach towards constructing a DNA biosensor allows studies on the hybridization even with crude DNA fragments and also to analyze sample obtained from real samples. The results show that the DNA biosensor has the potential for sensitive detection of a specific sequence of the Trichoderma harzianum gene and provides a quick, sensitive and convenient method for the study of microorganisms. (author)

Impact of a native Streptomyces flavovirens from mushroom compost on green mold control and yield of Agaricus bisporus.

Science.gov (United States)

Šantrić, Ljiljana; Potočnik, Ivana; Radivojević, Ljiljana; Umiljendić, Jelena Gajić; Rekanović, Emil; Duduk, Bojan; Milijašević-Marčić, Svetlana

2018-05-18

Thirty-five actinobacterial isolates, obtained from button mushroom (Agaricus bisporus) substrates (i.e., compost in different phases of composting, black peat or casing layer) in Serbia in 2014-2016 were tested in vitro against the causal agents of green mold in cultivated mushroom. Out of six most promising isolates, A06 induced 42.4% in vitro growth inhibition of Trichoderma harzianum T54, and 27.6% inhibition of T. aggressivum f. europaeum T77. The novel strain A06 was identified as Streptomyces flavovirens based on macroscopic and cultural characteristics and 16S rDNA sequence and used in mushroom growing room experiments. Actinobacteria had no negative influence on mycelial growth of the cultivated mushroom in compost in situ. Isolate S. flavovirens A06 enhanced mushroom yield significantly, up to 31.5%. The A06 isolate was more efficient in enhancing yield after inoculation with the compost mold T. aggressivum (26.1%), compared to casing mold T. harzianum (8%). Considering disease incidence, actinobacteria significantly prevented green mold in compost caused by T. aggressivum (6.8%). However, fungicide prochloraz-Mn had a more significant role in reducing symptoms of casing mold, T. harzianum, in comparison with actinobacteria (24.2 and 11.8%, respectively). No significant differences between efficacies of S. flavovirens A06 and the fungicide prochloraz-Mn against T. aggressivum were revealed. These results imply that S. flavovirens A06 can be used to increase mushroom yield and contribute to disease control against the aggressive compost green mold disease caused by Trichoderma aggressivum.

Competition in artifical plant growth media by Trichoderma spp

DEFF Research Database (Denmark)

Sarocco, Sabrina; Lübeck, Mette; Vannacci, Giovanni

of the reason why more biocontrol agents are reaching the market place. A comparative evaluation of life strategies of both the pathogen and its antagonists is required to predict the fate of a biopesticide in agricultural systems.The objectives of this work have been: 1) to screen a collection of Trichoderma......The key to achieve successful, reproducible biological control is the gradual appreciation that knowledge of the ecological interactions taking place in soil and root environments is required to predict the condition under which biological control can be achieved and, indeed, it may be part...... inoculation procedures were employed. Four potential biocontrol agents, all identified as T. harzianum according to the ITS sequences, were selected according to their ability of reducing radish damping off, and were employed in a Competitive Saprophytic Ability (CSA) test, in order to investigate the role...

Marine Isolates of Trichoderma spp. as Potential Halotolerant Agents of Biological Control for Arid-Zone Agriculture ▿ †

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Gal-Hemed, Inbal; Atanasova, Lea; Komon-Zelazowska, Monika; Druzhinina, Irina S.; Viterbo, Ada; Yarden, Oded

2011-01-01

The scarcity of fresh water in the Mediterranean region necessitates the search for halotolerant agents of biological control of plant diseases that can be applied in arid-zone agriculture irrigated with saline water. Among 29 Trichoderma strains previously isolated from Mediterranean Psammocinia sp. sponges, the greatest number of isolates belong to the Trichoderma longibrachiatum-Hypocrea orientalis species pair (9), H. atroviridis/T. atroviride (9), and T. harzianum species complex (7), all of which are known for high mycoparasitic potential. In addition, one isolate of T. asperelloides and two putative new species, Trichoderma sp. O.Y. 14707 and O.Y. 2407, from Longibrachiatum and Strictipilosa clades, respectively, have been identified. In vitro salinity assays showed that the ability to tolerate increasing osmotic pressure (halotolerance) is a strain- or clade-specific property rather than a feature of a species. Only a few isolates were found to be sensitive to increased salinity, while others either were halotolerant or even demonstrated improved growth in increasingly saline conditions. In vitro antibiosis assays revealed strong antagonistic activity toward phytopathogens due to the production of both soluble and volatile metabolites. Two marine-derived Trichoderma isolates, identified as T. atroviride and T. asperelloides, respectively, effectively reduced Rhizoctonia solani damping-off disease on beans and also induced defense responses in cucumber seedlings against Pseudomonas syringae pv. lachrimans. This is the first inclusive evaluation of marine fungi as potential biocontrol agents. PMID:21666030

Evaluation of Trichoderma spp. and Acibenzolar-S-Methyl (Bion® as resistance inducers in garlic (Allium sativum L. agains white rot Sclerotium cepivorum Berk. under field conditions

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Jiménez María A

2012-08-01

Full Text Available Inducers of biotic and abiotic resistance emerge as an alternative for the management of garlic white rot caused by the fungus Sclerotium cepivorum Berk. To evaluate the effect of these inducers on crop yield variables, final disease incidence (DI, the area under the curve of disease progress (AUDPC and variables associated with plant growth and yield of garlic, and experiment was set, using the fungi Trichoderma harzianum and T. koningiopsis with Bion® under field conditions in a randomized block design with eight treatments and four replications were incorporated. The treatments were conformed as follows: control, Trichoderma harzianum under commercial formulas Tricobiol® (Tri, Subiol® (S, T. koningiopsis (Tk, Bion® (B, T. koningiopsis + Bion (Tk+ B, Subiol Bion (S+B and Bion +Tricobiol (Tri+ B. The higher yields and growth variables correspond to those plants treated with Tk+ B treatment and coincided with some of the lowest AUDPC. The lower (DI was observed when applied Tk, Tk +B and Tri. These results indicated that the greatest protection against the disease was achieved with the combination of the antagonist with the chemical reflected in a decrease of AUDPC and a delay in the peak of infection. The total yield of garlic increased in the treatments in which both inducers and that stimulated plant productivity and thus the development and defense.

The Removal of Confidor Pesticide by Different Species of Trichoderma Fungi from Contaminated Waters

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Hossein Banejad

2017-01-01

Full Text Available Introduction: Pesticides are considered as the most important pollutants in surface water and groundwater. Neonicotinoids are new group of insecticides, derived from nicotine. Their physicochemical properties render them useful for a wide range of application techniques, including foliar, seed treatment, soil drench and stem applications. Confidor, the representative of the first generation of neonicotinoid insecticides, was patented in 1985 by Bayer and was placed on the market in 1991. The Canadian Pest Management Regulatory Agency considers confidor to have high potential for surface water contamination, leaching to groundwater and persistence in soils. Biodegradation is one of the most effective ways to destroy pesticides in the environment. The application of Bioremediation techniques is taken into consideration as an option to reduce or remove pollutants from the environment due to their low cost, high efficiency and environmentally friendly features. Bioremediation by using microorganisms has not any adverse effect after cleanup. The accumulator microorganism species, haven’t pathogenic properties and aren’t the cause of disease on the other organisms. The selection of a biomass for using in bioremediation is very important, it should be abundant in environment and adapted to environmental conditions. The aim of this study was to investigate the ability of various species of Trichoderma fungi to remove Confidor from contaminated water influenced by variables like pH, concentration of the confidor and time. Materials and Methods: In order to conduct this study three different fungal species belonging to the genus Trichoderma were used. The samples were transferred to PDA (Potato Dextrose Agar sterile solid media for in vitro testing usage. The samples were kept in refrigerator at 4◦C temperature, after the fungal biomass reached to maximal growth; the colonies were transferred to new media and used in our experiments as resources

IDENTIFICAÇÃO E UTILIZAÇÃO DE Trichoderma spp. ARMAZENADOS E NATIVOS NO BIOCONTROLE DE Sclerotinia sclerotiorum

OpenAIRE

GERARDA BEATRIZ PINTO DA SILVA; LEISE INÊS HECKLER; RICARDO FELICIANO DOS SANTOS; MIRIA ROSA DURIGO; ELENA BLUME

2015-01-01

The fungus Sclerotinia sclerotiorum , handles significant losses in lettuce production. Being a soil borne fungus, its management is difficult, and an alternative is the use of biological control using species of the Trichoderma genus. Thus, the objectives of this study were to identify native species of Trichoderma spp. presents in the soil with (CP) and without white mold (SP), evaluate the growth rate and in vitro antagonism of Trichoderma spp. against S. sclerotiorum and to ver...

Fungal diversity in the rhizosphere of endemic plant species of Tenerife (Canary Islands): relationship to vegetation zones and environmental factors

DEFF Research Database (Denmark)

Zachow, Christin; Berg, Christian; Müller, Henry

2008-01-01

, molecular analysis of fungal communities was determined by single-strand conformation polymorphism (SSCP) analysis using universal and specific primers for Trichoderma. The highly diverse fungal communities were mainly characterized by ectomycorrhiza-forming Basidiomycota and a high proportion of yet......-unidentified species. Besides, Trichoderma-specific SSCP resulted in low diversity of mainly cosmopolitan species, for example Hypocrea lixii/T. harzianum. The dominance of T. harzianum was confirmed by cultivation. All Trichoderma isolates show an extraordinarily high antagonistic potential towards different groups...... of plant pathogens, supporting the hypothesis of extensive colonization by highly competitive Trichoderma species from the continent. In contrast, biodiversity patterns of the whole fungal and plant communities follow the same ecological rules. Furthermore, a high statistical correlation between fungal...

Introducing a sustainable soil fertility system for chickpea ( Cicer ...

African Journals Online (AJOL)

): Trichoderma harzianum; (B3): Phosphate solubilizing bacteria + T. harzianum; and (B4): without biofertilizers were arranged in sub-sub plots. Results showed that green manure increased pod number and number of fertile pods per plant.

Regulation of Botrytis cinerea virulence genes in interaction with Trichoderma arundinaceum is mediated by the sesquiterpene harzianum A

Science.gov (United States)

Trichoderma includes a great diversity of species, some of them with the ability to control the growth of fungal phytopathogens. Many of these strains produce secondary metabolites that are able to inhibit the growth of their fungal preys. However, pathogens can also produce metabolites that in some...

Microbiological destruction of composite polymeric materials in soils

Science.gov (United States)

Legonkova, O. A.; Selitskaya, O. V.

2009-01-01

Representatives of the same species of microscopic fungi developed on composite materials with similar polymeric matrices independently from the type of soils, in which the incubation was performed. Trichoderma harzianum, Penicillium auranthiogriseum, and Clonostachys solani were isolated from the samples of polyurethane. Fusarium solani, Clonostachys rosea, and Trichoderma harzianum predominated on the surface of ultrathene samples. Ulocladium botrytis, Penicillium auranthiogriseum, and Fusarium solani predominated in the variants with polyamide. Trichoderma harzianum, Penicillium chrysogenum, Aspergillus ochraceus, and Acremonium strictum were isolated from Lentex-based composite materials. Mucor circinelloides, Trichoderma harzianum, and Penicillium auranthiogriseum were isolated from composite materials based on polyvinyl alcohol. Electron microscopy demonstrated changes in the structure of polymer surface (loosening and an increase in porosity) under the impact of fungi. The physicochemical properties of polymers, including their strength, also changed. The following substances were identified as primary products of the destruction of composite materials: stearic acid for polyurethane-based materials; imide of dithiocarbonic acid and 1-nonadecen in variants with ultrathene; and tetraaminopyrimidine and isocyanatodecan in variants with polyamide. N,N-dimethyldodecan amide, 2-methyloximundecanon and 2-nonacosane were identified for composites on the base of Lentex A4-1. Allyl methyl sulfide and imide of dithiocarbonic acid were found in variants with the samples of composites based on polyvinyl alcohol. The identified primary products of the destruction of composite materials belong to nontoxic compounds.

In vitro Antagonistic Mechanisms of Trichoderma spp. and Talaromyces flavus to Control Gaeumannomyces graminis var. tritici the Causal Agent of Wheat Take-all Disease

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Seddighe Mohammadi

2015-07-01

Full Text Available Wheat take-all disease caused by Gaeumannomyces graminis var. tritici has recently been detected in different regions of Iran. With respect to biocontrol effect of Trichoderma spp. on many pathogenic fungi, seven isolates of Trichoderma and four isolates of Talaromyces were in vitro evaluated in terms of their biological control against the disease causal agent. In dual culture test the five isolates showed efficient competition for colonization against pathogenic fungus and the highest percentages of inhibition belonging to Talaromyces flavus 60 and Talaromyces flavus 136 were 59.52 and 57.61%, respectively. Microscopic investigations showed that in regions where antagonistic isolates and Gaeumannomyces graminis var. tritici coincide, hyphal contact, penetration and fragmentation of Gaeumannomyces graminis var. tritici were observed. Investigating the effect of volatile and non-volatile compounds at 10 ml concentration showed that the highest inhibition percentage on mycelium growth of the pathogen caused by T. harzianum (44.76% and T. longibrachiatum (52.38% respectively.

Induced systemic resistance in cucumber and Arabidopsis thaliana by the combination of Trichoderma harzianum Tr6 and Pseudomonas sp.

NARCIS (Netherlands)

Alizadeh, H.; Behboudi, K.; Amadzadeh, M.; Javan-Nikkhah, M.; Zamioudis, C; Pieterse, C.M.J.; Bakker, P.A.H.M.

2013-01-01

Trichoderma species and fluorescent Pseudomonas spp. have been reported to induce systemic resistance in plants. In this study the effectiveness of a combination of these biological control agents on the efficacy of induced resistance was investigated in cucumber and the model plant Arabidopsis

Secretômica de Trichoderma atroviride e Trichoderma harzianum frente a Guignardia e citricarpa, agente etiológico da Pinta Preta dos Citros

OpenAIRE

Lima, Fernanda Blauth de

2016-01-01

Os agentes de controle biológico têm recebido grande reconhecimento, e o seu uso tem contribuído como um complemento ou substituição de agroquímicos. No entanto, existem poucos estudos sobre o controle biológico da Pinta Preta dos Citros, causada pelo fungo Guignardia citricarpa, o que impede o seu mercado in natura, além de prejudicar a sua exportação pelo uso intensivo de insumos químicos para controlar este patógeno. Os fungos do gênero Trichoderma são agentes de controle utilizados em tod...

White mold intensity on common bean in response to plant density, irrigation frequency, grass mulching, Trichoderma spp., and fungicide Intensidade do mofobranco em feijão em função de densidade de plantas, freqüência de irrigação, cobertura vegetal do solo, Trichoderma spp. e fungicida

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Trazilbo José de Paula Júnior

2009-02-01

Full Text Available The purpose of this study was to evaluate the efficiency of integrated managements on white mold control on common bean. Initially, in vitro testing was made to assess the antagonism of 11 Trichoderma isolates against Sclerotinia sclerotiorum and to investigate fungicides (fluazinam and procymidone inhibitory effects on those fungi. In two field experiments the following combinations were tested: irrigation frequencies (seven or 14 days, plant densities (six or 12 plants per meter, and three disease controls (untreated control, fungicide or Trichoderma spp.. In a third experiment plant densities were replaced by grass mulching treatments (with or without mulching. Fluazinam was applied at 45 and 55 days after emergence (DAE. The antagonists T. harzianum (experiments 1 and 3 and T. stromatica (experiment 2 were applied through sprinkler irrigation at 10 and 25 DAE, respectively. Most of the Trichoderma spp. were effective against the pathogen in vitro. Fluazinam was more toxic than procymidone to both the pathogen and the antagonist. Fungicide applications increased yield between 32 % and 41 %. In field one application of Trichoderma spp. did not reduce disease intensity and did not increase yield. The reduction from 12 to six plants per meter did not decrease yield, and disease severity diminished in one of the two experiments. It is concluded that of the strategies for white mold control just reduction of plant density and applications of fungicide were efficient.Objetivou-se estudar a eficácia de técnicas de manejo integrado no controle do mofo-branco em feijão. Inicialmente, foram feitos testes in vitro para avaliar o antagonismo de 11 isolados de Trichoderma contra Sclerotinia sclerotiorum e investigar os efeitos de fungicidas (fluazinam e procymidone sobre esses fungos. Em dois ensaios de campo, foram testadas estas combinações: freqüências de irrigação (sete ou 14 dias, densidades de plantas (seis ou 12 por metro e três controles

ISOLASI, KARAKTERISASI, DAN IDENTIFIKASI MIKOFLORA DARI RIZOSFER TANAH PERTANIAN TEBU (SACCHARUM OFFICINARUM L. SEBAGAI BAHAN AJAR KINGDOM FUNGI UNTUK SISWA KELAS X SMA

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Yesy Maulina Nadhifah

2016-10-01

Full Text Available Agricultural area, particularly in the rhizosfer area there are various species of mold which lives mainly in the soil. The students have known molds which live in nature but they have not recognized mold which lives in the soil. It is required to develop a teaching material in the form of module which is created based on a research result. The results of this study were; (1 it has been discovered 7 species of mold, which are Trichoderma harzianum, Penicillium nalgiovense, Penicillium corylophilum, Penicillium funiculosum, Penicillium camemberti, Aspergillus ochraceus, dan Penicillium citrinum, (2 the most dominant mold species was Trichoderma harzianum with the total number 1,8 x 104 cfu for every gram soil sample, and (3 there has been arranged of the learning module about fungi kingdom for students in high school level. Tanah pertanian, khususnya di daerah rizosfer, merupakan habitat dari berbagai spesies kapang yang hidup di dalamnya. Para siswa telah mengenal kapang yang tumbuh di alam tetapi mereka belum mengenal kapang yang hidup di dalam tanah pertanian. Perlu dikembangkan sebuah bahan ajar berupa modul yang dibuat berdasarkan hasil penelitian. Hasil penelitian ini, meliputi (1 ditemukan 7 spesies kapang, yaitu Trichoderma harzianum, Penicillium nalgiovense, Penicillium corylophilum, Penicillium funiculosum, Penicillium camemberti, Aspergillus ochraceus, dan Penicillium citrinum, (2 spesies kapang yang paling dominan adalah Trichoderma harzianum dengan total jumlah 1,8 x 104 cfu/g sampel tanah, dan (3 telah tersusun bahan ajar Biologi berupa modul Kingdom Fungi para siswa SMA.

Impacts of biocontrol products on Rhizoctonia disease of potato and soil microbial communities, and their persistence in soil

Science.gov (United States)

Four commercial biocontrol formulations (Bacillus subtilis GB03, Burkholderia ambifaria type Wisconsin isolate J82, Trichoderma virens Gl-21, and Trichoderma harzianum strain T-22), a chemical seed treatment (Topsin, mancozeb, and cymoxanil mixture, TMC), and a combination chemical/biological treatm...

Evaluation of antagonistic fungi against charcoal rot of sunflower ...

African Journals Online (AJOL)

In vitro, sensitivity of Macrophomina phaseolina (Tassi) Goid determined through inhibition zone technique to various antagonistic fungi viz., Aspergillus niger, Aspergillus flavus, Trichoderma viride, Trichoderma harzianum and Penicillium capsulatum amended into PDA medium. All the antagonists reduced the colony ...

Effect of Trichoderma-enriched organic charcoal in the integrated wood protection strategy.

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Javier Ribera

Full Text Available The gradual elimination of chromium from wood preservative formulations results in higher Cu leaching and increased susceptibility to wood decay fungi. Finding a sustainable strategy in wood protection has become of great interest among researchers. The objective of these in vitro studies was to demonstrate the effect of T-720-enriched organic charcoal (biochar against five wood decay basidiomycetes isolated from strongly damaged poles. For this purpose, the antagonistic potential of Trichoderma harzianum (strain T-720 was confirmed among other four Trichoderma spp. against five brown-rot basidiomycetes in dual culture tests. T-720 was genetically transformed and tagged with the green fluorescent protein (GFP in order to study its antagonistic mechanism against wood decay basidiomycetes. It was also demonstrated that T-720 inhibits the oxalic acid production by basidiomycetes, a well-known mechanism used by brown-rot fungi to detoxify Cu from impregnated wood. Additionally, this study evaluated the effect of biochar, alone or in combination with T-720, on Cu leaching by different preservatives, pH stabilization and prevention of wood decay caused by five basidiomycetes. Addition of biochar resulted in a significant Cu binding released from impregnated wood specimens. T-720-enriched biochar showed a significant reduction of wood decay caused by four basidiomycetes. The addition of T-720-enriched biochar to the soil into which utility poles are placed may improve the efficiency of Cr-free wood preservatives.

Inhibition coefficient and molecular diversity of multi stress tolerant Trichoderma as potential biocontrol agent against Sclerotium rolfsii Sacc.

Science.gov (United States)

Hirpara, Darshna G; Gajera, H P; Hirapara, Jaydeep G; Golakiya, B A

2017-11-01

Trichoderma is one of the most exploited biocontrol agent for the management of plant diseases. Twenty strains of Trichoderma (six of T. harzianum, four of T. viride, three of T. virens, three of T. koningii, each one of T. hamatum, T. reesei, T. parceramosum and Trichoderma spp.) subjected to in vitro antagonism up to 12days after inoculation against Sclerotium rolfsii Sacc. causing stem rot in groundnut. A new concept was developed to determine inhibition coefficient representing pathogen biology and biocontrol related biophysical variables. Results explained differential inhibition coefficient of test pathogen by Trichoderma antagonists. The inhibition coefficient of test pathogen was examined highest (91.13%) by T. virens NBAII Tvs12 followed by T. virens MTCC 794 (89.33%) and T. koningii MTCC 796 (62.39%). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs12 and antibiosis for T. koningii MTCC 796. The sclerotial biogenesis of test pathogen was elevated during weak antagonism and diminished in interactions with strong antagonists. The inhibition coefficient of S. rolfsii was significantly negatively correlated with sclerotia formation and lipid peroxidation during the antagonism. Trichoderma strains were screened for fungicides (carbendazim and tebuconazole, thiram and mancozeb) and abiotic stress (drought and salt) tolerance. Results indicated that T. koningii MTCC 796 efficiently grew better than the other strains with maximum radial growth under adverse conditions. The genetic variability among the Trichoderma was determined using 34 gene specific markers which amplified 146 alleles. The SSR similarities explained substantial diversity (15 to 87%) across Trichoderma strains and pathogen S. rolfsii. Principal coordinates analysis (PCA) were comparable to the cluster analysis and first three most informative PC components explained 64.45% of the total variation. In PCA, potent antagonists appear to be distinct from other strains. Five

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

Science.gov (United States)

2011-01-01

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. PMID:22070776

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

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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.

Evaluation of some bioagents and botanicals in in vitro control of ...

African Journals Online (AJOL)

Collectotrichum destructivum has not been effectively controlled. This led to trials on the use of bioagents and botanicals to control the pathogen. The bio-agents such as Pseudomonas fluorescens, Bacillus subtilis, Trichoderma harzianum and Trichoderma pseudokoningii were inoculated as dixenic culture with the ...

Análise da associação e do secretoma da interação entre Trichoderma spp. de solo do Cerrado com feijoeiro comum, Phaseolus vulgaris L.

OpenAIRE

Silva, Francilene Lopes da

2014-01-01

Algumas espécies de Trichoderma quando em íntima associação com plantas hospedeiras podem desencadear sua resposta de defesa e, desta forma, induzir resistência contra subsequentes infecções fúngicas. No presente trabalho foi analisado a expressão dos genes codificadores de proteínas de defesa (Glu, Chit 1, LOX, PER e PAL) em feijoeiro comum, Phaseolus vulgaris, em resposta à asssociação com os isolados das espécies T. asperellum (468/02) e T. harzianum (475/2 e 303/2), após 24, 48 e 72 horas...

Induced systemic resistance in cucumber and Arabidopsis thaliana by the combination of Trichoderma harzianum Tr6 and Pseudomonas sp. Ps14

NARCIS (Netherlands)

Alizadeh, H.; Behboudi, K.; Ahmadzadeh, M.; Javan-Nikkhah, M.; Zamioudis, C.; Pieterse, C.M.J.; Bakker, P.A.H.M.

2013-01-01

Trichoderma species and fluorescent Pseudomonas spp. have been reported to induce systemic resistance in plants. In this study the effectiveness of a combination of these biological control agents on the efficacy of induced resistance was investigated in cucumber and the model plant Arabidopsis

Genetic relatedness of Trichoderma isolates antagonistic against Fusarium oxysporum f.sp. dianthi inflicting carnation wilt.

Science.gov (United States)

Shanmugam, V; Sharma, Vivek; Ananthapadmanaban

2008-01-01

Twenty-eight isolates of Trichoderma belonging to four different species were screened in vitro for their antagonistic ability against Fusarium oxysporum f.sp. dianthi causing carnation wilt. Three different levels of antagonism observed in dual plate assay were further confirmed by cell-free culture filtrate experiments. Isolates showing class I level of antagonism produced maximum lytic enzymes, chitinases and beta-1,3-glucanases. Genetic variability of 25 selected isolates was assessed by random amplified polymorphic DNA technique and the amplified products were correlated for their level of antagonism. Unweighed pair-group method with arithmetical averages cluster analysis revealed prominent inter-and intraspecific genetic variation among the isolates. Based on their genetic relationship, the isolates were mainly distributed into 3 major groups representing T. atroviride, T. pseudokoningii and T. harzianum, with 20-35% interspecific dissimilarity. However, the polymorphism shown by the isolates did not correlate to their level of antagonism.

Evaluation of some bioagents and botanicals in in vitro control of ...

African Journals Online (AJOL)

SERVER

2008-04-03

Apr 3, 2008 ... Bacillus subtilis, Trichoderma harzianum and Trichoderma pseudokoningii were inoculated as dixenic culture with the ... The fungi toxicity of four plant extracts was compared with the ... extract in the Petri plates and swirled round five times for even dispersion .... microbes by antibiosis (Prescott et al., 2002).

Microplate-Based Evaluation of the Sugar Yield from Giant Reed, Giant Miscanthus and Switchgrass after Mild Chemical Pre-Treatments and Hydrolysis with Tailored Trichoderma Enzymatic Blends.

Science.gov (United States)

Cianchetta, Stefano; Bregoli, Luca; Galletti, Stefania

2017-11-01

Giant reed, miscanthus, and switchgrass are considered prominent lignocellulosic feedstocks to obtain fermentable sugars for biofuel production. The bioconversion into sugars requires a delignifying pre-treatment step followed by hydrolysis with cellulase and other accessory enzymes like xylanase, especially in the case of alkali pre-treatments, which retain the hemicellulose fraction. Blends richer in accessory enzymes than commercial mix can be obtained growing fungi on feedstock-based substrates, thus ten selected Trichoderma isolates, including the hypercellulolytic strain Trichoderma reesei Rut-C30, were grown on giant reed, miscanthus, or switchgrass-based substrates. The produced enzymes were used to saccharify the corresponding feedstocks, compared to a commercial enzymatic mix (6 FPU/g). Feedstocks were acid (H 2 SO 4 0.2-2%, w/v) or alkali (NaOH 0.02-0.2%, w/v) pre-treated. A microplate-based approach was chosen for most of the experimental steps due to the large number of samples. The highest bioconversion was generally obtained with Trichoderma harzianum Or4/99 enzymes (78, 89, and 94% final sugar yields at 48 h for giant reed, miscanthus, and switchgrass, respectively), with significant increases compared to the commercial mix, especially with alkaline pre-treatments. The differences in bioconversion yields were only partially caused by xylanases (maximum R 2  = 0.5), indicating a role for other accessory enzymes.

Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion.

Science.gov (United States)

Hatvani, Lóránt; Manczinger, László; Kredics, László; Szekeres, András; Antal, Zsuzsanna; Vágvölgyi, Csaba

2006-01-01

The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T. atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 microg/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 microg/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T. atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.

Biocontrol del moho foliar del tomate Cladosporium fulvum empleando cuatro hongos antagonistas Biocontrole do mofo foliar em tomateiro empregando quatro fungos antagonistas Biocontrol of leaf mold of tomato Cladosporium fulvum (Cooke 1883 employing four antagonistic fungi

Directory of Open Access Journals (Sweden)

Enrique Torres

2008-01-01

Full Text Available El moho foliar Cladosporium fulvum (Cooke 1883 produce una enfermedad que ataca el cultivo de tomate en invernadero en el Perú. Como alternativa al control de este patógeno se probó la eficiencia bajo condiciones in vitro e invernadero de cuatro hongos antagonistas: Hansfordia pulvinata (Berk y Curt 1958, Trichoderma harzianum (Rifai 1969, T. viride (Persoon 1821 y T. virens (Miller, Giddens y Foster 1963. El cultivo y crecimiento de los hongos se realizó en Agar Sabouraud para Trichoderma spp. y en Agar Jugo V8 para H. pulvinata. El bioensayo empleado fue el de placas precolonizadas propuesto por Krauss. En condiciones in vitro se observó que a 24 ºC y 72 h, el crecimiento de T. harzianum fue estadísticamente diferente de las otras tres especies ensayadas, y a 28 ºC fue similar a T. viride. A 24 ºC y a las 120 h, T. virens fue diferente de todas las especies evaluadas. A las 168 h el crecimiento de las tres especies de Trichoderma fue estadísticamente similar, diferenciándose de H. pulvinata. A 28 ºC a las 120 h y 168 h los resultados fueron estadísticamente similares para las tres especies de Trichoderma, pero diferentes de H. pulvinata. El mecanismo de acción más común para los hongos antagonistas fue el micoparasitismo. Finalmente, T. harzianum fue más eficiente debido a que redujo la severidad de la enfermedad en un 19,35 % bajo condiciones de invernadero.O mofo foliar Cladosporium fulvum (Cooke, 1883 produz uma enfermidade que ataca o cultivo do tomateiro em casa de vegetação no Peru. Como alternativa ao controle desse patógeno, testou-se a eficiência, sob condições in vitro e casa de vegetatção, de quatro fungos antagonistas: Hansfordia pulvinata (Berk y Curt 1958, Trichoderma harzianum (Rifai 1969, T. viride (Persoon 1821 y T. virens (Miller, Giddens y Foster 1963. O cultivo e crescimento dos fungos foi realizado em Agar Sabouraud para Trichoderma spp. e em Agar Suco V8 para H. pulvinata. O bioensaio empregado

Phytoremediation of Alberta oil sand tailings using native plants and fungal endophytes

Science.gov (United States)

Repas, T.; Germida, J.; Kaminskyj, S.

2012-04-01

Fungal endophytes colonize host plants without causing disease. Some endophytes confer plant tolerance to harsh environments. One such endophyte, Trichoderma harzianum strain TSTh20-1, was isolated from a plant growing on Athabasca oil sand tailings. Tailing sands are a high volume waste product from oil sand extraction that the industry is required to remediate. Tailing sands are low in organic carbon and mineral nutrients, and are hydrophobic due to residual polyaromatic hydrocarbons. Typically, tailing sands are remediated by planting young trees in large quantities of mulch plus mineral fertilizer, which is costly and labour intensive. In greenhouse trials, TSTh20-1 supports growth of tomato seedlings on tailing sands without fertilizer. The potential use of TSTh20-1 in combination with native grasses and forbs to remediate under field conditions is being assessed. Twenty-three commercially available plant species are being screened for seed germination and growth on tailing sands in the presence of TSTh20-1. The best candidates from this group will be used in greenhouse and small scale field trials. Potential mechanisms that contribute to endophyte-induced plant growth promotion, such as plant hormone production, stress tolerance, mineral solubilization, and uptake are also being assessed. As well, TSTh20-1 appears to be remarkably frugal in its nutrient requirements and the possibility that this attribute is characteristic of other plant-fungal endophytes from harsh environments is under study.

Correlation of structure, function and protein dynamics in GH7 cellobiohydrolases from Trichoderma atroviride, T. reesei and T. harzianum

DEFF Research Database (Denmark)

Borisova, Anna S.; Eneyskaya, Elena V.; Jana, Suvamay

2018-01-01

analyses and molecular dynamics (MD) simulations were performed to elucidate important structure/function correlations. Moreover, reverse conservation analysis (RCA) of sequence diversity revealed divergent regions of interest located outside the cellulose-binding tunnel of Trichoderma spp. GH7 CBHs. We...... that, for industrial use, it would be beneficial to combine loop motifs from TatCel7A with the thermostability features of TreCel7A. Furthermore, one region implicated in thermal unfolding is suggested as a primary target for protein engineering...

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

Energy Technology Data Exchange (ETDEWEB)

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

2008-01-10

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

Effect of gamma irradiation and environmental factors on the production of extracellular cellulase enzyme by trichoderma Spp. using banana waste under solid state bio processing

International Nuclear Information System (INIS)

El-Shafey, H.M.; Matar, Z.A.I.; Ghanem, S.M.A.

2007-01-01

Fungal strains were isolated from degraded banana waste including leaves, pseudo stems and skins. Many isolated strains showed cellulolytic activities using the plate screening medium. The hyper cellulolytic isolates were selected on the basis of the diameter of the hydrolysis zone surrounding the colonies and identified to the genus level. The identified strains were found to belong to one of the genera Trichoderma, Aspergillus, Pleurotus or Penicillium. The strain with the larger diameter of the hydrolysis zone was found to belong to the genus Trichoderma. It was further identified to be Trichoderma harzianum, which was selected to be studied. Banana waste including leaves and pseudo stems were inoculated by the selected fungus and the production of the carboxymethyl cellulase (CMCase) and filter paper cellulase (FPCase) was followed during changes of the growth conditions under solid state fermentation. It was found that the two enzymes shared the same incubation temperature (25 degree C) and incubation period (18 days) for the maximum enzyme production. The gamma radiation dose of 1.5 KGy increased the production of CMCase produced on leaves by 4.0% and on pseudo stems by 5.6% and the production of FPCase produced on leaves by 2.4% and on pseudo stems by 2.3%. The results also suggest that FPCase and CMCase enzymes produced on leaves were higher than those produced from pseudo stems and the level of CMCase enzyme produced was higher than that of FPCase

Integrated options for the management of black root rot of strawberry caused by Rhizoctonia solani Kuhn.

Science.gov (United States)

Asad-Uz-Zaman, Md; Bhuiyan, Mohammad Rejwan; Khan, Mohammad Ashik Iqbal; Alam Bhuiyan, Md Khurshed; Latif, Mohammad Abdul

2015-02-01

An investigation was made to manage strawberry black root rot caused by Rhizoctonia solani (R. solani) through the integration of Trichoderma harzianum (T. harzianum) isolate STA7, mustard oil cake and Provax 200. A series of preliminary experiments were conducted to select a virulent isolate of R. solani, an effective isolate of T. harzianum, a suitable organic amendment, and a suitable fungicide before setting the experiment for integration. The pathogenicity of the selected four isolates of R. solani was evaluated against strawberry and isolate SR1 was selected as the test pathogen due to its highest virulent (95.47% mortality) characteristics. Among the 20 isolates of T. harzianum, isolate STA7 showed maximum inhibition (71.97%) against the test pathogen (R. solani). Among the fungicides, Provax-200 was found to be more effective at lowest concentration (100 ppm) and highly compatible with Trichoderma isolates STA7. In the case of organic amendments, maximum inhibition (59.66%) of R. solani was obtained through mustard oil cake at the highest concentration (3%), which was significantly superior to other amendments. Minimum percentages of diseased roots were obtained with pathogen (R. solani)+Trichoderma+mustard oil cake+Provax-200 treatment, while the highest was observed with healthy seedlings with a pathogen-inoculated soil. In the case of leaf and fruit rot diseases, significantly lowest infected leaves as well as fruit rot were observed with a pathogen+Trichoderma+mustard oil cake+Provax-200 treatment in comparison with the control. A similar trend of high effectiveness was observed by the integration of Trichoderma, fungicide and organic amendments in controlling root rot and fruit diseases of strawberry. Single application of Trichoderma isolate STA7, Provax 200 or mustard oil cake did not show satisfactory performance in terms of disease-free plants, but when they were applied in combination, the number of healthy plants increased significantly. The

Ação antagônica e de metabólitos bioativos de Trichoderma spp. contra os patógenos Sclerotium rolfsii e Verticillium dahliae

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Carolina Oliveira Isaias

2014-03-01

Full Text Available O efeito antagônico exercido por fungos do gênero Trichoderma foi avaliado contra os fitopatógenos Sclerotium rolfsii e Verticillium dahliae, isolados de áster ornamental e morangueiro, respectivamente. Utilizaram-se 20 isolados do antagonista, pertencentes às espécies T. asperellum, T. hamatum, T. harzianum, T. koningiopsis, T. spirale e Trichoderma sp. O antagonismo foi verificado por meio de testes de culturas pareadas e produção de metabólitos bioativos pelo método de sobreposição de placas. A maioria dos isolados exibiu efeito inibitório sobre o crescimento micelial de S. rolfsii e V. dahliae quando comparado com as testemunhas. Excetuando-se os testes de produção de metabólitos não-voláteis, verificou-se maior ação inibitória contra V. dahliae. Mesmo os metabólitos não-voláteis termoestáveis apresentaram efeito inibitório contra ambos os patógenos testados, mostrando que os metabólitos secundários do antagonista exercem efeito significativo sobre essas espécies patogênicas.

Inhibitory activity of Beauveria bassiana and Trichoderma spp. on the insect pests Xylotrechus arvicola (Coleoptera: Cerambycidae) and Acanthoscelides obtectus (Coleoptera: Chrisomelidae: Bruchinae).

Science.gov (United States)

Rodríguez-González, Álvaro; Mayo, Sara; González-López, Óscar; Reinoso, Bonifacio; Gutierrez, Santiago; Casquero, Pedro Antonio

2017-01-01

Xylotrechus arvicola is an important pest in vineyards (Vitis vinifera) in the main Iberian wine-producing regions, and Acanthoscelides obtectus causes severe post-harvest losses in the common bean (Phaseolus vulgaris). Under laboratory conditions with a spray tower, the susceptibility of the immature stages of X. arvicola and A. obtectus against the entomopathogenic fungi Beauveria bassiana and four strains of Trichoderma spp. was evaluated. Both insect pests T. harzianum and B. bassiana showed a good inhibitory activity, accumulating an inhibition on the eggs of values above 85 and 82%, respectively. T. atroviride and T. citrinoviride had a lower inhibitory activity, with inhibition values of 74.1 and 73.3% respectively. These fungi can be considered a highly effective tool for the control during the immature stages of these species.

Inhibition of in vitro growth of soil-borne pathogens by compost-inhabiting indigenous bacteria and fungi

International Nuclear Information System (INIS)

Ramzan, N.; Noreen, N.; Shahzad, S.

2014-01-01

During the present studies, compost-inhabiting microorganisms including 44 fungi and 15 bacteria isolated from different compost samples were evaluated for their in vitro efficacy against soil-borne pathogens viz., Fusarium solani, Macrophomina phaseolina, Pythium aphanidermatum, Rhizoctonia solani, and Sclerotium rolfsii. Compost inhabiting microbes like Trichoderma harzianum, T. virens, Bacillus cereus, B. pumilus, B. subtilis, Micrococcus varians and Pseudomonas fluorescens were found to inhibit all the test pathogens. Acrophialophora fusispora and Penicillium citrinum reduced the mycelial growth of all the test pathogens except Sclerotium rolfsii. Bacillus licheniformis and Bacillus megaterium showed biocontrol activity against all the pathogens except Rhizoctonia solani. Trichoderma harzianum parasitized mycelia of all the tested pathogens and produced coiling around the mycelium. (author)

Growth response and nutrient uptake of blue pine (Pinus wallichiana seedlings inoculated with rhizosphere microorganisms under temperate nursery conditions

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M.A. Ahangar

2012-11-01

Full Text Available Microbial inoculants (Trichoderma harzianum, Pseudomonas fluorescens,Laccaria laccata inoculated either individually or in combinationsignificantly improved the growth and biomass of blue pine seedlings. The ECM fungus Laccaria laccata, when inoculated individually, showed significantly higher plant growth, followed by Pseudomonas fluorescens and Trichoderma harzianum. The combined inoculation of rhizosphere microorganisms showed synergistic growth promoting action and proved superior in enhancing the growth of blue pine than individual inoculation. Co-inoculation of L. laccata with P. fluorescens resulted in higher ectomycorrhizal root colonization. Uptake of nutrients (N, P, K was significantly improved by microbial inoculants, tested individually or in combination. Combined inoculation of L. laccata with T. harzianum and P. fluorescens significantly increased in N, P and K contents in blue pine seedlings as compared to control. Acid phosphatase activity in the rhizosphere of blue pine seedlings was also enhanced by these microorganisms. L. laccata exhibited higher acid phosphatase activity followed by P. fluorescens.

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

Science.gov (United States)

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

2015-06-01

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

Trichoderma stromaticum and its overseas relatives

Science.gov (United States)

Trichoderma stromaticum, T. rossicum and newly discovered species form a new lineage in Trichoderma. Phylogenetic and phenotypic diversity in Trichoderma stromaticum are examined in light of reported differences in ecological parameters and AFLP patterns. Multilocus phylogenetic analysis using 4 gen...

isolated from Trichoderma harzianum

African Journals Online (AJOL)

SAM

2014-05-21

May 21, 2014 ... segmentation, branching and hyphal bursting at the concentration of 200 µg ml-1. Key words: ... family 18 and can be further grouped into class III and class V. Many .... for their ability to act as a biocontrol agents against plant.

Biological Control of Fusarium oxysporum f.sp. cumini with Aspergillus versicolor

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S. Israel

2005-04-01

Full Text Available A native heat-tolerant strain of Aspergillus versicolor (Vuill. Tirab. highly antagonistic to Fusarium oxysporum f. sp. cumini (Foc was isolated from arid soils. In tests performed to ascertain its antagonistic activity against Foc as compared to Trichoderma harzianum, a 99.2 and 96.4% reduction in Foc propagules was achieved in A. versicolor and T. harzianum infested soil respectively. The reduction of Foc propagules in Foc and A. versicolorinfested soil was also determined. In a liquid-culture test, even at a low concentration of 0.5 ml cell-free filtrate, A. versicolor inhibited mycelial growth of Foc. Population changes of A. versicolor were examined at different soil moisture gradients, where maximum survival and multiplication of A. versicolor was estimated at 50% of moisture holding capacity. In general, with increasing concentrations of A. versicolor inoculum, soil population densities of Foc went down. Studies on thermal resistance showed that A. versicolor survived and multiplied even at 65°C. Soil amended with A. versicolor alone, or with a combination of T. harzianum and Verbisina enceloides residues was significantly better at reducing Foc than was non-amended control soil. A marked increase in the root length of cumin was observed in soil amended with A. versicolor or T. harzianum or both. The results suggest that A. versicolor has a potential value for use against Fusarium in hot arid soils because it can survive under dry and high-temperature conditions.

EVALUATION OF TRICHODERMA SPP. ON BEAN CULTURE, IN ANTHRACNOSE, WEB BLIGHT AND ROOT-KNOT NEMATODE

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P. E. V. Aguiar

2014-09-01

Full Text Available Mato Grosso is the third largest producer of bean from Brazil, being the third harvest (irrigated the most productive, but diseases such as anthracnose, web blight and nematodes of galls cause losses to producers. In addition, a measure widely used and little studied for the control of diseases and nematodes in Mato Grosso is the biological control, which consists of the action of other microorganisms on phytopathogens. Thus, the objective of the present study was to evaluate the effect of Trichoderma harzianum and T. asperellum in the development (height of plants, chlorophyll and number of pods of culture of bean, in the control of anthracnose (Colletotrichum lindemuthianum, web blight (Rhizoctonia solani and in the population of Meloidogyne spp. in the soil. The experiment was accomplished in area experimental of University Federal of Mato Grosso/Campus Sinop. The experimental design was of entirely randomized with 12 parcels of 5m² each, with 3 treatments and 4 replications. The cultivar used was Whitey, carioca group, and the seed treatment performed with product Pyraclostrobin + Thiophanate Methyl + Fipronil and after drying of the inoculation of biocontrol agents and manual seeding. It was observed that the application of T. harzianum and T. asperellum, not promoted increase of chlorophyll, height of plants in bean culture, without reducing the population of Meloidogyne spp.. However, biocontrol agents have reduced the severity of anthracnose and web blight and promoted an increase in the average number of plant pods-1. It is therefore concluded that biocontrol agents show potential for application in bean culture in the North of Mato Grosso.

Copper tolerance of Trichoderma species

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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

Trichoderma as an endophyte

Science.gov (United States)

Trichoderma species have been studied for many years for their usefulness in plant disease management. For much of this time, studies focused on the attributes of Trichoderma as a soil saprophyte possessing abilities such as mycoparasitism and antibiosis that directly impact pathogens. The ability...

Evaluation of bio-agent formulations to control Fusarium wilt of tomato

African Journals Online (AJOL)

ONOS

2010-09-06

Sep 6, 2010 ... studied with emphasis on biological control using fungi or ... mechanisms such as antibiosis, competition, suppression, ... Fusarium wilt pathogen such as Trichoderma harzianum, .... sphere around soil and above plants.

de Fusarium isolé du fruit de tomate (Solanum lycopersicum L ...

African Journals Online (AJOL)

Dina

A phytopathogenic fungus Fusarium F-02 is isolated from rotten tomato fruit. ..... between Trichoderma harzianum and Pythium ultimum”. .... [31] - HAAS, and C. KEEL, “Regulation of antibiotic production in root-colonizing Pseudomonas spp.

Microbial agents against Helicoverpa armigera: Where are we and ...

African Journals Online (AJOL)

SAM

2014-04-30

Apr 30, 2014 ... feed and fibre around the world. Different approaches namely: .... biocontrol agent from the fungus Trichoderma harzianum was also evaluated .... the antibiotic biosynthetic gene transfer (Baltz, 2005). Overall 23,000 bioactive ...

Extracellular proteases of Trichoderma species. A review.

Science.gov (United States)

Kredics, L; Antal, Zsuzsanna; Szekeres, A; Hatvani, L; Manczinger, L; Vágvölgyi, Cs; Nagy, Erzsébet

2005-01-01

Cellulolytic, xylanolytic, chitinolytic and beta-1,3-glucanolytic enzyme systems of species belonging to the filamentous fungal genus Trichoderma have been investigated in details and are well characterised. The ability of Trichoderma strains to produce extracellular proteases has also been known for a long time, however, the proteolytic enzyme system is relatively unknown in this genus. Fortunately, in the recent years more and more attention is focused on the research in this field. The role of Trichoderma proteases in the biological control of plant pathogenic fungi and nematodes has been demonstrated, and it is also suspected that they may be important for the competitive saprophytic ability of green mould isolates and may represent potential virulence factors of Trichoderma strains as emerging fungal pathogens of clinical importance. The aim of this review is to summarize the information available about the extracellular proteases of Trichoderma. Numerous studies are available about the extracellular proteolytic enzyme profiles of Trichoderma strains and about the effect of abiotic environmental factors on protease activities. A number of protease enzymes have been purified to homogeneity and some protease encoding genes have been cloned and characterized. These results will be reviewed and the role of Trichoderma proteases in biological control as well as their advantages and disadvantages in biotechnology will be discussed.

Bioprospecting for culturable actinobacteria with antimicrobial ...

African Journals Online (AJOL)

Strains of Fusarium sp. H24, Trichoderma harzianum H5 and Colletotrichum ... Antibiosis was indicated by visually observable growth inhibition of the ... Table 1: Antimicrobial activity of seven selected strains against fungi and bacterial strains.

Trichoderma sp. dalam Pengendalian Penyakit Layu Fusarium pada Tanaman Tomat

OpenAIRE

Novita, Trias

2013-01-01

Penelitian ini bertujuan untuk mengetahui peran Trichoderma sp dalam pengendalianpenyakit layu fusarium pada tanaman tomat. Penelitian dilaksanakan di Rumah Kaca FakultasPertanian Universitas Jambi, perlakuannya terdiri dari : t0 = tanpa Trichoderma sp; t1 = 25 gTrichoderma sp/8 kg media; t2 = 50 g Trichoderma sp/8 kg media; t3 = 75 g Trichoderma sp/8 kgmedia; dan t4 = 100 g Trichoderma sp /8 kg media. Hasil penelitian menunjukkan bahwa Trichodermasp berperan dalam mengendalikan penyakit layu...

Reducción de poblaciones de Ralstonia solanacearum raza 2 (Smith en plátano (Musa AAB Simmonds con aplicación de extractos de Trichoderma sp. (Alexopoulus y Mims y bacterias antagonistas

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Germán Ceballos

2014-01-01

Full Text Available El moko, enfermedad causada por Ralstonia solanacearum raza 2, afecta la producción de plátano (Musa AAB Simmonds en Colombia, ocasionando pérdidas hasta de 100%. In vitro, se evaluó el efecto de filtrados crudos de cepas de Trichoderma sp. y dos productos comerciales como posibles reductores de poblaciones de R. solanacearum. Para cada cepa de Trichoderma spp., se empleó filtrado (40% en solución acuosa y para R. solanacearum se utilizó una dilución de 10-4 por caja de petri, en un diseño completamente al azar con tres repeticiones para determinar el número de unidades formadoras de colonia de R. solanacearum. En invernadero, se aplicaron suspensiones de T. harzianum (Agroguard®, T. viride 14PDA3A y Ecoterra® en plantas de plátano Dominico Hartón y 30 días después se inoculó R. solanacearum en un diseño de bloques completos al azar con cuatro repeticiones. La severidad del ataque de R. solanacearum se evaluó utilizando una escala de síntomas foliares de 0 a 6. In vitro, los extractos crudos de dos cepas y dos productos comerciales de Trichoderma spp. inhibieron el 100% de R. solanacearum. Los tratamientos con T. viride y Ecoterra® mostraron bajos niveles de control de R. solanacearum en plantas (0.63 y 1.88, respectivamente.

BIOREMEDIATION PERSPECTIVES USING AUTOCHTHONOUS SPECIES OF Trichoderma sp. FOR DEGRADATION OF ATRAZINE IN AGRICULTURAL SOIL FROM THE TULANCINGO VALLEY, HIDALGO, MEXICO

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Margarita Islas Pelcastre

2013-08-01

Full Text Available The objective of this study was to show an in vitro bioremediation methodology for atrazine-contaminated soils through the use of local strains of native fungi isolated from the Phaseolus vulgaris L rhizosphere present in cultivable soils as well as to evaluate its resistance and capacity for atrazine degradation. A Trichoderma sp. species was identified in three cultivable soils from the Tulancingo Hidalgo, México region (contaminated with and without atrazine, which resisted atrazine concentrations of 10,000 mg L-1.  Tests showed that the strain grows exponentially in atrazine-contaminated soil over a range of 105-106 CFU g-1 in 15 days using atrazine as the only carbon and nitrogen source, while the control and witress showed a decrease of 100-103 UFC g-1 in the same period of time. For the atrazine degradation experiments, a treatment of the application of Trichoderma (104 - 105 CFU mL-1 was applied to sterilized and non-sterilized soil contaminated with 500 mg Kg-1 of atrazine, evaluated at four time intervals (5, 10, 20 and 40 days. Statistical differences were found (α=0.050, Tukey among treatments with the fungi and the test days. The native Trichoderma strain degraded 89% of the atrazine in 40 days. It showed that it is viable and cultivable in soil bioremediation.

Biological control of chickpea wilt caused by fusarium oxysporum f.sp.ciceris

International Nuclear Information System (INIS)

Yousif, F. A.; Suliman, W. S.

2010-01-01

This study was conducted in an attempt to control chickpea (Cicer arietinum L.) wilt, caused by fusarium oxysporum f.sp. ciceris, using antagonistic properties of soil microorganisms. It also aimed at avoiding problems resulting from the use of chemical fungicides. A trichoderma sp. was isolated from the rhizosphere of a resistant chickpea variety (ICCV-2) and a bacillus sp. from the rhizosphere and rhizoplane of the same variety. Both microorganisms proved to be effective in controlling the disease. In addition, trichoderma harzianum, which was obtained from Giza Research Station in Egypt, was also antagonistic to fusarium oxysporum f. sp. ciceris Wilt incidence was significantly reduced when chickpea was grown in posts containing soil mixed with any of the three antagonists or when chickpea seeds were initially treated with the seed-dressing fungicide vincit at 2 ml/kg seeds. Trichoderma harzianum proved to be the best bioagent as it gave the lowest disease incidence. In the field, the two trichoderma spp. were as effective as vincit in causing reduction in the wilt incidence. At the higher concentration of 140 g/m''2, the two antagonists were effective throughout the growth period, but they were less effective at the lower concentration of 70 g/m''2 particularly at the seedling stage.(Author)

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

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Paola Mendes Milanesi

2013-12-01

Full Text Available 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 isolates of Trichoderma spp. with better performance in the direct confrontation. The sampling of soil and roots was performed in an experimental area located in Cruz Alta, RS, Brazil. In the laboratory, serial dilutions of soil samples, counting of the number of Colony Forming Units (UFCs/g-1 of rhizospheric soil were performed as well as isolation for identification of isolates of Fusarium spp. and Trichoderma spp. and testing of direct confrontation. There were significant differences between the population of Trichoderma spp. in the rhizosphere of plants with and without symptoms of SDS. For the population of Fusarium spp., significant difference was observed only in the rhizosphere of plants without symptoms of SDS. In diseased roots the following species were identified: F. solani, F. avenaceum, F. graminearum, F. oxysporum and F. verticillioides. In the test of direct confrontation, eight isolates of Trichoderma spp. achieved the best performance in the antagonism to Fusarium spp. and Trichoderma spp. from areas with symptoms of SDS had a higher control efficiency in vitro. These isolates showed high similarity to the species of T. koningii agregate.

Short communication

African Journals Online (AJOL)

Human01

concentrations (0.5; 1.0 and 1.5%) of Funginil (Trichoderma harzianum formulation) against Botrytis gladiolorum .... places of India viz., in and around Delhi, Ghazia- bad (Uttar ..... fungi are assumed as an antibiosis, mycopara- sitism and ...

Molecular Identification, Enzyme Assay, and Metabolic Profiling of Trichoderma spp.

Science.gov (United States)

Bae, Soo-Jung; Park, Young-Hwan; Bae, Hyeun-Jong; Jeon, Junhyun; Bae, Hanhong

2017-06-28

The goal of this study was to identify and characterize selected Trichoderma isolates by metabolic profiling and enzyme assay for evaluation of their potential as biocontrol agents against plant pathogens. Trichoderma isolates were obtained from the Rural Development Administration Genebank Information Center (Wanju, Republic of Korea). Eleven Trichoderma isolates were re-identified using ribosomal DNA internal transcribed spacer (ITS) regions. ITS sequence results showed new identification of Trichoderma isolates. In addition, metabolic profiling of the ethyl acetate extracts of the liquid cultures of five Trichoderma isolates that showed the best anti- Phytophthora activities was conducted using gas chromatography-mass spectrometry. Metabolic profiling revealed that Trichoderma isolates shared common metabolites with well-known antifungal activities. Enzyme assays indicated strong cell walldegrading enzyme activities of Trichoderma isolates. Overall, our results indicated that the selected Trichoderma isolates have great potential for use as biocontrol agents against plant pathogens.

Xenobiotic Compounds Degradation by Heterologous Expression of a Trametes sanguineus Laccase in Trichoderma atroviride.

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Edgar Balcázar-López

Full Text Available Fungal laccases are enzymes that have been studied because of their ability to decolorize and detoxify effluents; they are also used in paper bleaching, synthesis of polymers, bioremediation, etc. In this work we were able to express a laccase from Trametes (Pycnoporus sanguineus in the filamentous fungus Trichoderma atroviride. For this purpose, a transformation vector was designed to integrate the gene of interest in an intergenic locus near the blu17 terminator region. Although monosporic selection was still necessary, stable integration at the desired locus was achieved. The native signal peptide from T. sanguineus laccase was successful to secrete the recombinant protein into the culture medium. The purified, heterologously expressed laccase maintained similar properties to those observed in the native enzyme (Km and kcat and kcat/km values for ABTS, thermostability, substrate range, pH optimum, etc. To determine the bioremediation potential of this modified strain, the laccase-overexpressing Trichoderma strain was used to remove xenobiotic compounds. Phenolic compounds present in industrial wastewater and bisphenol A (an endocrine disruptor from the culture medium were more efficiently removed by this modified strain than with the wild type. In addition, the heterologously expressed laccase was able to decolorize different dyes as well as remove benzo[α]pyrene and phenanthrene in vitro, showing its potential for xenobiotic compound degradation.

In-vitro evaluation of fungicides, plant extracts and bio-control agents against rice blast pathogen magnaporthe oryzae couch

International Nuclear Information System (INIS)

Hajano, J.U.D.; Lodhi, M.; Pathan, M.A.; Khanzada, A.; Shah, G.S.

2012-01-01

Among 5 fungicides viz., Thiophanate-methyl, Carbendazim, Fosetyl-aluminium, Mancozeb and Copper oxychloride, used against the Magnaporthe oryzae, only Mancozeb appeared as the highly effective fungicide that completely inhibited the mycelial growth of the fungus. All other fungicides showed little effect at higher concentrations. The extracts of garlic (Allium sativum L.), neem (Azadirachta indica L.) and calatropis (Calotropis procera L.) when used against M. oryzae by food poisoning method, only higher dose of garlic completely inhibited the mycelial growth of the test fungus. Six bio-control agents viz., Trichoderma harzianum, Trichoderma polysporum, Trichoderma pseudokoningii, Gliocladium virens, Paecilomyces variotii and Paecilomyces lilacinus were used. Maximum mycelial inhibition of M. oryzae was provided by P. lilacinus followed by Trichoderma spp. (author)

Suitability of various plant derived gelling agents as agar substitute ...

African Journals Online (AJOL)

USER

2012-06-05

Jun 5, 2012 ... of three test fungi (Trichoderma harzianum, Alternaria alternata and Alternaria solani) as good as agar. ... used for cell culture, derived from plants or animals and .... and used to jell various foods, drugs and cosmetics) and rice.

Three-way interactions between Fusarium species, their plant hosts and biocontrol organisms

DEFF Research Database (Denmark)

Kosawang, Chatchai

of ABC transporters in mycoparasitic fungi focused on C. rosea and Trichoderma spp. We showed that expression of zhd101 depended on concentrations of ZEA and that the gene was not induced by other agents, suggesting specificity of the enzyme towards ZEA and its derivates. To investigate effects of ZEA...... identified expansion of ABC transporter families in C. rosea as in Trichoderma virens and T. harzianum, but not in T. reesei and T. atroviridae. The expansion was profoundly observed in the subfamily G of C. rosea ABC transporters and the subfamily C of Trichoderma spp, repectively. The subfamilies C and G...

Use of biology agents for the control phytosanitary in the minitubers production of potato in the acclimatization conditions

Directory of Open Access Journals (Sweden)

Felipe Alberto Jimenez-Terry

2002-10-01

Full Text Available The employment of biological controls of pests and diseases is a technology with trends to increase at present for being economic, lasting and of beneficial results, related to the stability that they offer the ecosystem; avoiding the pollution of the environment produced by the application of pesticides. The present work was realized to evaluate the phytosanitary control on pests and diseases in the production of potato minitubers in acclimatization conditions. Combinations of agents Beauveria bassiana, Bacillus turhingiensis, Verticillum lecanii, Thrichoderma harzianum and Gliocladium viridens, were applied comparatively with the utilization of chemical products and a control without application. The results gave an efficient control of the chemical products and on the other hand, the biocontrol agents also exercised their role with valuable performance for the combinations of Trichoderma harzianum + Bacillus thuringiensis + Beauveria bassiana and Trichoderma harzianum + Bacillus thuringiensis + Verticillum lecanii, in which the yields of minitubers per plant were superior with significant differences to the control. Evidently these results allowed the proposal of using these biocontrol agents for the acclimatization of potato vitroplants. Key words: Beauveria, control phytosanitary, in vitro plants, Ttrichoderma, Verticillum

Species diversity of Trichoderma in Poland

Science.gov (United States)

Fifteen species of Trichoderma were identified from among 118 strains originating from different regions and ecological niches in Poland. This low number indicates low species diversity of Trichoderma in this Central European region. Using the ITS1-ITS2 regions, 64 strains were positively identified...

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.

Discoloration and tyrosinase activity in Agaricus bisporus fruit bodies infected with various pathogens

NARCIS (Netherlands)

Soler-Rivas, C.; Arpin, N.; Olivier, J.M.; Wichers, H.J.

2000-01-01

The discoloration of A. bisporus after infection by Pseudomonas tolaasii, P. gingeri, P. agarici, P. reactans, Verticillium fungicola and Trichoderma harzianum can be distinguished by chromametric measurements. Infection with P. tolaasii caused a specific colour change, independent of the bacterial

Preventing an increase in Verticillium wilt incidence in spinach seed production

DEFF Research Database (Denmark)

Olesen, Merete Halkjær; Deleuran, Lise Christina; Gislum, René

2014-01-01

A semifield assay was conducted from 2009 to 2011 to distinguish between different preventive methods of reducing Verticillium spp. in spinach seed production. The seed treatments for controlling seed infection levels included Thiram, Signum, Trichoderma harzianum, Gliocladium roseum and Natural II...

Formation and cell wall regeneration of protoplasts from Schizophyllum commune

NARCIS (Netherlands)

de Vries, Onno Minne Hotze

1974-01-01

Osmotically sensitive protoplasts were released from the mycelium of the basidiomycete Schizophyllum commune through the action ofan extracellular enzyme preparation isolated from the culture filtrate of Trichoderma viride (recently renamed T. harzianum) grown on hyphal walls of the former organism.

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.

Trichodiene production in a Trichoderma harzianum erg1-silenced strain provides evidence of the importance of the sterol biosynthetic pathway in inducing plant defense-related gene expression

Science.gov (United States)

Trichoderma species are often used as biocontrol agents against plant-pathogenic fungi. A complex molecular interaction occurs among the biocontrol agent, the antagonistic fungus, and the plant. Terpenes and sterols produced by the biocontrol fungus have been found to affect gene expression in both ...

Antagonistic Potential of Native Trichoderma viride Strain against Potent Tea Fungal Pathogens in North East India.

Science.gov (United States)

Naglot, A; Goswami, S; Rahman, I; Shrimali, D D; Yadav, Kamlesh K; Gupta, Vikas K; Rabha, Aprana Jyoti; Gogoi, H K; Veer, Vijay

2015-09-01

Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, β-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

Antagonistic Potential of Native Trichoderma viride Strain against Potent Tea Fungal Pathogens in North East India

Directory of Open Access Journals (Sweden)

A. Naglot

2015-09-01

Full Text Available Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, β-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s in the inhibitory activity.

Evaluation of biocontrol ability of native strains of Trichoderma spp on Rhizoctonia and Fusarium sp in coffee (Coffea arabica in experimental conditions

Directory of Open Access Journals (Sweden)

Nina Rudy

2016-06-01

Full Text Available Due to the indiscriminate use agrochemicals in conventional agriculture, it is causing pollution problems in the environment (soil, air and water, hence the search for alternatives that contribute to agricultural production by agro-chemical free sustainable production. This paper studies the biological control of damping off in coffee (Coffea arabica by applying antagonistic fungus Trichoderma sp. Under experimental conditions at laboratory facilities of the Academic Unit Carmen Pampa Campesina, a community of Carmen Pampa, Township Coroico. The aim of this study was to biologically control the "damping off", they found two genera that cause damping off in seedbed of coffee: Rhizoctonia sp. and Fusarium sp.To determine the percentage of growth and control in the culture medium, we used the method of counting quarters, where they gave the mycelial growth of antagonistic fungus Trichoderma sp., And the fungal pathogens Rhizoctonia sp. and Fusarium sp. Statistically there was a highly significant difference in the variable growth rate of Trichoderma sp. on pathogenic fungi Rhizoctonia sp. and Fusarium sp. at 3, 6 and 9 days that announces the time factor and treatments are interdependent. The control variable showed a highly significant difference in the time factor and treatment, but the interaction shows no significant difference this makes known factors that are independent, so the fungus Trichoderma sp. not depend on time in treatment, thus showing its inhibitory power to Rhizoctonia sp. and Fusarium sp .. This test gives references that there is antagonistic fungus control on the fungal pathogens Rhizoctonia sp. and Fusarium sp.

In vitro screening of fungicides and antagonists against Sclerotium ...

African Journals Online (AJOL)

A study was conducted in the microbiology laboratory of Plant Pathology Department, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, during 2010 to 2011 to control Sclerotium rolfsii with fungicides and Trichoderma harzianum. Six fungicides namely Provax-200, Bavistin, Ridomil, Dithane M-45, ...

Ife Journal of Science - Vol 13, No 1 (2011)

African Journals Online (AJOL)

Cellulase production by wild strains of Aspergillus niger, Penicillium chrysogenum and Trichoderma harzianum grown on waste cellulosic materials · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. SN Chinedu, VI Okochi, O Omidiji, 57-62 ...

The growth promotion of mung bean (Phaseolus radiatus) by ...

African Journals Online (AJOL)

Jane

2011-10-17

Oct 17, 2011 ... HPP16, mung bean, abscisic acid, phosphate-solubilizing bacteria (PSB). ... solubilization, PSB may also produce other secondary metabolites to improve the plant productivity. ...... Funding from the Key Laboratory of Experimental Marine. Biology ... biocontrol fungus Trichoderma harzianum Rifai1295-22.

Transpiration rates of rice plants treated with Trichoderma spp.

Science.gov (United States)

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

2014-09-01

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

A Novel Fungal Metabolite with Beneficial Properties for Agricultural Applications

Directory of Open Access Journals (Sweden)

Francesco Vinale

2014-07-01

Full Text Available Trichoderma are ubiquitous soil fungi that include species widely used as biocontrol agents in agriculture. Many isolates are known to secrete several secondary metabolites with different biological activities towards plants and other microbes. Harzianic acid (HA is a T. harzianum metabolite able to promote plant growth and strongly bind iron. In this work, we isolated from the culture filtrate of a T. harzianum strain a new metabolite, named isoharzianic acid (iso-HA, a stereoisomer of HA. The structure and absolute configuration of this compound has been determined by spectroscopic methods, including UV-Vis, MS, 1D and 2D NMR analyses. In vitro applications of iso-HA inhibited the mycelium radial growth of Sclerotinia sclerotiorum and Rhizoctonia solani. Moreover, iso HA improved the germination of tomato seeds and induced disease resistance. HPLC-DAD experiments showed that the production of HA and iso HA was affected by the presence of plant tissue in the liquid medium. In particular, tomato tissue elicited the production of HA but negatively modulated the biosynthesis of its analogue iso-HA, suggesting that different forms of the same Trichoderma secondary metabolite have specific roles in the molecular mechanism regulating the Trichoderma plant interaction.

A novel fungal metabolite with beneficial properties for agricultural applications.

Science.gov (United States)

Vinale, Francesco; Manganiello, Gelsomina; Nigro, Marco; Mazzei, Pierluigi; Piccolo, Alessandro; Pascale, Alberto; Ruocco, Michelina; Marra, Roberta; Lombardi, Nadia; Lanzuise, Stefania; Varlese, Rosaria; Cavallo, Pierpaolo; Lorito, Matteo; Woo, Sheridan L

2014-07-08

Trichoderma are ubiquitous soil fungi that include species widely used as biocontrol agents in agriculture. Many isolates are known to secrete several secondary metabolites with different biological activities towards plants and other microbes. Harzianic acid (HA) is a T. harzianum metabolite able to promote plant growth and strongly bind iron. In this work, we isolated from the culture filtrate of a T. harzianum strain a new metabolite, named isoharzianic acid (iso-HA), a stereoisomer of HA. The structure and absolute configuration of this compound has been determined by spectroscopic methods, including UV-Vis, MS, 1D and 2D NMR analyses. In vitro applications of iso-HA inhibited the mycelium radial growth of Sclerotinia sclerotiorum and Rhizoctonia solani. Moreover, iso HA improved the germination of tomato seeds and induced disease resistance. HPLC-DAD experiments showed that the production of HA and iso HA was affected by the presence of plant tissue in the liquid medium. In particular, tomato tissue elicited the production of HA but negatively modulated the biosynthesis of its analogue iso-HA, suggesting that different forms of the same Trichoderma secondary metabolite have specific roles in the molecular mechanism regulating the Trichoderma plant interaction.

Potential of ensiling for efficient management of spent residue from solid state fermentation system

OpenAIRE

Roussos, Sevastianos; Raimbault, Maurice; Geoffroy, F.; Saucedo-Castaneda, G.; Lonsane, B.K.

1992-01-01

Studies on ensiling of spent solids from solid state fermentation process for production of cellulases by #Trichoderma harzianum$ showed that good quality ensiled solids can be obtained by using about 43% initial substrate dry matter with 0.3% ensiling additive. (Résumé d'auteur)

Control of Blumeria graminis f.sp. hordei by treatment with mycelial extracts from cultured fungi

DEFF Research Database (Denmark)

Haugaard, H.; Jørgensen, H.J.L.; Lyngkjær, M.F.

2001-01-01

, Trichoderma harzianum, Pythium ultimum and Rhizopus stolonifer). Three mycelial extracts from the taxonomically different fungi B. oryzae, A ultimum and R. stolonifer were selected for detailed study. In general the number of colonies formed was reduced by 70-98% compared with controls. Furthermore, the few...

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

Science.gov (United States)

2012-06-13

... Trichoderma reesei; Proposed Significant New Use Rule AGENCY: Environmental Protection Agency (EPA). ACTION... Control Act (TSCA) for the genetically modified microorganism identified generically as Trichoderma reesei...: Trichoderma reesei (MCAN J-10-2) (generic). Chemical Abstracts Service (CAS) Registry Number: Not available...

In vitro antagonism of Thielaviopsis paradoxa by Trichoderma longibrachiatum.

Science.gov (United States)

Sánchez, Vladimir; Rebolledo, Oscar; Picaso, Rosa M; Cárdenas, Elizabeth; Córdova, Jesús; González, Orfil; Samuels, Gary J

2007-01-01

Seventy-nine Trichoderma strains were isolated from soil taken from 28 commercial plantations of Agave tequilana cv. 'Azul' in the State of Jalisco, Mexico. Nine of these isolates produced nonvolatile metabolites that completely inhibited the growth of Thielaviopsis paradoxa on potato dextrose agar plates. These isolates were identified as Trichoderma longibrachiatum on the basis of their morphology and DNA sequence analysis of two genes (ITS rDNA and translation elongation factor EF-1alpha). Mycoparasitism of Th. paradoxa by T. longibrachiatum strains in dual cultures was examined by scanning electron microscopy. The Trichoderma hyphae grew alongside the Th. paradoxa hyphae, but penetration of Thielaviopsis hyphae by Trichoderma was no apparent. Aleurioconidia of Th. paradoxa were parasitized by Trichoderma. Both hyphae and aleurioconidia of Th. paradoxa lost turgor pressure, wrinkled, collapsed and finally disintegrated. In liquid cultures, all nine Trichoderma isolates produced proteases, beta-1,3-glucanases and chitinases that would be responsible for the degradation of Thielaviopsis hyphae. These results demonstrate that the modes of action of T. longibrachiatum involved against Th. paradoxa in vitro experiments are mycoparasitism and the production of nonvolatile toxic metabolites.

Biological control of fusarium wilt of tomato by antagonist fungi and ...

African Journals Online (AJOL)

Biological control of Fusarium oxysporum f. sp. lycopersici (FOL) causing wilt disease of tomato was studied in vitro as well as under pot conditions. Dual culture technique showed that Aspergillus niger, Penicillium citrinum, Penicillium sp. and Trichoderma harzianum inhibited the radial colony growth of the test pathogen.

Development of a molecular approach to describe the composition of Trichoderma communities.

Science.gov (United States)

Meincke, Remo; Weinert, Nicole; Radl, Viviane; Schloter, Michael; Smalla, Kornelia; Berg, Gabriele

2010-01-01

Trichoderma and its teleomorphic stage Hypocrea play a key role for ecosystem functioning in terrestrial habitats. However, little is known about the ecology of the fungus. In this study we developed a novel Trichoderma-specific primer pair for diversity analysis. Based on a broad range master alignment, specific Trichoderma primers (ITSTrF/ITSTrR) were designed that comprise an approximate 650bp fragment of the internal transcribed spacer region from all taxonomic clades of the genus Trichoderma. This amplicon is suitable for identification with TrichoKey and TrichoBLAST. Moreover, this primer system was successfully applied to study the Trichoderma communities in the rhizosphere of different potato genotypes grown at two field sites in Germany. Cloning and sequencing confirmed the specificity of the primer and revealed a site-dependent Trichoderma composition. Based on the new primer system a semi-nested approach was used to generate amplicons suitable for denaturing gradient gel electrophoresis (DGGE) analysis and applied to analyse Trichoderma communities in the rhizosphere of potatoes. High field heterogeneity of Trichoderma communities was revealed by both DGGE. Furthermore, qPCR showed significantly different Trichoderma copy numbers between the sites. Copyright 2009 Elsevier B.V. All rights reserved.

Increasing rice plant growth by Trichoderma sp.

Science.gov (United States)

Doni, Febri; Isahak, Anizan; Zain, Che Radziah Che Mohd; Sulaiman, Norela; Fathurahman, F.; Zain, Wan Nur Syazana Wan Mohd.; Kadhimi, Ahsan A.; Alhasnawi, Arshad Naji; Anhar, Azwir; Yusoff, Wan Mohtar Wan

2016-11-01

Trichoderma sp. is a plant growth promoting fungi in many crops. Initial observation on the ability to enhance rice germination and vigor have been reported. In this study, the effectiveness of a local isolate Trichoderma asprellum SL2 to enhance rice seedling growth was assessed experimentally under greenhouse condition using a completely randomized design. Results showed that inoculation of rice plants with Trichoderma asprellum SL2 significantly increase rice plants height, root length, wet weight, leaf number and biomass compared to untreated rice plants (control). The result of this study can serve as a reference for further work on the application of beneficial microorganisms to enhance rice production.

Antagonism of Trichoderma isolates against Leucoagaricus gongylophorus (Singer) Möller.

Science.gov (United States)

do Nascimento, Mariela Otoni; de Almeida Sarmento, Renato; Dos Santos, Gil Rodrigues; de Oliveira, Cléia Almeida; de Souza, Danival José

2017-08-01

Filamentous fungi from the genus Trichoderma are commonly found in soil. They are considered facultative mycoparasites, and are antagonists of other fungi such as the cultivar of leaf-cutting ants (Leucoagaricus gongylophorus). The aim of the present study was to bioprospect Trichoderma spp. from different soils collected from Gurupi, Tocantins, Brazil, for antagonistic effects against the mutualistic fungus of leaf-cutting ants. To isolate filamentous fungi, samples were collected from six locations. Preliminarily, isolates were identified by morphological analysis as belonging to Trichoderma. Trichoderma spp. had their internal transcribed spacer region (ITS) of ribosomal RNA genes (rRNA) sequenced to confirm species-level taxonomy. L. gongylophorus was isolated from a laboratory ant colony. Antagonistic properties of seven isolates of Trichoderma against L. gongylophorus were measured using paired disks in Petri dishes with potato dextrose agar medium (PDA). All Trichoderma isolates inhibited the growth of L. gongylophorus in Petri dishes. Isolate 2 of Trichoderma spirale group exhibited slow mycelial growth in the Petri dish, and a high rate of inhibition against L. gongylophorus. This isolate is a promising fungus for field tests of biological control methods for leaf-cutting ants. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Snuffelen aan Trichoderma

NARCIS (Netherlands)

Baars, J.J.P.

2011-01-01

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

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

Science.gov (United States)

2010-04-01

... Trichoderma longibrachiatum. 184.1250 Section 184.1250 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT....1250 Cellulase enzyme preparation derived from Trichoderma longibrachiatum. (a) Cellulase enzyme preparation is derived from a nonpathogenic, nontoxicogenic strain of Trichoderma longibrachiatum (formerly T...

Trichoderma for climate resilient agriculture.

Science.gov (United States)

Kashyap, Prem Lal; Rai, Pallavi; Srivastava, Alok Kumar; Kumar, Sudheer

2017-08-01

Climate change is one of the biggest challenges of the twenty-first century for sustainable agricultural production. Several reports highlighted the need for better agricultural practices and use of eco-friendly methods for sustainable crop production under such situations. In this context, Trichoderma species could be a model fungus to sustain crop productivity. Currently, these are widely used as inoculants for biocontrol, biofertilization, and phytostimulation. They are reported to improve photosynthetic efficiency, enhance nutrient uptake and increase nitrogen use efficiency in crops. Moreover, they can be used to produce bio-energy, facilitate plants for adaptation and mitigate adverse effect of climate change. The technological advancement in high throughput DNA sequencing and biotechnology provided deep insight into the complex and diverse biotic interactions established in nature by Trichoderma spp. and efforts are being made to translate this knowledge to enhance crop growth, resistance to disease and tolerance to abiotic stresses under field conditions. The discovery of several traits and genes that are involved in the beneficial effects of Trichoderma spp. has resulted in better understanding of the performance of bioinoculants in the field, and will lead to more efficient use of these strains and possibly to their improvement by genetic modification. The present mini-review is an effort to elucidate the molecular basis of plant growth promotion and defence activation by Trichoderma spp. to garner broad perspectives regarding their functioning and applicability for climate resilient agriculture.

Influence of Environmental Parameters on Trichoderma Strains with Biocontrol Potential

Directory of Open Access Journals (Sweden)

Zsuzsanna Antal

2003-01-01

Full Text Available 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 important parameters to be considered are the effects of temperature, water potential and pH, and the presence of pesticides, metal ions and antagonistic bacteria in the soil. Most of the Trichoderma strains are mesophilic. Low temperatures in winter may cause a problem during biological control by influencing the activity of the biocontrol agents. Another problem emerging during the application of Trichoderma strains as biocontrol agents is that they cannot tolerate dry conditions, however, we may need biocontrol agents against plant pathogenic fungi which are able to grow and cause disease even in dry soils. The pH characteristics of the soil also belong to the most important environmental parameters affecting the activities of mycoparasitic Trichoderma strains. Within the frames of a complex integrated plant protection strategy, we may have to combine Trichoderma strains with chemical pesticides or metal compounds, therefore it is important to collect information about the effects of pesticides and metal ions on the biocontrol strains. Antagonistic soil bacteria may also have negative effects on the biocontrol abilities of Trichoderma strains, therefore it may be advantageous if a biocontrol strain possesses bacterium- degrading abilities as well. This review will discuss the literature about the influence of temperature, water potential, pH, pesticides, metal ions and antagonistic bacteria on mycoparasitic Trichoderma strains including the results of our

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

International Nuclear Information System (INIS)

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

1986-01-01

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

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

International Nuclear Information System (INIS)

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

1986-01-01

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

Comparative study of impact of Azotobacter and Trichoderma with other fertilizers on maize growth

Directory of Open Access Journals (Sweden)

Sanjay Mahato

2017-12-01

Full Text Available Biofertilizers may be a better eco-friendly option to maintain soil fertility. The study was conducted to investigate the effect of Azotobacter and Trichoderma on the vegetative growth of maize (Zea mays L. plants. The experiment was carried out in medium sized pots, at IAAS, Lamjung (Feb 2017 - May 2017 in completely randomized design (CRD, consisting eight treatments and three replications. Treatments were namely T1 (control, T2 (Azotobacter, T3 (Trichoderma, T4 (Azotobacter + Trichoderma, T5 (NPK, T6 (Azotobacter + Trichoderma + FYM, T7 (Azotobacter + Trichoderma + FYM + NPK, T8 (FYM. Azotobacter showed a positive increase in plant height, stem girth, dry shoot weight, root length and width, and root weight while Trichoderma displayed either negative or minimal impact. Effect of FYM was lower than Azotobacter but considerably higher than Trichoderma. Trichoderma seriously inhibited the expression of Azotobacter when used together. Trichoderma even suppressed the outcome (except shoot weight of FYM when used together. Root length was the longest in Azotobacter inoculation. The highest number of leaves was in T7 followed by Azotobacter (T2 and NPK (T5. Unlike leaf width, Azotobacter showed a negligible increase in leaves length while Trichoderma wherever present showed the negative impact. Minimum chlorophyll content was found in Azotobacter or Trichoderma after 73 days. Azotobacter treatment showed early tasseling than Trichoderma. The association of Azotobacter and Trichoderma increased the biomass. Azotobacter has significant effects on growth parameters of maize and can supplement chemical fertilizer, while Trichoderma was found to inhibit most of the growth parameters.

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.

Trichoderma (Hypocrea) species with green ascospores from China

NARCIS (Netherlands)

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

2015-01-01

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

Use of gamma radiation for increasing plant disease control efficacy

International Nuclear Information System (INIS)

Chamswarng, Chiradej; Intanoo, Wanwilai; Piadang, Nattayana

2006-09-01

Irradiation of Trichoderma harzianum with 0.5-8.0 k Gay of gamma ray revealed 41 strains resistant to 10 ppm propiconazole or benomyl fungicides and two strains (23/03-7 and 27/08-1) were resistant to 10 ppm of both fungicides. After these two mutant strains were repeatedly irradiated with gamma ray, 74 mutant strains were obtained. Among these, three mutant strains, used as seed treatment effectively protected tomato seedlings from the infection of Pythium apanidermatum with significantly higher surviving seedlings than the Pythium inoculated control. The higher root colonization of mutant strains was obtains from strains 03/7-113, 03-/7-114 and 08/1-11. Rice seeds (RD 17), previously soaked in water for 24 ht were placed in spore suspension of T. harzianum prepared from 1 kg of fresh culture of 81 mutant isolated derived from single or double irradiation with gamma ray in 50 1 of water for 30 min. Two mutants including 23/03-7 (derived from single irradiation) and 03/7-134 (derived from double irradiation) provided percentages of root colonization by 29.63 and 25.93, while growth-promoted roots were 12.48 and 12.65 cm in length. respectively. These two strains were tested in rice field by treating pre-soaked seeds with Trichoderma suspensions for 30 min and incubated for 24 hr before sowing. Detection of root colonization by T. harzianum at 35, 45 and 102 days after sowing revealed that all Trichoderma strains effectively colonized rice roots at all stages of growth, particularly two mutants completely colonized rice root at 102 days after sowing. After harvesting, a mutant strain 30/7-134 increased rice yield to the maximum level at 29.65% over a control, while the percentage of fertile-seeds and it's seed weight, total seed weight, fertile-seed weight were significantly higher than a control. However, all Trichoderma strains provided the potential increases of rice yield over a control. Strain 03/7-134 significantly reduced percentage of dirty-panicle diseased

Trichoderma genes

Science.gov (United States)

Foreman, Pamela [Los Altos, CA; Goedegebuur, Frits [Vlaardingen, NL; Van Solingen, Pieter [Naaldwijk, NL; Ward, Michael [San Francisco, CA

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.

Isolation and identification of two marine-derived Streptomyces from ...

African Journals Online (AJOL)

Administrator

2011-09-26

Sep 26, 2011 ... evaluated against the test phytopathogenic fungi; the results showed that the FBE of S. ... largest antibiotic-producing actinomycetes (Imada et al.,. 2005; Raja et al., 2010). ... sediments within the neritic zone in the sea area round. Zhuhai city of ..... effects of the ethyl acetate extract of Trichoderma harzianum.

Biology and biotechnology of Trichoderma.

Science.gov (United States)

Schuster, André; Schmoll, Monika

2010-07-01

Fungi of the genus Trichoderma are soilborne, green-spored ascomycetes that can be found all over the world. They have been studied with respect to various characteristics and applications and are known as successful colonizers of their habitats, efficiently fighting their competitors. Once established, they launch their potent degradative machinery for decomposition of the often heterogeneous substrate at hand. Therefore, distribution and phylogeny, defense mechanisms, beneficial as well as deleterious interaction with hosts, enzyme production and secretion, sexual development, and response to environmental conditions such as nutrients and light have been studied in great detail with many species of this genus, thus rendering Trichoderma one of the best studied fungi with the genome of three species currently available. Efficient biocontrol strains of the genus are being developed as promising biological fungicides, and their weaponry for this function also includes secondary metabolites with potential applications as novel antibiotics. The cellulases produced by Trichoderma reesei, the biotechnological workhorse of the genus, are important industrial products, especially with respect to production of second generation biofuels from cellulosic waste. Genetic engineering not only led to significant improvements in industrial processes but also to intriguing insights into the biology of these fungi and is now complemented by the availability of a sexual cycle in T. reesei/Hypocrea jecorina, which significantly facilitates both industrial and basic research. This review aims to give a broad overview on the qualities and versatility of the best studied Trichoderma species and to highlight intriguing findings as well as promising applications.

Effect of metabolites produced by Trichoderma species against ...

African Journals Online (AJOL)

Metabolites released from Trichoderma viride, T. polysporum, T. hamatum and T. aureoviride were tested in culture medium against Ceratocystis paradoxa, which causes black seed rot in oil palm sprouted seeds. The Trichoderma metabolites had similar fungistatic effects on the growth of C. paradoxa except those from T.

Linkage mapping of candidate genes for induce resistance and growth promotion by trichoderma koningiopsis (th003) in tomato solanum lycopersicum

International Nuclear Information System (INIS)

Simbaqueba, Jaime; Cotes, Alba Marina; Barrero, Luz Stella

2011-01-01

Induced systemic resistance (ISR) is a mechanism by which plants enhance defenses against any stress condition. ISR and growth promotion are enhanced when tomato (Solanum lycopersicum) is inoculated with several strains of Trichoderma ssp. this study aims to genetically map tomato candidate genes involved in ISR and growth promotion induced by the Colombian native isolate Trichoderma koningiopsis th003. Forty-nine candidate genes previously identified on tomato plants treated with th003 and T. hamatum T382 strains were evaluated for polymorphisms and 16 of them were integrated on the highly saturated genetic linkage map named TOMATO EXPEN 2000. The location of six unigenes was similar to the location of resistance gene analogs (RGAS), defense related ests and resistance QTLs previously reported, suggesting new possible candidates for these quantitative trait loci (QTL) regions. The candidate gene-markers may be used for future ISR or growth promotion assisted selection in tomato.

Trichoderma-plant-pathogen interactions: advances in genetics of biological control.

Science.gov (United States)

Mukherjee, Mala; Mukherjee, Prasun K; Horwitz, Benjamin A; Zachow, Christin; Berg, Gabriele; Zeilinger, Susanne

2012-12-01

Trichoderma spp. are widely used in agriculture as biofungicides. Induction of plant defense and mycoparasitism (killing of one fungus by another) are considered to be the most important mechanisms of Trichoderma-mediated biological control. Understanding these mechanisms at the molecular level would help in developing strains with superior biocontrol properties. In this article, we review our current understanding of the genetics of interactions of Trichoderma with plants and plant pathogens.

77 FR 4903 - Trichoderma

Science.gov (United States)

2012-02-01

...-0245- 0008;oldLink=false. 18. U.S. EPA. 2010a. Trichoderma asperellum strain ICC 012 Biopesticides... EPA consideration of voluntary consensus standards pursuant to section 12(d) of the National...

Postharvest biological control of brown rot in peaches after cold storage preceded by preharvest chemical control 1

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Elizandra Pivotto Pavanello

2015-12-01

Full Text Available ABSTRACT Pathogenic fungi cause skin darkening and peach quality depreciation in post harvest. Therefore, alternative techniques to chemical treatment are necessary in order to reduce risks to human health. The aim of this study was to evaluate the effect of the application of Trichoderma harzianum in association with different fungicides applied before harvest to 'Eldorado' peaches for brown rot control and other quality parameters during storage. The treatments consisted of five preharvest fungicide applications (control, captan, iprodione, iminoctadine and tebuconazole associated with postharvest application of T. harzianum, after cold storage (with and without application, in three evaluation times (zero, two and four days at 20 °C, resulting in a 5x2x3 factorial design. The application of T. harzianum only brought benefits to the control of brown rot when combined with the fungicide captan, at zero day shelf life. After two days, there was a greater skin darkening in peaches treated with T. harzianum compared with peaches without the treatment, except for peaches treated with the fungicide iprodione and T. harzianum The application of T. harzianum during postharvest showed no benefits for the control of brown rot, however, the association with fungicides reduced the incidence of Rhizopus stolonifer during the shelf life.

Can Tomato Inoculation with Trichoderma Compensate Yield and Soil Health Deficiency due to Soil Salinity?

Science.gov (United States)

Wagner, Karl; Apostolakis, Antonios; Daliakopoulos, Ioannis; Tsanis, Ioannis

2016-04-01

Soil salinity is a major soil degradation threat, especially for arid coastal environments where it hinders agricultural production and soil health. Protected horticultural crops in the Mediterranean region, typically under deficit irrigation and intensive cultivation practices, have to cope with increasing irrigation water and soil salinization. This study quantifies the beneficial effects of the Trichoderma harzianum (TH) on the sustainable production of Solanum lycopersicum (tomato), a major greenhouse crop of the RECARE project Case Study in Greece, the semi-arid coastal Timpaki basin in south-central Crete. 20 vigorous 20-day-old Solanum lycopersicum L. cv Elpida seedlings are treated with TH fungi (T) or without (N) and transplanted into 35 L pots under greenhouse conditions. Use of local planting soil with initial Electrical Conductivity (ECe) 1.8 dS m-1 and local cultivation practices aim to simulate the prevailing conditions at the Case Study. In order to simulate seawater intrusion affected irrigation, plants are drip irrigated with two NaCl treatments: slightly (S) saline (ECw = 1.1 dS m-1) and moderately (M) saline water (ECw = 3.5 dS m-1), resulting to very high and excessively high ECe, respectively. Preliminary analysis of below and aboveground biomass, soil quality, salinity, and biodiversity indicators, suggest that TH pre-inoculation of tomato plants at both S and M treatments improve yield, soil biodiversity and overall soil health.

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...

Evaluation of quality parameters of strawberry fruits in modified ...

African Journals Online (AJOL)

STORAGESEVER

2009-03-06

Mar 6, 2009 ... postharvest physiology of strawberry. In: Luby JJ, Dale A (Eds.), The. Strawberry into the 21th Century. Timber Press, Portland, OR, pp. 145-152. Kovach J, Petzoldt R, Harman GE (2000). Use honey bees and bumble bees to disseminate Trichoderma harzianum 1295-22 to straw- berries for botrytis control.

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

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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

Pengendalian Hayati Penyakit Layu Fusarium Pisang (Fusarium oxysporum f.sp. cubense dengan Trichoderma sp.

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Albertus Sudirman

2011-07-01

Full Text Available The aim of this research was to study the inhibiting ability of Trichoderma sp. to control fusarium wilt of banana in greenhouse condition. The experiments consisted of the antagonism test between Trichoderma sp. and Fusarium oxysporum f.sp. cubense (Foc in vitro using dual culture method and glass house experiment which was arranged in 3×3 Factorial Complete Randomized Design. First factor of the latter experiment was the dose of Trichoderma sp. culture (0, 25, and 50 g per polybag, second factor was time of Trichoderma culture application (2 weeks before Foc inoculation, at same time with Foc inoculation and 2 weeks after Foc inoculation. Trichoderma sp. was cultured in mixed rice brand and chaff medium. The disease intensity was observed with scoring system of wilting leaves (0–4. The results showed that Trichoderma sp. was antagonistic against Foc in vitro and inhibited 86% of Foc colony development. Mechanism of antagonism between Trichoderma sp. and Foc was hyperparasitism. Trichoderma hyphae coiled around Foc hyphae. Lysis of Foc hyphae was occurred at the attached site of Trichoderma hyphae on Foc hyphae. Added banana seedling with Trichoderma sp. Culture reduced disease intensity of Fusarium wilt. Suggested dose of Trichoderma culture application in glass house was 25 g/polybag, given at the same time with Foc inoculation. Penelitian ini bertujuan untuk mengetahui kemampuan Trichoderma sp. untuk pengendalian penyakit layu fusarium pisang di rumah kaca. Penelitian meliputi pengujian daya hambat Fusarium oxysporum f.sp. cubense (Foc in vitro dan kemampuan menekan intensitas penyakit di rumah kaca. Penelitian in vitro meliputi uji antagonisme dan mekanismenya yang dilakukan secara dual culture. Uji pengaruh Trichoderma sp. terhadap penyakit layu Fusarium dilakukan di rumah kaca dengan Rancangan Acak Lengkap Faktorial. Faktor pertama adalah dosis biakan Trichoderma sp., dengan tiga aras (0, 25, 50 g/per bibit dalam polibag. Faktor kedua

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

International Nuclear Information System (INIS)

Naher, L.; Ismail, A.

2014-01-01

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)

Cellulase production by two mutant strain of Trichoderma longibrachiatum Qm9414 and Rut C30; Produccion de celulasas a partir de dos cepas hiperproductoras de trichoderma longibrachiatum Qm9414 y Rut C30

Energy Technology Data Exchange (ETDEWEB)

Blanco, M.J.

1991-12-31

Native or pretreated biomass from Onopordum nervosum boiss, has been examined as candidate feedstock for cellulase production by two mutant strain of trichoderma longibrachiatum QM9414 and Rut C30. Batch cultivation methods were evaluated and compared with previous experiments using ball-milled, crystalline cellulose (Solka floc). Batch cultivation of T. longibrachiatum Rut C30 on 55% (W/V) acid pretreated O. nervosum biomass yielded enzyme productivities and activities comparable to those obtained on Solka floc. However, the overall enzyme production performance was lower than on Solka floc at comparable cellulose concentrations. This fact may be due to the accumulation of pretreated by products and lignin in the fermentor.(author)

Cellulase production by two mutant strain of Trichoderma longibrachiatum Qm9414 and Rut C30. Produccion de celulasas a partir de dos cepas hiperproductoras de trichoderma longibrachiatum Qm9414 y Rut C30

Energy Technology Data Exchange (ETDEWEB)

Blanco, M.J.

1991-01-01

Native or pretreated biomass from Onopordum nervosum boiss, has been examined as candidate feedstock for cellulase production by two mutant strain of trichoderma longibrachiatum QM9414 and Rut C30. Batch cultivation methods were evaluated and compared with previous experiments using ball-milled, crystalline cellulose (Solka floc). Batch cultivation of T. longibrachiatum Rut C30 on 55% (W/V) acid pretreated O. nervosum biomass yielded enzyme productivities and activities comparable to those obtained on Solka floc. However, the overall enzyme production performance was lower than on Solka floc at comparable cellulose concentrations. This fact may be due to the accumulation of pretreated by products and lignin in the fermentor.(author)

Proteomic response of the biological control fungus Trichoderma atroviride to growth on the cell walls of Rhizoctonia solani.

Science.gov (United States)

Grinyer, Jasmine; Hunt, Sybille; McKay, Matthew; Herbert, Ben R; Nevalainen, Helena

2005-06-01

Trichoderma atroviride has a natural ability to parasitise phytopathogenic fungi such as Rhizoctonia solani and Botrytis cinerea, therefore providing an environmentally sound alternative to chemical fungicides in the management of these pathogens. Two-dimensional electrophoresis was used to display cellular protein patterns of T. atroviride (T. harzianum P1) grown on media containing either glucose or R. solani cell walls. Protein profiles were compared to identify T. atroviride proteins up-regulated in the presence of the R. solani cell walls. Twenty-four protein spots were identified using matrix-assisted laser desorption ionisation mass spectrometry, liquid chromatography mass spectrometry and N-terminal sequencing. Identified up-regulated proteins include known fungal cell wall-degrading enzymes such as N-acetyl-beta-D: -glucosaminidase and 42-kDa endochitinase. Three novel proteases of T. atroviride were identified, containing sequence similarity to vacuolar serine protease, vacuolar protease A and a trypsin-like protease from known fungal proteins. Eukaryotic initiation factor 4a, superoxide dismutase and a hypothetical protein from Neurospora crassa were also up-regulated as a response to R. solani cell walls. Several cell wall-degrading enzymes were identified from the T. atroviride culture supernatant, providing further evidence that a cellular response indicative of biological control had occurred.

Isolation and identification of Bacillus spp. from compost material, compost and mushroom casing soil active against Trichoderma spp.

Directory of Open Access Journals (Sweden)

Stanojević Olja

2016-01-01

Full Text Available The isolation of bacteria was carried out from samples of straw and chicken manure, compost at various stages of the composting process and casing soil used for growing button mushrooms. A preliminary screening of 108 bacterial isolates for antagonistic activity against Trichoderma aggressivum f. europaeum showed that 23 tested isolates inhibited mycelial growth of the pathogenic fungus. Further screening with four indicator isolates of fungi revealed that all 23 bacterial isolates inhibited the growth of T. aggressivum f. europaeum, T. harzianum and T. koningii, while only 13 isolates inhibited the growth of T. atroviride. T. aggressivum f. europaeum proved to be the most sensitive, with many bacterial isolates generating a high percentage of growth inhibition. Only two bacterial isolates (B-129 and B-268 were successful in inhibiting the growth of all 4 tested pathogens. All 23 bacterial isolates were characterized as Gram-positive and catalase-positive and were subjected to molecular identification based on the partial sequence, the hypervariant region of the 16S rDNA. It was shown that the obtained bacterial strains belong to Bacillus subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus species. [Projekat Ministarstva nauke Republike Srbije, br. 31043 i br. 173026

Three new species of Trichoderma with hyaline ascospores from China.

Science.gov (United States)

Zhu, Z X; Zhuang, W Y

2015-01-01

Collections of Trichoderma having hyaline ascospores from different areas of China were examined. Using combined analyses of morphological data, culture characters and phylogenetic information based on rDNA sequences of partial nuc translation elongation factor 1-α encoding gene (TEF1-α) and the gene encoding the second largest nuc RNA polymerase subunit (RPB2), three new species, Trichoderma applanatum, T. oligosporum and T. sinoluteum, were discovered and are described. Trichoderma applanatum produces continuous flat to pulvinate, white to cream stromata with dense orange or pale brown ostioles, and simple acremonium-like to verticillium-like conidiophores, belongs to the Hypocreanum clade and is closely related to T. decipiens. Trichoderma oligosporum forms reddish brown stromata with a downy surface, hyaline conidia and gliocladium-like conidiophores, and is closely related to but distinct from T. crystalligenum in the Psychrophila clade. Trichoderma sinoluteum, as a member of the Polysporum clade, is characterized by pale yellow stromata, white pustulate conidiomata, pachybasium-like conidiophores, and hyaline conidia. Differences between the new species and their close relatives are discussed. © 2015 by The Mycological Society of America.

[Antagonism of Trichoderma spp. to fungi caused root rot of Sophora tonkinensis].

Science.gov (United States)

Qin, Liu-yan; Jiang, Ni; Tang, Mei-qiong; Miao, Jian-hua; Li, Lin-xuan

2011-04-01

To study the antagonism of Trichoderma spp. to fungi S9(Fusarium solani)which caused root rot of Sophora tonkinensis and discuss the further develop prospects of microbial biological control in soil-borne diseases on Chinese herbal medicines. Antagonism of H2 (Trichoderma harsianum), M6 (Trichoderma viride) and K1 (Trichoderma koningii) to Fusarium solani were researched by growth rate and confront culture. And their mechanisms were discussed. H2 and M6 had obvious competitive advantage, the growth rate of which were 1.43-2.72 times and 1.43-1.95 times as S9 respectively. The space competitive advantage of K1 was relatively weak; the growth rate was slower than S9. The antagonism of three species of Trichoderma spp. to S9 was in varying degrees. The antagonism to S9 of M6 and H2 was better,the inhibition rate were 100% and 82.35% respectively, even cultivated S9 for three days in advance. And their inhibition indexes were both reached class I. The inhibition index and inhibition rate of K1 was respectively 46.36% and class IV. The Trichoderma spp. could cause S9 mycelium to appear some phenomenon just like fracture, constriction reduced, digestion, etc. which were observed under the microscope. Trichoderma harsianum and Trichoderma viride showed the further develop prospects in the fight against soil-borne disease on Chinese herbal medicines.

Synthesis of enzymes connected with mycoparasitism by ectomycorrhizal fungi.

Science.gov (United States)

Mucha, Joanna; Dahm, Hanna; Strzelczyk, Edmund; Werner, Antoni

2006-03-01

The production of enzymes involved in mycoparasitism by several strains of ectomycorrhizal fungi: Amanita muscaria (16-3), Laccaria laccata (9-12), L. laccata (9-1), Suillus bovinus (15-4), S. bovinus (15-3), S. luteus (14-7) on different substrates such as colloidal chitin, mycelia of Trichoderma harzianum, T. virens and Mucor hiemalis was examined. Chitinases and beta-1,3-glucanases were assayed spectrophotometrically by measuring the amount of reducing sugars releasing from suitable substrate by means of Miller's method. Beta-glucosidases were determined by measuring the amount of p-nitrophenol released from p-nitrophenyl-beta-D-glucopyranoside. It was observed that A. muscaria (16-3) and L. laccata (9-12) biosynthesized the highest activity of enzymes in contrast to the strains of S. bovinus and S. luteus. The mycelium of T. harzianum turned out to be the best substrate for the induction of beta-1,3-glucanases and beta-glucosidases for both strains of L. laccata, although the difference in the induction of chitinases in the presence of mycelia of different species of Trichoderma was not indicated.

Biocontrol evaluation of wheat take-all disease by Trichoderma ...

African Journals Online (AJOL)

Wheat take-all disease, caused by Gaeumannomyces graminis var tritici (Ggt), has been observed in different areas of Iran in recent years. Current biocontrol studies have confirmed the effectiveness of the. Trichoderma species against many fungal phytopathogens. In this study, biocontrol effects of Trichoderma isolates ...

Biodiversity of Trichoderma (Hypocreaceae) in Southern Europe and Macaronesia

Science.gov (United States)

Jaklitsch, W.M.; Voglmayr, H.

2015-01-01

The first large-scale survey of sexual and asexual Trichoderma morphs collected from plant and fungal materials conducted in Southern Europe and Macaronesia including a few collections from French islands east of Africa yielded more than 650 specimens identified to the species level. Routine sequencing of tef1 revealed a genetic variation among these isolates that exceeds previous experience and ca. 90 species were recognized, of which 74 are named and 17 species newly described. Aphysiostroma stercorarium is combined in Trichoderma. For the first time a sexual morph is described for T. hamatum. The hitherto most complete phylogenetic tree is presented for the entire genus Trichoderma, based on rpb2 sequences. For the first time also a genus-wide phylogenetic tree based on acl1 sequences is shown. Detailed phylogenetic analyses using tef1 sequences are presented in four separate trees representing major clades of Trichoderma. Discussions involve species composition of clades and ecological and biogeographic considerations including distribution of species. PMID:26955191

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

Science.gov (United States)

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

2014-10-01

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