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Sample records for biocontrol fungus trichoderma

  1. [Improvement of Trichoderma strains for biocontrol].

    Science.gov (United States)

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

    2000-03-01

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

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

    Institute of Scientific and Technical Information of China (English)

    HUANG Caihong; YANG Qian

    2007-01-01

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

  3. Suppression of the biocontrol agent trichoderma harzianum by mycelium of the arbuscular mycorrhizal fungus glomus intraradices in root-free soil

    Science.gov (United States)

    Green; Larsen; Olsson; Jensen; Jakobsen

    1999-04-01

    Trichoderma harzianum is an effective biocontrol agent against several fungal soilborne plant pathogens. However, possible adverse effects of this fungus on arbuscular mycorrhizal fungi might be a drawback in its use in plant protection. The objective of the present work was to examine the interaction between Glomus intraradices and T. harzianum in soil. The use of a compartmented growth system with root-free soil compartments enabled us to study fungal interactions without the interfering effects of roots. Growth of the fungi was monitored by measuring hyphal length and population densities, while specific fatty acid signatures were used as indicators of living fungal biomass. Hyphal 33P transport and beta-glucuronidase (GUS) activity were used to monitor activity of G. intraradices and a GUS-transformed strain of T. harzianum, respectively. As growth and metabolism of T. harzianum are requirements for antagonism, the impact of wheat bran, added as an organic nutrient source for T. harzianum, was investigated. The presence of T. harzianum in root-free soil reduced root colonization by G. intraradices. The external hyphal length density of G. intraradices was reduced by the presence of T. harzianum in combination with wheat bran, but the living hyphal biomass, measured as the content of a membrane fatty acid, was not reduced. Hyphal 33P transport by G. intraradices also was not affected by T. harzianum. This suggests that T. harzianum exploited the dead mycelium but not the living biomass of G. intraradices. The presence of external mycelium of G. intraradices suppressed T. harzianum population development and GUS activity. Stimulation of the hyphal biomass of G. intraradices by organic amendment suggests that nutrient competition is a likely means of interaction. In conclusion, it seemed that growth of and phosphorus uptake by the external mycelium of G. intraradices were not affected by the antagonistic fungus T. harzianum; in contrast, T. harzianum was adversely

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

    Directory of Open Access Journals (Sweden)

    Susanne Zeilinger

    2007-01-01

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

  5. Use of a non-homologous end-joining-deficient strain (delta-ku70) of the biocontrol fungus Trichoderma virens to investigate the function of the laccase gene lcc1 in sclerotia degradation.

    Science.gov (United States)

    Catalano, Valentina; Vergara, Mariarosaria; Hauzenberger, Jasmin R; Seiboth, Bernhard; Sarrocco, Sabrina; Vannacci, Giovanni; Kubicek, Christian P; Seidl-Seiboth, Verena

    2011-02-01

    The aim of this study was to apply a generated Δtku70 strain with increased homologous recombination efficiency from the mycoparasitic fungus Trichoderma virens for studying the involvement of laccases in the degradation of sclerotia of plant pathogenic fungi. Inactivation of the non-homologous end-joining pathway has become a successful tool in filamentous fungi to overcome poor targeting efficiencies for genetic engineering. Here, we applied this principle to the biocontrol fungus T. virens, strain I10, by deleting its tku70 gene. This strain was subsequently used to delete the laccase gene lcc1, which we found to be expressed after interaction of T. virens with sclerotia of the plant pathogenic fungi Botrytis cinerea and Sclerotinia sclerotiorum. Lcc1 was strongly upregulated at early colonization of B. cinerea sclerotia and steadily induced during colonization of S. sclerotiorum sclerotia. The Δtku70Δlcc1 mutant was altered in its ability to degrade the sclerotia of B. cinerea and S. sclerotiorum. Interestingly, while the decaying ability for B. cinerea sclerotia was significantly decreased, that to degrade S. sclerotiorum sclerotia was even enhanced, suggesting the operation of different mechanisms in the mycoparasitism of these two types of sclerotia by the laccase LCC1. PMID:20872221

  6. Biocontrol of Rhizoctonia solani with Trichoderma Spp.

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

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

  7.   The fungus Trichoderma spp. on vegetables

    DEFF Research Database (Denmark)

    Winding, Anne

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

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Institute of Scientific and Technical Information of China (English)

    Sanz L; Hermosa M R; González F J; Monte E

    2004-01-01

    @@ Species of the fungus Trichoderma, a genus of Hyphomycetes, are ubiquitous in the environment, but especially in soil. They have been used in a wide range of commercial applications including the production of hydrolases and in the biological control of plant diseases. A fundamental part of the Trichoderma antifungal system consists of a series of genes coding for a surprising variety of extracellular cell wall degrading enzymes (CWDE).Characterisation and identification of strains at the species level is the first step in utilizing the full potential of fungi in specific applications. One aim when isolating Trichoderma strains is to identify those which can be used in new agricultural and industrial applications. In the past it was not uncommon that biocontrol strains were defined as T. harzianum Rifai, due to the limited classification system of the genus Trichoderma. In recent years, several PCR-based molecular techniques have been used to detect and discriminate among microorganisms. Sequence analysis of the ITS regions of the ribosomal DNA and gene fragments as those corresponding to tef1 gene have been helpful in the neotypification, description and characterization of species in the genus Trichoderna.Another useful method for the identification of Trichoderma strains is the randomly amplified polymorphic DNA (RAPD) technique.Isozyme polymorphisms evaluation of five putative extracellular lytic enzymes loci (β-1,3-glucanase, β-1,6-glucanase, cellulase, chitinase and protease antivities) were carried out using representative strains of defined molecular groups. CWDE groupings obtained from biocontrol strains are discussed in relation to their phylogenetic location and antifungal activities.Compiling morphological, biochemical and sequence information data into a common database would provide a useful resource that could be used to accurately name new haplotypes identified in the future and correctly place them within the genus Trichoderma.

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  11. Trichoderma spp.capable of growing at low temperatures with biocontrol potential

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Tavaszi A

    2004-01-01

    @@ Though there are successful commercial biocontrol products with Trichoderma spp. In many countries including US, Israel and Europe, their usefulness is limited in cold environments such as mid-and northern part of Europe, US and Canada, especially in the late fall, winter and early spring period.Trichoderma isolates capable of growing at low temperatures (5-10 ℃) and showing good antagonistic properties against plant pathogenic fungi may have therefore both scientific and commercial value.

  12. Enhanced biocontrol activity of Trichoderma through inactivation of a mitogen-activated protein kinase

    OpenAIRE

    Mendoza-Mendoza, Artemio; Pozo, María J.; Grzegorski, Darlene; Martínez, Pedro; García, Juan M.; Olmedo-Monfil, Vianey; Cortés, Carlos; Kenerley, Charles; Herrera-Estrella, Alfredo

    2003-01-01

    The production of lytic enzymes in Trichoderma is considered determinant in its parasitic response against fungal species. A mitogen-activated protein kinase encoding gene, tvk1, from Trichoderma virens was cloned, and its role during the mycoparasitism, conidiation, and biocontrol was examined in tvk1 null mutants. These mutants showed a clear increase in the level of the expression of mycoparasitism-related genes under simulated mycoparasitism and during direct confrontation with the plant ...

  13. Draft Whole-Genome Sequence of the Biocontrol Agent Trichoderma harzianum T6776.

    Science.gov (United States)

    Baroncelli, Riccardo; Piaggeschi, Giulia; Fiorini, Lisa; Bertolini, Edoardo; Zapparata, Antonio; Pè, Mario Enrico; Sarrocco, Sabrina; Vannacci, Giovanni

    2015-01-01

    Trichoderma harzianum T6776 is a promising beneficial isolate whose effects consist of growth promotion, positive response of photosynthetic activity, hormonal signaling, and carbon partitioning in tomato, coupled with biocontrol of plant pathogens. Here, we present the first genome assembly of T6776, providing a useful platform for the scientific community. PMID:26067977

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:24813508

  16. Construction of new GFP-tagged fusants for Trichoderma harzianum with enhanced biocontrol activity

    Directory of Open Access Journals (Sweden)

    Kowsari Mojegan

    2014-07-01

    Full Text Available Trichoderma is one of the most exploited biocontrol agents for the management of plant diseases. In biocontrol ecology, the critical factors are detection, and the monitoring and recovery of specific biocontrol agents either naturally present or deliberately released into the environment. Protoplast fusion is an appropriate tool for the improvement of biocontrol Trichoderma strains. Protoplast isolation from Trichoderma harzianum was achieved using 24 h culture age, 6.6 mg/ml Novazym L 1412 at 30°C which resulted the maximum protoplast yield of 5 × 108/ml. The self-fused protoplasts were regenerated and 12 fusants were selected based on their growth rate on 2% colloidal chitin medium. Next, a comparison was done for chitinase and antagonistic activity. Transcriptomic analysis based on quantitative real-time RT-PCR, demonstrated that T8-05 fusant expressed 1.5 fold of chit42 transcript as compared with the parental line. This fusant with 7.02±0.15U chitinase activity showed a higher growth inhibition rate (100% than the parent strain, against Rhizoctonia solani. To obtain a genetically marked fusant that can be used as a biomonitor, the fusant was cotransformed with the gfp and amdS genes. The morphology and viability of selected cotransformant (FT8-7MK-05-2 was similar to the parent. Green fluorescing conidia were observed within the first 2 days of incubation in the soil, and this was followed by the formation of chlamydopores after 60 days. The colonisation of the gfp-tagged fusant was also monitored visually on R. solani sclerotia by scanning electron microscopy. Production of gfp-tagged fusant of Trichoderma spp. provides a potentially useful tool for monitoring hyphal growth patterns and the population of biocontrol agent isolates introduced into environmental systems.

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

    NARCIS (Netherlands)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A.; van Wees, Saskia C M

    2014-01-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the ph

  18. Utilization of winery wastes for Trichoderma viride biocontrol agent production by solid state fermentation

    Institute of Scientific and Technical Information of China (English)

    BAI Zhihui; JIN Bo; LI Yuejie; CHEN Jian; LI Zuming

    2008-01-01

    Biocontrol agents are safe and environmental friendly alternatives for pesticides in agriculture application.Trichoderma v/ride WEBL0703 performed a high level of antagonistic activity toward a broad spectrum of phytopathogens and was determined as a biocontrol agent,which was produced by solid state fermentation using grape marc and wine lees.The maximum yield of T.viride conidia was up to 6.65×109 CFU/g initial dry substrate (IDS) after 10 d fermentation.As important enzymes for protecting plants from disease,ehitinase,β-glucanase,and pectinase yields were 47.8 U/g IDS,8.32 U/g IDS and 9.83 U/g IDS,respectively.These results show that it is feasible to convert winery wastes to a value-added and environmental friendly biocontrol agent.

  19. 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 ...

  20. 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. PMID:26774866

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-03-17

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

  2. RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease.

    Science.gov (United States)

    Romão-Dumaresq, Aline S; de Araújo, Welington Luiz; Talbot, Nicholas J; Thornton, Christopher R

    2012-01-01

    The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-ß-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte. PMID:23110120

  3. RNA interference of endochitinases in the sugarcane endophyte Trichoderma virens 223 reduces its fitness as a biocontrol agent of pineapple disease.

    Directory of Open Access Journals (Sweden)

    Aline S Romão-Dumaresq

    Full Text Available The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of the fungus, and to determine their roles in the biocontrol of soil-borne plant pathogens. The loss of N-acetyl-ß-D-glucosaminidase activities was dispensable for biocontrol of the plurivorous damping-off pathogens Rhizoctonia solani and Sclerotinia sclerotiorum, and of the sugarcane pathogen Ceratocystis paradoxa, the causal agent of pineapple disease. Similarly, suppression of endochitinase activities had no effect on R. solani and S. sclerotiorum disease control, but had a pronounced effect on the ability of T. virens 223 to control pineapple disease. Our work demonstrates a critical requirement for T. virens 223 endochitinase activity in the biocontrol of C. paradoxa sugarcane disease, but not for general antagonism of other soil pathogens. This may reflect its lifestyle as a sugarcane root endophyte.

  4. Systematics of the Trichoderma harzianum species complex and the re-identification of commercial biocontrol strains.

    Science.gov (United States)

    Chaverri, Priscila; Branco-Rocha, Fabiano; Jaklitsch, Walter; Gazis, Romina; Degenkolb, Thomas; Samuels, Gary J

    2015-01-01

    Trichoderma harzianum is known as a cosmopolitan, ubiquitous species associated with a wide variety of substrates. It is possibly the most commonly used name in agricultural applications involving Trichoderma, including biological control of plant diseases. While various studies have suggested that T. harzianum is a species complex, only a few cryptic species are named. In the present study the taxonomy of the T. harzianum species complex is revised to include at least 14 species. Previously named species included in the complex are T. guizhouense, T. harzianum, and T. inhamatum. Two new combinations are proposed, T. lentiforme and T. lixii. Nine species are described as new, T. afarasin, T. afroharzianum, T. atrobrunneum, T. camerunense, T. endophyticum, T. neotropicale, T. pyramidale, T. rifaii and T. simmonsii. We isolated Trichoderma cultures from four commercial biocontrol products reported to contain T. harzianum. None of the biocontrol strains were identified as T. harzianum s. str. In addition, the widely applied culture 'T. harzianum T22' was determined to be T. afroharzianum. Some species in the T. harzianum complex appear to be exclusively endophytic, while others were only isolated from soil. Sexual states are rare. Descriptions and illustrations are provided. A secondary barcode, nuc translation elongation factor 1-α (TEF1) is needed to identify species in this complex.

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

    Science.gov (United States)

    Martínez-Medina, Ainhoa; Del Mar Alguacil, Maria; Pascual, Jose A; Van Wees, Saskia C M

    2014-07-01

    The application of Trichoderma strains with biocontrol and plant growth-promoting capacities to plant substrates can help reduce the input of chemical pesticides and fertilizers in agriculture. Some Trichoderma isolates can directly affect plant pathogens, but they also are known to influence the phytohormonal network of their host plant, thus leading to an improvement of plant growth and stress tolerance. In this study, we tested whether alterations in the phytohormone signature induced by different Trichoderma isolates correspond with their ability for biocontrol and growth promotion. Four Trichoderma isolates were collected from agricultural soils and were identified as the species Trichoderma harzianum (two isolates), Trichoderma ghanense, and Trichoderma hamatum. Their antagonistic activity against the plant pathogen Fusarium oxysporum f. sp. melonis was tested in vitro, and their plant growth-promoting and biocontrol activity against Fusarium wilt on melon plants was examined in vivo, and compared to that of the commercial strain T. harzianum T-22. Several growth- and defense-related phytohormones were analyzed in the shoots of plants that were root-colonized by the different Trichoderma isolates. An increase in auxin and a decrease in cytokinins and abscisic acid content were induced by the isolates that promoted the plant growth. Principal component analysis (PCA) was used to evaluate the relationship between the plant phenotypic and hormonal variables. PCA pointed to a strong association of auxin induction with plant growth stimulation by Trichoderma. Furthermore, the disease-protectant ability of the Trichoderma strains against F. oxysporum infection seems to be more related to their induced alterations in the content of the hormones abscisic acid, ethylene, and the cytokinin trans-zeatin riboside than to the in vitro antagonism activity against F. oxysporum.

  6. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea.

  7. The toolbox of Trichoderma spp. in the biocontrol of Botrytis cinerea disease.

    Science.gov (United States)

    Vos, Christine M F; De Cremer, Kaat; Cammue, Bruno P A; De Coninck, Barbara

    2015-05-01

    Botrytis cinerea is a necrotrophic fungal pathogen causing disease in many plant species, leading to economically important crop losses. So far, fungicides have been widely used to control this pathogen. However, in addition to their detrimental effects on the environment and potential risks for human health, increasing fungicide resistance has been observed in the B. cinerea population. Biological control, that is the application of microbial organisms to reduce disease, has gained importance as an alternative or complementary approach to fungicides. In this respect, the genus Trichoderma constitutes a promising pool of organisms with potential for B. cinerea control. In the first part of this article, we review the specific mechanisms involved in the direct interaction between the two fungi, including mycoparasitism, the production of antimicrobial compounds and enzymes (collectively called antagonism), and competition for nutrients and space. In addition, biocontrol has also been observed when Trichoderma is physically separated from the pathogen, thus implying an indirect systemic plant defence response. Therefore, in the second part, we describe the consecutive steps leading to induced systemic resistance (ISR), starting with the initial Trichoderma-plant interaction and followed by the activation of downstream signal transduction pathways and, ultimately, the defence response resulting in ISR (ISR-prime phase). Finally, we discuss the ISR-boost phase, representing the effect of ISR priming by Trichoderma spp. on plant responses after additional challenge with B. cinerea. PMID:25171761

  8. A new cytotoxic cytochalasin from the endophytic fungus Trichoderma harzianum.

    Science.gov (United States)

    Chen, Huiqin; Daletos, Georgios; Okoye, Festus; Lai, Daowan; Dai, Haofu; Proksch, Peter

    2015-04-01

    The new natural product 4]-hydroxy-deacetyl-18-deoxycytochalasin H (1), together with the known deacetyl-18-deoxycytochalasin H (2) and 18-deoxycytochalasin H (3) were obtained from the endophytic fungus Trichoderma harzianum isolated from leaves of Cola nitida. The structure of the new compound was unambiguously determined by 1D and 2D NMR spectroscopy, and by HRESIMS measurements, as well as by comparison with the literature. Compounds 1-3 showed potent cytotoxic activity against the murine lymphoma (L5178Y) cell line and against human ovarian cancer (A2780 sens and A2780 CisR) cell lines (IC50 0.19-6.97 µM). The A2780 cell lines included cisplatin-sensitive (sens) and -resistant (R) cells. PMID:25973482

  9. Variations on conserved signaling pathways in biocontrol and development:G protein and MAPK genes of Trichoderma.atroviride and T.virens

    Institute of Scientific and Technical Information of China (English)

    Benjiamin A Horwitz

    2004-01-01

    @@ Filamentous fungi employ conserved eukaryotic signaling pathway to detect and respond to environmental signals, including the presence of the host. Genetic experiment in which a particular signaling protein is lost, or its activity enhanced, have defined some of the function of heterotrimeric G proteins and MAP kinases in development and virulence. A hallmark of these studies is that orthologs in different species may have different functions. Antagonistic fungal-fungal interactions form the basis for biological control of plant disease. These interactions may employ novel modes of regulation by conserved signaling elements. Tag1, a G protein a subunit of Trichoderma. atroviride belonging to fungal Gi class, is involved in repression of sporulation and hyphal coiling(1). Deletion of ortholog of this gene, TgaA, in Trichoderma (Gliocladium) virens, however, did not affect sporulation and growth, yet tgaA mutants are unable to parasitize S. rolfsii sclerotia(2). Mutation of a second G αsubunit gene is now under study. TmkA, a MAPK gene of T. virens, is involved in biocontrol properties and repression of conidiation (3). Using suppression-subtraction hybridization and other approaches, we are beginning to identify additional elements of the signaling cascades and their downsteam targets. The role of G protein and MAPK genes are sometimes specific to a particular host fungus or to parasitism of mycelia or sclerotia (2, 3). Also of relevance to biocontrol, signal transduction pathway provide a means to alter the balance between sporulation, mycelial growth and hyphal coiling.

  10. Transcriptomic response of the mycoparasitic fungus Trichoderma atroviride to the presence of a fungal prey

    Energy Technology Data Exchange (ETDEWEB)

    Seidl, Verena; Song, Lifu; Lindquist, Erika; Gruber, Sabine; Koptchinskiy, Alexeji; Zeilinger, Susanne; Schmoll, Monika; Martinez, Pedro; Sun, Jibin; Grigoriev, Igor; Herrera-Estrella, Alfredo; Baker, Scott E; Kubicek, Christian P.

    2010-07-23

    BACKGROUND: Combating the action of plant pathogenic microorganisms by mycoparasitic fungi has been announced as an attractive biological alternative to the use of chemical fungicides since two decades. The fungal genus Trichoderma includes a high number of taxa which are able to recognize, combat and finally besiege and kill their prey. Only fragments of the biochemical processes related to this ability have been uncovered so far, however. RESULTS: We analyzed genome-wide gene expression changes during the begin of physical contact between Trichoderma atroviride and two plant pathogens Botrytis cinerea and Rhizoctonia solani, and compared with gene expression patterns of mycelial and conidiating cultures, respectively. About 3000 ESTs, representing about 900 genes, were obtained from each of these three growth conditions. 66 genes, represented by 442 ESTs, were specifically and significantly overexpressed during onset of mycoparasitism, and the expression of a subset thereof was verified by expression analysis. The upregulated genes comprised 18 KOG groups, but were most abundant from the groups representing posttranslational processing, and amino acid metabolism, and included components of the stress response, reaction to nitrogen shortage, signal transduction and lipid catabolism. Metabolic network analysis confirmed the upregulation of the genes for amino acid biosynthesis and of those involved in the catabolism of lipids and aminosugars. CONCLUSION: The analysis of the genes overexpressed during the onset of mycoparasitism in T. atroviride has revealed that the fungus reacts to this condition with several previously undetected physiological reactions. These data enable a new and more comprehensive interpretation of the physiology of mycoparasitism, and will aid in the selection of traits for improvement of biocontrol strains by recombinant techniques.

  11. Nutrient activation of Trichoderma fungal spores for improved biocontrol activity

    Institute of Scientific and Technical Information of China (English)

    Linda Gordon Hjeljord; Arne Tronsmo

    2004-01-01

    @@ The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum P1 (ThP1) to suppress Botrytis cinerea was investigated in laboratory, greenhouse and field trials. Preliminary nutrient activation at 21 ℃ accelerated subsequent germination of the antagonist at temperatures from 9 ℃ to 21 ℃; at ≥ 18 ℃ the germination time of preactivated ThP1conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea,concentrated inocula of preactivated but ungerminated ThP1 conidia reduced in vitro germination of the pathogen by ≥ 87 % at 12 ℃ to 25 ℃; initially-quiescent conidia achieved this level of suppression only at 25 ℃. Application of quiescent ThP1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by ≥85 % at 24 ℃ , but only by 35 % at 12 ℃. Preactivated conidia reduced infection by ≥60% at 12 ℃. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperture of 19 ℃, while only preactivated conidia were effective in the field at a mean temperature of 14 ℃ on the day of treatment application.

  12. Novel Trichoderma polysporum Strain for the Biocontrol of Pseudogymnoascus destructans, the Fungal Etiologic Agent of Bat White Nose Syndrome.

    Directory of Open Access Journals (Sweden)

    Tao Zhang

    Full Text Available White-nose syndrome (WNS, an emerging disease of hibernating bats, has rapidly spread across eastern North America killing millions of bats. Pseudogymnoascus destructans (Pd, the sole etiologic agent of WNS, is widespread and persistent in bat hibernacula. Control of Pd in the affected sites is urgently needed to break the transmission cycle while minimizing any adverse impact on the native organisms. We isolated a novel strain of Trichoderma polysporum (Tp from one of the caves at the epicenter of WNS zoonotic. Detailed experimental studies revealed: (1 Tp WPM 39143 was highly adapted to grow at temperatures simulating the cave environment (6°C-15°C, (2 Tp WPM 39143 restricted Pd colony growth in dual culture challenges, (3 Tp WPM 39143 caused four logs reduction of Pd colony forming units and genome copies in autoclaved soil samples from one of the WNS affected caves, (4 Tp WPM 39143 extract showed specific fungicidal activity against Pd in disk diffusion assay, but not against closely related fungus P. pannorum (Pp, (5 Tp WPM 39143 extract retained inhibitory activity after exposure to high temperatures, light and proteinase K, and (6 Inhibitory metabolites in Tp WPM 39143 extract comprised of water-soluble, high polarity compounds. These results suggest that Tp WPM 39143 is a promising candidate for further evaluation as a biocontrol agent of Pd in WNS affected sites.

  13. Enhanced biocontrol activity of Trichoderma through inactivation of a mitogen-activated protein kinase.

    Science.gov (United States)

    Mendoza-Mendoza, Artemio; Pozo, María J; Grzegorski, Darlene; Martínez, Pedro; García, Juan M; Olmedo-Monfil, Vianey; Cortés, Carlos; Kenerley, Charles; Herrera-Estrella, Alfredo

    2003-12-23

    The production of lytic enzymes in Trichoderma is considered determinant in its parasitic response against fungal species. A mitogen-activated protein kinase encoding gene, tvk1, from Trichoderma virens was cloned, and its role during the mycoparasitism, conidiation, and biocontrol was examined in tvk1 null mutants. These mutants showed a clear increase in the level of the expression of mycoparasitism-related genes under simulated mycoparasitism and during direct confrontation with the plant pathogen Rhizoctonia solani. The null mutants displayed an increased protein secretion phenotype as measured by the production of lytic enzymes in culture supernatant compared to the wild type. Consistently, biocontrol assays demonstrated that the null mutants were considerably more effective in disease control than the wild-type strain or a chemical fungicide. In addition, tvk1 gene disruptant strains sporulated abundantly in submerged cultures, a condition that is not conducive to sporulation in the wild type. These data suggest that Tvk1 acts as a negative modulator during host sensing and sporulation in T. virens. PMID:14673101

  14. Comparison of real-time PCR and microscopy to evaluate sclerotial colonisation by a biocontrol fungus.

    Science.gov (United States)

    Kim, Tae Gwan; Knudsen, Guy R

    2011-01-01

    The biocontrol agent Trichoderma harzianum colonises sclerotia of the plant pathogenic fungus Sclerotinia sclerotiorum. Plating of sclerotia typically has been used to determine the incidence of mycoparasitism, but does not quantify the extent to which individual sclerotia are colonised. We developed a specific PCR primer/probe set for the green fluorescent protein (GFP)-transformant T. harzianum ThzID1-M3, which exhibited high precision and reproducibility. Quantitative real-time PCR was evaluated along with epifluorescence microscopy and image analysis to investigate dynamics of colonisation of sclerotia in non-sterile soil. Amounts of ThzID1-M3 DNA and S. sclerotiorum DNA from entire individual sclerotia were quantified using real-time PCR. Epifluorescence micrographs were captured from sclerotial thin-section samples, and GFP fluorescence from these was quantified using computer image analysis in order to estimate colonisation on a per-sclerotium basis. As determined by either method, ThzID1-M3 colonised sclerotia in soil, and both methods quantified colonisation dynamics over time. In a separate experiment, colonisation of sclerotia on agar plates was observed using confocal laser scanning microscopy to view the GFP-fluorescing hyphae of ThzID1-M3. This method, while highly labour-intensive, provided high spatial resolution of colonisation dynamics. Thus, each method has advantages: microscopy combined with image analysis can provide useful information on the spatial and temporal dynamics of colonisation, while real-time PCR can provide a more precise assessment of the extent of sclerotial colonisation over time and can more easily be used to sample entire sclerotia. PMID:21530913

  15. Biocontrol mechanisms of Trichoderma harzianum against soybean charcoal rot caused by Macrophomina phaseolina

    Directory of Open Access Journals (Sweden)

    Khaledi Nima

    2016-01-01

    Full Text Available Throughout the world, charcoal rot, caused by Macrophomina phaseolina, is one of the most destructive and widespread diseases of crop plants such as soybean. In this study, the biological control capability of 11 Trichoderma spp. isolates against M. phaseolina was investigated using screening tests. Among all the tested Trichoderma spp. isolates, inhibition varied from 20.22 to 58.67% in dual culture tests. Dual culture, volatile and non-volatile tests revealed that two isolates of Trichoderma harzianum (including the isolates T7 and T14 best inhibited the growth of M. phaseolina in vitro. Therefore, these isolates were selected for biocontrol of M. phaseolina in vivo. The results of greenhouse experiments revealed that disease severity in the seed treatment with T. harzianum isolates was significantly lower than that of the soil treatment. In most of the cases, though, soil treatment with T. harzianum resulted in higher plant growth parameters, such as root and shoot weight. The effects of T. harzianum isolates on the activity of peroxidase enzyme and phenolic contents of the soybean root in the presence and absence of M. phaseolina were determined in greenhouse conditions. Our results suggested that a part of the inhibitory effect of T. harzianum isolates on soybean charcoal rot might be related to the indirect influence on M. phaseolina. Plant defense responses were activated as an elicitor in addition to the direct effect on the pathogen growth.

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

    DEFF Research Database (Denmark)

    Jørgensen, Mikael Skaanning

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

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

    Science.gov (United States)

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

    2014-06-01

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

  18. Trichoderma harzianum as a biocontrol agent against Alternaria alternata on tobacco

    Directory of Open Access Journals (Sweden)

    Jugoslav Ziberoski

    2012-06-01

    Full Text Available Trichoderma fungi are the most popular agents used in a biological control. Therefore, our aim was to determine an impact of Trichoderma harzianum on the fungus Alternaria alternata - a causing agent of the brown spot disease on tobacco. In vitro analyses were made in several variants of double culture, in order to study the effect of difusible and volatile metabolites. There was strong reducing effect on the development of A.alternata with various mechanisms of antagonistic influence. The volatile metabolites have also shown reducing effect. Some abnormalities were observed in the pathogen's morphology both in difusible and volatile metabolites. The strong reducing effect of T.harzianum towards A. alternata can be applied in biological control of this pathogen.

  19. Native isolate of Trichoderma: a biocontrol agent with unique stress tolerance properties.

    Science.gov (United States)

    Mishra, N; Khan, S S; Sundari, S Krishna

    2016-08-01

    Species of Trichoderma are widely recognized for their biocontrol abilities, but seldom studied collectively, for their plant growth promotion, abiotic stress tolerance and bioremediation properties. Our study is a concentrated effort to establish the potential of native isolate Trichoderma harzianum KSNM (T103) to tolerate biotic (root pathogens) and abiotic stresses [high salt (100-1000 mM); heavy metal (chromium, nickel and zinc: 1-10 mM); pesticides: malathion (100-600 ppm), carbofuran (100-600 ppb)], along with its ability to support plant growth. In vitro growth promotion assays with T103 treated Vigna radiata, Vigna mungo and Hordeum vulgare confirmed 'non-species specific' growth promotion effects of T103. At lower metal concentration, T103 treatment was found to completely negate the impact of metal stress [60 % increase in radicle length (RL) with no significant decrease in %germination (%G)]. Even at 10 mM metal, T103 inoculation gave 80 % increase in %G and >50 % increase in RL. In vitro experiments confirmed high metal reduction capacity (47 %-Cr, 35 %-Ni and 42 %-Zn) of T103 at concentrations as high as 4 mM. At maximum residual concentrations of malathion (440 ppm) and carbofuran (100 ppb) reported in agricultural soils, T103 maintained 80 and 100 % survivability, respectively. T103 treatment has improved %G and RL in all three hosts challenged with pesticide. Isolate T103 was found to effectively suppress growth of three major root pathogens: Macrophomina phaseolina (65.83 %) followed by Sclerotium rolfsii (19.33 %) and Fusarium oxysporum (19.18 %). In the light of these observations, native T. harzianum (T103) seems to be a competent biocontrol agent for tropical agricultural soils contaminated with residual pesticides and heavy metals. PMID:27339311

  20. The importance of chorismate mutase in the biocontrol potential of Trichoderma parareesei.

    Science.gov (United States)

    Pérez, Esclaudys; Rubio, M Belén; Cardoza, Rosa E; Gutiérrez, Santiago; Bettiol, Wagner; Monte, Enrique; Hermosa, Rosa

    2015-01-01

    Species of Trichoderma exert direct biocontrol activity against soil-borne plant pathogens due to their ability to compete for nutrients and to inhibit or kill their targets through the production of antibiotics and/or hydrolytic enzymes. In addition to these abilities, Trichoderma spp. have beneficial effects for plants, including the stimulation of defenses and the promotion of growth. Here we study the role in biocontrol of the T. parareesei Tparo7 gene, encoding a chorismate mutase (CM), a shikimate pathway branch point leading to the production of aromatic amino acids, which are not only essential components of protein synthesis but also the precursors of a wide range of secondary metabolites. We isolated T. parareesei transformants with the Tparo7 gene silenced. Compared with the wild-type, decreased levels of Tparo7 expression in the silenced transformants were accompanied by reduced CM activity, lower growth rates on different culture media, and reduced mycoparasitic behavior against the phytopathogenic fungi Rhizoctonia solani, Fusarium oxysporum and Botrytis cinerea in dual cultures. By contrast, higher amounts of the aromatic metabolites tyrosol, 2-phenylethanol and salicylic acid were detected in supernatants from the silenced transformants, which were able to inhibit the growth of F. oxysporum and B. cinerea. In in vitro plant assays, Tparo7-silenced transformants also showed a reduced capacity to colonize tomato roots. The effect of Tparo7-silencing on tomato plant responses was examined in greenhouse assays. The growth of plants colonized by the silenced transformants was reduced and the plants exhibited an increased susceptibility to B. cinerea in comparison with the responses observed for control plants. In addition, the plants turned yellowish and were defective in jasmonic acid- and ethylene-regulated signaling pathways which was seen by expression analysis of lipoxygenase 1 (LOX1), ethylene-insensitive protein 2 (EIN2) and pathogenesis

  1. The importance of chorismate mutase in the biocontrol potential of Trichoderma parareesei

    Directory of Open Access Journals (Sweden)

    Esclaudys ePérez

    2015-10-01

    Full Text Available Species of Trichoderma exert direct biocontrol activity against soil-borne plant pathogens due to their ability to compete for nutrients and to inhibit or kill their targets through the production of antibiotics and/or hydrolytic enzymes. In addition to these abilities, Trichoderma spp. have beneficial effects for plants, including the stimulation of defenses and the promotion of growth. Here we study the role in biocontrol of the T. parareesei Tparo7 gene, encoding a chorismate mutase (CM, a shikimate pathway branch point leading to the production of aromatic amino acids, which are not only essential components of protein synthesis but also the precursors of a wide range of secondary metabolites. We isolated T. parareesei transformants with the Tparo7 gene silenced. Compared with the wild-type, decreased levels of Tparo7 expression in the silenced transformants were accompanied by reduced CM activity, lower growth rates on different culture media, and reduced mycoparasitic behavior against the phytopathogenic fungi Rhizoctonia solani, Fusarium oxysporum and Botrytis cinerea in dual cultures. By contrast, higher amounts of the aromatic metabolites tyrosol, 2-phenylethanol and salicylic acid were detected in supernatants from the silenced transformants, which were able to inhibit the growth of F. oxysporum and B. cinerea. In in vitro plant assays, Tparo7-silenced transformants also showed a reduced capacity to colonize tomato roots. The effect of Tparo7-silencing on tomato plant responses was examined in greenhouse assays. The growth of plants colonized by the silenced transformants was reduced and the plants exhibited an increased susceptibility to B. cinerea in comparison with the responses observed for control plants. In addition, the plants turned yellowish and were defective in jasmonic acid- and ethylene-regulated signaling pathways which was seen by expression analysis of lipoxygenase 1 (LOX1, ethylene-insensitive protein 2 (EIN2 and

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

    Institute of Scientific and Technical Information of China (English)

    Saravanakumar Kandasamy; Kathiresan Kandasamy

    2014-01-01

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

  3. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture.

  4. Selection and characterization of Argentine isolates of Trichoderma harzianum for effective biocontrol of Septoria leaf blotch of wheat.

    Science.gov (United States)

    Stocco, Marina C; Mónaco, Cecilia I; Abramoff, Cecilia; Lampugnani, Gladys; Salerno, Graciela; Kripelz, Natalia; Cordo, Cristina A; Consolo, Verónica F

    2016-03-01

    Species of the genus Trichoderma are economically important as biocontrol agents, serving as a potential alternative to chemical control. The applicability of Trichoderma isolates to different ecozones will depend on the behavior of the strains selected from each zone. The present study was undertaken to isolate biocontrol populations of Trichoderma spp. from the Argentine wheat regions and to select and characterize the best strains of Trichoderma harzianum by means of molecular techniques. A total of 84 out of the 240 strains of Trichoderma were able to reduce the disease severity of the leaf blotch of wheat. Thirty-seven strains were selected for the reduction equal to or greater than 50% of the severity, compared with the control. The percentage values of reduction of the pycnidial coverage ranged between 45 and 80%. The same last strains were confirmed as T. harzianum by polymerase chain reaction amplification of internal transcribed spacers, followed by sequencing. Inter-simple sequence repeat was used to examine the genetic variability among isolates. This resulted in a total of 132 bands. Further numerical analysis revealed 19 haplotypes, grouped in three clusters (I, II, III). Shared strains, with different geographical origins and isolated in different years, were observed within each cluster. The origin of the isolates and the genetic group were partially related. All isolates from Paraná were in cluster I, all isolates from Lobería were in cluster II, and all isolates from Pergamino and Santa Fe were in cluster III. Our results suggest that the 37 native strains of T. harzianum are important in biocontrol programs and could be advantageous for the preparation of biopesticides adapted to the agroecological conditions of wheat culture. PMID:26873560

  5. Evaluation of the enzymatic activity of Trichoderma inhamatum (BOL-12QD as possible biocontroller

    Directory of Open Access Journals (Sweden)

    García-Espejo Cielo Noemí

    2016-02-01

    Full Text Available It is known that Trichoderma spp. acts as a natural biocontroller of pathogen fungi, is for this reason, that this research studies the potential of its hydrolytic enzyme activity. In this article, first we determined that the speed of growth of Trichoderma inhamatum cepa BOL-12QD is 9 hours. Later, we proposed a simple and sensitive method based in the use of basal media (BM with coloidal chitin as the only carbon resource and supplemented with bromocresol purple for the qualitative determination of chitinase activi-ty. On the other hand, it was determined the celullolytic and proteolytic activities of Trichoderma in-hamatum cepa BOL-12QD and it was observed that agitation, type and concentration of sustrate are determinant factors in enzymatic production. Then, we evaluated the cellulolytic activity of Trichoderma inhamatum cepa BOL-12QD in agitation and stationary using carboxymethylcellulose (CMC as sustrate, finding that using a 2% of sustrate the highest activity is registered at 8 days of incubation in agitation with a value of 99.23 IU/L. In relation to the results at stationary the optimal value is at the fourth day with a value of 92.76 IU/L. The protease activity it was determined taking in consideration variables of agitation and stationary, using different types and concentration of sustrate at 2%, 4% y 6% (w/v of meat extract, 1%, 3%, 5% (w/v of jelly and 1%, 2%, 4% (w/v of casein. The highest protease activity was obtained at the end of the sev-enth day with an enzymatic activity of 3075.45 IU/L at stationary using a concentration of 6% (w/v of meat extract, and using jelly at 3% (w/v at stationary it was found an activity of 568.36 IU/L on the tenth day and in agitation a value of 547.27 IU/L was reached on the twelveth day, while using casein at 1% (w/v at stationary an activity of 407.06 IU/L is reached in the fifth day, and in agitation at 4% (w/v of casein a value of 547.27 IU/L is obtained on the twelfth day, while using

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

    Science.gov (United States)

    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.

  7. 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...

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

    Science.gov (United States)

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

    2014-08-01

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

  9. Use of Trichoderma spp.for biological control of the livestock feed contaminant fungus Fusarium proliferatum

    Institute of Scientific and Technical Information of China (English)

    Ruocco M; Ferraioli S; Scala F; Lorito M; Pane F; Ritieni A; Lanzuise S; Ambrosino P; Marra R; Woo S L; Ciliento R; Soriente I

    2004-01-01

    @@ Fusarium spp. are pathogens of many important agricultural crops, and are often strong mycotoxin producers. Fusarium proliferatum, in particular, causes disease in cereals and secretes the toxin Beauvaricin that contaminates livestock feed and cereals, producing a variety of toxicity symptoms ranging from poor weight gain to mortality. Beauvaricin is a cyclodepsipeptide and acts as a potent mycotoxin known to have insecticidal properties. This compound is highly toxic to human cell lines,where it induces apoptosis and specifically inhibits cholesterol acetyltransferase. Nothing is known about the role of this mycotoxin during the interaction of F. proliferatum with other microorganisms, including the fungal antagonists Trichoderma spp. In vitro tests have demonstrated that the antagonistic and mycoparasitic activity of Trichoderma is not inhibited by the presence of Beauvaricin at concentrations up to 10 mg/kg in the substrate. In vivo biocontrol assays on barley and wheat with Trichoderma against F. proliferatum isolates, producing and non-producing Beauvaricin, confirmed the ability of the antagonist to control this pathogen in all cases. Also Trichoderma culture filtrates obtained in conditions that promote _Cell Wall _Degrading Enzyme (CWDE) secretion, were able to inhibit spore germination of different F. proliferatum isolates.These results suggest the possibility of using Trichoderma and/or its metabolites to control contaminants of livestock feed by mycotoxin-producing Fusarium.

  10. Biocontrol of seed pathogens and growth promotion of common bean seedlings by Trichoderma harzianum

    Directory of Open Access Journals (Sweden)

    Daniel Diego Costa Carvalho

    2011-08-01

    Full Text Available The objective of this work was to evaluate isolates of Trichoderma harzianum regarding biocontrol of common bean seed-borne pathogens, plant growth promotion, and rhizosphere competence. Five isolates of T. harzianum were evaluated and compared with commercial isolate (Ecotrich, Carboxin+Thiram, and an absolute control. Bean seeds of the cultivar Jalo Precoce, contaminated with Aspergillus, Cladosporium, and Sclerotinia sclerotiorum, were microbiolized with antagonists, and seed health tests were carried out. Isolates were evaluated on autoclaved substrate and in field conditions. Ten days after sowing (DAS, plant length was measured. To test rhizosphere competence, isolates were applied in boxes containing autoclaved washed sand, and root colonization was evaluated at 10 DAS, using five plants per box. The most effective isolates in the seed health tests were: CEN287 and CEN289 to control Aspergillus; the commercial isolate to control Cladosporium; and CEN287 and CEN316 to control S. sclerotiorum. Isolates CEN289 and CEN290 promoted bean growth in greenhouse and field. Seed treatment with T. harzianum reduces the incidence of Aspergillus, Cladosporium, and S. sclerotiorum in 'Jalo Precoce' common bean seeds.

  11. Research Progress of Biocontrol Effect of Trichoderma harzianum%哈茨木霉生物防治研究进展

    Institute of Scientific and Technical Information of China (English)

    吕黎; 许丽媛; 罗志威; 周艳; 郭帅; 丰来

    2013-01-01

    木霉被认为是最有希望的生物防治因子,而哈茨木霉是木霉属中应用最为广泛的菌株。从哈茨木霉的生防效果、生防作用机制以及基因水平上的抗生机制等方面的研究进展进行了综述,并阐述了哈茨木霉可能的应用前景。%Trichoderma is the most promising isolated soil fungi for their ability to protect plants and contain pathogen populations under different soil conditions. While Trichoderma harzianum is the most commonly applied strain among Trichoderma. The biocontrol effect, the biocontrol mechanism, and the antibiotic mechanism at the gene level of Trichoderma harzianum were summarized, and the applying future of it was elaborated.

  12. Antagonistic and Biocontrol Potential of Trichoderma asperellum ZJSX5003 Against the Maize Stalk Rot Pathogen Fusarium graminearum.

    Science.gov (United States)

    Li, Yaqian; Sun, Ruiyan; Yu, Jia; Saravanakumar, Kandasamy; Chen, Jie

    2016-09-01

    The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot.

  13. Antagonistic and Biocontrol Potential of Trichoderma asperellum ZJSX5003 Against the Maize Stalk Rot Pathogen Fusarium graminearum.

    Science.gov (United States)

    Li, Yaqian; Sun, Ruiyan; Yu, Jia; Saravanakumar, Kandasamy; Chen, Jie

    2016-09-01

    The efficacy of seven strains of Trichoderma asperellum collected from the fields in Southern China was assessed against Fusarium graminearum (FG) the causal agent of corn stalk rot of maize were in vitro for their antagonistic properties followed by statistical model of principal compound analysis to identify the beneficial antagonist T. asperellum strain. The key factors of antagonist activity were attributed to a total of 13 factors including cell wall degrading enzymes (chitnase, protease and β-glucanases), secondary metabolites and peptaibols and these were analyzed from eight strains of Trichoderma. A linear regression model demonstrated that interaction of enzymes and secondary metabolites of T. asperellum strain ZJSX5003 enhanced the antagonist activity against FG. Further, this strain displayed a disease reduction of 71 % in maize plants inoculated with FG compared to negative control. Pointing out that the T. asperellum strain ZJSX5003 is a potential source for the development of a biocontrol agent against corn stalk rot. PMID:27407296

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

    Science.gov (United States)

    Shida, Yosuke; Furukawa, Takanori; Ogasawara, Wataru

    2016-09-01

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

  15. Virulence and Experimental Treatment of Trichoderma longibrachiatum, a Fungus Refractory to Treatment.

    Science.gov (United States)

    Paredes, Katihuska; Capilla, Javier; Mayayo, Emilio; Guarro, Josep

    2016-08-01

    Different inocula of Trichoderma longibrachiatum were tested in a murine model, and only the highest one (1 × 10(7) CFU/animal) killed all of the mice at day 15 postinfection, with spleen and liver the most affected organs. The efficacies of amphotericin B deoxycholate, liposomal amphotericin B, voriconazole, and micafungin were evaluated in the same model, with very poor results. Our study demonstrated the low virulence but high resistance to antifungal compounds of this fungus. PMID:27216056

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

    Energy Technology Data Exchange (ETDEWEB)

    Kubicek, Christian P.; Herrera-Estrella, Alfredo; Seidl, Verena; Crom, St& #233; phane Le; Martinez, Diego A.; Druzhinina, Irina S.; Zeilinger, Susanne; Casas-Flores, Sergio; Horwitz, Benjamin A.; Mukherjee, Prasun K.; Mukherjee, Mala; Kredics, L& #225; szlo; Alcaraz, Luis David; Aerts, Andrea; Antal, Zsuzsanna; Atanasova, Lea; Cervantes-Badillo, Mayte Guadalupe; Challacombe, Jean; Chertkov, Olga; McCluskey, Kevin; Coulpier, Fanny; Deshpande, Nandan; D& #246; hren, Hans von; Ebbole, Daniel J.; Esquivel-Naranjo, Edgardo Ulises; Fekete, Erzs& #233; bet; Flipphi, Michel; Glaser, Fabian; Gomez-Rodriguez, Elida Yazmin; Gruber, Sabine; Han, Cliff; Henrissat, Bernard; Hermosa, Rosa; Hern& #225; ndez-O?ate, Miguel; Karaffa, Levente; Kosti, Idit; Lindquist, Erika; Lucas, Susan; L& #252; beck, Mette; L& #252; beck, Peter Stephensen; Margeot, Antoine; Metz, Benjamin; Misra, Monica; Nevalainen, Helena; Omann, Markus; Packer, Nicolle; Perrone, Giancarlo; Uresti-Rivera, Edith Elena; Salamov, Asaf; Schmoll, Monika; Seiboth, Bernhard; Shapiro, Harris; Sukno, Serenella; Tamayo-Ramos, Juan Antonio; Thon, Michael; Tisch, Doris; Wiest, Aric; Wilkinson, Heather H.; Zhang, Michael; Coutinho, Pedro M.; Kenerley, Charles M.; Monte, Enrique; Baker, Scott E.; Grigoriev, Igor V.

    2011-04-29

    Mycoparasitism, a lifestyle where one fungus is parasitic on another fungus has special relevance when the prey is a plant pathogen, providing a strategy for biological control of pests for plant protection. Probably, the most studied biocontrol agents are species of the genus Hypocrea/Trichoderma.

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

    Science.gov (United States)

    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. PMID:26631988

  18. The Trichoderma-plant interaction is mediated by avirulence proteins produced by this fungus

    Institute of Scientific and Technical Information of China (English)

    Ruocco M; Kip N; P J G M de Wit; Lorito M; Lanzuise S; Woo S L; Ambrosino P; Marra R; Turrà D; Gigante S; Formisno E; Scala F

    2004-01-01

    @@ The molecular basis of Trichoderma -plant interaction is very complex and still not completely understood. The colonization of the root system by rhizosphere competent strains of Trichoderma results in increased development of root/aerial systems, in improved yields and in plant disease control.Other beneficial effects, such as the induction of plant systemic resistance, have also been described.To understand the mechanisms involved we are using different approaches, including the making of transformants expressing genes that encode for compounds able to affect plant response to pathogens.Trichoderma transformants carrying the avirulence gene Avr4 from Cladosporium fulvum under the control of constitutive and inducible promoters were obtained and tested on tomato plants having the Cf4 resistance gene. Necrosis and suberification zones, similar to the symptoms appearing during Cladosporium-tomato interaction, were found when the roots of the Cf4 plants were treated with Avr4-Trichoderma. This demonstrates that selected Trichoderma strains are able to transfer to the plant molecules that may deeply affect metabolism, disease resistance etc. Therefore, these beneficial fungi can be regarded as biotechnological tools to provide a variety of crops with useful compounds.Moreover, in in vitro competition assays the transformants were found to be more effective as antagonists against Alternaria alternata than the wild type. Trichoderma sends a variety of biochemical signals to the plants including avirulence molecules; therefore the presence of avr-like proteins in the fungus proteome was investigated. Proteome analysis has permitted us to isolate and sequence many proteins potentially having this function. From the extraeellular protein extracts, we have purified and sequenced a protein with structural characteristics similar to Avr4 of C. fulvum.The protein, Hytra1, was found to be a hydrophobin with chitin binding activity, the typical 8cysteine residues, and 4

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

    OpenAIRE

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

    2011-01-01

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

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:25879509

  2. Biocontrol de la pudrición de raíz de nochebuena de interior con Trichoderma spp. Root rot biocontrol for indoor poinsettia with Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Felipe de Jesús Osuna-Canizalez

    2012-06-01

    Full Text Available En Morelos, la pudrición de la raíz causada por Fusarium spp., es una de las principales enfermedades de la nochebuena de interior. Por su efecto devastador, en su prevención o control se realizan aplicaciones frecuentes de productos químicos, con los riesgos inherentes a la salud humana y al ambiente. En la búsqueda de alternativas bioracionales al manejo de esta enfermedad, se realizó un ensayo en el que se evaluaron tres cepas comerciales de Trichoderma spp., en tres diferentes sustratos: S1= "tierra de hoja" (70% v/v+tezontle grueso (15% v/v+tezontle fino (10% v/v+agrolita (5% v/v; S2= turba (80% v/v+ fibra de coco (20% v/v; S3= "tierra de hoja" (70% + "tepojal" (30%, en las variedades comerciales más comunes, Freedom Red y Prestige Red. Se utilizó un diseño factorial de tratamientos 4 x 3 x 2 y los tratamientos resultantes se evaluaron en un diseño completamente al azar con seis repeticiones. Respecto a la incidencia de pudrición de la raíz, las cepas comerciales de Trichoderma spp., no mostraron diferencias entre sí ni con el testigo químico. La pudrición de la raíz estuvo asociada con S2, debido a una baja capacidad de aireación, y sólo se presentó en Prestige Red. La población (UFC g-¹ de Trichoderma spp., en el sustrato al término del ciclo, fue igual (pIn Morelos, root rot caused by Fusarium spp., is one of the main diseases of indoor poinsettia. In order to prevent or control its devastating effect, frequent applications of chemical products are performed, with inherent risks to human health and environment. In quest for alternative biorational options, an essay in which three commercial strains of Trichoderma spp., was done, in three different substrates: S1= "organic soil" (70% v/v+thick tezontle (15% v/v+thin tezontle (10% v/v+agrolita (5% v/v; S2= peat moss (80% v/v+ coconut fiber (20% v/v; S3= "organic soil" (70%+"tepojal" (30%, in most common commercial varieties, Freedom Red and Prestige Red. A factorial

  3. The Fungus Trichoderma Regulates Submerged Conidiation Using the Steroid Pregnenolone.

    Science.gov (United States)

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J

    2016-09-16

    In previous work, we evolved a population of Trichoderma citrinoviride in liquid cultures to speed up its asexual development cycle. The evolved population, called T-6, formed conidia 3 times sooner and in >1000-fold greater numbers. Here, we identify the steroid pregnenolone as a molecular signal for this different behavior. Media in which the ancestral T. citrinoviride population was grown (called ancestral spent media) contained a submerged conidiation inhibitor. Growing the evolved population T-6 in ancestral spent media eliminated the abundant formation of conidia. This inhibition depended on the amount and age of the ancestral spent medium and the time that the ancestral spent medium was added to the T-6 culture. Fractionation of the ancestral spent medium identified a hydrophobic inhibiting compound with a molecular weight less than 2000 g/mol. A combination of GC-MS, ELISA, and reaction with cholesterol oxidase identified it as pregnenolone. The addition of pregnenolone to cultures of T-6 inhibited submerged conidiation by inhibiting formation of conidiophores, while 10 other analogous steroids did not. Pregnenolone also inhibited submerged conidiation of Fusarium graminearum PH-1, a plant pathogen that causes head blight in wheat and barley. This identification of steroids as signal molecules in fungi creates opportunities to disrupt this signaling to control fungal behavior. PMID:27413801

  4. New C13 lipids from the marine-derived fungus Trichoderma harzianum.

    Science.gov (United States)

    Li, Bo; Huang, Qi-Xi; Gao, Du; Liu, Dong; Ji, Yu-Bin; Liu, Hua-Gang; Lin, Wen-Han

    2015-05-01

    Chemical examination of the fermentation broth of a sponge-associated fungus Trichoderma harzinum HMS-15-3 led to the isolation of four pairs of new C13 lipid enantiomers namely harzianumols A-H (1a-4b). Their structures were elucidated on the basis of extensive spectroscopic (IR, MS, 1D, and 2D NMR) data analysis, including the modified Mosher's method for the assignment of their absolute configurations. The new compounds were evaluated for antihyperlipidemic effects in HepG2 cells. PMID:26031203

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

    Directory of Open Access Journals (Sweden)

    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

  6. Biocontrol potential of salinity tolerant mutants of Trichoderma harzianum against Fusarium oxysporum Potencial de biocontrole de mutantes sal-tolerantes de Trichoderma harzianum contra Fusarium oxysporum

    Directory of Open Access Journals (Sweden)

    Hassan Abdel-Latif A. Mohamed

    2006-06-01

    Full Text Available Exposing a wild-type culture of Trichoderma harzianum to gamma irradiation induced two stable salt-tolerant mutants (Th50M6 and Th50M11. Under saline conditions, both mutants greatly surpassed their wild type strain in growth rate, sporulation and biological proficiency against Fusarium oxysporum, the causal agent of tomato wilt disease. Tolerant T. harzianum mutants detained a capability to grow and convinced sporulation in growth media containing up to 69 mM NaCl. In comparison with their parent strain, characterization of both mutants confirmed that they have reinforced contents of proline and hydroxyproline, relatively higher sodium content compared to potassium, calcium or magnesium contents, higher level of total phenols. Electrophoretic analysis of total soluble proteins in the salt tolerance mutant Th50M6 showed different bands accumulated in response to 69 mM NaCl. Data also showed that mutants produce certain active metabolites, such as chitinases, cellulases, beta-galactosidases, as well as, some antibiotics i.e., trichodermin, gliotoxin and gliovirin. Trichoderma mutants significantly reduced wilt disease incidence and improved yield and mineral contents of tomato plants under both saline and non-saline soil conditions, as well as, under infested and natural conditions. T. harzianum mutants were also more efficient in dropping the F. oxysporum growth in rhizosphere compared to the wild type strain. Population density of both mutants in rhizosphere far exceeded that of T. harzianum wild type strain.A exposição de uma cepa selvagem de Trichoderma harzianum à irradiação gama induziu dois mutantes tolerantes a sal (Th50M6 e Th50M11. Em condições salinas, os dois mutantes foram muito superiores à cepa selvagem em relação à velocidade de multiplicação, esporulação e eficiência contra Fusarium oxysporum, o agente causador da doença wilt do tomate. Os mutantes tolerantes foram capazes de multiplicação e esporulação em

  7. Identification of New Lactone Derivatives Isolated from Trichoderma sp., An Endophytic Fungus of Brotowali (Tinaspora crispa

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    Elfita

    2014-03-01

    Full Text Available Endophytic fungi is a rich source of novel organic compounds with interesting biological activities and a high level of structural diversity. As a part of our systematic search for new bioactive lead structures and specific profiles from endophytic fungi, an endophytic fungus was isolated from roots of brotowali (Tinaspora crispa, an important medicinal plant. Colonial morphological trait and microscopic observation revealed that the endophytic fungus was Trichoderma sp. The pure fungal strain was cultivated on 7 L Potatos Dextose Broth (PDB medium under room temperature (no shaking for 8 weeks. The ethyl acetate were added to cultur medium and left overnight to stop cell growth. The culture filtrates were collected and extracted with EtOAc and then taken to evaporation. Two new lactone derivatives, 5-hydroxy-4-hydroxymethyl-2H-pyran-2-one (1 and (5-hydroxy-2-oxo-2H pyran-4-yl methyl acetate (2 were obtained from the EtOAc extracts of Trichoderma sp. Their structures were determined on the basic of spectroscopic methods including UV, IR, 1H-NMR, 13C-NMR, HMQC, and HMBC.

  8. 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

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

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    S. Matei

    2011-12-01

    Full Text Available Various fungal strains belonging to genus Trichoderma act as biological control agents for soil born plant pathogens. Two new strains of Trichoderma harzianum (T.h. and Trichoderma viride (T.v. were isolated from forest soils in Ilfov county and their morphological aspects, enzymatic and antagonistic activity were examined. Current chemical fungicides had constantly, in time, less influence on pathogens due to their diversity, adaptability and increasing resistance.The paper present the morphological characterization of two strains of Trichoderma isolated from forest soils. Growth rate was higher in strain T.v.SP456 (0,675mm/h than in strain T.h.P8 (0,505mm/h when fungi were grown on Czapek culture medium.Morphological description is completed with photographs of colonies in Petri plates and microscopical aspects of fungal structures belonging to Trichoderma strains SP456 and P8.Comparative aspects concerning the level of main enzymes released by T.h. isolate P8 and T.v.SP456 in liquid culture media showed differences as a function of genetic structure of each fungal isolate. The optimum culture media for inducing peroxidase, polyphenol-oxidase, β-1,3-glucanase activity in T.v.SP456 isolate was Czapek and PDA for phenil-alanin-ammonium-oxidase and chitinase. T.v.SP456 was more efficient than T.h.P8 concerning enzymes activity.The interaction between Trichoderma fungal strains SP456 and P8 and strawberry plant pathogen strains, three belonging to Botrytis cinerea (S1, P1, P2 and one to Phytophtora spp. were examined, also. Both Trichoderma strains act as mycoparasites for plant pathogens. The inhibition percent of radial growth was higher for T.v.SP456 when compared with T.h.P8 for almost all pathogenic isolates.

  10. RNA Interference of Endochitinases in the Sugarcane Endophyte Trichoderma virens 223 Reduces Its Fitness as a Biocontrol Agent of Pineapple Disease

    OpenAIRE

    Aline S Romão-Dumaresq; Welington Luiz Araújo; Nicholas J Talbot; Thornton, Christopher R.

    2012-01-01

    The sugarcane root endophyte Trichoderma virens 223 holds enormous potential as a sustainable alternative to chemical pesticides in the control of sugarcane diseases. Its efficacy as a biocontrol agent is thought to be associated with its production of chitinase enzymes, including N-acetyl-ß-D-glucosaminidases, chitobiosidases and endochitinases. We used targeted gene deletion and RNA-dependent gene silencing strategies to disrupt N-acetyl-ß-D-glucosaminidase and endochitinase activities of t...

  11. The SRAP based molecular diversity related to antifungal and antioxidant bioactive constituents for biocontrol potentials of Trichoderma against Sclerotium rolfsii Scc.

    Science.gov (United States)

    Hirpara, Darshna G; Gajera, H P; Bhimani, R D; Golakiya, B A

    2016-08-01

    The study was performed to examine 11 isolates of Trichoderma for their bio-control potentials against Sclerotium rolfsii Sacc. causing stem rot in groundnut. The antagonists Trichoderma were subjected to sequence related amplified polymorphism (SRAP) based molecular diversity analysis and compared with their hardness to S. rolfsii with respect to secretary antifungal and antioxidant profile. T. virens NBAII Tvs 12 evident highest (87.91 %) growth inhibition of test pathogen followed by T. koningii MTCC 796 (67.03 %) at 7 days after inoculation (DAI). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs 12 and antibiosis for MTCC 796. The growth inhibition of test pathogen was significantly negatively correlated with sclerotia formation and lipid peroxidation during antagonism due to release of secretary bioactive antioxidants by antagonists to terminate oxidative burst generated by S. rolfsii and causing inhibition of sclerotium formation. The GC-MS profile identified antifungal and antioxidant constituents hexadecane, 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, 1-hexadecanesulfonyl chloride, and octadecane in potent antagonists Tvs 12; and nonacosane and octadecane in MTCC 796 along with two novel compounds 1-pentadecene and 1-heneicosyl formate for biocontrol activity. Molecular diversity of Trichoderma isolates associated with antagonistic activity was assessed by SRAP markers. The 115 primer combinations generate total 1328 amplified products of which, 1095 are shared polymorphic and 199 are unique polymorphic. The 15 SRAP combinations produced 18 bands to diagnose best antagonist Tvs 12 and 13 SRAP combinations generated 19 unique bands for identification of MTCC 796. The mycoparasitic antagonist Tvs 12 would be the best antagonist and released unique antifungal and antioxidant constituents to combat pathogen infection. The SRAP based genetic diversity indicates Tvs12 strain clustered with T. viride NBAII Tv23 and shared

  12. Patogenicidade de Pythium aphanidermatum a alface cultivada em hidroponia e seu biocontrole com Trichoderma Pythium aphanidermatum pathogenicity to hydroponics lettuce and its biocontrol with Trichoderma

    Directory of Open Access Journals (Sweden)

    Katya da Silva Patekoski

    2010-08-01

    Full Text Available O objetivo deste trabalho foi avaliar a patogenicidade de Pythium aphanidermatum a variedades de alface, e a ação do produto Biotrich, formulado com Trichoderma, no controle deste patógeno e na promoção do crescimento das plantas. Em experimento in vitro, plântulas recém-germinadas das variedades de alface Vera e Elisa foram colocadas em placas de Petri com ágar-água e 1 mL de suspensão do produto Biotrich (0,2 mL L-1 e, após 24 horas, em discos com micélio do isolado de Pythium. As avaliações foram realizadas após dez dias de incubação a 20 e 31ºC. Os testes in vivo foram realizados na primavera e verão, em sistema hidropônico "Nutrient Film Technique" (NFT, em delineamento inteiramente casualizado, em esquema fatorial 2x2x2, como segue: duas variedades; presença ou ausência do patógeno; e presença ou ausência de Biotrich. Ao final do cultivo, foram avaliadas as massas de matéria fresca e seca das plantas. No experimento in vitro, P. aphanidermatum apresentou maior agressividade a 31ºC. Contudo, não foi verificada patogenicidade nos testes in vivo. De modo geral, o Biotrich não promoveu o crescimento das plantas, mas foi efetivo no controle do patógeno in vitro. Pythium aphanidermatum é patogênico às variedades de alface Vera e Elisa, a 20 e 31ºC, e o Biotrich é efetivo para o controle desse patógeno nessas temperaturas.The objective of this work was to evaluate the pathogenicity of Pythium aphanidermatum to lettuce varieties, and the action of the product Biotrich, formulated with Trichoderma, in the control of this pathogen and its effect on plant growth promotion. In a in vitro experiment, germinated seedlings of Vera and Elisa lettuce varieties were placed in Petri dishes with water-agar and 1mL Biotrich suspension (0.2 mL L-1, and after 24 hours, on plugs with the Pythium isolate mycelium. The evaluations were done ten days after the incubation at 20 and 31ºC. In vivo experiments were carried out during

  13. Study on the Biocontrol Activities of Trichoderma species in Greengram with Infected Fungal Pathogens

    International Nuclear Information System (INIS)

    Seven species of Trichoderma were isolated from rhizospheric soil sources and studied by cultural morphology and microscopic examinations. In dual plate assay, antifungal effects of seven Trichoderma strains were screened against three plant pathogenic fungi (Fusarium oxysporum, Rhizoctonia solani and Pythium sp.) on PDA medium and T-5 isolate showed a wide percentage of inhibitory effects on target pathogens with PIRG value. All Trichoderma strains exhibited a clear zone formation on minimal synthetic medium supplemented with 1% colloidal chitin. T-2 and T-5 were the best chitinase producer strains. In vitro screening for protease activity, the highest protease producing activity of Trichoderma isolate (T-2) were observed in pH indicator medium after 7 days incubation. In pot trial experiment, only T-5 strain exhibited more fungal suppression efficiency on green gram plant than commercial fungicide, Trisan and the other strains. So, it can be said that the effective strain was T-5 strain only which have been more antifungal producing power on three fungal pathogens than Trisan and the resting strains.

  14. 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.

  15. Screening of bacterial biocontrols against sapstain fungus (Lasiodiplodia theobromae Pat.) of rubberwood (Hevea brasiliensis Muell.Arg.).

    Science.gov (United States)

    Sajitha, K L; Maria Florence, E J; Dev, Suma Arun

    2014-09-01

    Diverse bacterial biocontrol agents from various sources of aerobic composts against the sapstain fungus Lasiodiplodia theobromae in rubberwood (Hevea brasiliensis) were isolated, screened and identified by various morphological, biochemical and molecular techniques. The inhibitory effect of seventeen bacterial isolates was examined and seven exhibited inhibition towards the sapstain fungus. Among the seven antagonists, six were conclusively identified as Bacillus subtilis and one as Paenibacillus polymyxa using 16S rRNA-encoding gene sequencing. This is the first report on the occurrence of P. polymyxa, a potent biofertilizer and antagonist in vermicompost. HiCrome Bacillus agar was identified as an effective medium for differentiation of B. subtilis from other Bacillus species. The present work demonstrates the efficacy of the antagonistic property of B. subtilis strains against rubberwood sapstain fungus. Culture-based antagonistic inhibition displayed by B. subtilis can be extended to cater to the biocontrol requirements of wood-based industries against the stain fungus. The study showed the utility of an integrated approach, employing morphological, biochemical and molecular tools for conclusive identification of several bacterial isolates present in aerobic composts from diverse sources. PMID:25049165

  16. Effect of microelement and chemical fungicides on biocontrol effect of Trichoderma T23

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

    @@ Recently there have been many reports about soil diseases controlled by Trichoderma, but few could be applied on agriculture production in large areas. T23 isolated from soil around plant roots in the field by Biopesticide Engineering Center of Shenyang Agricultural University could control effectively Cucurbits Fusarium Wilt. The effects of 9 microelements which include copper, zinc, iron, boron,molybdenum, calcium, manganese, magnesium, potassium and frequently-used chemical fungicides,such as-carbendazim, thiram, thiophanate-methyl, chlorothalonil and hymexazol on the growth and the amounts of spores of Fusarium oxysporum FJ and Trichoderma T23 were studied. The effects of those factors on control effect of T23 to melon diseases were discussed and gave basis for the screening of synergistic agents and fungicides in controlling synergistically the pathogen.

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

    Directory of Open Access Journals (Sweden)

    Oskiera Michał

    2015-06-01

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

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

    Science.gov (United States)

    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. PMID:26349455

  19. Antifungal activity of metabolites of the endophytic fungus Trichoderma brevicompactum from garlic

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    Xuping Shentu

    2014-01-01

    Full Text Available The endophytic fungus strain 0248, isolated from garlic, was identified as Trichoderma brevicompactum based on morphological characteristics and the nucleotide sequences of ITS1-5.8SITS2 and tef1. The bioactive compound T2 was isolated from the culture extracts of this fungus by bioactivity-guided fractionation and identified as 4β-acetoxy-12,13-epoxy-Δ9-trichothecene (trichodermin by spectral analysis and mass spectrometry. Trichodermin has a marked inhibitory activity on Rhizoctonia solani, with an EC50 of 0.25 µgmL-1. Strong inhibition by trichodermin was also found for Botrytis cinerea, with an EC50 of 2.02 µgmL-1. However, a relatively poor inhibitory effect was observed for trichodermin against Colletotrichum lindemuthianum (EC50 = 25.60 µgmL-1. Compared with the positive control Carbendazim, trichodermin showed a strong antifungal activity on the above phytopathogens. There is little known about endophytes from garlic. This paper studied in detail the identification of endophytic T. brevicompactum from garlic and the characterization of its active metabolite trichodermin.

  20. Proteins Related to the Biocontrol of Pythium Damping-off in Maize with Trichoderma harzianum Rifai

    Institute of Scientific and Technical Information of China (English)

    Jie CHEN; Gary E HARMAN; Afio COMIS; Gen-Wu CHENG

    2005-01-01

    Induced resistance has been evidenced as one of mechanisms of Trichoderma to control plantdiseases, however, no study showed the change of host proteomics in Trichoderma-induced resistance ofmaize against damping-off caused by Pythium ultimum Trow. The mechanism of Trichoderma harzianumRifai for controlling maize seedling disease caused by Pythium ultimum Trow was investigated firstly byproteome technique and the result suggested that T. harzianum strain T22 was not only able to promoteseedling growth but also protein accumulation. One-dimensional electrophoresis assay showed that morebands appeared on the gel with T22 or T22 combined with P. ultimum (T22 + P. ultimum) treatment than withother treatments. Enzyme assay showed that two chitinases of the root sample were more activated in thetreatments with T22 than in the other treatments without T22. Proteins in the seedling roots from the varioustreatments were separated through protein extraction and 2-D electrophoresis technique. In the seedlingsproduced from the T22-treated seeds, there were 104 up-regulated proteins and 164 down-regulated pro-teins relative to the control, and 97 and 150, respectively, after treatment with T22 + P. ultimum; however,with P. ultimum alone the values were much lower than with the other two treatments. The correlationcoefficient values were 0.72, 0.51 and 0.49 for the comparison of protein spot distribution on gel amongcontrol with T22, P. ultimum and T22 + P. ultimum, respectively. So it seemed that P. ultimum infection wasmore effective than T22 in interfering with the host proteome profile. Furthermore, analysis with MALDI-TOF-MAS showed that some important proteins associated with defensive reactions were identified in T22or T22 + P. ultimum treatments, including endochitinase, pathogenesis-related protein PRMS (pathogenesis-related maize seed), GTP-binding protein, isoflavone reductase and other proteins related to respiration. Allthose proteins are probably part of the

  1. In Vitro Inhibition of Cellulolytic Enzymes of Fusarium Oxysporum by Trichoderma spp and Pseudomonas Fluorescens on Arachis Hypogaea L

    OpenAIRE

    P.Rajeswari

    2015-01-01

    In an attempt to develop biocontrol system for management of Fusarium wilt in groundnut, Trichoderma viride, Trichoderma harzianum,and Pseudomonas fluorescens were evaluated for their antagonistic activity against Fusarium oxysporum in vitro. .Fusarium wilt diseasescaused by the fungus Fusarium oxysporum lead to significant yield losses of crops. Experiments were conducted on the effect of culture filtratesof T.viride (1%), T. harzianum (1.5%), and P. fluorescens (2%) on the in vitro inhibiti...

  2. Functional analysis of the class II hydrophobin gene HFB2-6 from the biocontrol agent Trichoderma asperellum ACCC30536.

    Science.gov (United States)

    Huang, Ying; Mijiti, Gulijimila; Wang, Zhiying; Yu, Wenjing; Fan, Haijuan; Zhang, Rongshu; Liu, Zhihua

    2015-02-01

    A class II hydrophobin gene, HFB2-6, was cloned from Trichoderma asperellum ACCC30536 and its biocontrol function was studied. According to our previous transcriptome data, six of the eight class II hydrophobin genes were obviously differential expression in four inducing conditions, especially the gene HFB2-6. Moreover, HFB2-6 proven to be differentially transcribed under eight different treatments. HFB2-6 transcripts were up-regulated under 1% Alternaria alternata cell wall and 5% A. alternata fermentation liquid treatments, and by nutritional stress conditions, suggesting that HFB2-6 plays roles in interactions with both biotic and abiotic environmental conditions. HFB2-6 expression was down-regulated under 1% poplar leaf powder culture conditions, but its expression was up-regulated under 1% poplar root powder, indicating that HFB2-6 has a function in root colonization. Furthermore, the recombinant hydrophobin rHFB2-6 was successfully expressed in Escherichia coli BL21-HFB2-6 and purified from the recombinant strain. Genes related to both the jasmonic acid and salicylic acid signal transduction pathways were up-regulated by interaction with renatured rHFB2-6. The ORCA3 (octadecanoid-derivative responsive Catharanthus AP2-domain) gene of the poplar jasmonic acid signal transduction pathway showed a peak expression of 4.48 times at 2 h, and the peak expression of PR1 (pathogenesis-related protein gene) in the salicylic acid signal transduction pathway was 4.58 times at 72 h. Two genes, MP (monopteros) and GH3.17 (auxin original response gene), in the auxin signal transduction pathway were also up-regulated after induction with rHFB2-6, indicating that rHFB2-6 can promote poplar growth and confer broad-spectrum resistance to pathogens.

  3. Evaluation of biocontrol ability of native strains of Trichoderma spp on Rhizoctonia and Fusarium sp in coffee (Coffea arabica) in experimental conditions

    OpenAIRE

    Nina Rudy; Smeltekop Hugh; Almanza JC; Loza-Murguia Manuel

    2016-01-01

    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 Campe...

  4. Extracellular ATP activates MAPK and ROS signaling during injury response in the fungus Trichoderma atroviride

    Directory of Open Access Journals (Sweden)

    Elizabeth eMedina-Castellanos

    2014-11-01

    Full Text Available The response to mechanical damage is crucial for the survival of multicellular organisms, enabling their adaptation to hostile environments. Trichoderma atroviride, a filamentous fungus of great importance in the biological control of plant diseases, responds to mechanical damage by activating regenerative processes and asexual reproduction (conidiation. During this response, reactive oxygen species (ROS are produced by the NADPH oxidase (Nox1/NoxR complex. To understand the underlying early signaling events, we evaluated molecules such as extracellular ATP (eATP and Ca2+ that are known to trigger wound-induced responses in plants and animals. Concretely, we investigated the activation of mitogen-activated protein kinase (MAPK pathways by eATP, Ca2+ and ROS. Indeed, application of exogenous ATP and Ca2+ triggered conidiation. Furthermore, eATP promoted the Nox1-dependent production of ROS and activated a MAPK pathway. Mutants in the MAPK-encoding genes tmk1 and tmk3 were affected in wound-induced conidiation, and phosphorylation of both Tmk1 and Tmk3 was triggered by eATP. We conclude that in this fungus, eATP acts as a damage-associated molecular pattern (DAMP. Our data indicate the existence of an eATP receptor and suggest that in fungi, eATP triggers pathways that converge to regulate asexual reproduction genes that are required for injury-induced conidiation. By contrast, Ca2+ is more likely to act as a downstream second messenger. The early steps of mechanical damage response in T. atroviride share conserved elements with those known from plants and animals.

  5. Proteomics related to the biocontrol of Pythium damping off in maize with Trichoderma harzianum

    Institute of Scientific and Technical Information of China (English)

    CHEN Jie; Gary G Harman; Alfio Comis

    2004-01-01

    @@ Trichoderma harzianum strain T22 controls various diseases of maize and other crops, including seedling and root rots caused by Pythium ultimum. Seedlings of inbred line Mo17 were grown from T22-treated or untreated seeds in field soil or in field soil intested with the pathogen. Five days after planting, seedlings of Mo17 (5-days-old) were smaller in the presence of P. ultimum and larger in the presence of T22 relative to the control. The combination of T22 with P. ultimum (T22 + P.ultimum ) resulted in plants as large as T22 alone. Methods for protein extraction and 2-D gel electrophoresis were developed. Proteins in seedlings roots from the various treatments were separated on 2-D gels and analyzed using PDQuestTM 2-D software. With seedlings produced from T22-treated seeds, there were 104 unmatched proteins and 164 matched proteins relative to the control, and 97 and 150 from the treatment with T22 + P. ultimum, respectively, however, with P. ultimum alone the numbers were much lower than above two treatments. Comparatively, there was very lower similarity of proteome patterns of seedling roots with T22 or P. ultimum or both to control seedlings,the correlative coefficient values were 0.72, 0.51 and 0.49 for the comparisons among control with T22, P. ultimum and T22 + P. ultimum, respectively. Moreover, correlative coefficient of proteome patterns between T22 with P. ultimum was only 0.65, and T22 fungal proteome were also not same as any one of seedling roots with various treatments. Taken together, the components in seedling root proteome seemed to be mostly coming from Mo17 plants themselves and affected strongly by either microbes, but the effects appeared to be stronger by P. ultimum than by T22. 41 spots were selected for protein mass fingerprinting identification, and most detected-spots were intensified in abundance by T22 or T22 + P. ultimum treatments such as pathogenesis-related protein and endochitnase etc. SOD (Mn) was found to be involved in

  6. Water Deficit as a Driver of the Mutualistic Relationship between the Fungus Trichoderma harzianum and Two Wheat Genotypes▿

    Science.gov (United States)

    Donoso, Eduardo P.; Bustamante, Ramiro O.; Carú, Margarita; Niemeyer, Hermann M.

    2008-01-01

    The aim of this study was to assess the occurrence of mutualistic interactions between the fungus Trichoderma harzianum and two wheat genotypes, Triticum aestivum cv. Talhuén and T. turgidum subsp. durum cv. Alifén, and the extent to which water deficit affected these interactions. Two wheat genotypes were cultivated in the presence or absence of T. harzianum and in the presence or absence of water deficit. T. harzianum was in turn cultivated in the presence or absence of wheat plants and in the presence or absence of water deficit. To evaluate the plant-fungus interactions, the root volume, dry biomass, and fecundity of wheat were determined, as was the population growth rate of the fungus. Trichoderma harzianum exerted a positive effect only on plants subjected to water deficit. The population growth rate of T. harzianum was negative in the absence of wheat plants and reached its highest level in the presence of plants under conditions of water deficit. These results confirm the occurrence of a mutualistic interaction between wheat and T. harzianum and show that it is asymmetric and context dependent. PMID:18192424

  7. In-vitro evaluation of fungicides, plant extracts and bio-control agents against rice blast pathogen magnaporthe oryzae couch

    International Nuclear Information System (INIS)

    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)

  8. Screening the Biosphere: The Fungicolous Fungus Trichoderma phellinicola, a Prolific Source of Hypophellins, New 17-, 18-, 19-, and 20-Residue Peptaibiotics

    DEFF Research Database (Denmark)

    Röhrich, Christian René; Iversen, Anita; Jaklitsch, Walter Michael;

    2013-01-01

    To investigate the significance of antibiotics for the producing organism(s) in the natural habitat, we screened a specimen of the fungicolous fungus Trichoderma phellinicola (syn. Hypocrea phellinicola) growing on its natural host Phellinus ferruginosus. Results revealed that a particular group...

  9. Characterization of novel Trichoderma spp. isolates as a search for effective biocontrollers of fungal diseases of economically important crops in Argentina.

    Science.gov (United States)

    Consolo, Verónica Fabiana; Mónaco, Cecilia Inés; Cordo, Cristina Alicia; Salerno, Graciela Lidia

    2012-04-01

    Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens. PMID:22805919

  10. Characterization of novel Trichoderma spp. isolates as a search for effective biocontrollers of fungal diseases of economically important crops in Argentina.

    Science.gov (United States)

    Consolo, Verónica Fabiana; Mónaco, Cecilia Inés; Cordo, Cristina Alicia; Salerno, Graciela Lidia

    2012-04-01

    Monoconidial cultures of 33 isolates of Trichoderma from Buenos Aires Province, Argentina were characterized on the basis of twenty eight morphological, physiological and biochemical features. All of them were screened for proteinase, endochitinase and β-1,3 glucanase activity. Universally primed PCR (UP-PCR) and inter-simple sequence repeat (ISSR) techniques were used to examine the genetic variability among isolates, which resulted in 127 bands for the total number of isolates. These results were subjected to numerical analysis revealing 20 haplotypes grouped in five clusters. The ability of Trichoderma isolates to antogonize soil-borne fungal plant pathogens using a dual culture assay was done against five fungal species: Alternaria sp., Bipolaris sorokiniana, Fusarium graminearum, F. solani, and Pyricularia oryzae. The highest inhibition values (85% RI) were obtained against B. sorokiniana and P. oryzae. Three isolates of T. harzianum named as FCCT2, FCCT3 and FCCT9 were capable of causing a high growth inhibition on four of the fungal species assayed, which was in agreement with their higher extracellular hydrolytic activity. Our results suggest that these isolates have the potential to be effective agents for biocontrol of cereal and tomato fungal pathogens.

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

    Science.gov (United States)

    Strakowska, Judyta; Błaszczyk, Lidia; Chełkowski, Jerzy

    2014-07-01

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

  12. 保护地土壤生防木霉菌种群多样性研究%Study on Genetic Diversity of Biocontrol Trichoderma Population in Greenhouse Soils

    Institute of Scientific and Technical Information of China (English)

    贺字典; 高增贵; 高玉峰; 张洋

    2011-01-01

    采用传统的形态学特征和分子方法(ITS、TEF序列和UP-PCR)研究了蔬菜保护地土壤中木霉菌种群多样性及其影响因素.结果表明:提交genbank 11个木霉种的序列,分别为长枝木霉(Trichoderma longibrachiatum)、拟康氏木霉(Trichoderma pseudokoningii)、黄绿木霉(Trichoderma aureoviride)、棘孢木霉(Trichoderma asperellum)、深绿木霉(Trichoderma atroviride)、哈茨木霉(Trichoderma harzianum)、非钩木霉(Trichoderma inhamatum)、微孢木霉(Trichoderma minutisporum)、长孢木霉(Trichoderma longipile)和粘绿木霉(Trichoderma virens);木霉菌24个菌株经UP-PCR扩增,引物AS4、AS19、L45扩增出一条500 bp大小的木霉菌种的特征性谱带,其他谱带则为多态性谱带,多态性达93.5%;营养条件、杀菌剂及土壤因素对不同种木霉菌的影响不同,得到2株适应性较强的木霉菌株,有望成为生防菌株.%11 Trichoderma species were submitted to genbank by ITS ,TEF sequences analysis methods and effect factors of population diversity were studied in this paper, which included T. Longibrachiatum, T. Atroviride, T. Harziamun, T. Viren, T. Minutisporum, T. Pseudokoningii, T. Aureoviride, T. Inhamatum, T. Asperellum, T. Longipile and T. Helicum. At the same time, genetic diversity of Tnchoderma isolates were analyzed by Universally Primed PCR. A feature band ,500 bp, was the common one in 11 Trichoderma species,other bands were polymorphic,93.5 %. Futhermore, the effect of nutrition conditions, fungicides, soil factors etc. On Trichoderma was greatly different. 2 obtained strains which were strong adaptation, would be good bio-control strains.

  13. Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining

    Science.gov (United States)

    Li, Erfeng; Mao, Zhenchuan; Ling, Jian; Yang, Yuhong; Yin, Wen-Bing; Xie, Bingyan

    2016-01-01

    Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1) of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes), proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs) encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold) of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent. PMID:27416025

  14. Biosynthesis of Antibiotic Leucinostatins in Bio-control Fungus Purpureocillium lilacinum and Their Inhibition on Phytophthora Revealed by Genome Mining.

    Directory of Open Access Journals (Sweden)

    Gang Wang

    2016-07-01

    Full Text Available Purpureocillium lilacinum of Ophiocordycipitaceae is one of the most promising and commercialized agents for controlling plant parasitic nematodes, as well as other insects and plant pathogens. However, how the fungus functions at the molecular level remains unknown. Here, we sequenced two isolates (PLBJ-1 and PLFJ-1 of P. lilacinum from different places Beijing and Fujian. Genomic analysis showed high synteny of the two isolates, and the phylogenetic analysis indicated they were most related to the insect pathogen Tolypocladium inflatum. A comparison with other species revealed that this fungus was enriched in carbohydrate-active enzymes (CAZymes, proteases and pathogenesis related genes. Whole genome search revealed a rich repertoire of secondary metabolites (SMs encoding genes. The non-ribosomal peptide synthetase LcsA, which is comprised of ten C-A-PCP modules, was identified as the core biosynthetic gene of lipopeptide leucinostatins, which was specific to P. lilacinum and T. ophioglossoides, as confirmed by phylogenetic analysis. Furthermore, gene expression level was analyzed when PLBJ-1 was grown in leucinostatin-inducing and non-inducing medium, and 20 genes involved in the biosynthesis of leucionostatins were identified. Disruption mutants allowed us to propose a putative biosynthetic pathway of leucinostatin A. Moreover, overexpression of the transcription factor lcsF increased the production (1.5-fold of leucinostatins A and B compared to wild type. Bioassays explored a new bioactivity of leucinostatins and P. lilacinum: inhibiting the growth of Phytophthora infestans and P. capsici. These results contribute to our understanding of the biosynthetic mechanism of leucinostatins and may allow us to utilize P. lilacinum better as bio-control agent.

  15. Oviposition response of the mosquito, Culex quinquefasciatus to the secondary metabolite(s of the fungus, Trichoderma viride

    Directory of Open Access Journals (Sweden)

    Geetha I

    2003-01-01

    Full Text Available Secondary metabolites produced by Trichoderma viride, a deuteromycetes fungus, under submerged culture condition were formulated and evaluated for oviposition attractancy against gravid females of Culex quinquefasciatus mosquito. At a concentration of 10 µg ml-1 the formulation showed remarkable attractancy with an oviposition active index (OAI of +0.52. When the oviposition attractancy of the formulation was compared with a known oviposition attractant, p-cresol, both at 10 µg ml-1, the former was found to be more attractive to result in 70% egg laying than the later with 30% egg laying. Thin layer chromatography fractions of the secondary metabolites showed that a fraction with Rf value of 0.88 was highly active as oviposition attractant with an OAI of +0.65. Further work on identification of the active principle(s of the microbial formulation might lead to an oviposition attractant useful in mosquito vector management.

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

    Directory of Open Access Journals (Sweden)

    Alexandru Manoliu

    2012-03-01

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

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

    Science.gov (United States)

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

  18. 木霉菌生物防治作用机理与应用研究进展%Progress on Mechanism and Applications of Trichoderma as a Biocontrol Microbe

    Institute of Scientific and Technical Information of China (English)

    陈捷; 朱洁伟; 张婷; 王秉丽

    2011-01-01

    Trichoderma is widely used as a biocontrol microbe against plant diseases in the world, which is mainly applied for controlling a variety of soil bome plant diseases and some plant aboveground diseases. With the advent of MAMPs (microbe-associated molecular patterns) theory, the study on mechanism of plant disease control and improved plant resistance to stress factors with Trichoderma as a biocontrol agent has come into a new stage, especially as the development of technologies based on systems biology, it has become possible to understand the nature of interaction of plant- Trichoderma -pathogen from point of "omics" view, which therefore would intensively enrich the theory basis of Trichoderma as biocontrol agent. Meanwhile, a diversified technical system for applying Trichoderma products developed in national agricultural industry would provide great promotion to sustainable development of agriculture in the future. This review will focus on new research progress on biocontrol mechanism with Trichoderma against plant diseases at molecular level.%木霉菌Trichoderma是国际上应用非常普遍的生防真菌.随着植物免疫的MAMPs(微生物互作分子模式)理论的发展,利用木霉菌防治病害和提高作物抗逆性机理研究进入了一个新阶段,尤其是系统生物学方法的应用,使得人们在"组学"尺度上认识木霉菌与植物和病原菌互作的本质成为可能,这将极大丰富木霉菌生物防治植物病害的理论基础.在我国建立木霉菌多样化应用技术途径是发挥木霉菌在农业可持续发展中作用的重要发展方向.本文重点介绍木霉菌生物防治分子机理研究的新进展.

  19. Colonization and degradation of senescent flowers of zucchini squash by Trichoderma harzianum YC459, a biocontrol agent of gray mold, Botrytis cinerea

    Institute of Scientific and Technical Information of China (English)

    Geun Gon Kim; Young Ryun Chung

    2004-01-01

    @@ In commercial greenhouses, senescent flower petals or flowers of vegetables such as tomato,strawberry, hot pepper and zucchini squash were blighted to be removed from fruits within five days after spraying of Trichoderna harzianun YC459 (TORY() , JGreen Inc.), a biocontrol agent with good and consistent efficacy as chemical fungicides for the control of gray mold rot caused by B.cinerea. The mechanism for selective colonization of senescent floral tissues by T. harzianum YCA59was elucidated using fresh and senescent (4 days and 14 days after pollination, respectively) floral tissues of zucchini squash (Cucurbita moschata Duchesne) . The spores of T. harzianum YCA59were produced much more on water agar and liquid culture media supplemented with 5% dry powder of senescent floral tissues than with fresh tissues during 15 days incubation. Mycelial growth was also much better in the media with senescent tissues than with fresh tissues. Enzyme activities of carboxymethyl cellulase, amylase and polygalacturonase in the liquid media, which might be involved in the colonization and degradation of tissues by T. harzianum YCA59 were compared. The activities of three enzymes were significantly higher in the media with senescent floral tissues than with fresh floral tissues reaching to the maximum during 9 to 12 days of incubation. Especially, the activities of carboxymethyl cellulase and polygalacturonase of T. harzianum YC459 were much higher than those of other Trichoderma species, T. asperellum, T. viride and T. koningii in the liquid media with senescent floral tissues. Based on the results, the selective colonization and degradation of senescent floral tissues, an important habitat for B. cinerea, may be another mechanism for the biocontrol of gray mold rot of vegetables by T. harzianurm YC459.

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

    Directory of Open Access Journals (Sweden)

    José Luis Hernández Mendoza

    2015-12-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  2. 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.

  3. 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. PMID:26168138

  4. Three encochitinase-encoding genes identified in the biocontrol fungus Clonostachys rosea are differentially expressed

    DEFF Research Database (Denmark)

    Mamarabadi, Mojtaba; Jensen, Birgit; Lübeck, Mette

    2008-01-01

    . rosea were made by disruption of the endochitinase genes via Agrobacterium-mediated transformation and their biocontrol activity was evaluated. While in planta bioassays showed no significant difference in biocontrol efficacy between the disruptants and the wildtype, the real time RT-PCR analysis showed...

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

    OpenAIRE

    Jose A. Rodríguez-León; Domenech, F.; M. León; Méndez, T.; D. E. Rodríguez; Ashok Pandey

    1999-01-01

    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, (NH4)2SO4 , NH4H2PO4 and (NH4)2HPO4 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 substra...

  6. Bioremediation from wastewater and extracellular synthesis of copper nanoparticles by the fungus Trichoderma koningiopsis.

    Science.gov (United States)

    Salvadori, Marcia R; Ando, Rômulo A; Oller Do Nascimento, Cláudio A; Corrêa, Benedito

    2014-09-19

    This is the first study describing the rapid extracellular production of copper nanoparticles by dead biomass of Trichoderma koningiopsis. The production and uptake of copper nanoparticles by dead biomass of Trichoderma koningiopsis were characterized by investigating physicochemical factors, equilibrium concentrations and biosorption kinetics, combined with scanning electron microscopy (SEM), energy dispersive X-ray (EDS), transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). A successful route for the metallic copper nanoparticles synthesis was achieved, and followed a Langmuir isotherm where a high biosorption capacity was observed, 21.1 mg g(-1). The kinetic analysis showed that copper biosorption followed a pseudo-second-order model. The nanoparticles mainly exhibited a spherical shape, with an average size of 87.5 nm, and were synthesized extracellularly. The presence of proteins as stabilizing agents of the nanoparticles was demonstrated. The extracellular biosynthesis and uptake of copper nanoparticles using dead fungal biomass is a low-cost green processes, and bioremediation of impacted local.

  7. Draft genome sequence of a strain of cosmopolitan fungus Trichoderma atroviride

    NARCIS (Netherlands)

    Shi-Kunne, X.; Seidl, M.F.; Faino, L.; Thomma, B.P.H.J.

    2015-01-01

    An unknown fungus has been isolated as a contaminant of in vitro-grown fungal cultures. In an attempt to identify the contamination, we isolated the causal agent and performed whole-genome sequencing. BLAST analysis of the internal transcribed spacer (ITS) sequence against the NCBI database showed 1

  8. Biological pre-treatment: Enhancing biogas production using the highly cellulolytic fungus Trichoderma viride.

    Science.gov (United States)

    Mutschlechner, Mira; Illmer, Paul; Wagner, Andreas Otto

    2015-09-01

    With regard to renewable sources of energy, bioconversion of lignocellulosic biomass has long been recognized as a desirable endeavor. However, the highly heterogeneous structure of lignocellulose restricts the exploitation of its promising potential in biogas plants. Hence, effective pre-treatment methods are decisive prerequisites to overcome these challenges in order to improve the utilization ratio of (ligno) cellulosic substrates during fermentation. In the present study, the application of Trichoderma viride in an aerobic upstream process prior to anaerobic digestion led up to a threefold increase in the yield of methane and total gas in a lab-scale investigation. Due to its highly efficient cellulolytic activities, T. viride seemed to be responsible for an improved nutrient availability that positively influenced the anaerobic microbiocenosis. Aerobic pre-treatment of organic matter with T. viride is therefore a promising solution to achieve higher methane yields and degradation performances without any additional energy demand, nor undesired by-product inhibition. PMID:26013693

  9. Isolation and Mass Production of Trichoderma.

    Science.gov (United States)

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

    2016-01-01

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

  10. The ABC transporter ABCG29 is involved in H2O2 tolerance and biocontrol traits in the fungus Clonostachys rosea.

    Science.gov (United States)

    Dubey, Mukesh; Jensen, Dan Funck; Karlsson, Magnus

    2016-04-01

    For successful biocontrol interactions, biological control organisms must tolerate toxic metabolites produced by themselves or plant pathogens during mycoparasitic/antagonistic interactions, by host plant during colonization of the plant, and xenobiotics present in the environment. ATP-binding cassette (ABC) transporters can play a significant role in tolerance of toxic compounds by mediating active transport across the cellular membrane. This paper reports on functional characterization of an ABC transporter ABCG29 in the biocontrol fungus Clonostachys rosea strain IK726. Gene expression analysis showed induced expression of abcG29 during exposure to the Fusarium spp. mycotoxin zearalenone (ZEA) and the fungicides Cantus, Chipco Green and Apron. Expression of abcG29 in C. rosea was significantly higher during C. rosea-C. rosea (Cr-Cr) interaction or in exposure to C. rosea culture filtrate for 2 h, compared to interaction with Fusarium graminearum or 2 h exposure to F. graminearum culture filtrate. In contrast with gene expression data, ΔabcG29 strains did not display reduced tolerance towards ZEA, fungicides or chemical agents known for inducing oxidative, cell wall or osmotic stress, compared to C. rosea WT. The exception was a significant reduction in tolerance to H2O2 (10 mM) in ΔabcG29 strains when conidia were used as an inoculum. The antagonistic ability of ΔabcG29 strains towards F. graminearum, Fusarium oxysporum or Botrytis cinerea in dual plate assays were not different compared with WT. However, in biocontrol assays ΔabcG29 strains displayed reduced ability to protect Arabidopsis thaliana leaves from B. cinerea, and barley seedling from F. graminearum as measured by an A. thaliana detached leaf assay and a barley foot rot disease assay, respectively. These data show that the ABCG29 is dispensable for ZEA and fungicides tolerance, and antagonism but not H2O2 tolerance and biocontrol effects in C. rosea. PMID:26520102

  11. Disruption of the Eng18B ENGase gene in the fungal biocontrol agent Trichoderma atroviride affects growth, conidiation and antagonistic ability.

    Directory of Open Access Journals (Sweden)

    Mukesh K Dubey

    Full Text Available The recently identified phylogenetic subgroup B5 of fungal glycoside hydrolase family 18 genes encodes enzymes with mannosyl glycoprotein endo-N-acetyl-β-D-glucosaminidase (ENGase-type activity. Intracellular ENGase activity is associated with the endoplasmic reticulum associated protein degradation pathway (ERAD of misfolded glycoproteins, although the biological relevance in filamentous fungi is not known. Trichoderma atroviride is a mycoparasitic fungus that is used for biological control of plant pathogenic fungi. The present work is a functional study of the T. atroviride B5-group gene Eng18B, with emphasis on its role in fungal growth and antagonism. A homology model of T. atroviride Eng18B structure predicts a typical glycoside hydrolase family 18 (αβ(8 barrel architecture. Gene expression analysis shows that Eng18B is induced in dual cultures with the fungal plant pathogens Botrytis cinerea and Rhizoctonia solani, although a basal expression is observed in all growth conditions tested. Eng18B disruption strains had significantly reduced growth rates but higher conidiation rates compared to the wild-type strain. However, growth rates on abiotic stress media were significantly higher in Eng18B disruption strains compared to the wild-type strain. No difference in spore germination, germ-tube morphology or in hyphal branching was detected. Disruption strains produced less biomass in liquid cultures than the wild-type strain when grown with chitin as the sole carbon source. In addition, we determined that Eng18B is required for the antagonistic ability of T. atroviride against the grey mould fungus B. cinerea in dual cultures and that this reduction in antagonistic ability is partly connected to a secreted factor. The phenotypes were recovered by re-introduction of an intact Eng18B gene fragment in mutant strains. A putative role of Eng18B ENGase activity in the endoplasmic reticulum associated protein degradation pathway of endogenous

  12. Sequences of stilboflavin C: towards the peptaibiome of the filamentous fungus Stilbella (= Trichoderma) flavipes.

    Science.gov (United States)

    Degenkolb, Thomas; Götze, Lutz; von Döhren, Hans; Vilcinskas, Andreas; Brückner, Hans

    2016-08-01

    Filamentous fungi of the genus Stilbella are recognized as an abundant source of naturally occurring α-aminoisobutyric acid-containing peptides. The culture broth of Stilbella (Trichoderma) flavipes CBS 146.81 yielded a mixture of peptides named stilboflavins (SF), and these were isolated and separated by preparative TLC into groups named SF-A, SF-B, and SF-C. Although all three of these groups resolved as single spots on thin-layer chromatograms, HPLC analysis revealed that each of the groups represents very microheterogeneous mixtures of closely related peptides. Here, we report on the sequence analysis of SF-C peptides, formerly isolated by preparative TLC. HPLC coupled to QqTOF-ESI-HRMS provided the sequences of 10 16-residue peptides and five 19-residue peptides, all of which were N-terminally acetylated. In contrast to the previously described SF-A and SF-B peptaibols, SF-C peptaibols contain Ser-Alaol or Ser-Leuol, which are rarely found as C-termini, and repetitive Leu-Aib-Gly sequences, which have not been detected in peptaibols before. Taking the previously determined sequences of SF-A and SF-B into account, the entirety of peptides produced by S. flavipes (the 'peptaibiome') approaches or exceeds 100 non-ribosomally biosynthesized peptaibiotics. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. PMID:27443977

  13. EXTRACELLULAR PROTEINS PRODUCED BY DIFFERENT SPECIES OF THE FUNGUS TRICHODERMA ON SECONDARY PAPER MILL SLUDGE SUBSTRATE

    Directory of Open Access Journals (Sweden)

    Ida Vaskova,

    2012-01-01

    Full Text Available Kraft pulping is the most commonly used pulping process in the pulp and paper industry. In this process wood chips are chemically delignified using sodium sulfide and sodium hydroxide. Delignification is usually followed by mechanical fiberization and a bleaching process of the resulting wood pulp. In addition to lignin-free wood pulp, this process also produces waste that contains residues of used chemicals, lignin, cellulose, hemicelluloses, and small amounts of other wood components. Because of the worldwide large-scale production of paper, the sludge from paper mills contributes significantly to environmental pollution. Although there have been great efforts being made to utilize this lignin-rich material, sludge is mostly disposed in landfills or incinerated in a boiler. This research project used secondary sludge as a substrate for 7 wood-decay fungi taxonomically belonging to the genus Trichoderma. The examined fungi expressed the capability of consuming sludge components as a carbon source to produce extracellular proteins. The proteins were separated by gel electrophoresis. Before and after fungi cultivation, the sludge was analyzed by Fourier transform infrared spectroscopy (FTIR.

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

    Science.gov (United States)

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

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Penttilä Merja

    2006-10-01

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

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

    Directory of Open Access Journals (Sweden)

    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

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

    Science.gov (United States)

    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. PMID:24966041

  18. The plant pathogenic fungus Gaeumannomyces graminis var. tritici improves bacterial growth and triggers early gene regulations in the biocontrol strain Pseudomonas fluorescens Pf29Arp.

    Science.gov (United States)

    Barret, M; Frey-Klett, P; Boutin, M; Guillerm-Erckelboudt, A-Y; Martin, F; Guillot, L; Sarniguet, A

    2009-01-01

    In soil, some antagonistic rhizobacteria contribute to reduce root diseases caused by phytopathogenic fungi. Direct modes of action of these bacteria have been largely explored; however, commensal interaction also takes place between these microorganisms and little is known about the influence of filamentous fungi on bacteria. An in vitro confrontation bioassay between the pathogenic fungus Gaeumannomyces graminis var. tritici (Ggt) and the biocontrol bacterial strain Pseudomonas fluorescens Pf29Arp was set up to analyse bacterial transcriptional changes induced by the fungal mycelium at three time-points of the interaction before cell contact and up until contact. For this, a Pf29Arp shotgun DNA microarray was constructed. Specifity of Ggt effect was assessed in comparison with one of two other filamentous fungi, Laccaria bicolor and Magnaporthe grisea. During a commensal interaction, Ggt increased the growth rate of Pf29Arp. Before contact, Ggt induced bacterial genes involved in mycelium colonization. At contact, genes encoding protein of stress response and a patatin-like protein were up-regulated. Among all the bacterial genes identified, xseB was specifically up-regulated at contact by Ggt but down-regulated by the other fungi. Data showed that the bacterium sensed the presence of the fungus early, but the main gene alteration occurred during bacterial-fungal cell contact. PMID:19121038

  19. A mutant of the nematophagous fungus Paecilomyces lilacinus (Thom) is a novel biocontrol agent for Sclerotinia sclerotiorum.

    Science.gov (United States)

    Yang, Fan; Abdelnabby, Hazem; Xiao, Yannong

    2015-12-01

    Sclerotinia sclerotiorum causes severe stem rot and yield loss in oilseed rape (Brassica napus L.) and other crops worldwide. Extensive studies have been conducted on Paecilomyces lilacinus as a nematophagous bioagent. However, no reports stated the effect of P. lilacinus as a biocontrol agent against oilseed rape rot S. sclerotiorum. This study describes such effect in lab and field trials using the new transformant pt361 derived from the wild strain P. lilacinus 36-1. Unlike the wild-type strain, the mutant pt361 showed high antagonistic effect against S. Sclerotiorum A. Under lab conditions, the pt361 inhibited (65%) radial mycelial growth of S. sclerotiorum in dual culture test producing 5.9 mm inhibition zone IZ in front of the S. sclerotiorum colony. Moreover, the cell-free filtrate of pt361 culture showed strong inhibitory effects (60.3-100%) on mycelial growth of S. sclerotiorum. In leaf detached assay, pt361 significantly (p rape stem rot, promoted growth and increased yield compared to the control and exceeded, at dose 100%, the action of the fungicide procymidone(®). In conclusion, the mutant pt361 of P. lilacinus is a novel and promising biocontrol agent against oilseed rape Sclerotinia stem rot. PMID:26521137

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

    Institute of Scientific and Technical Information of China (English)

    孙奕; 吕阿丽; 田黎; 魏岚; 裴月湖

    2007-01-01

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

  1. Variabilidade genética na região its do rDNA de isolados de trichoderma spp. (Biocontrolador e Fusarium oxysporum f. sp. Chrysanthemi Genetic variability in rDNA ITS region of Trichoderma spp. (biocontrole agent and Fusarium oxysporum f. sp. chrysanthemi isolates

    Directory of Open Access Journals (Sweden)

    Josiane Pacheco Menezes

    2010-02-01

    Full Text Available A análise de características morfológicas e culturais podem não ser suficientes para uma caracterização precisa das espécies de Trichoderma e Fusarium. Objetivou-se, neste trabalho, caracterizar a região do Espaço Interno Transcrito (ITS do rDNA dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma spp. utilizados no biocontrole de Fusarium oxysporum f. sp. chrysanthemi (isolado UFSMF6. A extração de DNA de cada isolado foi realizada a partir de micélio produzido em meio líquido Batata-Dextrose. As amostras de DNA genômico foram submetidas à Reação em Cadeia da Polimerase (PCR com os oligonucleotídeos iniciadores universais ITS1 e ITS4 e o produto gerado foi sequenciado. Os fragmentos gerados pela amplificação da PCR foram tratados com as enzimas de restrição HaeIII, HinfI e MboI. As regiões ITS1, ITS2 e 5.8S do rDNA desses isolados fúngicos foram amplificadas com sucesso. A região ITS dos isolados UFSMT15.1, UFSMT16 e UFSMT17 de Trichoderma e o isolado UFSMF6 de Fusarium apresentaram uma banda simples com um fragmento de aproximadamente 600 pares de base (pb. As enzimas de restrição HaeIII, HinfI e MboI geraram polimorfismo de bandas entre os isolados. Com base nas análises da sequência de DNA, os isolados UFSMT15.1, UFSMT16, UFSMT17 e UFSMF6 apresentaram maior similaridade com as espécies Trichoderma koningiopsis, Hypocrea virens, Hypocrea lixii e Fusarium oxysporum, respectivamente.The analysis of morphological and cultural characteristics may not enough for the characterization of the species of Trichoderma and Fusarium. The aim of this work was to characterize the Internal Transcribed Spacer (ITS region of the rDNA of UFSMT15.1, UFSMT16 and UFSMT17 isolates of Trichoderma spp. used in the biocontrol of Fusarium oxysporum f. sp. chrysanthemi UFSMF6. DNA extraction of each isolate was accomplished starting from hyphae produced in liquid medium Potato-Dextrose-Agar. The samples of genomic DNA were submitted to

  2. The transcription factor Ste12 mediates the regulatory role of the Tmk1 MAP kinase in mycoparasitism and vegetative hyphal fusion in the filamentous fungus Trichoderma atroviride.

    Science.gov (United States)

    Gruber, Sabine; Zeilinger, Susanne

    2014-01-01

    Mycoparasitic species of the fungal genus Trichoderma are potent antagonists able to combat plant pathogenic fungi by direct parasitism. An essential step in this mycoparasitic fungus-fungus interaction is the detection of the fungal host followed by activation of molecular weapons in the mycoparasite by host-derived signals. The Trichoderma atroviride MAP kinase Tmk1, a homolog of yeast Fus3/Kss1, plays an essential role in regulating the mycoparasitic host attack, aerial hyphae formation and conidiation. However, the transcription factors acting downstream of Tmk1 are hitherto unknown. Here we analyzed the functions of the T. atroviride Ste12 transcription factor whose orthologue in yeast is targeted by the Fus3 and Kss1 MAP kinases. Deletion of the ste12 gene in T. atroviride not only resulted in reduced mycoparasitic overgrowth and lysis of host fungi but also led to loss of hyphal avoidance in the colony periphery and a severe reduction in conidial anastomosis tube formation and vegetative hyphal fusion events. The transcription of several orthologues of Neurospora crassa hyphal fusion genes was reduced upon ste12 deletion; however, the Δste12 mutant showed enhanced expression of mycoparasitism-relevant chitinolytic and proteolytic enzymes and of the cell wall integrity MAP kinase Tmk2. Based on the comparative analyses of Δste12 and Δtmk1 mutants, an essential role of the Ste12 transcriptional regulator in mediating outcomes of the Tmk1 MAPK pathway such as regulation of the mycoparasitic activity, hyphal fusion and carbon source-dependent vegetative growth is suggested. Aerial hyphae formation and conidiation, in contrast, were found to be independent of Ste12.

  3. Dichlorodiaportinol A - A new chlorine-containing isocoumarin from an endophytic fungus Trichoderma sp. 09 from Myoporum bontioides A. Gray and its cytotoxic activity

    Directory of Open Access Journals (Sweden)

    Chunyuan Li

    2014-01-01

    Full Text Available Background: Myoporum bontioides A. Gray, an evergreen shrub from the Myoporaceae family, is a commonly used medicinal plant. Many studies have been conducted on the biologically active constituents of whole parts of M. bontioides. However, the endophytes of M. bontioides have not been intensively investigated. A new chlorine-containing isocoumarin, named dichlorodiaportinol A (1 was isolated from the endophytic fungus Trichoderma sp. 09 isolated from the root of M. bontioides. Its cytotoxic activity against human breast cancer (MCF-7 and human liver cancer (HepG2 cell lines was evaluated. Materials and Methods: Different open silica gel column chromatographic techniques with different solvent systems were used for the separation of the constituents of the ethyl acetate extract of the culture broth of the endophytic fungus Trichoderma sp. 09. The structure of compound one was identified by analysis of spectroscopic data [one-dimensional (1D, two-dimensional (2D-nuclear magnetic resonance (NMR, ultraviolet (UV, infrared (IR and Mass spectrometry (MS]. 3-(4,5-Dimethylthiazol-2-yl-2,5-Diphenyltetrazolium Bromide (MTT assay method was used for the evaluation of cytotoxic activity of compound one against MCF-7 and HepG2 cell lines. Results: Compound one was identified as 3-(3,3-dichloro-2,3-dihydroxy-propyl-8-hydroxy-6- methoxy-isochromen-1-one. It inhibited MCF-7 and HepG2 cell lines, with half maximal inhibitory concentration (IC 50 values of 17.8 and 39.6 μg/mL, respectively. Conclusions: Compound one is a new chlorine-containing isocoumarin with moderate cytotoxic activity against MCF-7 and HepG2 cell lines. Thus, endophytes of M. bontioides are worthy of consideration for the development and research of antitumor agents.

  4. The transcription factor Ste12 mediates the regulatory role of the Tmk1 MAP kinase in mycoparasitism and vegetative hyphal fusion in the filamentous fungus Trichoderma atroviride.

    Science.gov (United States)

    Gruber, Sabine; Zeilinger, Susanne

    2014-01-01

    Mycoparasitic species of the fungal genus Trichoderma are potent antagonists able to combat plant pathogenic fungi by direct parasitism. An essential step in this mycoparasitic fungus-fungus interaction is the detection of the fungal host followed by activation of molecular weapons in the mycoparasite by host-derived signals. The Trichoderma atroviride MAP kinase Tmk1, a homolog of yeast Fus3/Kss1, plays an essential role in regulating the mycoparasitic host attack, aerial hyphae formation and conidiation. However, the transcription factors acting downstream of Tmk1 are hitherto unknown. Here we analyzed the functions of the T. atroviride Ste12 transcription factor whose orthologue in yeast is targeted by the Fus3 and Kss1 MAP kinases. Deletion of the ste12 gene in T. atroviride not only resulted in reduced mycoparasitic overgrowth and lysis of host fungi but also led to loss of hyphal avoidance in the colony periphery and a severe reduction in conidial anastomosis tube formation and vegetative hyphal fusion events. The transcription of several orthologues of Neurospora crassa hyphal fusion genes was reduced upon ste12 deletion; however, the Δste12 mutant showed enhanced expression of mycoparasitism-relevant chitinolytic and proteolytic enzymes and of the cell wall integrity MAP kinase Tmk2. Based on the comparative analyses of Δste12 and Δtmk1 mutants, an essential role of the Ste12 transcriptional regulator in mediating outcomes of the Tmk1 MAPK pathway such as regulation of the mycoparasitic activity, hyphal fusion and carbon source-dependent vegetative growth is suggested. Aerial hyphae formation and conidiation, in contrast, were found to be independent of Ste12. PMID:25356841

  5. The transcription factor Ste12 mediates the regulatory role of the Tmk1 MAP kinase in mycoparasitism and vegetative hyphal fusion in the filamentous fungus Trichoderma atroviride.

    Directory of Open Access Journals (Sweden)

    Sabine Gruber

    Full Text Available Mycoparasitic species of the fungal genus Trichoderma are potent antagonists able to combat plant pathogenic fungi by direct parasitism. An essential step in this mycoparasitic fungus-fungus interaction is the detection of the fungal host followed by activation of molecular weapons in the mycoparasite by host-derived signals. The Trichoderma atroviride MAP kinase Tmk1, a homolog of yeast Fus3/Kss1, plays an essential role in regulating the mycoparasitic host attack, aerial hyphae formation and conidiation. However, the transcription factors acting downstream of Tmk1 are hitherto unknown. Here we analyzed the functions of the T. atroviride Ste12 transcription factor whose orthologue in yeast is targeted by the Fus3 and Kss1 MAP kinases. Deletion of the ste12 gene in T. atroviride not only resulted in reduced mycoparasitic overgrowth and lysis of host fungi but also led to loss of hyphal avoidance in the colony periphery and a severe reduction in conidial anastomosis tube formation and vegetative hyphal fusion events. The transcription of several orthologues of Neurospora crassa hyphal fusion genes was reduced upon ste12 deletion; however, the Δste12 mutant showed enhanced expression of mycoparasitism-relevant chitinolytic and proteolytic enzymes and of the cell wall integrity MAP kinase Tmk2. Based on the comparative analyses of Δste12 and Δtmk1 mutants, an essential role of the Ste12 transcriptional regulator in mediating outcomes of the Tmk1 MAPK pathway such as regulation of the mycoparasitic activity, hyphal fusion and carbon source-dependent vegetative growth is suggested. Aerial hyphae formation and conidiation, in contrast, were found to be independent of Ste12.

  6. Occurrence and characterization of peptaibols from Trichoderma citrinoviride, an endophytic fungus of cork oak, using electrospray ionization quadrupole time-of-flight mass spectrometry.

    Science.gov (United States)

    Maddau, Lucia; Cabras, Annalisa; Franceschini, Antonio; Linaldeddu, Benedetto T; Crobu, Salvatore; Roggio, Tonina; Pagnozzi, Daniela

    2009-10-01

    A cork oak endophytic strain of Trichoderma citrinoviride, previously selected for its antagonistic potential against various fungal pathogens involved in oak decline, was screened for the production of bioactive secondary metabolites. From liquid culture a mixture of polypeptide antibiotics (peptaibols) belonging to the paracelsin family was isolated and characterized. This peptide mixture was purified by column chromatography and preparative TLC on silica gel, and separated by analytical HPLC. It was analysed by MALDI-TOF MS and nano-ESI-QTOF MS. Tandem mass experiments were performed to determine the amino acid sequences based on the fragmentation pattern of selected parent ions. The mixture comprised 20-residue peptides with C-terminal phenylalaninol and N-terminal acetylation. Twenty-eight amino acid sequences were identified, and amino acid exchanges were located in positions 6, 9, 12 and 17. Among them, seven sequences are new as compared to those reported in the database specifically for peptaibols and in the literature. In addition, we obtained experimental evidence suggesting the existence of non-covalent dimeric forms (homo- and hetero-) of the various peptaibol species. The peptide mixture showed strong antifungal activity toward seven important forest tree pathogens, and it was highly toxic in an Artemia salina (brine shrimp) bioassay. These results emphasize the cryptic role of endophytic fungi as a source of novel bioactive natural products and biocontrol agents. PMID:19574303

  7. Occurrence and characterization of peptaibols from Trichoderma citrinoviride, an endophytic fungus of cork oak, using electrospray ionization quadrupole time-of-flight mass spectrometry.

    Science.gov (United States)

    Maddau, Lucia; Cabras, Annalisa; Franceschini, Antonio; Linaldeddu, Benedetto T; Crobu, Salvatore; Roggio, Tonina; Pagnozzi, Daniela

    2009-10-01

    A cork oak endophytic strain of Trichoderma citrinoviride, previously selected for its antagonistic potential against various fungal pathogens involved in oak decline, was screened for the production of bioactive secondary metabolites. From liquid culture a mixture of polypeptide antibiotics (peptaibols) belonging to the paracelsin family was isolated and characterized. This peptide mixture was purified by column chromatography and preparative TLC on silica gel, and separated by analytical HPLC. It was analysed by MALDI-TOF MS and nano-ESI-QTOF MS. Tandem mass experiments were performed to determine the amino acid sequences based on the fragmentation pattern of selected parent ions. The mixture comprised 20-residue peptides with C-terminal phenylalaninol and N-terminal acetylation. Twenty-eight amino acid sequences were identified, and amino acid exchanges were located in positions 6, 9, 12 and 17. Among them, seven sequences are new as compared to those reported in the database specifically for peptaibols and in the literature. In addition, we obtained experimental evidence suggesting the existence of non-covalent dimeric forms (homo- and hetero-) of the various peptaibol species. The peptide mixture showed strong antifungal activity toward seven important forest tree pathogens, and it was highly toxic in an Artemia salina (brine shrimp) bioassay. These results emphasize the cryptic role of endophytic fungi as a source of novel bioactive natural products and biocontrol agents.

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

    Directory of Open Access Journals (Sweden)

    Mihaela Cristica

    2010-09-01

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

  9. Trichocitrin, a new fusicoccane diterpene from the marine brown alga-endophytic fungus Trichoderma citrinoviride cf-27.

    Science.gov (United States)

    Liang, Xiao-Rui; Miao, Feng-Ping; Song, Yin-Ping; Guo, Zhan-Yong; Ji, Nai-Yun

    2016-07-01

    One new diterpene, trichocitrin (1), and four known secondary metabolites, nafuredin (2), 5-hydroxy-2,3-dimethyl-7-methoxychromone (3), 24-methylenecycloartanol (4) and citrostadienol (5), were isolated from the culture of marine brown alga-endophytic Trichoderma citrinoviride cf-27. Trichocitrin (1) represents the first Trichoderma-derived and furan-bearing fusicoccane diterpene, and its structure and relative configuration were identified by analysis of 1D/2D NMR and mass spectroscopic data. Compounds 1 and 2 exhibited 8.0- and 9.5-mm inhibition zones, respectively, against Escherichia coli at 20 μg/disc and 54.1 and 36.7% growth inhibition, respectively, of Prorocentrum donghaiense at 80 μg/mL. PMID:26728965

  10. Trichocitrin, a new fusicoccane diterpene from the marine brown alga-endophytic fungus Trichoderma citrinoviride cf-27.

    Science.gov (United States)

    Liang, Xiao-Rui; Miao, Feng-Ping; Song, Yin-Ping; Guo, Zhan-Yong; Ji, Nai-Yun

    2016-07-01

    One new diterpene, trichocitrin (1), and four known secondary metabolites, nafuredin (2), 5-hydroxy-2,3-dimethyl-7-methoxychromone (3), 24-methylenecycloartanol (4) and citrostadienol (5), were isolated from the culture of marine brown alga-endophytic Trichoderma citrinoviride cf-27. Trichocitrin (1) represents the first Trichoderma-derived and furan-bearing fusicoccane diterpene, and its structure and relative configuration were identified by analysis of 1D/2D NMR and mass spectroscopic data. Compounds 1 and 2 exhibited 8.0- and 9.5-mm inhibition zones, respectively, against Escherichia coli at 20 μg/disc and 54.1 and 36.7% growth inhibition, respectively, of Prorocentrum donghaiense at 80 μg/mL.

  11. Antimicrobial activities of novel mannosyl lipids isolated from the biocontrol fungus Simplicillium lamellicola BCP against phytopathogenic bacteria.

    Science.gov (United States)

    Le Dang, Quang; Shin, Teak Soo; Park, Myung Soo; Choi, Yong Ho; Choi, Gyung Ja; Jang, Kyoung Soo; Kim, In Seon; Kim, Jin-Cheol

    2014-04-16

    The antagonistic fungus Simplicillium lamellicola BCP has been developed as a microbial biopesticide that effectively controls the development of various plant diseases caused by both pathogenic bacteria and pathogenic fungi. Antibacterial bioassay-directed fractionation was used to isolate mannosyl lipids from S. lamellicola BCP, and the structures of these compounds were elucidated using spectral analysis and chemical degradation. Three novel mannosyl lipids were characterized and identified as halymecins F and G and (3R,5R)-3-O-β-D-mannosyl-3,5-dihydrodecanoic acid. Massoia lactone and (3R, 5R)-3-hydroxydecan-5-olide were also isolated from S. lamellicola BCP. The three novel compounds inhibited the growth of the majority of phytopathogenic bacteria that were tested, and halymecin F displayed the strongest antibacterial activity. Agrobacterium tumefaciens was the most sensitive to the three novel compounds, with IC₅₀ values ranging from 1.58 to 24.8 μg/mL. The ethyl acetate extract of the fermentation broth from the antagonistic fungus effectively reduced the bacterial wilt caused by Ralstonia solanacearum on tomato seedlings. These results indicate that S. lamellicola BCP suppresses the development of plant bacterial diseases through the production of antibacterial metabolites. PMID:24660753

  12. Primary Study on Biological Control Potential of Trichoderma harzianum TL-1

    Institute of Scientific and Technical Information of China (English)

    Su; Zhenyu; Xiao; Man; Gao; Xinzheng; Tang; Libo; Li; Li

    2014-01-01

    Trichoderma harzianum is a widely used biocontrol fungus. The growth promoting effect of strain Trichoderma harzianum TL-1 on tomato and pepper and its biological control effects against tomato seedling damping-off and pepper blight were investigated through pot experiments. The results showed that the stain TL-1 had significant promotion effect on growth of pepper and tomato in sterilized and natural soils. With the application dose of 3. 0 and 0. 5g/ pot,their dry weight were increased up to 46% and 150% compared with control,respectively. In addition,TL-1 had good control effects against tomato seedling damping-off and pepper blight. Compared with fungicide treatment,TL-1 treatment could control diseases for long term,without repeat occurrence of diseases.

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

    Science.gov (United States)

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

    2015-01-01

    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. PMID:26647876

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

    Science.gov (United States)

    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

    2015-01-01

    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. PMID:26647876

  15. 木霉对玉米纹枯病的生物防治%Biocontrol of maize sheath blight with Trichoderma spp.

    Institute of Scientific and Technical Information of China (English)

    张广志; 文成敬

    2005-01-01

    为明确木霉菌在玉米纹枯病防治上的作用,研究了从雅安郊区大田玉米根际土壤中分离到的18个木霉菌株对立枯丝核菌的抑制作用及对玉米纹枯病的田间防治效果.对峙培养结果表明,这些木霉菌株对立枯丝核菌的抑制率为39.44%~84.98%;而木霉发酵滤液的抑菌作用为13.48%~85.81%.对峙培养和液体发酵滤液的抑菌测定相结合,筛选出木霉菌株T8、T2、T13,经鉴定分别为哈茨木霉Trichoderma harzianum、黄绿木霉Trichoderma aureoviride和长枝木霉Trichoderma Longibrachiatum.用这3种木霉的固体培养物作进一步的田间防治试验,防治效果分别为62.75%、64.48%、68.52%,比井冈霉素(49.68%)效果高;T13菌株的发酵滤液的防治效果仅为40.07%.3株木霉固体培养物对玉米产量影响显著,增产率依次为29.77%、43.37%、54.21%.

  16. 哈茨木霉对辣椒生长的影响%Effects of Biocontrol Agents Trichoderma harzianum on Plant Growth in Capsicum

    Institute of Scientific and Technical Information of China (English)

    刘云龙; 何永宏; 张旭东

    2002-01-01

    在田间用哈茨木霉(Trichoderma harzianum)制剂处理土壤,栽培辣椒,结果表明:哈茨木霉不但对辣椒疫霉(Phytophthora capsici)和尖镰孢(Fusarium oxysporum)引起的根腐病具有一定的防病作用,而且能促进植株生长,提前开花、挂果,提高产量15%左右.

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  18. 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. PMID:15856359

  19. Screening of Trichoderma strains tolerant to benzimidazole

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  20. Effect of plant growth regulators on in vitro biological control of Fusarium oxysporum by Trichoderma harzianum (T8).

    Science.gov (United States)

    Badri, M; Zamani, M R; Motallebi, M

    2007-09-01

    In this study the effect of two plant growth regulators (indolacetic acid, IAA and gibberellic acid, GA3) and also Trichoderma harzianum (T8) on the phytopathogen fungus Fusarium oxysporium (F15) was investigated. IAA and GA3 with 15 and 30 ppm concentration have no significant effect on T. harzianum (T8) growth. The biocontrol activity of T. harzianum on F. oxysporum was slightly decreased by the presence of IAA and/or GA3. Addition of 40 ppm of GA3 to the culture medium of F. oxsporum increased polygalacturonase activity about 100%. A strong increasing effect on chitinase activity (60%) by T. harzianum (T8) was observed in the presence of phytopathogenic fungus F. oxysporum, but 40 ppm IAA and/or GA3 decreased about 47% of chitinase activity of T. harzianum.

  1. Study of signal transduction factors involved in mycoparasitic response of Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

    Lorito M; Zeilinger S; Ambrosino P; Brunner K; Reithner B; Mach R L; Woo S L; Cristilli M; Scala F

    2004-01-01

    @@ Numerous Trichoderma spp. are mycoparasites and commercially applied as biological control agents against a large number of plant pathogenic fungi. The mycoparasitic interaction is host-specific and several research strategies have been applied to identify the main genes and compounds involved in the antagonist-plant-pathogen three-way interaction. During mycoparasitism, signals from the host fungus are recognised by Trichoderma, stimulating antifungal activities that are accompanied by morphological changes and the secretion of hydrolytic enzymes and antibiotics. Interestingly some morphological changes appeared highly conserved in the strategy of pathogenicity within the fungal world, i.e. the formation of appressoria as well as the secretion of hydrolytic enzymes seem to be general mechanisms of attack both for plant pathogens and mycoparasitic antagonists. This knowledge is being used to identify receptors and key components of signalling pathways involved in fungus-fungus interaction. For this purpose we have cloned the first genes (tmk1 , tga1 , tga3) from T. atroviride showing a high similarity to MAP kinase and G protein subunits (see abstract by Zeilinger et al.),which have been found to have an important role in pathogenicity by Magnaporthe grisea. To identify the function and involvement of these factors in mycoparasitism by T. atroviride, tmk1, tga1, tga3disruptant strains were produced. The knock-out mutants were tested by in vivo biocontrol assays for their ability to inhibit soil and foliar plant pathogens such as Rhizoctonia solani, Pythium ultimum and Botrytis cinerea . Disruption of these genes corresponded to a complete loss of biocontrol ability,suggesting a significant role in mycoparasitism. In particular, it has been suggested that tga3 regulates the expression of chitinase-encoding genes, the secretion of the corresponding enzymes and the process of conidiation. Comparative proteome analysis of wild type and disruptants supported this

  2. Método para preservação da viabilidade e atividade antagônica de Trichoderma stromaticum , agente de biocontrole da vassoura-de-bruxa do cacaueiro Method for preservation of viability and antagonic activity of Trichoderma stromaticum, biocontrol agent of witches´broom disease of cocoa

    Directory of Open Access Journals (Sweden)

    Cleber Novais Bastos

    2008-09-01

    Full Text Available Avaliou-se a viabilidade da massa esporógena de Trichoderma stromaticum, através do crescimento micelial em meio de cultivo e a atividade antagônica (parasitária em vassouras secas de cacaueiro, após a preservação de quatro isolados (Ts1606, Ts3107, Ts0108, Ts2705 do antagonista por quatro anos em fragmentos de vassoura secas, acondicionados em tubos de ensaio e mantidos em refrigerador com temperatura aproximada de 5 °C. Todos os isolados preservados apresentaram-se viáveis, com crescimento e esporulação normais e continuavam antagônicos a Crinipellis perniciosa. Os resultados obtidos indicam a eficiência do método, que é capaz de manter os isolados de T. stromaticum viáveis por longos períodos de tempo, preservando características morfológicas, fisiológicas e antagônicas.After four years of preservation by using dried cocoa brooms in fridge at 5 °C, the viability of four isolates (Ts1606, Ts3107, Ts0108, Ts2705 of Trichoderma stromaticum was evaluated based on germination of conidia, mycelium growth in a culture medium and antagonistic activity on dry cocoa brooms. All the isolates preserved maintained viability, showing normal growth, sporulation and antagonistic activity against Crinipellis perniciosa.. These results indicate the efficience of the method for the long term preservation of T. stromaticum, maintaining their morphological, physiological, and antagonistic characteristics.

  3. Agrobacterium tumefaciens-mediated transformation of CryⅠA(b) gene to Trichoderma harzianum

    Institute of Scientific and Technical Information of China (English)

    GAO Xingxi; YANG Qian

    2004-01-01

    In this study, CryⅠA(b) gene was successfully transferred into the biocontrol fungus Trichoderma harzianum with an efficiency of 60-180 transformants per 106 spores by using Agrobacterium tumefaciens-mediated trans- formation. Putative transformants were analyzed to test the presence of CryⅠA(b) gene by Southern blot. Most transformants contained a single T-DNA copy. RT-PCR analysis showed that the CryⅠA(b) gene was transcribed. Antifungal activities and insecticidal activities of the transformants were examined. There was no obvious difference in antifungal activities between the transformants and their wild strains. The modified mortalities of the transformants T1 and T2 were 69.57% and 91.30%, respectively. The tranformation system mediated by A. tumefaciens proved to be a powerful tool for the filamentous fungi transformation and functional genomic study with its high transformation frequency, simplicity of T-DNA integration, and genetic stability of transformants.

  4. Microencapsulation Technique with Organic Additives for Biocontrol Agents

    Directory of Open Access Journals (Sweden)

    Szczech Magdalena

    2016-06-01

    Full Text Available The aim of these studies was to develop an effective formulation for microbial strains active in plant protection. Emulsification technique in oil was used to produce alginate microcapsules supplemented with organic additives to improve production process and stability of the preparations during storage. The release of microorganisms from the capsules in soil and their effectiveness in biocontrol of Fusarium wilt were evaluated. Three bacterial strains Burkholderia cepacia strain CAT5, Bacillus spp. strains PZ9 and SZ61, and fungus Trichoderma virens TRS106 were immobilized separately in calcium alginate supplemented with chitosan, peat powder, or skim milk. The productivity of microcapsules was enhanced by 60% when peat was added to the alginate matrix. Peat reduced also contamination of the capsules during storage, significantly enhancing their quality. By contrast, the addition of skim milk reduced quality of the microcapsules. The additives did not influence the viability of entrapped microorganisms and their release in soil. The survival of the microbial cells was mainly related to the kind of microorganism used, and the highest viability showed Bacillus sp. PZ9 and Trichoderma TRS106. Lyophilization of the microcapsules appeared to be unfavorable by reducing microbial viability in the capsules and in the soil after application. The best properties: good storage ability and sufficient microbial release in the soil, exhibited wet microcapsules amended with peat. These capsules were used to control Fusarium wilt in tomato plants. The protective effect was obtained when the microcapsule-entrapped bacteria PZ9 were used. The effectiveness of this bacterium was comparable with fungicide. Peat-amended microcapsules entrapping Bacillus PZ9 showed the best quality and may have potential for commercial use.

  5. The Psychrotolerant Antarctic Fungus Lecanicillium muscarium CCFEE 5003: A Powerful Producer of Cold-Tolerant Chitinolytic Enzymes.

    Science.gov (United States)

    Fenice, Massimiliano

    2016-01-01

    Lecanicillium muscarium CCFEE 5003, isolated in Continental Antarctica, is a powerful producer of extracellular cold-tolerant enzymes. Chitin-hydrolyzing enzymes seems to be the principal extracellular catalytic activities of this psychrotolerant fungus. The production of chitinolytic activities is induced by chitin and other polysaccharides and is submitted to catabolite repression. The chitinolytic system of L. muscarium consists of a number of different proteins having various molecular weights and diverse biochemical characteristics, but their most significant trait is the marked cold-tolerance. L. muscarium and selected strains of the biocontrol agent of pathogenic fungi Trichoderma harzianum, have been compared for their ability to produce chitinolytic enzymes at different temperatures. At low temperatures the Antarctic strain was definitely much more efficient. Moreover, the fungus was able to exert a strong mycoparasitic action against various other fungi and oomycetes at low temperatures. The parasitic role of this organism appeared related to the production of cell wall degrading enzymes being the release of extracellular chitinolytic enzymes a key event in the mycoparasitic process. Due to the mentioned characteristics, L. muscarium could have an important role for potential applications such as the degradation of chitin-rich materials at low temperature and the biocontrol of pathogenic organisms in cold environments. For these reasons and in view of future industrial application, the production of chitinolytic enzymes by the Antarctic fungus has been up-scaled and optimised in bench-top bioreactor. PMID:27058517

  6. Biocontrol con Trichoderma spp. de Fusarium oxysporum causal del “mal de almácigos” en pre y post emergencia en cebolla

    OpenAIRE

    Sánchez, Aixa D.; Barrera, Viviana; Reybet, Graciela; Sosa, M. Cristina

    2015-01-01

    En Argentina, la “podredumbre basal” y el “mal de almácigos” causados por el complejo Fusarium spp. constituyen uno de los problemas más serios en cebolla. En este trabajo se estudió el uso de especies del género Trichoderma como agentes de control biológico (ACB) de F. oxysporum, causal del “mal de almácigos” en cebolla. Se identificaron y caracterizaron morfológica, cultural y fisiológicamente a 10 aislamientos de Trichoderma spp. del cepario de Fitopatología. Por secuenciación del gen TEF-...

  7. 真菌Trichoderma viride Pers.ex Fr降解三苯废气性能实验%PERFORMANCE EXPERIMENT ON AEROBIC BIODEGRADATION OF BENZENE,TOLUENE AND XYLENE BY FUNGUS TRICHODERMA VIRIDE PERS.EX FR

    Institute of Scientific and Technical Information of China (English)

    侯晨涛; 马广大

    2007-01-01

    利用固相静态试验考察前期实验驯化出的真菌Trichoderma viride Pers.ex Fr对苯、甲苯、二甲苯的好氧生物降解性能.结果表明,在本研究的气相浓度范围内(苯47.25~1677.47g/m3,甲苯64.70~1258.50g/m3、二甲苯20.59~1707.85g/m3),3种气体的降解速率随其初始浓度的增大而增加,降解规律苯、甲苯符合Monod方程,二甲苯符合一级反应.

  8. EFECTO SOBRE LA SARNA POLVOSA DE CUATRO AISLAMIENTOS DEL HONGO Trichoderma asperellum EN TRES TIPOS DE SUELO EFFECT ON THE POWDERY SCAB OF FOUR ISOLATES OF THE FUNGUS Trichoderma asperellum IN THREE TYPES OF SOILS

    Directory of Open Access Journals (Sweden)

    Elizabeth Gilchrist

    2009-06-01

    Full Text Available El protozoario Spongospora subterranea f. sp. subterranea es un parásito obligado que replica en raíces y tubérculos de papa causando la sarna polvosa. Esta enfermedad se ha difundido a escala global debido al comercio de tubérculos-semilla infectados y a la ausencia de tratamientos efectivos. En este trabajo se investigó la influencia sobre la sarna polvosa de cuatro aislamientos de Trichoderma asperellum en tres tipos de suelo: Andisol, Entisol e Inceptisol. Tubérculos de Solanum tuberosum variedad Diacol Capiro fueron plantados en parcelas infectadas con 1942 ± 226 quistosoros por gramo de suelo. No se observaron diferencias entre las plantas tratadas y no tratadas con T. asperellum. La superficie de raíz ocupada por agallas fue en promedio 2,2%, sin encontrarse diferencias significativas (PThe protozoan Spongospora subterranea f. sp. subterranea is an obligate parasite that replicates in roots and tubers of potato causing the powdery scab. This disease has disseminated on a global scale, because the commercial traffic of infected tubers used as seed and the absence of effective treatments. The present work investigated the effects of four isolates of Trichoderma asperellum in three soil types: Andisol, Entisol and Inceptisol. The soils were infected with 1942 ± 226 cystosori per gram of soil. It was used Solanum tuberosum variety Diacol Capiro. No differences were observed between treated and non treated plants with T. asperellum. The root surface affected by galls was an average of 2.2 %, and no significant differences (p<0.05 were observed among the soils. The reduction of plant growth was of 41, 32 and 28 % for the Andisol, Entisol and Inceptisol, respectively. The reduction in tubers production was of 40, 28 and 0,1% for the Andisol, Entisol and Inceptisol, respectively. Similar percent of root surface covered with galls resulted in production reduction of 40%, but also of 0,1%, indicating the necessity of other methods to

  9. 20-Residue and 11-residue peptaibols from the fungus Trichoderma longibrachiatum are synergistic in forming Na+/K+ -permeable channels and adverse action towards mammalian cells.

    Science.gov (United States)

    Mikkola, Raimo; Andersson, Maria A; Kredics, László; Grigoriev, Pavel A; Sundell, Nina; Salkinoja-Salonen, Mirja S

    2012-11-01

    Certain species of the filamentous fungal genus Trichoderma (e.g. Trichoderma longibrachiatum and Trichoderma citrinoviride) are among the emerging clinical pathogens and also the most common species in the indoor space of mould-damaged buildings. The molecules involved in its pathology are not known. In the present study, we report that 0.5-2.6 wt% of the T. longibrachiatum mycelial biomass consisted of thermostable secondary metabolites mitochondriotoxic to mammalian cells. These were identified by LC/MS as one 11-residue and eight 20-residue peptaibols, AcAib-Asn-Leu/Ile-Leu/Ile-Aib-Pro-Leu/Ile-Leu/Ile-Aib-Pro-Leuol/Ileol (1175 Da) and AcAib-Ala-Aib-Ala-Aib-Ala/Aib-Gln-Aib-Val/Iva-Aib-Gly-Leu/Ile-Aib-Pro-Val/Iva-Aib-Val/Iva/Aib-Gln/Glu-Gln-Pheol(1936-1965 Da) (Aib, α-aminoisobutyric acid; Ac, acetyl; Ileol, isoleucinol; Iva, isovaline; Leuol, leucinol; Pheol, phenylalaninol). The toxic effects on boar sperm cells depended on these peptaibols, named trilongins. The trilongins formed voltage dependent, Na(+)/K(+) permeable channels in biomembranes. The permeability ratios for Na(+) ions, relative to K(+), of the 11-residue trilongin channel (0.95 : 1) and the 20-residue trilongin channel (0.8 : 1) were higher than those of alamethicin. The combined 11-residue and 20-residue trilongins generated channels that remained in an open state for a longer time than those formed by either one of the peptaibols alone. Corresponding synergy was observed in toxicokinetics. With 11-residue and 20-residue trilongins combined 1 : 2 w/w, an effective median concentration (EC(50) ) of 0.6 μg·mL(-1) was reached within 30 min, and the EC(50) shifted down to 0.2 μg·mL(-1) upon extended exposure. By contrast, with 11-residue or 20-residue trilonging separately in 30 min of exposure, the EC(50) values were 15 and 3 μg·mL(-1) , respectively, and shifted down to 1.5 and 0.4 μg·mL(-1) upon extended exposure. This is the first report on ion-channel forming peptaibols with

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

    OpenAIRE

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

    1996-01-01

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

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

    Science.gov (United States)

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

    2014-07-10

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

  12. Applications of Trichoderma formulations in crop protection

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

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

    Institute of Scientific and Technical Information of China (English)

    袁志林; 陈益存; 章初龙; 林福呈; 陈连庆

    2008-01-01

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

  14. In vitro interactions between Armillaria species and potential biocontrol fungi

    Directory of Open Access Journals (Sweden)

    Keča Nenad

    2009-01-01

    Full Text Available Interaction between Armillaria species and seven other fungi were tested in vitro. Tree antagonistic (Trichoderma viride, Trichotecium roseum and Penicillium sp. and four decaying (Hypholoma fasciculare¸ Hypholoma capnoides, Phlebiopsis gigantea, and Pleurotus ostreatus fungi were chosen for this study. The best results were noted for Trichoderma viride, because fungus was able to kill both mycelia and rhizomorphs of Armillaria species, while Hypholoma spp. inhibited both growth of Armillaria colonies and rhizomorph production.

  15. Cyclonerol derivatives from Trichoderma longibrachiatum YM311505.

    Science.gov (United States)

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

    2014-03-01

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

  16. Using of green fluorescent reporter gene (GFP) to monitor the fate of Fusarium moniliforme mycoparasitized by Trichoderma viride

    Institute of Scientific and Technical Information of China (English)

    ZHU Ting-heng; WANG Wei-xia; WANG Chang-chun; YANG Rui-qin; CAI Xin-zhong

    2004-01-01

    @@ Fusarium moniliforme Sheld. is a rice pathogenic fungus and causes the disease called Bakanae,which has increasingly damaged rice production in the recent years. Trichoderma spp. has been one of the most widely used biological control agent of plant disease. By geneticaly labelling F. moniliforme with the GFP reporter gene, we have studied the antagonistic action of Trichoderma viride against this pathogenic fungus.

  17. Role of gliotoxin in the symbiotic and pathogenic interactions of Trichoderma virens.

    Science.gov (United States)

    Vargas, Walter A; Mukherjee, Prasun K; Laughlin, David; Wiest, Aric; Moran-Diez, Maria E; Kenerley, Charles M

    2014-10-01

    Using a gene disruption strategy, we generated mutants in the gliP locus of the plant-beneficial fungus Trichoderma virens that were no longer capable of producing gliotoxin. Phenotypic assays demonstrated that the gliP-disrupted mutants grew faster, were more sensitive to oxidative stress and exhibited a sparse colony edge compared with the WT strain. In a plate confrontation assay, the mutants deficient in gliotoxin production were ineffective as mycoparasites against the oomycete, Pythium ultimum, and the necrotrophic fungal pathogen, Sclerotinia sclerotiorum, but retained mycoparasitic ability against Rhizoctonia solani. Biocontrol assays in soil showed that the mutants were incapable of protecting cotton seedlings from attack by P. ultimum, against which the WT strain was highly effective. The mutants, however, were as effective as the WT strain in protecting cotton seedlings against R. solani. Loss of gliotoxin production also resulted in a reduced ability of the mutants to attack the sclerotia of S. sclerotiorum compared with the WT. The addition of exogenous gliotoxin to the sclerotia colonized by the mutants partially restored their degradative abilities. Interestingly, as in Aspergillus fumigatus, an opportunistic human pathogen, gliotoxin was found to be involved in pathogenicity of T. virens against larvae of the wax moth, Galleria mellonella. The loss of gliotoxin production in T. virens was restored by complementation with the gliP gene from A. fumigatus. We have, thus, demonstrated that the putative gliP cluster of T. virens is responsible for the biosynthesis of gliotoxin, and gliotoxin is involved in mycoparasitism and biocontrol properties of this plant-beneficial fungus. PMID:25082950

  18. Development of a biocontrol agent for plant disease control with special emphasis on the near commercial fungal antagonist Clonostachys rosea strain "IK726"

    DEFF Research Database (Denmark)

    Jensen, Dan Funck; Knudsen, Inge M.B.; Lübeck, Mette;

    2007-01-01

    . Among the success stories for control of seed- and soilborne diseases are fungal biocontrol agents based on Trichoderma harzianum, Clonostachys rosea and Conithyrium minitans, and bacterial biocontrol agents based on strains of Agrobacterium, Pseudomonas and Streptomyces. We have developed C. rosea...

  19. Novel understanding of Trichoderma interaction mechanisms

    Institute of Scientific and Technical Information of China (English)

    Matteo Lorito

    2004-01-01

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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  2. Trichoderma genes

    Science.gov (United States)

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

    2012-06-19

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

  3. Characterization and gene cloning of a novel serine protease with nematicidal activity from Trichoderma pseudokoningii SMF2.

    Science.gov (United States)

    Chen, Lei-Lei; Liu, Li-Jun; Shi, Mei; Song, Xiao-Yan; Zheng, Chang-Ying; Chen, Xiu-Lan; Zhang, Yu-Zhong

    2009-10-01

    Trichoderma pseudokoningii SMF2 is a biocontrol fungus with inhibitory ability against phytopathogenic fungi. Here, a crude extract of strain SMF2 in a solid ferment exhibited strong nematicidal activity against Meloidogyne incognita, and a novel serine protease SprT with nematicidal activity was purified from the crude extract. Protease SprT has a molecular mass of 31 kDa, a pH optimum of 8.5, and a temperature optimum of 60-65 degrees C. It had good thermostability, and was stable in an alkaline environment. SprT could degrade bovine serum albumin, lysozyme, and gelatin, and its activity was enhanced by many metal ions. The cuticles of nematodes treated by protease SprT obviously crimpled. Purified protease SprT could kill juveniles of M. incognita and inhibit egg hatch, suggesting that it is involved in the nematicidal process of T. pseudokoningii SMF2. The full-length cDNA gene-encoding protease SprT was cloned by rapid amplification of cDNA ends. Sequence analysis showed that SprT is a monodomain subtilase containing 284 amino acid residues. It had higher identities and a closer relation to the nematicidal serine proteases (59-69%) from nematode parasitic fungi than to the serine proteases (activity from Trichoderma. PMID:19702879

  4. Enhancing rice resistance to fungal pathogens by transformation with cell wall degrading enzyme genes from Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

    刘梅; 孙宗修; 朱洁; 徐同; HARMANGaryE; LORITOMatteo

    2004-01-01

    Three genes encoding for fungal cell wall degrading enzymes (CWDEs), ech42, nag70 and gluc78 from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305.2 singly and in all possible combinations and transformed to rice plants. More than 1800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had lesser effect. The expression level of endochitinase but exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the effect of endochitinase on disease resistance when the two genes co-expressed in transgenics. Resistance to Magnaporthe grisea was found in all kinds of regenerated plants including that with single gluc78. A few lines expressing either ech42 or nag70 gene were immune to the disease. Transgenic plants are being tested to further evaluate disease resistance at field level. This is the first report of multiple of expression of genes encoding CWDEs from Trichoderma atroviride that result in resistance to blast and sheath blight in rice.

  5. Genome-wide characterization of ISR induced in Arabidopsis thaliana by Trichoderma hamatum T382 against Botrytis cinerea infection

    Directory of Open Access Journals (Sweden)

    Janick eMathys

    2012-05-01

    Full Text Available In this study, the molecular basis of the induced systemic resistance (ISR in Arabidopsis thaliana by the biocontrol fungus Trichoderma hamatum T382 against the phytopathogen Botrytis cinerea B05-10 was unraveled by microarray analysis both before (ISR-prime and after (ISR-boost additional pathogen inoculation. The observed high numbers of differentially expressed genes allowed us to classify them according to the biological pathways in which they are involved. By focusing on pathways instead of genes, a holistic picture of the mechanisms underlying ISR emerged. In general, a close resemblance is observed between ISR-prime and systemic acquired resistance (SAR, the systemic defense response that is triggered in plants upon pathogen infection leading to increased resistance towards secondary infections. Treatment with Trichoderma hamatum T382 primes the plant (ISR-prime, resulting in an accelerated activation of the defense response against Botrytis cinerea during ISR-boost and a subsequent moderation of the Botrytis cinerea induced defense response (BIDR. Microarray results were confirmed for representative genes by qRT-PCR, by analysis of transgenic plants expressing relevant promoter-GUS constructs and by phenotypic analysis of mutants affected in various defense-related pathways, thereby proving the validity of our approach.

  6. Evaluation of Trichoderma spp. and Clonostachys spp. Strains to Control Fusarium circinatum in Pinus radiata Seedlings Evaluación de Cepas de Trichoderma spp. y Clonostachys spp. para Controlar Fusarium circinatum en Plántulas de Pinus radiata

    Directory of Open Access Journals (Sweden)

    Priscila Moraga-Suazo

    2011-09-01

    Full Text Available The fungus Fusarium circinatum Nirenberg & O’Donnell causes pine pitch canker, an important disease for conifers worldwide. F. circinatum was first detected in Chile in 2001 and to date is present in nurseries and clonal hedges from Libertador General Bernardo O’Higgins Region to Los Rios Region. The purpose of this study was to evaluate the potential of Trichoderma spp. and Clonostachys spp. strains to control F. circinatum in Pinus radiata D. Don seedlings in the absence of other effective control methods. Eighty-one Trichoderma spp. and Clonostachys spp. strains were evaluated through in vitro assays to determine their ability to act as antagonists of F. circinatum and 21 strains were tested for their ability to reduce post-emergence mortality and increase P. radiata survival under greenhouse conditions. During in vitro experiments, 15 strains of Trichoderma inhibited mycelial growth of the pathogen by more than 60% and one strain of Clonostachys showed parasitism of F. circinatum hyphae. Greenhouse experiments showed no control of the disease when the antagonists were added to substrate after the pathogen. However, when the antagonists were added before the pathogen, four strains (Clonostachys UDC-32 and UDC-222 and Trichoderma UDC-23 and UDC-408 reduced post-emergence mortality between 80 and 100%. Among these strains, only Clonostachys UDC-222 significantly increased the survival of P. radiata seedlings. These results showed that Clonostachys UDC-222 has the potential to be used as a biocontrol agent against F. circinatum in the production of P. radiata plants.Fusarium circinatum Nirenberg & O’Donnell es el hongo que causa el cancro resinoso del pino, una enfermedad de importancia mundial en coníferas. En Chile, F. cicirnatum fue detectado por primera vez el año 2001 y a la fecha se encuentra presente en algunos viveros y huertos clonales desde la Región del Libertador General Bernardo O’Higgins hasta la Región de Los R

  7. Genetic regulation of conidiation in Trichoderma hamatun

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  8. Genetic diversity of Trichoderma atroviride strains collected in Poland and identification of loci useful in detection of within-species diversity.

    Science.gov (United States)

    Skoneczny, Dominik; Oskiera, Michał; Szczech, Magdalena; Bartoszewski, Grzegorz

    2015-07-01

    Molecular markers that enable monitoring of fungi in their natural environment or assist in the identification of specific strains would facilitate Trichoderma utilization, particularly as an agricultural biocontrol agent (BCA). In this study, sequence analysis of internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) of the ribosomal RNA (rRNA) gene cluster, a fragment of the translation elongation factor 1-alpha (tef1) gene, and random amplified polymorphic DNA (RAPD) markers were applied to determine the genetic diversity of Trichoderma atroviride strains collected in Poland, and also in order to identify loci and PCR-based molecular markers useful in genetic variation assessment of that fungus. Although tef1 and RAPD analysis showed limited genetic diversity among T. atroviride strains collected in Poland, it was possible to distinguish major groups that clustered most of the analyzed strains. Polymorphic RAPD amplicons were cloned and sequenced, yielding sequences representing 13 T. atroviride loci. Based on these sequences, a set of PCR-based markers specific to T. atroviride was developed and examined. Three cleaved amplified polymorphic sequence (CAPS) markers could assist in distinguishing T. atroviride strains. The genomic regions identified may be useful for further exploration and development of more precise markers suitable for T. atroviride identification and monitoring, especially in environmental samples.

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

    Directory of Open Access Journals (Sweden)

    Seddighe Mohammadi

    2015-08-01

    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.

  10. Development of a qPCR strategy to select bean genes involved in plant defence response and regulated by the Trichoderma velutinum - Rhizoctonia solani interaction

    Directory of Open Access Journals (Sweden)

    Sara Mayo

    2016-08-01

    Full Text Available Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defence response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defence-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from From the 48 bean genes initially analysed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34 or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected.As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defence, which respond to the presence of a biocontrol agent or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defence genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani.

  11. Identification of novel Trichoderma hamatum genes expressed during mycoparasitism

    Institute of Scientific and Technical Information of China (English)

    Margaret Carpenter; Alison Stewart; Hayley Ridgway

    2004-01-01

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

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

    Science.gov (United States)

    Zafra, German; Cortés-Espinosa, Diana V

    2015-12-01

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

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

    Science.gov (United States)

    Zafra, German; Cortés-Espinosa, Diana V

    2015-12-01

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

  14. A paralog of the proteinaceous elicitor sm1 affects colonization of maize roots by Trichoderma virens

    Science.gov (United States)

    The biocontrol agent, Trichoderma virens, has the ability to protect plants from pathogens by eliciting plant defense responses, involvement in mycoparasitism, or secreting antagonistic secondary metabolites. SM1, an elicitor of induced systemic resistance (ISR), was found to have three paralogs wi...

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

    Science.gov (United States)

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

    2015-02-01

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

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

    Science.gov (United States)

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

    2015-02-01

    Trichoderma spp. have been reported to aid in imparting biotic as well as abiotic tolerance to plants. However, there are only few reports unfolding the differential ability of separate species of Trichoderma genera generally exploited for their biocontrol potential in this framework. A study was undertaken to evaluate the biocontrol potential of different Trichoderma species namely T. harzianum, T. asperellum, T. koningiopsis, T. longibrachiatum, and T. aureoviride as identified in the group of indigenous isolates from the agricultural soils of Eastern Uttar Pradesh, India. Their biocontrol potential against three major soilborne phytopathogens, i.e., Sclerotium rolfsii, Sclerotinia sclerotiorum, and Colletotrichum capsici was confirmed by dual culture plate technique. Efficient mycoparasitic ability was further assessed in all the isolates in relation to chitinase, β-1,3 glucanase, pectinase, lipase, amylase, and cellulase production while equally consistent results were obtained for their probable phosphate solubilization and indole acetic acid (IAA) production abilities. The selected isolates were further subjected to test their ability to promote plant growth, to reduce disease incidence and to tolerate biotic stress in terms of lignification pattern against S. rolfsii in chickpea plants. Among the identified Trichoderma species, excellent results were observed for T. harzianum and T. koningiopsis indicating better biocontrol potential of these species in the group and thus exhibiting perspective for their commercial exploitation. PMID:25205162

  17. 77 FR 4903 - Trichoderma

    Science.gov (United States)

    2012-02-01

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

  18. Biological control of banana black Sigatoka disease with Trichoderma

    Directory of Open Access Journals (Sweden)

    Poholl Adan Sagratzki Cavero

    2015-06-01

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

  19. 生防木霉对草坪土壤微生物区系的影响及定殖能力研究%Impact and colonisation ability of Trichoderma biocontrol on lawn soil microflora

    Institute of Scientific and Technical Information of China (English)

    古丽君; 徐秉良; 梁巧兰; 尹婷

    2013-01-01

    为了筛选出适宜培养土壤细菌、真菌、放线菌的定向选择性培养基,对19种培养基进行了比较试验,筛选出的优良培养基分别为牛肉膏蛋白胨培养基+甲霜·霜霉、马丁氏培养基+1 mL1‰孟加拉红、高氏1号培养基+重铬酸钾0.1g.将生防木霉施入草坪土壤中,测定了其对土壤中细菌、放线菌、真菌3种微生物数量的影响.结果表明,施入深绿木霉菌后能够明显引起微生物群落结构发生改变.施入深绿木霉17d后,真菌的数量比对照减少了4 001 cfu/g;土壤中细菌的数量增加了1.27×1010cfu/g;放线菌数量没有明显变化.定殖测定试验结果表明,木霉在施人草坪草土壤1个月后能够很好的定殖.木霉在施入25 d时,相对含量达到最低,但是30 d以后木霉的含量略有所上升,并基本保持稳定.%To screen for a selective medium suitable for cultivation of bacteria,fungi and actinomycetes in the soil,19 types of medium were tested.Beef extract peptone medium +Jia Shuang Shuang Mei,Mading medium+ 1 mL 1% rose bengal and Martin medium+0.1 g potassium dichromate were best for culturing bacteria,fungi and actinomycetes.The numbers of bacteria,fungi and actinomycetes were counted after Trichoderma aureoviride T2 strain was applied to the lawn soil.T.aureoviride cause significant changes in the microbial community structure.Seventeen days after T.aureoviride was applied,the number of fungi decreased by 4 001 cfu/g but the number of bacteria increased by 1.27 × 1010 cfu/g compared with the control,while the number of actinomyces did not change significantly.The ability of T.aureoviride colonisation demonstrated that T.aureoviride could colonise the soil of turf grass within one month.Twenty five days after T.aureoviride was applied,the relative content of T.aureoviride was the lowest,but 5 days later it increased and then remained stable.

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

    Science.gov (United States)

    Al-Hazmi, Ahmad Saad; TariqJaveed, Muhammad

    2016-03-01

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

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

    NARCIS (Netherlands)

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

    2011-01-01

    Background: Mycoparasitism, a lifestyle where one fungus is parasitic on another fungus, has special relevance when the prey is a plant pathogen, providing a strategy for biological control of pests for plant protection. Probably, the most studied biocontrol agents are species of the genus Hypocrea/

  2. Commercial development of Trichoderma species for control of soil-borne vegetable diseases and their integration into standard crop management practices

    Institute of Scientific and Technical Information of China (English)

    Kirstin L McLean; John S Hunt; Alison Stewart

    2004-01-01

    @@ A 10 year research programme at Lincoln University, investigating the use of Trichoderma species for biological control of soil-borne diseases of vegetable crops, has resulted in the development of two commercial products. TrichodryTM 6S and TrichoflowTM 6S based upon Trichoderma hamatum isolate 6SR4, are used to control Sclerotinia lettuce drop disease. The Trichodry 6S product is formulated as a dry flake, which is incorporated into nursery seedling mix and the Trichoflow 6S is a wettable powder which is used as a top-up drench before planting. The treatment stimulates seedling establishment and vigour and protects the developing seedling from Sclerotinia minor infection after transplanting in the field. The second commercial product is TrichopelTM Ali 52. Based upon Trichoderma atroviride isolate C52, which is used to control Sclerotium cepivorum, the causal agent of Allium white rot disease. The product is formulated as a granule and applied into the furrow at planting time. The fungus proliferates in the rhizosphere region and protects the growing seedling from pathogen attack by a combination of nutrient competition, antibiosis and mycoparasitism. The use of Trichopel Ali 52 under low to medium disease pressure in Pukekohe, the main vegetable growing region of New Zealand, gave a three fold cost benefit through yield increases in the 2003-2004 season. Current field development work involves the use of a wettable powder formulation of T. Atroviride distributed via a T-tape irrigation system to target mid-season applications of the product to the onion roots. Both products perform well under low to moderate disease pressure but, when there is high disease pressure, an integrated programme is required to give satisfactory control. Current research is focused on gaining a greater understanding of the biotic and abiotic factors, which influence biocontrol activity under field conditions as a means to enhance integrated control approaches. For example, T

  3. Functional genomic approach to the study of biodiversitywithin Trichoderma

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  4. Genetic improvement of Trichoderma ability to induce systemic resistance

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

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

    Institute of Scientific and Technical Information of China (English)

    Elham Siassi; YANG Qian

    2005-01-01

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

  6. Genome-Wide Characterization of ISR Induced in Arabidopsis thaliana by Trichoderma hamatum T382 Against Botrytis cinerea Infection

    Science.gov (United States)

    Mathys, Janick; De Cremer, Kaat; Timmermans, Pieter; Van Kerckhove, Stefan; Lievens, Bart; Vanhaecke, Mieke; Cammue, Bruno P. A.; De Coninck, Barbara

    2012-01-01

    In this study, the molecular basis of the induced systemic resistance (ISR) in Arabidopsis thaliana by the biocontrol fungus Trichoderma hamatum T382 against the phytopathogen Botrytis cinerea B05-10 was unraveled by microarray analysis both before (ISR-prime) and after (ISR-boost) additional pathogen inoculation. The observed high numbers of differentially expressed genes allowed us to classify them according to the biological pathways in which they are involved. By focusing on pathways instead of genes, a holistic picture of the mechanisms underlying ISR emerged. In general, a close resemblance is observed between ISR-prime and systemic acquired resistance, the systemic defense response that is triggered in plants upon pathogen infection leading to increased resistance toward secondary infections. Treatment with T. hamatum T382 primes the plant (ISR-prime), resulting in an accelerated activation of the defense response against B. cinerea during ISR-boost and a subsequent moderation of the B. cinerea induced defense response. Microarray results were validated for representative genes by qRT-PCR. The involvement of various defense-related pathways was confirmed by phenotypic analysis of mutants affected in these pathways, thereby proving the validity of our approach. Combined with additional anthocyanin analysis data these results all point to the involvement of the phenylpropanoid pathway in T. hamatum T382-induced ISR. PMID:22661981

  7. SSR Markers for Trichoderma virens: Their Evaluation and Application to Identify and Quantify Root-Endophytic Strains

    Directory of Open Access Journals (Sweden)

    Joerg Geistlinger

    2015-11-01

    Full Text Available Using biological fertilizers and pesticides based on beneficial soil microbes in order to reduce mineral fertilizers and chemical pesticides in conventional agriculture is still a matter of debate. In this regard, a European research project seeks to elucidate the role of root-endophytic fungi and to develop molecular tools to trace and quantify these fungi in the rhizosphere and root tissue. To do this, the draft genome sequence of the biocontrol fungus Trichoderma virens (T. virens was screened for simple sequence repeats (SSRs and primers were developed for 12 distinct loci. Primers were evaluated using a global collection of ten isolates where an average of 7.42 alleles per locus was detected. Nei’s standard genetic distance ranged from 0.18 to 0.27 among the isolates, and the grand mean of haploid diversity in AMOVA analysis was 0.693 ± 0.019. Roots of tomato plants were inoculated with different strains and harvested six weeks later. Subsequent PCR amplification identified root-endophytic strains and co-colonization of roots by different strains. Markers were applied to qPCR to quantify T. virens strains in root tissue and to determine their identity using allele-specific melting curve analysis. Thus, the root-endophytic lifestyle of T. virens was confirmed, strains in roots were quantified and simultaneous colonization of roots by different strains was observed.

  8. 生防菌哈茨木霉T4对黄瓜根围土壤细菌群落的影响%Effects of the Introduction of Biocontrol Agent Trichoderma harzianum T4 on the Bacterial Community in Cucumber Rhizosphere

    Institute of Scientific and Technical Information of China (English)

    尹丹韩; 高观朋; 夏飞; 王伟

    2012-01-01

    [目的]评价生防菌哈茨木霉(Trichoderma harzianum)T4对根围土壤细菌群落的影响,为生防菌田间应用的生态安全提供依据.[方法]结合DGGE技术与T-RFLP方法,综合考察引入生防菌哈茨木霉T4后黄瓜根围土壤细菌群落变化,[结果]在引入生防菌的14-56 d内(黄瓜整个生育期),木霉对根围土壤细菌群落可产生显著影响,并持续一段的时间,随后逐渐减弱.表现为对蓝细菌、β-变形细菌、葡萄球菌、伯克霍尔德菌、沙雷氏菌、酸杆菌和一些不可培养细菌的有效抑制作用,对芽孢杆菌、土壤杆菌、芽单胞菌的明显促进作用.但到70d时,即收获后2周,检测不到木霉对细菌群落影响.[结论]DGGE和T-RFLP序列分析技术相结合研究生防菌对土壤细菌群落的影响,既能全面描述土壤细菌群落结构变化,还可以初步定性描述变化细菌的种类信息.生防菌哈茨木霉T4的引入对土壤细菌群落产生短期影响,不会长期对土壤细菌群落稳定构成威胁.%[ Objective ] The objective of this study is to analysis the impact of T. Harzianum T4 introduction on the indigenous bacterial community of rhizosphere soil, and to provide objective biosafety evaluations for the field application of biocontrol agents.[ Method 1 Comprised molecular tools DGGE and T-RFLP were used in this study to assess the modifications of the bacterial community in cucumber rhizosphere after inoculation with T. Harzianum T4. [Result] Introduction of T. Harzianum T4 in the soil resulted in significant variations of the bacterial community structures from 14 to 56 days post-inoculation. During this period the impact of T4 was sustainable and weakened gradually. T. Harzianum T4 could restrain effectively Cyanobacterium, Beta-proteobacterium, Staphylococcus, Burkholderia, Serratia, Acidobacterium and some kinds of soil uncultured bacterium, while it improved the growth of Bacillus, Agrobacterium and Gemmatimonadetes observably

  9. Enhancement of biocontrol efficacy against Botrytis cinerea through the manipulation of nitrogen fertilization of tomato plants

    OpenAIRE

    Abro, Manzoor Ali; Lecompte, François; Bardin, Marc; Duffaud, Magali

    2013-01-01

    Although nitrogen fertilization is known to affect plant susceptibility to certain pathogens, little is known on its possible effect on the efficacy of biological control. In the present study we examined the effect of five levels of NO3- nutrition on the efficacy of two biocontrol agents (Trichoderma harzianum and Microdochium dimerum) to protect pruning wounds of tomato against Botrytis cinerea. Plants were grown for two months in a greenhouse with a soil-less drip-irrigationsystem. Differe...

  10. Trichoderma species from China

    Institute of Scientific and Technical Information of China (English)

    ZHANG Chu-long; XU Tong

    2004-01-01

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

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

    DEFF Research Database (Denmark)

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

    2015-01-01

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

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

    Science.gov (United States)

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

    2015-08-01

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

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

    Directory of Open Access Journals (Sweden)

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

    2012-06-01

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

  14. AM真菌摩西球囊霉对2种花生叶斑病的生防及促生作用%Biocontrol Effects of AM Fungus Glomus mosseae on Two Peanut Leaf Spots and its Growth-promoting Effects on Peanut

    Institute of Scientific and Technical Information of China (English)

    鄢洪海; 张茹琴; 安佰国

    2011-01-01

    为了研究丛枝菌根(Arbuscular mycorrhiza,AM)真菌对2种花生叶部病害黑斑病(Cercosporium personatuma)和网斑病(Phoma arachidicola)的生物防治作用以及对花生生长的促进作用,为开发绿色菌肥奠定基础,在盆栽条件下对花生品种‘花育22号’接种AM真菌摩西球囊霉(Glomus mosseae),在自然发病情况下,测定摩西球囊霉接种对花生黑斑病和网斑病的防治效果以及对花生株高、分枝、果荚数、果荚干重、地上部干重、地下部干重等生长指标的影响.结果表明,接种AM真菌摩西球囊霉使花生株高、分枝、荚果数、荚果干重、地上部干重等生长指标显著地增加;对花生黑斑病防治效果为5.4%,对网斑病防治效果为10.2%.AM通过促进花生生长而补偿了病害造成的损失是其抗病机制之一.表明分离的摩西球囊霉是一株具有促进花生生长,对2种叶部病害具有一定生防作用的AM菌株.%In order to study biocontrol effects of arbuscular mycorrhizal (AM) on two leaf spots, peanut (Arachis hypogaea L.) black spot (Cercosporium personatuma) and peanut net blotch {Phoma arachidicola), the growth-promoting effects of AM fungus on peanut, and as a result, laying a foundation for the development of green manure, peanut cultivar ' Huayu 22' inoculated with AM fungus Glomus mosseae in a pot experiment, under natural infection conditions, biocontrol effects of G. Mosseae on peanut black spot and peanut net blotch, growth indices including plant height, the numbers of branches or pods, dry mass of pods, aboveground or underground dry mass of peanut were investigated. The results indicated that, inoculation peanut with G. Mosseae significantly improved plant height, the numbers of branches or pods, dry mass of pods, aboveground dry mass of peanut, and induced systemic acquired resistance to the two leaf spots. Control effect on peanut black spot was 5.4%, whereas on net peanut blotch was 10.2%. One of

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

    Institute of Scientific and Technical Information of China (English)

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

    2008-01-01

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

  16. Competition in artifical plant growth media by Trichoderma spp

    DEFF Research Database (Denmark)

    Sarocco, Sabrina; Lübeck, Mette; Vannacci, Giovanni

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

  17. Identifying beneficial qualities of Trichoderma parareesei for plants.

    Science.gov (United States)

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

    2014-03-01

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

  18. Snuffelen aan Trichoderma

    NARCIS (Netherlands)

    Baars, J.J.P.

    2011-01-01

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

  19. Phylogeny and biodiversity of Trichoderma and Hypocrea and its implications on taxonomy

    Institute of Scientific and Technical Information of China (English)

    Christian P Kubicek

    2004-01-01

    @@ Safe strain identification and species recognition is an important issue for Trichoderma and Hypocrea,because members of the genus are economically important producers of industrial enzymes and antibiotics, have application as biocontrol agents against plant pathogens, whereas some have become known as opportunistic pathogens of immunocompromised mammals and humans. However, classical approaches based on the use of morphological and phenetic characters have been difficult to apply, due to the plasticity of characters and the discordance of morphological and molecular evolution.

  20. Novel plant bio-protectants based on Trichoderma spp. strains with superior characteristics

    OpenAIRE

    Abadi, Khalid M.

    2008-01-01

    Global warming caused by the greenhouse effect will alter the geographical distribution of host and pathogen populations, thus affecting the natural physiology of their interaction and reducing the efficacy of chemical and biological control strategies presently used in agriculture. In perspective to this scenario, new management practices will be required. The main task of this thesis was to isolate and characterize new biocontrol agents of the genus Trichoderma originating from Libya, where...

  1. Development of a qPCR Strategy to Select Bean Genes Involved in Plant Defense Response and Regulated by the Trichoderma velutinum - Rhizoctonia solani Interaction.

    Science.gov (United States)

    Mayo, Sara; Cominelli, Eleonora; Sparvoli, Francesca; González-López, Oscar; Rodríguez-González, Alvaro; Gutiérrez, Santiago; Casquero, Pedro A

    2016-01-01

    Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani. PMID:27540382

  2. Development of a qPCR Strategy to Select Bean Genes Involved in Plant Defense Response and Regulated by the Trichoderma velutinum - Rhizoctonia solani Interaction.

    Science.gov (United States)

    Mayo, Sara; Cominelli, Eleonora; Sparvoli, Francesca; González-López, Oscar; Rodríguez-González, Alvaro; Gutiérrez, Santiago; Casquero, Pedro A

    2016-01-01

    Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani.

  3. Development of a qPCR Strategy to Select Bean Genes Involved in Plant Defense Response and Regulated by the Trichoderma velutinum – Rhizoctonia solani Interaction

    Science.gov (United States)

    Mayo, Sara; Cominelli, Eleonora; Sparvoli, Francesca; González-López, Oscar; Rodríguez-González, Alvaro; Gutiérrez, Santiago; Casquero, Pedro A.

    2016-01-01

    Bean production is affected by a wide diversity of fungal pathogens, among them Rhizoctonia solani is one of the most important. A strategy to control bean infectious diseases, mainly those caused by fungi, is based on the use of biocontrol agents (BCAs) that can reduce the negative effects of plant pathogens and also can promote positive responses in the plant. Trichoderma is a fungal genus that is able to induce the expression of genes involved in plant defense response and also to promote plant growth, root development and nutrient uptake. In this article, a strategy that combines in silico analysis and real time PCR to detect additional bean defense-related genes, regulated by the presence of Trichoderma velutinum and/or R. solani has been applied. Based in this strategy, from the 48 bean genes initially analyzed, 14 were selected, and only WRKY33, CH5b and hGS showed an up-regulatory response in the presence of T. velutinum. The other genes were or not affected (OSM34) or down-regulated by the presence of this fungus. R. solani infection resulted in a down-regulation of most of the genes analyzed, except PR1, OSM34 and CNGC2 that were not affected, and the presence of both, T. velutinum and R. solani, up-regulates hGS and down-regulates all the other genes analyzed, except CH5b which was not significantly affected. As conclusion, the strategy described in the present work has been shown to be effective to detect genes involved in plant defense, which respond to the presence of a BCA or to a pathogen and also to the presence of both. The selected genes show significant homology with previously described plant defense genes and they are expressed in bean leaves of plants treated with T. velutinum and/or infected with R. solani. PMID:27540382

  4. Characterisation of a Trichoderma hamatum monooxygenase gene involved in antagonistic activity against fungal plant pathogens.

    Science.gov (United States)

    Carpenter, Margaret A; Ridgway, Hayley J; Stringer, Alison M; Hay, Amanda J; Stewart, Alison

    2008-04-01

    A monooxygenase gene was isolated from a biocontrol strain of Trichoderma hamatum and its role in biocontrol was investigated. The gene had homologues in other fungal genomes, but was not closely related to any fully characterised gene. The T. hamatum monooxygenase gene was expressed specifically in response to the plant pathogens Sclerotinia sclerotiorum, Sclerotinia minor and Sclerotium cepivorum, but not in response to Botrytis cinerea or T. hamatum. Expression of the gene did not occur until contact had been made between the two fungal species. Homologues in T. atroviride and T. virens showed similar expression patterns. Expression of the gene in response to S. sclerotiorum was influenced by pH, with a peak of expression at pH 4, and was subject to nitrogen catabolite repression. Disruption of the monooxygenase gene did not affect the growth or morphology of T. hamatum, but caused a decrease in its ability to inhibit the growth and sclerotial production of S. sclerotiorum. The monooxygenase gene had a role in the antagonistic activity of Trichoderma species against specific fungal plant pathogens and is therefore a potentially important factor in biocontrol by Trichoderma species. PMID:18231791

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

    Science.gov (United States)

    Zavala-González, Ernesto A; Lopez-Moya, Federico; Aranda-Martinez, Almudena; Cruz-Valerio, Mayra; Lopez-Llorca, Luis Vicente; Ramírez-Lepe, Mario

    2016-07-01

    The effect of chitosan on growth of Trichoderma spp., a cosmopolitan genus widely exploited for their biocontrol properties was evaluated. Based on genotypic (ITS of 18S rDNA) characters, four isolates of Trichoderma were identified as T. pseudokoningii FLM16, T. citrinoviride FLM17, T. harzianum EZG47, and T. koningiopsis VSL185. Chitosan reduces radial growth of Trichoderma isolates in concentration-wise manner. T. koningiopsis VSL185 was the most chitosan tolerant isolate in all culture media amended with chitosan (0.5-2.0 mg ml(-1) ). Minimal Inhibitory Concentration (MIC) and Minimal Fungicidal Concentration (MFC) were determined showing that T. koningiopsis VSL185 displays higher chitosan tolerance with MIC value >2000 μg ml(-1) while for other Trichoderma isolates MIC values were around 10 μg ml(-1) . Finally, free fatty acid composition reveals that T. koningiopsis VSL185, chitosan tolerant isolate, displays lower linolenic acid (C18:3) content than chitosan sensitive Trichoderma isolates. Our findings suggest that low membrane fluidity is associated with chitosan tolerance in Trichoderma spp. PMID:27213758

  6. Fun Microbiology: How To Measure Growth of a Fungus.

    Science.gov (United States)

    Mitchell, James K.; And Others

    1997-01-01

    Describes an experiment to demonstrate a simple method for measuring fungus growth by monitoring the effect of temperature on the growth of Trichoderma viride. Among the advantages that this experimental model provides is introducing students to the importance of using the computer as a scientific tool for analyzing and presenting data. (AIM)

  7. Are mycoparasitism and chitinase production species or isolate dependent in Trichoderma ?

    Institute of Scientific and Technical Information of China (English)

    Szakacs G; Nagy V; Kovacs K

    2004-01-01

    @@ The relationship between taxonomic status of Trichoderma spp., chitinase production in solid substrate fermentation (SSF) on four media and mycoparasitism in dual culture (confrontation assay)against four plant pathogenic fungi was studied. Seventy five Trichoderma isolates belonging to 35species have been screened. The plant pathogenic fungi used in confrontation assay were Botrytis cinerea , Fusarium oxysporum f. sp. dianthi , Rhizoctonia solani and Sclerotinia sclerotiorum . The SSF media contained wheat bran, crude chitin (from crab shells, SIGMA) and salt solutions. The best performing isolates in mycoparasitism tests were Trichoderma flavofuscum, T. harzianum, T.inhamatum, T. koningii and T. strigosum. Some isolates exhibiting good mycoparasitism produced chitinase in SSF only at low or medium level. In contrary there were isolates with excellent extracellular chitinase production but their biocontrol potential did not belong to the leading group.Statistical methods have been used to evaluate the data.

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

    Science.gov (United States)

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

    2012-12-01

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

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

    Science.gov (United States)

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

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

    DEFF Research Database (Denmark)

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

    2014-01-01

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

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

    NARCIS (Netherlands)

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

    2016-01-01

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

  12. Trichoderma L-Lysine-α-Oxidase Producer Strain Culture Fluid Inhibits Impatiens Necrotic Spot Virus.

    Science.gov (United States)

    Smirnova, I P; Shneider, Yu A; Karimova, E V

    2016-01-01

    A method for PCR diagnosis of impatiens necrotic spot virus is developed. Concentrated culture fluid with active L-lysine-α-oxidase (0.54 U/ml) from Trichoderma harzianum Rifai fungus producer strain F-180 inhibits vitally hazardous impatiens necrotic spot phytovirus. PMID:26742733

  13. Trichoderma volatiles effecting Arabidopsis

    DEFF Research Database (Denmark)

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

    2015-01-01

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

  14. Extracellular proteome of Trichoderma harzianum to identify proteins with biotechnological value

    Institute of Scientific and Technical Information of China (English)

    Ambrosino P; Lorito M; Scala V; Marra R; Vinale F; Soriente I; Ferraioli S; Carbone V; Ruocco M; Woo S L

    2004-01-01

    @@ Trichoderma harzianum strain T22 parasitizes and controls many phytopatogenic fungi and is applied commercially as biological control agent. The production of hydrolitic enzymes appears to be a key factor in the parasitic process. We tested the endo-esochitinolitic and glucanolitic activities of culture filtrates of T22 grown under carbon and nitrogen starvation or in presence of biomass or cell walls of the phytopathogenic fungi Botrytis cinerea , Rhizoctonia solani and Pythium ultimum. The highest level of enzimatic activities was found in culture where the mycoparasite interacted with a phytopathogenic fungus. Therefore we used a proteomic approach to investigate changes in the complex mixture of extracellular proteins secreted by T. harzianum strain T22 in order to identify proteins of potential biotechnology value for commercial and industrial use. Proteome technology has greatly enhanced our ability to conduct functional genomics studies. Nevertheless only a few studies have been published so far on the fungal extracellular proteome. Sample preparation remains the most critical step in analyses based on two-dimensional gel electrophoresis (2-DE), and it requires to be optimized for each specific application. In this study, our first aim was to set up the extraction protocol of the extracellular proteins secreted by T . harzianum strain T22 when it was grown in vitro . The secreted proteins were analysed by two-dimensional electrophoresis (2-DE) and substantial changes in the extracellular proteome of the mycoparasite have been observed. Comparing the 2D maps of the fungus grown in minimal medium with glycerol as carbon source (used as control condition) with those obtained in inducing conditions, a lot of novel proteins appeared. The higher number of novel and upregulated spots was obtained in the presence of Rhizoctonia solani biomass. Other spots were specifically up-regulated by the interaction with different plant pathogens. Differentially expressed

  15. 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.

  16. Effects of nitrogen availability and spore concentration on the biocontrol activity of Ascochyta caulina in common lambsquarters (Chenopodium album)

    NARCIS (Netherlands)

    Ghorbani, R.; Scheepens, P.C.; Zweerde, van der W.; Leifert, C.; McDonald, A.J.S.; Seel, W.

    2002-01-01

    Common lambsquarters is an annual weed of many important crops. Ascochyta caulina is a plant pathogenic fungus that causes necrotic lesions on the leaves and stems of common lambsquarters. The objective of the present study was to estimate the effect of plant N supply on the biocontrol activity of A

  17. Xylanase production by Trichoderma harzianum E58

    Energy Technology Data Exchange (ETDEWEB)

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

    1989-12-01

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

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

    Science.gov (United States)

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

    2016-01-01

    application of Trichoderma biocontrol strains. PMID:27482910

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

    Science.gov (United States)

    Chen, Jing; Mao, Li-Juan; Feng, Xiao-Xiao; Zhang, Chu-Long; Lin, Fu-Cheng

    2016-01-01

    application of Trichoderma biocontrol strains. PMID:27482910

  20. 与油菜菌核病菌在土壤中存活有关的木霉及其生物防治潜在能力%Trichoderma Species Associated with Survival of Sclerotia of Sclerotinia Sclerotiorum of Rape in Soil and Their Potential in Biocontrol

    Institute of Scientific and Technical Information of China (English)

    文成敬; 牛应泽; 王迎春; 马炳田

    2001-01-01

    Fifty two Trichoderma isolates, which are associated with survival of sclerotia of Sclerotinia sclerotiorum, are obtained from the different areas of province Sichuan. They are 25 T.aureoviride (48.1% of the total), 17 T.hamatum (32.7%), 7 T.harzianum (13.3%), 2 T.koningii (4.0%) and 1 T.pseudokoningii (2.0%). The dual culture of Trichoderma spp. And S.sclerotiorum on a cellophane strip indicate that Trichoderma hypha parallel and touch to those of S.sclerotiorum, sometimes coiling around the later, two days after the dual culture is made. The hypha adjacent to Trichoderma is to be broken and their cytoplasms are collapsed. The results of dual culture made directly on PDA plates, however, indicate when the colonies contact each other, those of S.sclerotiorum stop extending. In most cases, an yellowish-brown antagonistic zone is produced in the side of colony of S.sclerotiorum. In this zone, sclerotinia hypha becomes brown, swollen and broken. Furthermore, 84% of Trichoderma isolates can overran the colonies of S.sclerotiorum and make sclerotia formed by S.sclerotiorum decrease. 5 tested isolates of Trichoderma spp. Can suppress the production of sclerotia completely. The non-volatile substances of Trichoderma have the obvious inhibitions to the hyphal growth, number and size of sclerotia of S.sclerotiorum. But the volatiles have only an inhibition to the size of sclerotia. When the cultures of Trichoderma isolates which possess the stronger antagonistic ability to S.sclerotiorum are introduced into the soil, the sclerotia burried in this soil is to decay six months later, the carpogenic germination of sclerotia that has not decayed reduces sharply.%将油菜菌核病菌(S.sclerotiorum)之菌核作为诱饵埋入油菜土壤中,二个月后从菌核表面和内部分离到52株木霉,其中黄绿木霉(T.aureoviride)25株,占总数48.1%;钩状木霉(T.hamatum)17株,32.7%;哈茨木霉(T.harzianum)7株,13.3%;康氏木霉(T.koningii)2株,4.0%,

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

    Directory of Open Access Journals (Sweden)

    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.

  2. 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

    characterization of Trichoderma isolates parasitic to Pleurotus indicated that they are different from both T. aggressivum forms parasitic to Agaricus, and the majority of the isolates probably belong to the species T. harzianum. In vitro confrontation plates were performed with 26 isolates of aggressive Trichoderma obtained from compost, three Trichoderma isolates used in biological control and 12 varieties of Pleurotus. No inhibitory effect was observed between any of the Trichoderma isolates with Pleurotus, although some growth inhibition was caused by the biocontrol isolates of Trichoderma on some of the aggressive isolates. The temperature optimum for Pleurotus growth was at 28 ℃, whereas Trichoderma grew well at a wider range (20-28℃), and exceeded the growth rate of Pleurotus by three times at 25℃.The pH optimum for the growth of Pleurotus was alkaline (pH 8-9) whereas Trichoderma preferred acidic-neutral pH (5-7) . Various commercial fungicides used in agriculture (procloraz, thiabendazole,dichloran, benomyl, propiconazole, thiofanatomethyl) were tested against the aggressive and biocontrol isolates of Trichoderma, as well as the different varieties of Pleurotus to determine dose response curves and combinations that would inhibit spore germination, mycelial growth and subsequent sporulation. Both procloraz and thiabendazole, which are pesticides allowed in edible mushroom production, were found to control the growth of the aggressive Trichoderma isolates and did not have a negative effect on Pleurotus.

  3. Gate crashing arbuscular mycorrhizas: in vivo imaging shows the extensive colonization of both symbionts by Trichoderma atroviride.

    Science.gov (United States)

    Lace, Beatrice; Genre, Andrea; Woo, Sheridan; Faccio, Antonella; Lorito, Matteo; Bonfante, Paola

    2015-02-01

    Plant growth-promoting fungi include strains of Trichoderma species that are used in biocontrol, and arbuscular mycorrhizal (AM) fungi, that enhance plant nutrition and stress resistance. The concurrent interaction of plants with these two groups of fungi affects crop performance but has only been occasionally studied so far. Using in vivo imaging of green fluorescent protein-tagged lines, we investigated the cellular interactions occurring between Trichoderma atroviride PKI1, Medicago truncatula and two Gigaspora species under in vitro culture conditions. Trichoderma atroviride did not activate symbiotic-like responses in the plant cells, such as nuclear calcium spiking or cytoplasmic aggregations at hyphal contact sites. Furthermore, T. atroviride parasitized G. gigantea and G. margarita hyphae through localized wall breaking and degradation - although this was not associated with significant chitin lysis nor the upregulation of two major chitinase genes. Trichoderma atroviride colonized broad areas of the root epidermis, in association with localized cell death. The infection of both symbionts was also observed when T. atroviride was applied to a pre-established AM symbiosis. We conclude that - although this triple interaction is known to improve plant growth in agricultural environments - in vitro culture demonstrate a particularly aggressive mycoparasitic and plant-colonizing behaviour of a biocontrol strain of Trichoderma.

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

    Science.gov (United States)

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

    2013-08-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  6. MORPHOLOGICAL CHARACTERIZATION AND MASS PRODUCTION OF NEMATOPHAGOUS FUNGUS NEMATOCTONUS ROBUSTUS

    OpenAIRE

    Ramesh babu S,; Anirudh; R. K. Singh

    2014-01-01

    The plant parasitic nematodes infect the root tissues of the plant causing root galls that lead to reduced water and mineral uptake in the plant root system. Nematophagous fungus are used as biocontrol for the nematodes. Among those Nematoctonus are one of the species used as bioagent. Nematoctonus species produces an extensive mycelium and capture many nematodes with hour glass shaped adhesive knobs on the hyphae. Nematodes become attached to these adhesive knobs and the cuticle of nematode ...

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

    Institute of Scientific and Technical Information of China (English)

    DRUZHININA Irina; KUBICEK Christian P.

    2005-01-01

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

  8. Antagonistic potentiality of Trichoderma harzianum towards seed-borne fungal pathogens of winter wheat cv. Protiva in vitro and in vivo.

    Science.gov (United States)

    Hasan, M M; Rahman, S M E; Kim, Gwang-Hee; Abdallah, Elgorban; Oh, Deog-Hwan

    2012-05-01

    The antagonistic effect of Trichoderma harzianum on a range of seed-borne fungal pathogens of wheat (viz. Fusarium graminearum, Bipolaris sorokiniana, Aspergillus spp., and Penicillium spp.) was assessed. The potential of T. harzianum as a biocontrol agent was tested in vitro and under field conditions. Coculture of the pathogens and Trichoderma under laboratory conditions clearly showed dominance of T. harzianum. Under natural conditions, biocontrol effects were also obtained against the test fungi. One month after sowing, field emergence (plant stand) was increased by 15.93% over that obtained with the control treatment, and seedling infection was reduced significantly. Leaf blight severity was decreased from 22 to 11 at the heading stage, 35 to 31 at the flowering stage, and 86 to 74 at the grain filling stage. At harvest, the number of tillers per plant was increased by 50%, the yield was increased by 31.58%, and the 1,000-seed weight was increased by 21%.

  9. Bacteriophage biocontrol of foodborne pathogens.

    Science.gov (United States)

    Kazi, Mustafa; Annapure, Uday S

    2016-03-01

    Bacteriophages are viruses that only infect bacterial cells. Phages are categorized based on the type of their life cycle, the lytic cycle cause lysis of the bacterium with the release of multiple phage particles where as in lysogenic phase the phage DNA is incorporated into the bacterial genome. Lysogeny does not result in lysis of the host. Lytic phages have several potential applications in the food industry as biocontrol agents, biopreservatives and as tools for detecting pathogens. They have also been proposed as alternatives to antibiotics in animal health. Two unique features of phage relevant for food safety are that they are harmless to mammalian cells and high host specificity, keeping the natural microbiota undisturbed. However, the recent approval of bacteriophages as food additives has opened the discussion about 'edible viruses'. This article reviews in detail the application of phages for the control of foodborne pathogens in a process known as "biocontrol". PMID:27570260

  10. Rice transformation with cell wall degrading enzyme genes from Trichoderma atroviride and its effect on plant growth and resistance to fungal pathogens

    Institute of Scientific and Technical Information of China (English)

    Liu Mei; Sun Zong-Xiu; Zhu Jie; Xu Tong; Gary E Harman; Matteo Lorito; Sheri Woo

    2004-01-01

    @@ Three genes encoding for fungal cell wall degrading enzymes (CWDE), ech42, nag70 and gluc78from the biocontrol fungus Trichoderma atroviride were inserted into the binary vector pCAMBIA1305. 2 singly and in all possible combinations. The coding sequences were placed downstream of the rice actin promoter and all vectors were used to transform rice plants. A total of more than 1,800 independently regenerated plantlets in seven different populations (for each of the three genes and each of the four gene combinations) were obtained. Expression in plant was obtained for all the fungal genes used singly or in combinations. The ech42 gene encoding for an endochitinase increased resistance to sheath blight caused by Rhizoctonia solani, while the exochitinase-encoding gene, nag70, had a lesser effect. The expression level of endochitinase but not of the exochitinase was correlated with disease resistance. Nevertheless, exochitinase enhanced the positive effect of endochitinase on disease resistance when two genes were co-expressed in transgenic rice. Improved resistance to Magnaporthe grisea was found in all types of regenerated plants, including those with the gluc78 gene alone, while a few lines expressing either ech42 or nag70 appeared to be immune to this pathogen. Transgenic plants expressing the gluc78 gene alone were stunted and only few of them survived, even though they showed resistance to M. grisea. However, combination with either one of the two other genes ( ech42, nag70 ) as included in the same T-DNA region, reduced the negative effect of gluc78 on plant growth. This is the first report of single or multiple of expression of transgens encoding CWDEs that results in resistance to blast and sheath blight in rice.

  11. Production of a newly isolated Paenibacillus polymyxa biocontrol agent using monosodium glutamate wastewater and potato wastewater.

    Science.gov (United States)

    Gu, Likun; Bai, Zhihui; Jin, Bo; Zhang, Jianyun; Li, Wenying; Zhuang, Guoqiang; Zhang, Hongxun

    2010-01-01

    A phyllosphere bacterial strain EBL-06 was isolated from wheat leaves. The morphology, cultural characteristics, phospholipid fatty acids, physiological and antagonistic fungus activities of this strain were investigated. A phylogenetic tree was constructed by comparing with the published 16S rDNA sequences of the relevant bacteria. The results showed that the isolate EBL-06 was a strain of Paenibacillus polymyxa; this strain performed a high level of antagonistic fungus activity toward a broad spectrum of phytopathogens, such as Botrytis cinerea, Cladosporium cucumerinum, Fusarium spp. The isolate EBL-06 can grow well using monosodium glutamate wastewater (MGW) and potato wastewater (PW) as culture medium. The maximum yield of 6.5 x 10(9) CFU/mL of the isolate EBL-06 anti-fungus biocontrol agent was reached in 15 hr cultivation at 28 degrees C, pH 6.0-7.5 using the mixture of MGW and PW (1:9). PMID:21174972

  12. Copper tolerance of Trichoderma species

    Directory of Open Access Journals (Sweden)

    Jovičić-Petrović Jelena

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Goldman Maria Helena S.

    1998-01-01

    Full Text Available The mycoparasite Trichoderma harzianum produces an alkaline proteinase that may be specifically involved in mycoparasitism. We have constructed transformant strains of this fungus that overexpress this alkaline proteinase. Some of the transformants were assessed for alkaline proteinase activity, and those with higher activity than the wild type were selected for further studies. One of these transformant strains produced an elevated and constitutive pbr1 mRNA level during mycoparasitic interactions with Rhizoctonia solani.

  14. Optimum Concentrations of Trichoderma longibrachiatum and Cadusafos for Controlling Meloidogyne javanica on Zucchini Plants

    OpenAIRE

    Sokhandani, Zahra; Moosavi, Mohammad Reza; Basirnia, Tahereh

    2016-01-01

    A factorial experiment was established in a completely randomized design to verify the effect of different inoculum levels of an Iranian isolate of Trichoderma longibrachiatum separately and in combination with various concentrations of cadusafos against Meloidogyne javanica in the greenhouse. Zucchini seeds were soaked for 12 hr in five densities (0, 105, 106, 107, and 108 spores/ml suspension) of the fungus prior to planting in pots containing four concentrations of cadusafos (0, 0.5, 1, an...

  15. Transformation of Cyclaneusma minus with Green Fluorescent Protein (GFP to Enable Screening of Fungi for Biocontrol Activity

    Directory of Open Access Journals (Sweden)

    Alison Stewart

    2012-02-01

    Full Text Available Cyclaneusma needle-cast has a major impact on the New Zealand forest industry. The causal agent, Cyclaneusma minus, causes most severe damage to 11–20 year-old trees and currently there are no economically viable procedures for control of the disease in New Zealand. Here we present a method for genetic transformation of C. minus using protoplasts generated by incubation with Glucanex™ enzyme. C. minus was transformed with a gene encoding green fluorescent protein (GFP and expression was stable after successive sub-culturing of the strain in the absence of selective pressure. Expression of the gfp gene allowed us to utilize an in vitro GFP-based screening method to identify strains of Trichoderma with potential for biocontrol of this disease. The strain that showed the most promise as a potential biocontrol candidate exhibited a low level of inhibition by uncharacterized metabolite(s that C. minus secretes into the medium, and consistently caused a loss of GFP expression from the GFP-labeled C. minus strain. The interaction between C. minus and the biocontrol strain, in the interaction zone where GFP expression was lost, was determined to be fungicidal. The utility of such biocontrol strains is discussed. This study represents the first genetic manipulation of C. minus and will pave the way for further studies of the life cycle and infection biology of this organism.

  16. Biocontrol potential of endophytes harbored in Radula marginata (liverwort) from the New Zealand ecosystem.

    Science.gov (United States)

    Kusari, Parijat; Kusari, Souvik; Spiteller, Michael; Kayser, Oliver

    2014-10-01

    Radula marginata and Cannabis sativa L. are two phylogenetically unrelated plant species containing structurally similar secondary metabolites like cannabinoids. The major objective of our work was the isolation, identification, biocontrol efficacies, biofilm forming potential and anti-biofilm ability of endophytic microbial community of the liverwort R. marginata, as compared to bacterial endophytic isolates harbored in C. sativa plants. A total of 15 endophytic fungal and 4 endophytic bacterial isolates were identified, including the presence of a bacterial endosymbiont within an endophytic fungal isolate. The endosymbiont was visible only when the fungus containing it was challenged with two phytopathogens Botrytis cinerea and Trichothecium roseum, highlighting a tripartite microbe-microbe interaction and biocontrol potency of endophytes under biotic stress. We also observed sixteen types of endophytic fungal-pathogen and twelve types of endophytic bacterial-pathogen interactions coupled to varying degree of growth inhibitions of either the pathogen or endophyte or both. This showed the magnitude of biocontrol efficacies of endophytes in aiding plant fitness benefits under different media (environmental) conditions. Additionally, it was ecologically noteworthy to find the presence of similar endophytic bacterial genera in both Radula and Cannabis plants, which exhibited similar functional traits like biofilm formation and general anti-biofilm activities. Thus far, our work underlines the biocontrol potency and defensive functional traits (in terms of antagonism and biofilm formation) of endophytes harbored in liverwort R. marginata as compared to the endophytic community of phylogenetically unrelated but phytochemically similar plant C. sativa. PMID:25100187

  17. Preliminary in vitro insights into the use of natural fungal pathogens of leaf-cutting ants as biocontrol agents.

    Science.gov (United States)

    Folgarait, Patricia; Gorosito, Norma; Poulsen, Michael; Currie, Cameron R

    2011-09-01

    Leaf-cutting ants are one of the main herbivores of the Neotropics, where they represent an important agricultural pest. These ants are particularly difficult to control because of the complex network of microbial symbionts. Leaf-cutting ants have traditionally been controlled through pesticide application, but there is a need for alternative, more environmentally friendly, control methods such as biological control. Potential promising biocontrol candidates include the microfungi Escovopsis spp. (anamorphic Hypocreales), which are specialized pathogens of the fungi the ants cultivate for food. These pathogens are suppressed through ant behaviors and ant-associated antibiotic-producing Actinobacteria. In order to be an effective biocontrol agent, Escovopsis has to overcome these defenses. Here, we evaluate, using microbial in vitro assays, whether defenses in the ant-cultivated fungus strain (Leucoagaricus sp.) and Actinobacteria from the ant pest Acromyrmex lundii have the potential to limit the use of Escovopsis in biocontrol. We also explore, for the first time, possible synergistic biocontrol between Escovopsis and the entomopathogenic fungus Lecanicillium lecanii. All strains of Escovopsis proved to overgrow A. lundii cultivar in less than 7 days, with the Escovopsis strain isolated from a different leaf-cutting ant species being the most efficient. Escovopsis challenged with a Streptomyces strain isolated from A. lundii did not exhibit significant growth inhibition. Both results are encouraging for the use of Escovopsis as a biocontrol agent. Although we found that L. lecanii can suppress the growth of the cultivar, it also had a negative impact on Escovopsis, making the success of simultaneous use of these two fungi for biocontrol of A. lundii questionable. PMID:21739253

  18. Studies on strains of Trichoderma spp. from high altitude of Garhwal Himalayan region.

    Science.gov (United States)

    Joshi, B B; Vishwakarma, M P; Bahukhandi, D; Bhatt, R P

    2012-09-01

    Two species of Trichoderma i.e. T. harzianum and T. viride have been isolated from the soil samples collected from the higher altitude (2000-3500 m) of Garhwal Himalayan region in India. The two species were grown in Petri plates on TSM agar media and it was also observed that the optimum temperature and pH for Trichoderma growth was 30 degrees C and 5.5 respectively. When incubated on TSM agar medium at 4 degrees C, the fungus grew normally with heavy induced sporulation within three weeks of incubation. Induction of sporulation on exposure to low temperature appeared to be strategies for survival of these species in extreme cold environment temperature 4 to 5 degrees C. Antifungal activities of the two species of Trichoderma were demonstrated with phytopathogenic fungi in dual cultures. The antifungal metabolites produced by Trichoderma spp., diffusible as well as volatile, caused abnormalities in pathogenic fungi. Plant growth promotion of Trichoderma spp. was also shown through plant analysis in greenhouse.

  19. Arabidopsis thaliana polyamine content is modified by the interaction with different Trichoderma species.

    Science.gov (United States)

    Salazar-Badillo, Fatima Berenice; Sánchez-Rangel, Diana; Becerra-Flora, Alicia; López-Gómez, Miguel; Nieto-Jacobo, Fernanda; Mendoza-Mendoza, Artemio; Jiménez-Bremont, Juan Francisco

    2015-10-01

    Plants are associated with a wide range of microorganisms throughout their life cycle, and some interactions result on plant benefits. Trichoderma species are plant beneficial fungi that enhance plant growth and development, contribute to plant nutrition and induce defense responses. Nevertheless, the molecules involved in these beneficial effects still need to be identify. Polyamines are ubiquitous molecules implicated in plant growth and development, and in the establishment of plant microbe interactions. In this study, we assessed the polyamine profile in Arabidopsis plants during the interaction with Trichoderma virens and Trichoderma atroviride, using a system that allows direct plant-fungal contact or avoids their physical interaction (split system). The plantlets that grew in the split system exhibited higher biomass than the ones in direct contact with Trichoderma species. After 3 days of interaction, a significant decrease in Arabidopsis polyamine levels was observed in both systems (direct contact and split). After 5 days of interaction polyamine levels were increased. The highest levels were observed with T. atroviride (split system), and with T. virens (direct contact). The expression levels of Arabidopsis ADC1 and ADC2 genes during the interaction with the fungi were also assessed. We observed a time dependent regulation of ADC1 and ADC2 genes, which correlates with polyamine levels. Our data show an evident change in polyamine profile during Arabidopsis - Trichoderma interaction, accompanied by evident alterations in plant root architecture. Polyamines could be involved in the changes undergone by plant during the interaction with this beneficial fungus.

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

    Science.gov (United States)

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

    2014-10-01

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

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

    Directory of Open Access Journals (Sweden)

    Felix Carlos

    2002-01-01

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

  2. Postharvest Biocontrol: Introspection and Paradigm Shifts

    Science.gov (United States)

    The use of postharvest biocontrol agents as an alternative to the synthetic, chemical fungicides on a widespread basis has many constraints. During the last twenty years, the field of postharvest biocontrol research has significantly grown and developed and seen the creation of several products. Des...

  3. 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

  4. 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. PMID:27487807

  5. Ecological impacts of Phlebiopsis gigantea biocontrol treatment against Heterobasidion spp. as revealed by fungal community profiling and population analyses

    OpenAIRE

    Vainio, Eeva Johanna

    2008-01-01

    Wood decay fungi belonging to the species complex Heterobasidion annosum sensu lato are among the most common and economically important species causing root rot and stem decay in conifers of the northern temperate regions. New infections by these pathogens can be suppressed by tree stump treatments using chemical or biological control agents. In Finland, the corticiaceous fungus Phlebiopsis gigantea has been formulated into a commercial biocontrol agent called Rotstop (Verdera Ltd.). Th...

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

    Science.gov (United States)

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

    2016-03-01

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

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

    Science.gov (United States)

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

    2016-03-01

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

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

    DEFF Research Database (Denmark)

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

    2012-01-01

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

  9. BIOETHANOL PRODUCTION FROM CELLULOSE IN RED ALGAE Gracilaria verrucosa BY SEPARATED HYDROLYSIS AND FERMENTATION SYSTEM USING Trichoderma viride AND Zymomonas mobilis

    OpenAIRE

    Ahmad, Ahyar

    2014-01-01

    In this study, renewable marine cellulose from red algae Gracilaria verrucosa was utilized for the production of bioethanol. Bioethanol from the red alga cellulose was produced by the enzymatic hydrolysis and fermentation methods and the conversion value of the cellulose in Gracilaria verrucosa was estimated. Trichoderma viride fungus and Zymomonas mobilis bacterium were used for enzymatic hydrolysis and bioethanol fermentation, r...

  10. Autodissemination of the entomopathogenic fungus Metarhizium anisopliae amongst adults of the malaria vector anopheles gambiae s.s.

    NARCIS (Netherlands)

    Scholte, E.J.; Knols, B.G.J.; Takken, W.

    2004-01-01

    Background - The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent for adult African malaria vectors. In the laboratory, work was carried out to assess whether horizontal transmission of the pathogen can take place during copulation, as this would enhance the i

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  12. Biological Action of Plant Extracts on a Fungal Plant Biostimulant Strain of Trichoderma Viride

    Directory of Open Access Journals (Sweden)

    Şesan Tatiana Eugenia

    2015-11-01

    Full Text Available The antifungal activity of nine plant extracts manufactured by Hofigal Import Export S.A. Romania against the biocontrol fungal agent Trichoderma viride Pers. (isolate Tv 82 was assessed in vitro for the first time in Romania. In general, the development (mycelial growth and sporulation was not inhibited by the six plant extracts (Satureja hortensis, Achillea millefolium, Allium sativum, Mentha sp., Hyssopus officinalis, Artemisia dracunculus 'Sativa', with three exceptions (Rosmarinus officinalis, Valeriana officinalis, Tagetes patula, applied in a concentration of 20%. Among these, the extract of Tagetes patula has inhibited the Tv 82 development, applied as lower concentrations (10% and 5%, efficacy being 54.3% and 50%, respectively. In addition, the tested plant extracts of Satureja hortensis, Achillea millefolium, Mentha sp. proved stimulative effect on Tv 82 development. This approach add to the early studies on the selectivity of Trichoderma spp. to chemicals used in plant protection, new data referring to the use of antagonistic fungi, like Trichoderma spp., as a protective mean against phytopathogens. Also, these data sustain the possibility of applying plant extracts as an alternative in plant protection or to apply together chemical (pesticides and biological means (plant extracts especially to protect ecological crops, as vegetables, medicinal plants a.o.

  13. Trichoderma spp. alleviate phytotoxicity in lettuce plants (Lactuca sativa L.) irrigated with arsenic-contaminated water.

    Science.gov (United States)

    Caporale, Antonio G; Sommella, Alessia; Lorito, Matteo; Lombardi, Nadia; Azam, Shah M G G; Pigna, Massimo; Ruocco, Michelina

    2014-09-15

    The influence of two strains of Trichoderma (T. harzianum strain T22 and T. atroviride strain P1) on the growth of lettuce plants (Lactuca sativa L.) irrigated with As-contaminated water, and their effect on the uptake and accumulation of the contaminant in the plant roots and leaves, were studied. Accumulation of this non-essential element occurred mainly into the root system and reduced both biomass development and net photosynthesis rate (while altering the plant P status). Plant growth-promoting fungi (PGPF) of both Trichoderma species alleviated, at least in part, the phytotoxicity of As, essentially by decreasing its accumulation in the tissues and enhancing plant growth, P status and net photosynthesis rate. Our results indicate that inoculation of lettuce with selected Trichoderma strains may be helpful, beside the classical biocontrol application, in alleviating abiotic stresses such as that caused by irrigation with As-contaminated water, and in reducing the concentration of this metalloid in the edible part of the plant.

  14. Marine isolates of Trichoderma spp. as potential halotolerant agents of biological control for arid-zone agriculture.

    Science.gov (United States)

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

    2011-08-01

    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.

  15. Identification of growth stage molecular markers in Trichoderma sp. 'atroviride type B' and their potential application in monitoring fungal growth and development in soil.

    Science.gov (United States)

    Mendoza-Mendoza, Artemio; Steyaert, Johanna; Nieto-Jacobo, Maria Fernanda; Holyoake, Andrew; Braithwaite, Mark; Stewart, Alison

    2015-11-01

    Several members of the genus Trichoderma are biocontrol agents of soil-borne fungal plant pathogens. The effectiveness of biocontrol agents depends heavily on how they perform in the complex field environment. Therefore, the ability to monitor and track Trichoderma within the environment is essential to understanding biocontrol efficacy. The objectives of this work were to: (a) identify key genes involved in Trichoderma sp. 'atroviride type B' morphogenesis; (b) develop a robust RNA isolation method from soil; and (c) develop molecular marker assays for characterizing morphogenesis whilst in the soil environment. Four cDNA libraries corresponding to conidia, germination, vegetative growth and conidiogenesis were created, and the genes identified by sequencing. Stage specificity of the different genes was confirmed by either Northern blot or quantitative reverse-transcriptase PCR (qRT-PCR) analysis using RNA from the four stages. con10, a conidial-specific gene, was observed in conidia, as well as one gene also involved in subsequent stages of germination (L-lactate/malate dehydrogenase encoding gene). The germination stage revealed high expression rates of genes involved in amino acid and protein biosynthesis, while in the vegetative-growth stage, genes involved in differentiation, including the mitogen-activated protein kinase kinase similar to Kpp7 from Ustilago maydis and the orthologue to stuA from Aspergillus nidulans, were preferentially expressed. Genes involved in cell-wall synthesis were expressed during conidiogenesis. We standardized total RNA isolation from Trichoderma sp. 'atroviride type B' growing in soil and then examined the expression profiles of selected genes using qRT-PCR. The results suggested that the relative expression patterns were cyclic and not accumulative.

  16. Disentangling the Trichoderma viridescens complex

    NARCIS (Netherlands)

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

    2013-01-01

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

  17. In vitro sensitivity of antagonistic Trichoderma atroviride to herbicides

    Directory of Open Access Journals (Sweden)

    Patricia Helena Santoro

    2014-04-01

    Full Text Available Trichoderma atroviride was tested in vitro for its sensitivity to different herbicides. The dosages tested were recommended dosage (RD, half dosage (½RD, and double dosage (2RD. Germination, colony-forming units (CFU, radial growth, and spore production were evaluated. Carfentrazone-ethyl and sulfentrazone inhibited the germination at RD and 2RD. A reduction in the CFU was observed for glufosinate-ammonium, atrazine, carfentrazone-ethyl, diuron + paraquat dichloride, imazapyr, oxyfluorfen, and sulfentrazone at each of the tested dosages. Radial growth was influenced by ametryn, atrazine, carfentrazone-ethyl, oxyfluorfen, and sulfentrazone herbicides, with an 80% reduction of the colonial area. Spore production was affected by carfentrazone-ethyl, oxyfluorfen, and sulfentrazone with colonial area reductions of over 70%. It was concluded that 2,4 D, clomazone, and imazapyr herbicides showed the least toxicity to T. atroviride and should be used in the crops where the fungus has been applied for phytopathogen control.

  18. BIOTECHNOLOGY OF TRICHODERMA-BASED FEED ADDITIVE WITH CELLULOLYTIC PROPERTIES

    Directory of Open Access Journals (Sweden)

    Koshchayev A. G.

    2013-11-01

    Full Text Available In the work, we have presented the information of elaboration of a manufacturing process of Mycocel feed additive with the cellulolytic activity for poul-try industry. Manufacturing process includes follow-ing steps: stock culture maintenance and storage of Trichoderma lignorum 81-17, growing fluid culture of microscopic fungus in sucrose yeast extract me-dium, feed additive with cellulolytic properties out-put and quality control, packaging, storage and disposal of waste. We have shown that the Mycocel is non-toxic feed additive for protozoa and warm-blooded animals (laboratory mice and quails. This study demonstrated total population livability in the experimental group with feed additive. Quail body-weight of experimental group was higher by 6% as compared to the control and feed consumption per 1 kg of live weight of bird was 3,58 kg, 7,5% lower than the control

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

    OpenAIRE

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

    2014-01-01

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

  20. Taxon-specific metagenomics of Trichoderma reveals a narrow community of opportunistic species that regulate each other’s development

    OpenAIRE

    Friedl, Martina A; Druzhinina, Irina S.

    2012-01-01

    In this paper, we report on the in situ diversity of the mycotrophic fungus Trichoderma (teleomorph Hypocrea, Ascomycota, Dikarya) revealed by a taxon-specific metagenomic approach. We designed a set of genus-specific internal transcribed spacer (ITS)1 and ITS2 rRNA primers and constructed a clone library containing 411 molecular operational taxonomic units (MOTUs). The overall species composition in the soil of the two distinct ecosystems in the Danube floodplain consisted of 15 known specie...

  1. Anthracnose of lucky bamboo Dracaena sanderiana caused by the fungus Colletotrichum dracaenophilum in Egypt.

    Science.gov (United States)

    Morsy, Ahmed A; Elshahawy, Ibrahim E

    2016-05-01

    Dracaena sanderiana, of the family Liliaceae, is among the ornamental plants most frequently imported into Egypt. Typical anthracnose symptoms were observed on the stems of imported D. sanderiana samples. The pathogen was isolated, demonstrated to be pathogenic based on Koch's rule and identified as Colletotrichum dracaenophilum. The optimum temperature for its growth ranges from 25 to 30 °C, maintained for 8 days. Kemazed 50% wettable powder (WP) was the most effective fungicide against the pathogen, as no fungal growth was observed over 100 ppm. The biocontrol agents Trichoderma harzianum and Trichoderma viride followed by Bacillus subtilis and Bacillus pumilus caused the highest reduction in fungal growth. To the best of our knowledge, this report describes the first time that this pathogen was observed on D. sanderiana in Egypt. PMID:27222738

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

    Science.gov (United States)

    Lange, Claudia; Weld, Richard J; Cox, Murray P; Bradshaw, Rosie E; McLean, Kirstin L; Stewart, Alison; Steyaert, Johanna M

    2016-01-01

    Biological control agents (BCA) are beneficial organisms that are applied to protect plants from pests. Many fungi of the genus Trichoderma are successful BCAs but the underlying mechanisms are not yet fully understood. Trichoderma cf. atroviride strain LU132 is a remarkably effective BCA compared to T. cf. atroviride strain LU140 but these strains were found to be highly similar at the DNA sequence level. This unusual combination of phenotypic variability and high DNA sequence similarity between separately isolated strains prompted us to undertake a genome comparison study in order to identify DNA polymorphisms. We further investigated if the polymorphisms had functional effects on the phenotypes. The two strains were clearly identified as individuals, exhibiting different growth rates, conidiation and metabolism. Superior pathogen control demonstrated by LU132 depended on its faster growth, which is a prerequisite for successful distribution and competition. Genome sequencing identified only one non-synonymous single nucleotide polymorphism (SNP) between the strains. Based on this SNP, we successfully designed and validated an RFLP protocol that can be used to differentiate LU132 from LU140 and other Trichoderma strains. This SNP changed the amino acid sequence of SERF, encoded by the previously undescribed single copy gene "small EDRK-rich factor" (serf). A deletion of serf in the two strains did not lead to identical phenotypes, suggesting that, in addition to the single functional SNP between the nearly clonal Trichoderma cf. atroviride strains, other non-genomic factors contribute to their phenotypic variation. This finding is significant as it shows that genomics is an extremely useful but not exhaustive tool for the study of biocontrol complexity and for strain typing. PMID:27190719

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

    Science.gov (United States)

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

    2016-06-01

    Trichoderma spp. are well-known bioagents for the plant growth promotion and pathogen suppression. The beneficial activities of the fungus Trichoderma spp. are attributed to their ability to produce and secrete certain secondary metabolites such as trichodermin that belongs to trichothecene family of molecules. The initial steps of trichodermin biosynthetic pathway in Trichoderma are similar to the trichothecenes from Fusarium sporotrichioides. Trichodiene synthase (TS) encoded by tri5 gene in Trichoderma catalyses the conversion of farnesyl pyrophosphate to trichodiene as reported earlier. In this study, we have carried out a comprehensive comparative sequence and structural analysis of the TS, which revealed the conserved residues involved in catalytic activity of the protein. In silico, modelled tertiary structure of TS protein showed stable structural behaviour during simulations. Two single-substitution mutants, i.e. D109E, D248Y and one double-substitution mutant (D109E and D248Y) of TS with potentially higher activities are screened out. The mutant proteins showed more stability than the wild type, an increased number of electrostatic interactions and better binding energies with the ligand, which further elucidates the amino acid residues involved in the reaction mechanism. These results will lead to devise strategies for higher TS activity to ultimately enhance the trichodermin production by Trichoderma spp. for its better exploitation in the sustainable agricultural practices.

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

    Science.gov (United States)

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

    2016-06-01

    Trichoderma spp. are well-known bioagents for the plant growth promotion and pathogen suppression. The beneficial activities of the fungus Trichoderma spp. are attributed to their ability to produce and secrete certain secondary metabolites such as trichodermin that belongs to trichothecene family of molecules. The initial steps of trichodermin biosynthetic pathway in Trichoderma are similar to the trichothecenes from Fusarium sporotrichioides. Trichodiene synthase (TS) encoded by tri5 gene in Trichoderma catalyses the conversion of farnesyl pyrophosphate to trichodiene as reported earlier. In this study, we have carried out a comprehensive comparative sequence and structural analysis of the TS, which revealed the conserved residues involved in catalytic activity of the protein. In silico, modelled tertiary structure of TS protein showed stable structural behaviour during simulations. Two single-substitution mutants, i.e. D109E, D248Y and one double-substitution mutant (D109E and D248Y) of TS with potentially higher activities are screened out. The mutant proteins showed more stability than the wild type, an increased number of electrostatic interactions and better binding energies with the ligand, which further elucidates the amino acid residues involved in the reaction mechanism. These results will lead to devise strategies for higher TS activity to ultimately enhance the trichodermin production by Trichoderma spp. for its better exploitation in the sustainable agricultural practices. PMID:26207800

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

    Directory of Open Access Journals (Sweden)

    Maria Angélica G. Barbosa

    2001-06-01

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

  6. Effect of Seed Quality and Combination Fungicide-Trichoderma spp. Seed Treatments on Pre- and Postemergence Damping-Off in Cotton.

    Science.gov (United States)

    Howell, Charles R

    2007-01-01

    ABSTRACT Good quality seeds of cotton cultivars often escaped pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae, and they were resistant to postemergence damping-off incited by Rhizoctonia solani. Poor quality seeds, however, were highly susceptible to both phases of seedling disease and required seed treatment in order to survive. Pre-emergence damping-off incited by Pythium spp. and Rhizopus oryzae could be controlled by seed treatment with biocontrol preparations of a number of Trichoderma spp., but these treatments were much less effective in controlling postemergence disease incited by Rhizoctonia solani. Postemergence seedling disease can be controlled by fungicides, but they were much less effective in controlling the pre-emergence phase of the disease. Combination seed treatments of poor quality cotton seeds with fungicides and Trichoderma spp. preparations, followed by planting in pathogen-infested soil, indicated that this technique will control both phases of seedling disease. Seed treatment with either the fungicides or the biocontrol agents alone did not achieve this goal. The optimum combination treatment for disease control was that of chloroneb plus Trichoderma spp., followed by chloroneb plus metalaxyl (Deltacoat AD) plus T. virens strain G-6. PMID:18942938

  7. A fitful fungus from a hot, arid climate increases grain yield in cool-cultivated barley

    OpenAIRE

    Murphy, Brian R.; Fiona M Doohan; HODKINSON, TREVOR R.

    2014-01-01

    Purpose The fungus Piriformospora indica was first isolated from plants growing in arid, hot desert conditions and has been shown to have significant potential as a biocontrol and biofertilising organism in barley under optimal growth conditions. However, it was not thought to be effective in plants grown in low temperatures and has consequently not been well tested in cold-stressed crops. This study sought to determine the effects of inoculating barley plants with this f...

  8. Role of 2-hexyl, 5-propyl resorcinol production by Pseudomonas chlororaphis PCL1606 in the multitrophic interactions in the avocado rhizosphere during the biocontrol process.

    Science.gov (United States)

    Calderón, Claudia E; de Vicente, Antonio; Cazorla, Francisco M

    2014-07-01

    Different bacterial traits can contribute to the biocontrol of soilborne phytopathogenic fungus. Among others, (1) antagonism, (2) competition for nutrients and niches, (3) induction of systemic resistance of the plants and (4) predation and parasitism are the most studied. Pseudomonas chlororaphis PCL1606 is an antagonistic rhizobacterium that produces the antifungal metabolite 2-hexyl, 5-propyl resorcinol (HPR). This bacterium can biologically control the avocado white root rot caused by Rosellinia necatrix. Confocal laser scanning microscopy of the avocado rhizosphere revealed that this biocontrol bacterium and the fungal pathogen compete for the same niche and presumably also for root exudate nutrients. The use of derivative mutants in the geners related to HPR biosynthesis (dar genes) revealed that the lack of HPR production by P. chlororaphis PCL1606 negatively influences the bacterial colonisation of the avocado root surface. Microscopical analysis showed that P. chlororaphis PCL1606 closely interacts and colonises the fungal hyphae, which may represent a novel biocontrol mechanism in this pseudomonad. Additionally, the presence of HPR-producing biocontrol bacteria negatively affects the ability of the fungi to infect the avocado root. HPR production negatively affects hyphal growth, leading to alterations in the R. necatrix physiology visible under microscopy, including the curling, vacuolisation and branching of hyphae, which presumably affects the colonisation and infection abilities of the fungus. This study provides the first report of multitrophic interactions in the avocado rhizosphere, advancing our understanding of the role of HPR production in those interactions.

  9. Trichoderma species collected from Iran

    Institute of Scientific and Technical Information of China (English)

    Doostmorad Zafari

    2004-01-01

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

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

    Science.gov (United States)

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

    2016-07-01

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

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

    Science.gov (United States)

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

    2016-07-01

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

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

    Directory of Open Access Journals (Sweden)

    Itamar Soares de Melo

    2000-03-01

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

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

    Science.gov (United States)

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

    2014-08-01

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

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

    Science.gov (United States)

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

    2014-08-01

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

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

    Directory of Open Access Journals (Sweden)

    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.

  16. 木霉重寄生过程分子机制的研究进展%Research Progress on Molecular Mechanism of Trichoderma Mycoparasitism

    Institute of Scientific and Technical Information of China (English)

    杨萍; 杨谦

    2012-01-01

    木霉(Trichoderma spp.)是一种广谱性拮抗菌,能够拮抗多种病原真菌,重寄生作用是木霉生物防治的主要机制,掌握其分子机制有利于对木霉进行合理应用和改造.为此,综述了木霉重寄生过程的分子机制研究进展,包括木霉重寄生过程的信号转导途径和产生水解病原菌细胞壁的水解酶的研究概况.木霉重寄生过程中的信号转导途径主要是异源三聚体G蛋白信号和MAPK基序,分泌的细胞壁水解酶主要有几丁质酶,葡聚糖酶和蛋白酶.%Trichoderma spp.was a broad spectrum antagonistic fungus which could antagonized a variety of pathogenic fungi,mycoparasitism was main mechanism of Trichoderma biological control. To master the molecular mechanisms was in favor of reasonable application and transformation of Trichoderma,therefore,this article reviewed the molecular mechanism research progress of Trichoderma mycoparasitism,including the signal transduction of Trichoderma mycoparasitism and the hydrolytic enzymes producing in the process of mycoparasitism which hydrolysis of pathogenic fungi cell wall. Trichoderma signal transduction pathways in the process of mycoparasitism was heterotrimeric G-proteins and mitogen-activated protein kinase (MAPK) cascades,the cell wall-degrading enzymes of Trichoderma included chitinase,glucanase and protease.

  17. 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

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

    Institute of Scientific and Technical Information of China (English)

    Sarah L Dodd; Alison Stewart

    2004-01-01

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

  19. Commercialization of postharvest biocontrol: barriers and opportunities

    Science.gov (United States)

    The past twenty years has seen the field of postharvest biocontrol evolve from an unknown entity with one or two novel reports in the literature to a sophisticated science with strong research programs worldwide, hundreds of publications, patented technologies, and now several commercial products. ...

  20. Heterologous expression of the Hsp24 from Trichoderma asperellum improves antifungal ability of Populus transformant Pdpap-Hsp24 s to Cytospora chrysosperma and Alternaria alternate.

    Science.gov (United States)

    Ji, S D; Wang, Z Y; Fan, H J; Zhang, R S; Yu, Z Y; Wang, J J; Liu, Z H

    2016-09-01

    The tolerance of plants to biotic and abiotic stresses could be improved by transforming with fungal resistance-related genes. In this study, the cDNA sequence (GenBank Acc. No. KP337939) of the resistance-related gene Hsp24 encoding the 24 kD heat shock protein was obtained from the biocontrol fungus Trichoderma asperellum ACCC30536. The promoter region of Hsp24 contained many cis-regulators related to stresses response, such as "GCN4" and "GCR1" etc. Hsp24 transcription in T. asperellum was up-regulated under six different environmental stresses, compared with the control. Furthermore, following heterologous transformation into Populus davidiana × P. alba var. Pyramidalis (Pdpap), Hsp24 was successfully transcribed in transformant Pdpap-Hsp24s. Pathogen-related genes (PRs) in four Pdpap-Hsp24s were up-regulated compared with those in the control Pdpap (Pdpap-Con). After co-culture of Pdpap-Hsp24s with the weak parasite Cytospora chrysosperma, the transcription of genes related to hormone signal pathway (JA and SA) were up-regulated in Pdpap-Hsp24s, and ethidium bromide (EtBr) and Nitro-blue tetrazolium (NBT) staining assays indicated that the cell membrane permeability and the active oxygen content of Pdpap-Hsp24s leaves were lower than that of the control Pdpap-Con. And when the Pdpap-Hsp24s were under the Alternaria alternate stress, the activities of superoxide dismutase (SOD) and peroxidase (POD) got higher in Pdpap-Hsp24s than that in Pdpap-Con, and the disease spots in Pdpap-Con leaves were obviously larger than those in Pdpap-Hsp24s leaves. In summary, Hsp24 of T. asperellum ACCC30536 is an important defense response gene, and its heterologous expression improved the resistance of transformant Pdpap-Hsp24s to C. chrysosperma and A. alternate. PMID:27193371

  1. Antioxidant defense response induced by Trichoderma viride against Aspergillus niger Van Tieghem causing collar rot in groundnut (Arachis hypogaea L.).

    Science.gov (United States)

    Gajera, H P; Katakpara, Zinkal A; Patel, S V; Golakiya, B A

    2016-02-01

    The study was conducted to examine the antioxidant enzymes induced by Trichoderma viride JAU60 as initial defense response during invasion of rot pathogen Aspergillus niger Van Tieghem in five groundnut varieties under pot culture. Seed treatment of T. viride JAU60 reduced 51-58% collar rot disease incidence in different groundnut varieties under pathogen infected soil culture. The activities of the antioxidant enzymes, viz., superoxide dismutase (SOD, EC 1.15.1.1), guaiacol peroxidase (GPX, EC 1.11.1.7) and ascorbate peroxidase (APX, EC 1.11.1.11), elevated in response to pathogen infection, in higher rate by tolerant varieties (J-11 and GG-2) compared with susceptible (GAUG-10, GG-13, GG-20) and further induced by T. viride treatment. Trichoderma treatment remarkably increased the 2.3 fold SOD, 5 fold GPX and 2.5 fold APX activities during disease development in tolerant varieties and the same was found about 1.2, 1.5 and 2.0 folds, respectively, in susceptible varieties. Overall, T. viride JAU60 treated seedlings (T3) witnessed higher activities of SOD (1.5 fold), GPX (3.25 fold) and APX (1.25 fold) than pathogen treatment (T2) possibly suggest the induction of antioxidant defense response by Trichoderma bio-controller to combat oxidative burst produced by invading pathogen.

  2. Biodegradation of poly(lactic acid) and some of its based systems with Trichoderma viride.

    Science.gov (United States)

    Lipsa, Rodica; Tudorachi, Nita; Darie-Nita, Raluca Nicoleta; Oprică, Lacramioara; Vasile, Cornelia; Chiriac, Aurica

    2016-07-01

    The purpose of this study was to assess the biodegradation of poly(lactic acid) (PLA) and some plasticized PLA based systems by Trichoderma viride fungus, in liquid medium and controlled laboratory conditions. The studied systems were achieved using PLA, hydrolyzed collagen (HC) as biological macromolecules and other additives by melt processing procedure. PLA and the systems' biodegradability was examined by the weight losses of the samples (after 7 and 21 days of exposure) and FTIR-ATR, GPC, SEM analyses (after 21 fungus inoculation days). The thermogravimetry (TG-DTG) study showed that the thermostability of the samples decreased after biodegradation and was influenced by the chemical structure of the systems' components. PMID:27064086

  3. 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.

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

    Science.gov (United States)

    Zhang, J D; Yang, Q

    2015-01-01

    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. PMID:25867321

  5. The isolation and identification of a P.polymyxa sp.serain and its bio-control function against pathogenic fungus%一株具有抗菌作用的P. polymyxa sp.菌株的分离鉴定

    Institute of Scientific and Technical Information of China (English)

    陈林; 董兆麟; 段康民

    2011-01-01

    produce phosphatase, cellu-lase or lipase activity. There is a mutualistic relationship between Paenibacillus polymyxa J and the host. Therefore, this strain is a potential biocontrol agent.

  6. Functions of thga1 Gene in Trichoderma harzianum Based on Transcriptome Analysis

    Science.gov (United States)

    Sun, Qing; Pang, Li; Wang, Lirong

    2016-01-01

    Trichoderma spp. are important biocontrol filamentous fungi, which are widely used for their adaptability, broad antimicrobial spectrum, and various antagonistic mechanisms. In our previous studies, we cloned thga1 gene encoding GαI protein from Trichoderma harzianum Th-33. Its knockout mutant showed that the growth rate, conidial yield, cAMP level, antagonistic action, and hydrophobicity decreased. Therefore, Illumina RNA-seq technology (RNA-seq) was used to determine transcriptomic differences between the wild-type strain and thga1 mutant. A total of 888 genes were identified as differentially expressed genes (DEGs), including 427 upregulated and 461 downregulated genes. All DEGs were assigned to KEGG pathway databases, and 318 genes were annotated in 184 individual pathways. KEGG analysis revealed that these unigenes were significantly enriched in metabolism and degradation pathways. GO analysis suggested that the majority of DEGs were associated with catalytic activities and metabolism processes that encode carbohydrate-active enzymes, secondary metabolites, secreted proteins, or transcription factors. According to the functional annotation of these DEGs by KOG, the most abundant group was “secondary metabolite biosynthesis, transport, and catabolism.” Further studies for functional characterization of candidate genes and pathways reported in this paper are necessary to further define the G protein signaling system in T. harzianum. PMID:27672660

  7. Investigating the beneficial traits of Trichoderma hamatum GD12 for sustainable agriculture – insights from genomics.

    Directory of Open Access Journals (Sweden)

    David J Studholme

    2013-07-01

    Full Text Available Trichoderma hamatum strain GD12 is unique in that it can promote plant growth, activate biocontrol against pre- and post-emergence soil pathogens and can induce systemic resistance to foliar pathogens. This study extends previous work in lettuce to demonstrate that GD12 can confer beneficial agronomic traits to other plants, providing examples of plant growth promotion in the model dicot, Arabidopsis thaliana and induced foliar resistance to Magnaporthe oryzae in the model monocot rice. We further characterize the lettuce-T. hamatum interaction to show that bran extracts from GD12 and a N-acetyl-β-D-glucosamindase-deficient mutant differentially promote growth in a concentration dependent manner, and these differences correlate with differences in the small molecule secretome. We show that GD12 mycoparasitises a range of isolates of the pre-emergent soil pathogen Sclerotinia sclerotiorum and that this interaction induces a further increase in plant growth promotion above that conferred by GD12. To understand the genetic potential encoded by T. hamatum GD12 and to facilitate its use as a model beneficial to study plant growth promotion, induced systemic resistance and mycoparasitism we present de novo genome sequence data. We compare GD12 with other published Trichoderma genomes and show that T. hamatum GD12 contains unique genomic regions with the potential to encode novel bioactive metabolites that may contribute to GD12’s agrochemically important traits.

  8. Investigating the beneficial traits of Trichoderma hamatum GD12 for sustainable agriculture-insights from genomics.

    Science.gov (United States)

    Studholme, David J; Harris, Beverley; Le Cocq, Kate; Winsbury, Rebecca; Perera, Venura; Ryder, Lauren; Ward, Jane L; Beale, Michael H; Thornton, Chris R; Grant, Murray

    2013-01-01

    Trichoderma hamatum strain GD12 is unique in that it can promote plant growth, activate biocontrol against pre- and post-emergence soil pathogens and can induce systemic resistance to foliar pathogens. This study extends previous work in lettuce to demonstrate that GD12 can confer beneficial agronomic traits to other plants, providing examples of plant growth promotion in the model dicot, Arabidopsis thaliana and induced foliar resistance to Magnaporthe oryzae in the model monocot rice. We further characterize the lettuce-T. hamatum interaction to show that bran extracts from GD12 and an N-acetyl-β-D-glucosamindase-deficient mutant differentially promote growth in a concentration dependent manner, and these differences correlate with differences in the small molecule secretome. We show that GD12 mycoparasitises a range of isolates of the pre-emergence soil pathogen Sclerotinia sclerotiorum and that this interaction induces a further increase in plant growth promotion above that conferred by GD12. To understand the genetic potential encoded by T. hamatum GD12 and to facilitate its use as a model beneficial organism to study plant growth promotion, induced systemic resistance and mycoparasitism we present de novo genome sequence data. We compare GD12 with other published Trichoderma genomes and show that T. hamatum GD12 contains unique genomic regions with the potential to encode novel bioactive metabolites that may contribute to GD12's agrochemically important traits.

  9. TRICHODERMA HARZIANUM AND PSEUDOMONAS SP. MEDIATED MANAGEMENT OF SCLEROTIUM ROLFSII ROT IN TOMATO (LYCOPERSICON ESCULENTUM MILL.

    Directory of Open Access Journals (Sweden)

    S. P. SINGH

    2013-01-01

    Full Text Available Trichoderma spp. and Pseudomonas spp. are known for their biocontrol potential and growth promotionactivity. Combination of these two microbes was used to assess the synergistic effect of compatible isolates forplant growth promotion and management of S. rolfsii. Trichoderma harzianum and Pseudomonas and theircombination were applied as seed and seedlings treatment in tomato. Consortium treatment showed the greaterplant growth promotion activity in comparison to solo treatment. Consortium treatment showed highest chlorophyllcontent (1.05mgg-1 fresh weight of leaves while in the pathogen inoculated control lowest amount was recorded(0.8mgg-1 fresh weight of leaves. The lowest mean disease rating (MDR 1.96 and maximum percent diseasereduction (PDR, 53.23% recorded in consortium treatment. The yield was also significantly higher in bioagentstreated treatments, T3 (37.41 tons h-1, T4 (36.60 tons h-1, T5 (43.84 tons h-1 and untreated unchallengedtreatment (45.29 tons h-1 than untreated challenged control (24.07 tons h-1. From these results it can beconcluded that the application of consortium of compatible bioagents will enhance the plant growth andbiological control of phytopathogens in contrast to treatment with single bioagent.

  10. Optimum Concentrations of Trichoderma longibrachiatum and Cadusafos for Controlling Meloidogyne javanica on Zucchini Plants.

    Science.gov (United States)

    Sokhandani, Zahra; Moosavi, Mohammad Reza; Basirnia, Tahereh

    2016-03-01

    A factorial experiment was established in a completely randomized design to verify the effect of different inoculum levels of an Iranian isolate of Trichoderma longibrachiatum separately and in combination with various concentrations of cadusafos against Meloidogyne javanica in the greenhouse. Zucchini seeds were soaked for 12 hr in five densities (0, 10(5), 10(6), 10(7), and 10(8) spores/ml suspension) of the fungus prior to planting in pots containing four concentrations of cadusafos (0, 0.5, 1, and 2 mg a.i./kg soil). The data were analyzed using a custom response surface regression model and the response surface curve and contour plots were drawn. Reliability of the model was examined by comparing the result of new experimental treatments with the predicted results. The optimal levels of these two variables also were calculated. The interactive effects of concentrations of Trichoderma and cadusafos were insignificant for several responses such as the total number of eggs per gram soil, the number of intact eggs per gram soil, nematode reproduction factor, and control percent. Closeness of experimental mean values with the expected values proved the validity of the model. The optimal levels of the cadusafos concentration and Trichoderma concentration that caused the best plant growth and lowest nematode reproduction were 1.7 mg a.i./kg soil and 10(8) conidia/ml suspension, respectively. PMID:27168653

  11. EVALUATION OF TRICHODERMA SPP. ON BEAN CULTURE, IN ANTHRACNOSE, WEB BLIGHT AND ROOT-KNOT NEMATODE

    Directory of Open Access Journals (Sweden)

    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.

  12. Taxon-specific metagenomics of Trichoderma reveals a narrow community of opportunistic species that regulate each other's development.

    Science.gov (United States)

    Friedl, Martina A; Druzhinina, Irina S

    2012-01-01

    In this paper, we report on the in situ diversity of the mycotrophic fungus Trichoderma (teleomorph Hypocrea, Ascomycota, Dikarya) revealed by a taxon-specific metagenomic approach. We designed a set of genus-specific internal transcribed spacer (ITS)1 and ITS2 rRNA primers and constructed a clone library containing 411 molecular operational taxonomic units (MOTUs). The overall species composition in the soil of the two distinct ecosystems in the Danube floodplain consisted of 15 known species and two potentially novel taxa. The latter taxa accounted for only 1.5 % of all MOTUs, suggesting that almost no hidden or uncultivable Hypocrea/Trichoderma species are present at least in these temperate forest soils. The species were unevenly distributed in vertical soil profiles although no universal factors controlling the distribution of all of them (chemical soil properties, vegetation type and affinity to rhizosphere) were revealed. In vitro experiments simulating infrageneric interactions between the pairs of species that were detected in the same soil horizon showed a broad spectrum of reactions from very strong competition over neutral coexistence to the pronounced synergism. Our data suggest that only a relatively small portion of Hypocrea/Trichoderma species is adapted to soil as a habitat and that the interaction between these species should be considered in a screening for Hypocrea/Trichoderma as an agent(s) of biological control of pests. PMID:22075025

  13. IN-VITRO EVALUATION OF FUNGICIDES, BIOCONTROL AGENTS AND PLANT EXTRACTS AGAINST RICE SHEATH BLIGHT PATHOGEN RHIZOCTONIA SOLANI

    Directory of Open Access Journals (Sweden)

    P. Srinivas

    2014-02-01

    Full Text Available Of the fourteen fungicides of different groups evaluated in-vitro against Rhizoctonia solani , Metalaxyl (0.1%, Mancozeb (0.1%, Tricyclazole (0.1%, Thiophenate methyl (0.1%, Carbendizm+ Mancozeb (0.1% were proved to be most effective in inhibiting the growth of the fungus. Among the bio-agents screened, Trichoderma viride was most effective in restricting the growth of Rhizoctonia solani followed by Penicillium notatum where as Aspergillus niger was proved least effective. Among the thirteen plant extracts evaluated garlic extract (10% was most effective in inhibiting the growth of fungus followed by calotropis (10%. Datura leaf extract (10% was found to be least effective in inhibiting the growth of Rhizoctonia solani.

  14. Inoculação de sementes de milho utilizando o Trichoderma harzianum como promotor de crescimento Corn seed inoculation using Trichoderma harzianum as a growth promoter

    Directory of Open Access Journals (Sweden)

    Maria de Lourdes Resende

    2004-08-01

    Full Text Available Conduziu-se este trabalho com o objetivo de estudar o efeito do fungo Trichoderma harzianum como promotor de crescimento de sementes de milho tratadas com diferentes fungicidas. O experimento foi realizado em vasos em casa-de-vegetação, adotando o delineamento experimental inteiramente casualizado em esquema fatorial (3x2, utilizando os tratamentos com fungicidas: Captan®, Maxim® e testemunha e inoculação (com e sem inóculo em quatro repetições. As características agronômicas avaliadas foram: altura de planta, peso da matéria seca da parte aérea e de raízes e também a técnica de isolamento do fungo nas raízes aos 45 dias após semeadura. Observou-se diferença significativa somente para o fator inóculo na característica peso de matéria seca de raízes. Pelos resultados obtidos, verificou-se que o fungo Trichoderma harzianum estimulou maior acúmulo de matéria seca nas raízes das plantas de milho e também estava presente nas raízes das plantas oriundas de sementes inoculadas, independentemente do tratamento fungicida das sementes.The objective of this work was to study the effect of the fungus Trichoderma harzianum as a growth promoter on the corn seeds treated with different fungicides. This experiment was carried out in pots in a greenhouse by adopting the experimental completely randomized design in (3 X 2 factorial scheme, by utilizing the treatments with fungicides: Captan®, Maximim® and control and inoculation (with and without an inoculum with four replicates. The agronomic characteristics evaluated were: plant height, shoot and root dry matter weight and also the inoculum isolation technique on the rots after sowing. Significant differences were found only for the factor inoculum in the characteristic root dry matter weight. The results obtained revealed that the fungus Trichoderma harzianum stimulated greatest dry matter accumulation in the roots of corn plants and also it was present in the roots of plants from

  15. Tremella with Edible Fungus

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    (Meiwei Shuang’er)Remove the tremella and edible fungus roots, clean and drain. Slice green peppers and carrots.Heat some oil in a wok, add tremella, edible fungus, green peppers and carrots, and clear stock, salt and sugar. Simmer for two minutes. Add MSG and pepper, remove to a plate, and serve.Features: Attractively black and white.Taste: Crisp and savory.

  16. IMPROVMENT BIOCONTROL OF DAMPING-OFF AND ROOT ROT/WILT OF FABA BEAN BY SALICYLIC ACID AND HYDROGEN PEROXIDE

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    Montaser F. Abdel-Monaim

    2013-04-01

    Full Text Available Rhizoctonia solani, Fusarium solani, F. oxysporum and Macrophomina phaseolina were found to be associated with root rot and wilt symptoms of faba bean plants collected from different fields in New Valley governorate, Egypt. All the obtained isolates were able to attack faba bean plants (cv. Giza 40 causing damping-off and root rot/wilt diseases. R. solani isolates 2 and 5, F. solani isolate 8, F. oxysporum isolate 12 and M. phaseolina isolate 14 were the more virulent ones in the pathogenicity tests. Biocontrol agents (Trichoderma viride, Bacillus megaterium and chemical inducers (salicylic acid and hydrogen peroxide individually or in combination were examined for biological control of damping-off and root rot/wilt and growth promotion of faba bean plants in vitro and in vivo. Both antagonistic biocontrol agents and chemical inducers either individually or in combination inhibited growth of the tested pathogenic fungi.Biocontrol agents combined with chemical inducers recorded the highest inhibited growth especially in case SA + T. viride and SA+ B. megaterium. Under green house and field conditions, all treatments significantly reduced damping-off and root rot/ wilt severity and increased survival of plants. Also, these treatments increased fresh and dry weights of the survived plants in pots compared with control.  The combination of biocontrol agents and chemical inducers were more effective than using them individually and SA+ T. viride was the best treatment in this respect. Under field conditions, all these treatments significantly increased growth parameters (plant height and No. of branches plant-1 and yield components (No. of pods and seedsplant-1, weight of 100 seeds and total yield feddan-1 and protein content in both seasons (2010-2011 and 2011-2012. Faba bean seeds soaked in SA+ T. viride and SA+ B. megaterium were recorded the highest growth parameters and yield components. Generally, the combination of biocontrol agents and

  17. Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

    Science.gov (United States)

    Nor, Nurafifah Mohd; Hadibarata, Tony; Zubir, Meor Mohd Fikri Ahmad; Lazim, Zainab Mat; Adnan, Liyana Amalina; Fulazzaky, Mohamad Ali

    2015-11-01

    Cresol Red belongs to the triphenylmethane (TPM) class of dyes which are potentially carcinogenic or mutagenic. However, very few studies on biodegradation of Cresol Red were investigated as compared to other type dyes such as azo and anthraquinone dye. The aim of this work is to evaluate triphenylmethane dye Cresol Red degradation by fungal strain isolated from the decayed wood in Johor Bahru, Malaysia. Detailed taxonomic studies identified the organisms as Trichoderma species and designated as strain Trichoderma harzianum M06. In this study, Cresol Red was decolorized up to 88% within 30 days under agitation condition by Trichoderma harzianum M06. Data analysis revealed that a pH value of 3 yielded a highest degradation rate among pH concentrations (73%), salinity concentrations of 100 g/L (73%), and a volume of 0.1 mL of Tween 80 (79%). Induction in the enzyme activities of manganese peroxidase, lignin peroxidase, laccase, 1,2- and 2,3-dioxygenase indicates their involvement in Cresol Red removal. Various analytical studies such as Thin-Layer Chromatography (TLC), UV-Vis spectrophotometer, and Gas chromatography mass spectrometry (GC-MS) confirmed the biotransformation of Cresol Red by the fungus. Two metabolites were identified in the treated medium: 2,4-dihydroxybenzoic acid (t R 7.3 min and m/z 355) and 2-hydroxybenzoic acid (t R 8.6 min and m/z 267). Based on these products, a probable pathway has been proposed for the degradation of Cresol Red by Trichoderma harzianum M06.

  18. Mechanism of triphenylmethane Cresol Red degradation by Trichoderma harzianum M06.

    Science.gov (United States)

    Nor, Nurafifah Mohd; Hadibarata, Tony; Zubir, Meor Mohd Fikri Ahmad; Lazim, Zainab Mat; Adnan, Liyana Amalina; Fulazzaky, Mohamad Ali

    2015-11-01

    Cresol Red belongs to the triphenylmethane (TPM) class of dyes which are potentially carcinogenic or mutagenic. However, very few studies on biodegradation of Cresol Red were investigated as compared to other type dyes such as azo and anthraquinone dye. The aim of this work is to evaluate triphenylmethane dye Cresol Red degradation by fungal strain isolated from the decayed wood in Johor Bahru, Malaysia. Detailed taxonomic studies identified the organisms as Trichoderma species and designated as strain Trichoderma harzianum M06. In this study, Cresol Red was decolorized up to 88% within 30 days under agitation condition by Trichoderma harzianum M06. Data analysis revealed that a pH value of 3 yielded a highest degradation rate among pH concentrations (73%), salinity concentrations of 100 g/L (73%), and a volume of 0.1 mL of Tween 80 (79%). Induction in the enzyme activities of manganese peroxidase, lignin peroxidase, laccase, 1,2- and 2,3-dioxygenase indicates their involvement in Cresol Red removal. Various analytical studies such as Thin-Layer Chromatography (TLC), UV-Vis spectrophotometer, and Gas chromatography mass spectrometry (GC-MS) confirmed the biotransformation of Cresol Red by the fungus. Two metabolites were identified in the treated medium: 2,4-dihydroxybenzoic acid (t R 7.3 min and m/z 355) and 2-hydroxybenzoic acid (t R 8.6 min and m/z 267). Based on these products, a probable pathway has been proposed for the degradation of Cresol Red by Trichoderma harzianum M06. PMID:26275435

  19. Strain-level diversity of secondary metabolism in the biocontrol species Aneurinibacillus migulanus.

    Science.gov (United States)

    Alenezi, Faizah N; Rekik, Imen; Bełka, Marta; Ibrahim, Abrar F; Luptakova, Lenka; Jaspars, Marcel; Woodward, Steve; Belbahri, Lassaad

    2016-01-01

    Aneurinibacillus migulanus strains Nagano and NCTC 7096 show potential in biocontrol against fungal and fungus-like plant pathogens, including a wide range of Ascomycota, Basidiomycota, and Oomycetes. Differences in terms of the range of pathogens that each strain inhibits, however, suggested that production of a single antibiotic cyclic peptide, gramicidin S (GS), by the two strains, is not the sole mechanism of inhibition. The availability of four sequenced genomes of Aneurinibacillus prompted us to apply genome mining techniques to identify the bioactive potential of A. migulanus and to provide insights into the secondary metabolite arsenal of the genus Aneurinibacillus. Up to eleven secondary metabolite biosynthetic gene clusters were present in the three Aneurinibacillus species. Biosynthetic gene clusters specifying bacteriocins, microcins, non-ribosomal peptides, polyketides, terpenes, phosphonates, lasso peptides and linaridins were identified. Chitinolytic potential and iron metabolism regulation were also investigated. With increasing numbers of biocontrol bacterial genomes being sequenced and mined, the use of approaches similar to those described in this paper will lead to an increase in the numbers of environmentally friendly natural products available to use against plant diseases. PMID:26686620

  20. Bio-control trials of Chaetomium spirale ND35 against apple canker

    Institute of Scientific and Technical Information of China (English)

    XINYa-fen; SHANGJin-jie

    2005-01-01

    A new endophytic antagonistic fungus, Chaetomium spirale ND35 from Populus tomentosa, was reported. The bio-control trials of C. spirale ND35 against the Valsa Canker of apple were preliminarily investigated. The results of dual culture on PDA plate showed that C. spirale ND35 was capable of strong antagonism against Valsa ceratosperma, and for inhibiting the mycelial growth of V. ceratosperma,.the crude extract of liquid culture of corn steep powder broth was more effective than that one of malt extract broth (MEB). The results of bio-control in greenhouse and field indicated that the disease incidence of apple tree treated with C. spirale ND35 was lower significantly than that treated by other methods. The re-isolation experiment suggested that C. spirale ND35 could colonize in stems and branches of apple trees successfully, and the ND35 colonization rate of the treatment with solid wheat bran culture was higher than that of corn steep powder broth, but the field experiment result the control effect of liquid culture of C. spirale ND35 was better than that of solid culture.

  1. EFEITO ANTAGÔNICO DE Trichoderma sp. NO DESENVOLVIMENTO DE Beauveria bassiana (Bals. Vuill. e Metarhizium anisopliae (Metsch. Sorok ANTAGONISTIC EFFECT OF Trichoderma sp. ON THE DEVELOPMENT OF Beauveria bassiana (Bals. Vuill. AND Metarhizium anisopliae (Metsch. SOROK

    Directory of Open Access Journals (Sweden)

    Alcides Moino Jr.

    1999-01-01

    Full Text Available Este trabalho teve por objetivo avaliar o efeito de Trichoderma sp. no desenvolvimento de Beauveria bassiana e Metarhizium anisopliae. Trichoderma sp., B. bassiana (isolado 634 e M. anisopliae (isolado E-9 foram inoculados em meio BDA, com intervalos de 0, 48, 120 e 168 horas entre a inoculação de Trichoderma sp. e dos entomopatógenos. Avaliou-se o crescimento radial das colônias nos períodos de 48 e 120 horas após a inoculação de Trichoderma sp., sendo que este afetou o desenvolvimento dos entomopatógenos quando inoculado simultaneamente ou após 48 horas. B. bassiana e M. anisopliae desenvolveram-se normalmente quando inoculados 168 horas antes de Trichoderma sp.. Também foi avaliado o efeito de um extrato de Trichoderma sp. sobre os entomopatógenos, com a adição de 0,1; 0,5; 1,0 e 5,0 ml de extrato/100,0 ml de meio, onde foram inoculados os entomopatógenos. Foram medidos os diâmetros de colônias e o número de conídios produzidos por B. bassiana e M. anisopliae na presença do extrato. A concentração de 5,0 ml de extrato/100,0 ml de meio alterou o crescimento e a conidiogênese de B. bassiana. O fungo M. anisopliae foi afetado a partir da adição de 1,0 ml de extrato/100,0 ml de meio.The objective of this work was to evaluate the effect of Trichoderma sp. on the development of Beauveria bassiana and Metarhizium anisopliae. The fungus Trichoderma sp. was inoculated on PDA culture medium, 0, 48, 120 and 168 hours after inoculation of the same plates with either B. bassiana (isolate 634 or M. anisopliae (isolate E-9. The radial growth of fungal colonies was measured 48 and 120 hours after Trichoderma sp. inoculation. Trichoderma sp. affected the development of both entomopathogenic fungi when inoculated simultaneously or 48 hours later. B. bassiana and M. anisopliae had normal development when inoculated 168 hours before Trichoderma sp. The effect of a toxic extract from Trichoderma sp. on the entomopathogenic fungi was also

  2. Use of Trichoderma fungi in spray solutions to reduce Moniliophthora roreri infection of Theobroma cacao fruits in Northeastern Costa Rica.

    Science.gov (United States)

    Seng, John; Herrera, Geovanny; Vaughan, Christopher S; McCoy, Michael B

    2014-09-01

    Cacao (Theobroma cacao) is an important cash crop in tropical climates such as that of Latin America. Over the past several decades, the infection of cultivated cacao by Moniliophthllora roreri, known commonly as "monilia", has significantly hindered cacao production in Latin America. Studies have proposed the use of Trichoderma sp. fungi in biocontrol treatments to prevent and reduce monilia infection, yet tests of Trichoderma-containing spray treatments on cacao agroforests have produced mixed results. Researchers and agricultural workers have suggested that addition of soil, fly ash, or other carbon sources to a Trichoderma spray may improve its efficacy in fighting monilia. To test these suggestions, we designed a series of spray mixtures including Thichoderma cultures, soil, and all necessary controls. We applied the spray mixtures to 80 cacao trees (20 trees for each of four resistant-selected clones to monilia) at the FINMAC organic cacao plantation in Pueblo Nuevo de Guacimo, Limón Province, in northeastern Costa Rica in March-April 2013. Five treatments were applied (control, water, water plus sterilized soil, water plus Trichoderma, and water plus sterilized soil plus Trichoderma). Each treatment was applied to four trees of each clone. We monitored the incidence of monilia infection under each spray treatment over the course of 35d. We found that spraying entire cacao trees two times with a mixture containing Trichoderma and sterilized soil significantly reduced the incidence of monilia infection by 11% (p ≤ 0.05) in only 35d, as compared to the control. This reduction in loss of cacao pods translates into an increase of plantation mean productivity of 1,500 kg dried beans/ha by 198 kg/ha up to 1,698 kg/ha or by a total increase over the whole 110 ha plantation by 21,780 kg. We propose that using such an antifungal spray over the whole course of a crop cycle (120 days) would decrease infection incidence even more. Application of this fungal control

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

    Science.gov (United States)

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

    2009-09-01

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

  4. 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...

  5. Generation, annotation and analysis of ESTs from Trichoderma harzianum CECT 2413

    Directory of Open Access Journals (Sweden)

    Gutiérrez Santiago

    2006-07-01

    Full Text Available Abstract Background The filamentous fungus Trichoderma harzianum is used as biological control agent of several plant-pathogenic fungi. In order to study the genome of this fungus, a functional genomics project called "TrichoEST" was developed to give insights into genes involved in biological control activities using an approach based on the generation of expressed sequence tags (ESTs. Results Eight different cDNA libraries from T. harzianum strain CECT 2413 were constructed. Different growth conditions involving mainly different nutrient conditions and/or stresses were used. We here present the analysis of the 8,710 ESTs generated. A total of 3,478 unique sequences were identified of which 81.4% had sequence similarity with GenBank entries, using the BLASTX algorithm. Using the Gene Ontology hierarchy, we performed the annotation of 51.1% of the unique sequences and compared its distribution among the gene libraries. Additionally, the InterProScan algorithm was used in order to further characterize the sequences. The identification of the putatively secreted proteins was also carried out. Later, based on the EST abundance, we examined the highly expressed genes and a hydrophobin was identified as the gene expressed at the highest level. We compared our collection of ESTs with the previous collections obtained from Trichoderma species and we also compared our sequence set with different complete eukaryotic genomes from several animals, plants and fungi. Accordingly, the presence of similar sequences in different kingdoms was also studied. Conclusion This EST collection and its annotation provide a significant resource for basic and applied research on T. harzianum, a fungus with a high biotechnological interest.

  6. Complete Genome Sequence of Biocontrol Strain Pseudomonas fluorescens LBUM 223

    OpenAIRE

    Roquigny, Roxane; Arseneault, Tanya; Gadkar, Vijay J.; Novinscak, Amy; Joly, David L.; Filion, Martin

    2015-01-01

    Pseudomonas fluorescens LBUM 223 is a plant growth-promoting rhizobacterium (PGPR) with biocontrol activity against various plant pathogens. It produces the antimicrobial metabolite phenazine-1-carboxylic acid, which is involved in the biocontrol of Streptomyces scabies, the causal agent of common scab of potato. Here, we report the complete genome sequence of P. fluorescens LBUM 223.

  7. THE INSECT PATHOGENIC FUNGUS Verticillium lecanii (Zimm. Viegas AND ITS USE FOR PESTS CONTROL: A REVIEW

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    Thiery B C ALAVO

    2015-08-01

    Full Text Available Chemical insecticides play an important role in the control of plant damage and plant diseases. However, extensive use of these products has led to the disruption of ecosystems because of several reasons such as death of non-target species, accumulation of pesticide residues in the environment and food, and buildup of pesticide resistance in the target species. Biological control is one of the alternatives to chemical pesticides and it can be described as the limitation of the abundance of living organisms and their products by other living organisms. Predators, parasitoids, fungi and other beneficial organisms can be used for the biocontrol of insect pests. The fungus Verticillium lecanii is one of the members of Deuteromycetes and it can be used for crop protection. This paper is a review of the international literature related to V. lecanii for the bio-control of insects of agricultural importance.

  8. Gliocladium and Trichoderma in agricultural soil

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

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

    Science.gov (United States)

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

    2016-08-01

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

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

    OpenAIRE

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

    2011-01-01

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

  11. Complete genome sequence of Pseudomonas fluorescens strain PICF7, an indigenous root endophyte from olive (Olea europaea L.) and effective biocontrol agent against Verticillium dahliae.

    Science.gov (United States)

    Martínez-García, Pedro Manuel; Ruano-Rosa, David; Schilirò, Elisabetta; Prieto, Pilar; Ramos, Cayo; Rodríguez-Palenzuela, Pablo; Mercado-Blanco, Jesús

    2015-01-01

    Pseudomonas fluorescens strain PICF7 is a native endophyte of olive roots. Previous studies have shown this motile, Gram-negative, non-sporulating bacterium is an effective biocontrol agent against the soil-borne fungus Verticillium dahliae, the causal agent of one of the most devastating diseases for olive (Olea europaea L.) cultivation. Here, we announce and describe the complete genome sequence of Pseudomonas fluorescens strain PICF7 consisting of a circular chromosome of 6,136,735 bp that encodes 5,567 protein-coding genes and 88 RNA-only encoding genes. Genome analysis revealed genes predicting factors such as secretion systems, siderophores, detoxifying compounds or volatile components. Further analysis of the genome sequence of PICF7 will help in gaining insights into biocontrol and endophytism.

  12. Screening of Mutation High-Yielding Biocontrol Bacterium BJ1 by Ion Beam Irradiation and Effect of Controlling Fusarium oxysporum cucunerinum Disease

    International Nuclear Information System (INIS)

    BJ1 of Bacillus subtilis is an important biocontrol factor in control of fungus disease. In order to improve the antagonistic ability of the strain,and obtain high-efficiency strains, 12C6+ of different doses and linear energy transfer (LET) was used to irradiate the biocontrol bacterium BJ1. The optimum dose and LET of ion beam irradiation for the BJ1 are 200-400 Gy and 60 keV/μm,respectively. The antagonistic ability is increased by 2%-21%. The control effect of mutation to Fusarium oxysporum f. sp. cucunerinum is increased by 17.48% over that of BJ1, and mutation also has better plant growth-promoting effect. (authors)

  13. The Terpenoid Biosynthesis Toolkit of Trichoderma.

    Science.gov (United States)

    Bansal, Ravindra; Mukherjee, Prasun Kumar

    2016-04-01

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

  14. The Terpenoid Biosynthesis Toolkit of Trichoderma.

    Science.gov (United States)

    Bansal, Ravindra; Mukherjee, Prasun Kumar

    2016-04-01

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

  15. 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.

  16. Identification of potential marker genes for Trichoderma harzianum strains with high antagonistic potential against Rhizoctonia solani by a rapid subtraction hybridization approach.

    Science.gov (United States)

    Scherm, Barbara; Schmoll, Monika; Balmas, Virgilio; Kubicek, Christian P; Migheli, Quirico

    2009-02-01

    A rapid subtraction hybridization approach was used to isolate genes differentially expressed during mycelial contact between Trichoderma harzianum (Hypocrea lixii) and Rhizoctonia solani, and could serve as marker genes for selection of superior biocontrol strains. Putatively positive clones were evaluated by transcription analysis during mycelial contact with R. solani versus growth on glucose, and for their differential transcription between two strains with either strong or poor biocontrol capability before, at, and after contact with R. solani. Besides four clones, which had similarity to putative but as yet uncharacterized proteins, they comprised ribosomal proteins, proteins involved in transcriptional switch and regulation, amino acid and energy catabolism, multidrug resistance, and degradation of proteins and glucans. Transcription of three clones was evaluated in five T. harzianum strains under confrontation conditions with R. solani. Two clones-acetyl-xylane esterase AXE1 and endoglucanase Cel61b-showed significant upregulation during in vivo confrontation of a T. harzianum strain that successively demonstrated a very high antagonistic capability towards R. solani, while expression was progressively lower in a series of T. harzianum strains with intermediate to poor antagonistic activity. These clones are promising candidates for use as markers in the screening of improved T. harzianum biocontrol strains.

  17. Repression of tropolone production and induction of a Burkholderia plantarii pseudo-biofilm by carot-4-en-9,10-diol, a cell-to-cell signaling disrupter produced by Trichoderma virens.

    Directory of Open Access Journals (Sweden)

    Mengcen Wang

    Full Text Available BACKGROUND: The tropolone-tolerant Trichoderma virens PS1-7 is a biocontrol agent against Burkholderia plantarii, causative of rice seedling blight. When exposed to catechol, this fungus dose-dependently produced carot-4-en-9,10-diol, a sesquiterpene-type autoregulatory signal molecule that promotes self-conidiation of T. virens PS1-7 mycelia. It was, however, uncertain why T. virens PS1-7 attenuates the symptom development of the rice seedlings infested with B. plantarii. METHODOLOGY/PRINCIPAL FINDINGS: To reveal the antagonism by T. virens PS1-7 against B. plantarii leading to repression of tropolone production in a coculture system, bioassay-guided screening for active compounds from a 3-d culture of T. virens PS1-7 was conducted. As a result, carot-4-en-9,10-diol was identified and found to repress tropolone production of B. plantarii from 10 to 200 µM in a dose-dependent manner as well as attenuate virulence of B. plantarii on rice seedlings. Quantitative RT-PCR analysis revealed that transcriptional suppression of N-acyl-L-homoserine lactone synthase plaI in B. plantarii was the main mode of action by which carot-4-en-9,10-diol mediated the quorum quenching responsible for repression of tropolone production. In addition, the unique response of B. plantarii to carot-4-en-9,10-diol in the biofilm formed in the static culture system was also found. Although the initial stage of B. plantarii biofilm formation was induced by both tropolone and carot-4-en-9,10-diol, it was induced in different states. Moreover, the B. plantarii biofilm that was induced by carot-4-en-9,10-diol at the late stage showed defects not only in matrix structure but also cell viability. CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate that carot-4-en-9,10-diol released by T. virens PS1-7 acts as an interkingdom cell-to-cell signaling molecule against B. plantarii to repress tropolone production and induces pseudo-biofilm to the cells. This observation also led to

  18. Lectins of fungal pathogens as potential tools in selecting promising biological control agents of Trichoderma spp%植物病原菌凝集素用于筛选生防木霉菌

    Institute of Scientific and Technical Information of China (English)

    杨合同; 唐文华; Maarten Ryder; 李纪顺; 郭勇; 周红姿

    2004-01-01

    In this article, lectins of eight pathogen strains were extracted and purified. By testing agglutination lectins with 19 Trichoderma spp. srains, all lectins from fungi agglutinated the conidia of Trichoderma spp. with different titers, except lectin from Alternaria solani Ms. Result indicated that the agglutination of Trichoderma conidia by fungal lectins was related to their effectiveness caused by the corresponding pathogens. Statistical analysis showed significant correlation between titers of Fusarium lectin and the effectiveness of Trichoderma against the disease.The agglutination test might be applied to predict potential effectiveness of a given Trichoderma strain against the corresponding disease and the lectins could be employed in screening Trichoderma spp as biocontrol agents efficiently.%提取并纯化了番茄早疫病、黑根霉、棉花立枯病菌和黄瓜灰霉病菌等8个植物病原菌菌株的凝集素,试验观察凝集素与供试19个木霉菌株的凝集反应效价,结果表明除番茄早疫病外,木霉菌分生孢子不同浓度悬浮液与植物病原菌产生的凝集素均有凝集反应,并且木霉菌分生孢子与病原菌凝集素的凝集反应效价与木霉菌对其防效有显著相关性,统计分析表明镰刀菌产生的凝集素与木霉菌的凝集反应效价与木霉菌对它的防效具有显著相关性.因此可以利用凝集素筛选木霉高效生防菌株.

  19. Cloning and characteristic analysis of a novel aspartic protease gene Asp55 from Trichoderma asperellum ACCC30536.

    Science.gov (United States)

    Dou, Kai; Wang, Zhiying; Zhang, Rongshu; Wang, Na; Fan, Haijuan; Diao, Guiping; Liu, Zhihua

    2014-12-01

    Proteases secreted by fungi belonging to the genus Trichoderma play important roles in biocontrol. In this study, the coding sequence and promoter region of the novel aspartic protease gene Asp55 were cloned from strain Trichoderma asperellum ACCC30536. Many cis-elements involved in phytopathogenic and environmental stress responses were identified in the Asp55 promoter region and may be recognized by MYB or WRKY transcription factors. The expression pattern of Asp55 under eight culture conditions was investigated by RT-qPCR. The expression level of Asp55 was up-regulated by poplar stem powder, Alternaria alternata cell wall fragments and A. alternata fermentation liquid, while it was down-regulated by carbon and nitrogen source starvation, and by powdered poplar leaves and roots. Additionally, the expression patterns of 15 genes encoding MYB transcription factors (Myb1 to Myb15) were also analyzed by RT-qPCR. Myb2 showed the most similar expression pattern with Asp55. The cDNA of Asp55 was expressed in Escherichia coli BL21, and recombinant ASP55 (rASP55) was purified. The purified rASP55 was evaluated for enzymatic activity and showed inhibitory effect on phytopathogenic A. alternata.

  20. Antagonismo de Trichoderma SPP. E Bacillus subtilis (UFV3918 a Fusarium sambucinum em Pinus elliottii engelm

    Directory of Open Access Journals (Sweden)

    Caciara Gonzatto Maciel

    2014-06-01

    Full Text Available Pinus elliottii é uma espécie de importância no setor florestal e apresenta vulnerabilidade na qualidade sanitária de suas sementes, especialmente pela associação de Fusarium spp., responsável por perdas de plântulas no viveiro. Este trabalho teve como objetivo avaliar a ação antagonista in vitro e in vivo dos agentes Trichoderma spp. e Bacillus subtilis (UFV3918 no controle de Fusarium sambucinum, responsável por danos em plântulas de Pinus elliottii. O controle in vitro foi avaliado através da inibição do crescimento micelial (confronto pareado de culturas, após a incubação a 25±2 ºC e fotoperíodo de 12 h. Para os testes in vivo (desenvolvidos em condições de viveiro, as sementes inicialmente foram inoculadas com o patógeno e, na sequência, microbiolizadas com os agentes antagônicos, para posterior semeadura. Utilizaram-se as técnicas de contato com o biocontrolador em meio BDA por 48 h e peliculização, como formas de microbiolização. Tanto Trichoderma spp. quanto Bacillus subtilis (UFV3918 foram eficientes no controle in vitro de F. sambucinum, e no teste de biocontrole in vivo o produto Bacillus subtilis (UFV3918 destacou-se, reduzindo as perdas de plântulas causadas pelo patógeno, assim como potencializando as variáveis de comprimento de plântula, massa verde e massa seca.

  1. 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.

  2. Microecological Relationship Between Metarizium and Trichoderma in Artificial Medium and Soil%培养基和土壤中木霉菌与绿僵菌微生态关系

    Institute of Scientific and Technical Information of China (English)

    衣思瑶; 吕淑霞; 陈捷; 宋磊; 黄磊; 李雅乾

    2012-01-01

    The relationship between Trichoderma and Metarhizium cultured on the plate and in soil were studied respectively. The growth of Trichoderma and Metarhizium and the interacting effect between the two strains were observed according to the morphological colony confrontation on plate and re-isolation test from soil when the strains of Trichoderma and Metarhizium were inoculated at different proportion simultaneously. The similar observation was conducted when Metarhizium was inoculated and cultured first followed by inoculating Trichoderma at different intervals. The results showed that when Trichoderma and Metarhizium were inoculated simultaneously at the ratio of 1:100, the average growth distance of Trichoderma was as 6 times as that of Metarhizium on the plate. Metarhizium hardly isolated from Trichoderma after culturing for 4 days in soil. Therefore, Trichoderma inhibited to Metarhizium growth significantly, and it was hard for Metarhizium to grow due to the dominant growth of Trichoderma. When Metarhizium was inoculated and cultured for 7 days before inoculating Trichoderma, the growth distance of Metarhizium was lmm lower than that of Trichoderma, and the spores quantity of Metarhizium was higher than that of Trichoderma after the two strains were in a stable growth. The growth of Trichoderma turned to be weak and the growth rate of Metarhizium increased, consequently the growth of the two strains tended to be balanced, and it could achieve a certain level of co-existence. The thorough research need to be done further more in terms of the complex microbial ecological relationships between Trichoderma and Metarhizium in soil, in order to obtain the key techniques for avoiding the direct ecologic competition of the two strains and to realize the application of the two bio-control agents together synergistically.%为研究木霉菌与绿僵菌在平板及土壤中相互关系,以不同配比同时接种2株菌,采用平板对峙和土壤再分离试验,观察2种

  3. G protein signalling involved in host recognition and mycoparasitismrelated chitinase expression in Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

    Susanne Zeilinger; Barbara Reithner; Kurt Brunner; Valeria Scala; Isabel Peiβl; Matteo Lorito; Robert L Mach

    2004-01-01

    @@ Mycoparasitic species of Trichoderma are commercially applied as biological control agents against various fungal pathogens. The mycoparasitic interaction is host specific and includes recognition,attack and subsequent penetration and killing of the host. Investigations on the underlying events revealed that Trichoderma responds to multiple signals from the host (e. g. lectins or other ligands such as low molecular weight components released from the host's cell wall) and host attack is accompanied by morphological changes and the secretion of hydrolytic enzymes and antibiotics.Degradation of the cell wall of the host fungus is-besides glucanases and proteases-mainly achieved by chitinases. In vivo studies showed that the ech42 gene (encoding endochitinase 42) is expressed before physical contact of Trichoderma with its host, probably representing one of the earliest events in mycoparasitism, whereas Nag1 (N-acetylglucosaminidase) plays a key role in the general induction of the chitinolytic enzyme system of T. atroviride . Investigations on the responsible signal transduction pathways of T. atroviride led to the isolation of several genes encoding key components of the cAMP and MAP kinase signaling pathways, as alpha and β subunits of heterotrimeric G proteins, the regulatory subunit of cAMP-dependent protein kinase,adenylate cyclase, and three MAP kinases. Analysis of knockout mutants, generated by Agrobacterium-mediated transformation, revealed that at least two alpha-subunits of heterotrimeric G proteins are participating in mycoparasitism-related signal transduction. The Tga1 G alpha subunit was shown to be involved in mycoparasitism-related processes such as chitinase expression and overproduction of toxic secondary metabolites, whereas Tga3 was found to be completely avirulent showing defects in chitinase formation and host recognition.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1986-06-01

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

  5. 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.

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

    Directory of Open Access Journals (Sweden)

    Maria Augusta Crivelente Horta

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Aline Machado de Castro

    2010-01-01

    Full Text Available The low-cost production of cellulolytic complexes presenting high action at mild conditions and well-balanced cellulase activities is one of the major bottlenecks for the economical viability of the production of cellulosic ethanol. In the present paper, the filamentous fungus Trichoderma harzianum IOC-3844 was used for the production of cellulases from a pretreated sugarcane bagasse (namely, cellulignin, by submerged fermentation. This fungal strain produced high contents of endoglucanase activity (6,358 U·L−1 after 72 hours of process, and further relevant β-glucosidase and FPase activities (742 and 445 U·L−1, resp.. The crude enzyme extract demonstrated appropriate characteristics for its application in cellulose hydrolysis, such as high thermal stability at up to 50∘C, accessory xylanase activity, and absence of proteolytic activity towards azocasein. This strain showed, therefore, potential for the production of complete cellulolytic complexes aiming at the saccharification of lignocellulosic materials.

  8. Absorbing film assisted laser induced forward transfer of fungi (Trichoderma conidia)

    Science.gov (United States)

    Hopp, B.; Smausz, T.; Antal, Zs.; Kresz, N.; Bor, Zs.; Chrisey, D.

    2004-09-01

    We present an investigation on absorbing film assisted laser induced forward transfer (AFA-LIFT) of fungus (Trichoderma) conidia. A KrF excimer laser beam [λ =248nm,FWHM=30ns (FWHM, full width at half maximum)] was directed through a quartz plate and focused onto its silver coated surface where conidia of the Trichoderma strain were uniformly spread. The laser fluence was varied in the range of 0-2600mJ/cm2 and each laser pulse transferred a pixel of target material. The average irradiated area was 8×10-2mm2. After the transfer procedure, the yeast extract medium covered glass slide and the transferred conidia patterns were incubated for 20 h and then observed using an optical microscope. The transferred conidia pixels were germinated and the areas of the culture medium surfaces covered by the pixels were evaluated as a function of laser fluence. As the laser fluence was increased from 0 to 355mJ/cm2 the transferred and germinated pixel area increased from 0 to 0.25mm2. Further increase in fluence resulted in a drastic decrease down to an approximately constant value of 0.06mm2. The yield of successful transfer by AFA-LIFT and germination was as much as 75% at 355mJ/cm2. The results prove that AFA-LIFT can successfully be applied for the controlled transfer of biological objects.

  9. Xenobiotic Compounds Degradation by Heterologous Expression of a Trametes sanguineus Laccase in Trichoderma atroviride.

    Directory of Open Access Journals (Sweden)

    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.

  10. Xenobiotic Compounds Degradation by Heterologous Expression of a Trametes sanguineus Laccase in Trichoderma atroviride

    Science.gov (United States)

    Balcázar-López, Edgar; Méndez-Lorenzo, Luz Helena; Batista-García, Ramón Alberto; Esquivel-Naranjo, Ulises; Ayala, Marcela; Kumar, Vaidyanathan Vinoth; Savary, Olivier; Cabana, Hubert; Herrera-Estrella, Alfredo; Folch-Mallol, Jorge Luis

    2016-01-01

    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. PMID:26849129

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

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  12. Identification and Characterization of Lipopeptides from Bacillus subtilis B1 Against Sapstain Fungus of Rubberwood Through MALDI-TOF-MS and RT-PCR.

    Science.gov (United States)

    Sajitha, K L; Dev, Suma Arun; Maria Florence, E J

    2016-07-01

    Bacillus subtilis is a potent biocontrol agent producing a wide array of antifungal lipopeptides for the inhibition of fungal growth. B. subtilis B1 isolated from market-available compost provided an efficient control of rubberwood sapstain fungus, Lasiodiplodia theobromae. The current study is aimed to identify and characterize the lipopeptides responsible for the biocontrol of rubberwood sapstain fungus by Bacillus subtilis B1. The bacterial whole-cell surface extract from the dual culture of B. subtilis B1 and sapstain fungus (L. theobromae) was analysed using MALDI-TOF-MS. The protonated as well as sodium, potassium adducts of homologues of iturin C, surfactin, bacillomycin D and fengycin A and B were identified and expression of the lipopeptide biosynthetic genes could be confirmed through RT-PCR. This is the first report of mycobacillin and trimethylsilyl derivative of bacilysin during antagonism through MALDI-TOF-MS. MALDI-TOF-MS with RT-PCR offered easy platforms to characterize the antifungal lipopeptides. The identification of antifungal lipopeptides can lead to the formulation of prospective biocontrol by-products which have wide-scale utility. PMID:27004481

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

    OpenAIRE

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

    2014-01-01

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

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

    OpenAIRE

    Majid, Muhammad

    2016-01-01

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

  15. MORPHOLOGICAL CHARACTERIZATION AND MASS PRODUCTION OF NEMATOPHAGOUS FUNGUS NEMATOCTONUS ROBUSTUS

    Directory of Open Access Journals (Sweden)

    Ramesh babu S,

    2014-06-01

    Full Text Available The plant parasitic nematodes infect the root tissues of the plant causing root galls that lead to reduced water and mineral uptake in the plant root system. Nematophagous fungus are used as biocontrol for the nematodes. Among those Nematoctonus are one of the species used as bioagent. Nematoctonus species produces an extensive mycelium and capture many nematodes with hour glass shaped adhesive knobs on the hyphae. Nematodes become attached to these adhesive knobs and the cuticle of nematode is penetrated by the infective hyphae. This isolate of Nematoctonus robustus is characterized by hyaline mycelium, dikaryotic in nature containing genetically two different nuclei in each cell, having distinct clamp connection. The fungus has better colonizing ability on natural solid substrates like wheat straw and rice straw. It also show good ability to colonize on different cereal grains and various other waste products like coconut coir and FYM etc. This species is one of the best used for mass production and effective for control of plant parasitic nematodes.

  16. 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.

  17. CONTROLE DE Monosporascus cannonballus POR TIAZOLIDINA-2,4-DIONA E EFEITO SOBRE O AGENTE DE CONTROLE BIOLÓGICO Trichoderma spp

    Directory of Open Access Journals (Sweden)

    Erika Valente de Medeiros

    2006-01-01

    Full Text Available The objective of this study was to evaluate the effect of the synthetic compound Thiazolidines-2,4- dione on the ¿in vitro¿ development of Monosporascus cannonballus, the causal agent of the melon sudden wilt (vine decline and Trichoderma sp., the biocontroller agent of the referred pathogen. The work was done through two experiments. In the first experiment the following concentrations of the composition were used: 0, 25, 50, 75, 100, 125, 150, 175 ìg.mL-1, combined with four isolates of M. cannonballus. The evaluated variables were mycelial growth inhibition (MGI, mycelial growth rate (MGR and area below mycelial growth curve (ABMGC. In the second experiment the effect of the same concentrations of the synthetic compound were analyzed regarding a Trichoderma sp isolate. In the first experiment, there was a significant interaction between concentrations andisolates. High correlation coefficients confirmed the agreement of combination among the variables. The progress curves of the variables, according to compound concentrations, were adjusted by polynomial models. The most efficient concentration was 75 ìg.mL-1,, for inhibiting the mycelia growth until the experiment end, except for the Mc4 that showed TCM of 5.07. Regarding other isolates, Mc 3 was the least resistant, as with concentration of 50 g.mL- 1showed TCM of 2.36, while Mc1, Mc2 and Mc4 presented a higher growth rate, being 8.48; 8.08 and 8.97, respectively. The recommended dosage of the compound for the inhibition of M. cannonballus development when measured in vitro is 153 ug.mL .Neither concentrations influenced the mycelial development of Trichoderma sp.,as it didn¿t differ from control (P=0,05, demonstrating the potential of this synthetic compound as a complementary form of Monosporascus cannonballus control, together to the Trichoderma spp.

  18. 绿色木霉菌对喷气燃料理化性质影响的研究%Study on Influence of Trichoderma viride contamination on Physiochemical Properties of Jet Fuel

    Institute of Scientific and Technical Information of China (English)

    殷鹤; 黄雪峰; 杨浩

    2016-01-01

    influence of Trichoderma viride contamination on physiochemical properties of jet fuel was studied. The results show that, induction period, acid number and surface tension of jet fuel decrease with increasing of the trichoderma viride culture time, however, the level of particle contamination has a significant increase.In addition, the growth of trichoderma viride in jet fuel can contribute to the steel corrosion, which will cause the great damage to oil tanks and pipes. These results indicate that trichoderma viride as one of the dominate contaminating fungus has a significant influence on physiochemical properties of jet fuel .%对绿色木霉菌(Trichoderma viride)油水体系进行培养,并对喷气燃料理化性质进行分析。试验结果表明,随着培养时间的延长,喷气燃料中颗粒数明显增加,而诱导时间以及表面张力减小。另外,喷气燃料中绿色木霉菌的存在加速了铜片腐蚀,这对储存油罐以及输油管道构成巨大威胁。说明木霉菌作为一个主导污染真菌对喷气燃料的理化性质有重要影响。

  19. Improvement of Biocontrol of Damping-off and Root Rot/Wilt of Faba Bean by Salicylic Acid and Hydrogen Peroxide.

    Science.gov (United States)

    Abdel-Monaim, Montaser Fawzy

    2013-03-01

    Rhizoctonia solani, Fusarium solani, F. oxysporum, and Macrophomina phaseolina were found to be associated with root rott and wilt symptoms of faba bean plants collected from different fieldes in New Valley governorate, Egypt. All the obtained isolates were able to attack faba bean plants (cv. Giza 40) causing damping-off and root rot/wilt diseases. R. solani isolates 2 and 5, F. solani isolate 8, F. oxysporum isolate 12 and M. phaseolina isolate 14 were the more virulent ones in the pathogenicity tests. Biocontrol agents (Trichoderma viride and Bacillus megaterium) and chemical inducers (salicylic acid [SA] and hydrogen peroxide) individually or in combination were examined for biological control of damping-off and root rot/wilt and growth promoting of faba bean plants in vitro and in vivo. Both antagonistic biocontrol agents and chemical inducers either individually or in combination inhibited growth of the tested pathogenic fungi. Biocontrol agents combined with chemical inducers recorded the highest inhibited growth especially in case SA + T. viride and SA + B. megaterium. Under green house and field conditions, all treatments significantly reduced damping-off and root rot/wilt severity and increased of survival plants. Also, these treatments increased fresh and weights of the survival plants in pots compared with control. The combination between biocontrol agents and chemical inducers were more effective than used of them individually and SA + T. viride was the best treatment in this respect. Also, under field conditions, all these treatments significantly increased growth parameters (plant height and number of branches per plant) and yield components (number of pods per plant and number of seeds per plant, weight of 100 seeds and total yield per feddan) and protein content in both seasons (2010~2011 and 2011~2012). Faba bean seeds soaked in SA + T. viride and SA + B. megaterium were recorded the highest growth parameters and yield components. Generally, the

  20. Diversity of Trichoderma in greenhouse soil

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  1. Universally Primed PCR (UP-PCR) and its applications for taxonomy in Trichoderma

    Institute of Scientific and Technical Information of China (English)

    Mette Lübeck

    2004-01-01

    @@ Universally Primed PCR (UP-PCR) is a PCR fingerprinting method that has demonstrated its applicability in different aspects of mycology. These applications constitute analysis of genome structures, identification of species, analysis of population and species diversity, revealing of genetic relatedness at infra-and inter-species level, and identification of UP-PCR markers at different taxonomic levels (strain, group and/or species) . A further development of the UP-PCR technique is an UP-PCR product cross hybridisation assay that facilitates investigation of sequence similarity (homology) of UP-PCR products and grouping of strains into UP-PCR hybridisation groups. This separates the strains into entities with high genetic similarity (DNA homology) . UP-PCR has been used as an aid in taxonomy and species delineation, and to monitor biocontrol strains following their release into the environment by fingerprint characterisation of pure cultures and through direct detection in soil by amplification of UP-PCR-derived SCAR markers. The technique has been applied to Trichoderma strains in particularly with the aims of strain recognition and classification.

  2. EVALUATION OF BIOLOGICAL EFFICACY OF TRICHODERMA SPECIES ISOLATES AGAINST ALTERNARIA LEAF SPOT DISEASE OF SESAME

    Directory of Open Access Journals (Sweden)

    A. S. Lubaina

    2015-03-01

    Full Text Available Alternaria leaf spot disease is a major threat to sesame (Sesamum orientale L. caused by Alternari asesami. Induced resistance is an alternative to systemic disease resistance response of plants. The present study aims to evaluate Trichoderma species efficacy as biocontrol via induction of resistance against A. sesami in sesame species. During in vitro bio control test, T. harzianum colonize and parallely inhibit the growth of the fungal pathogen. Expression of various defence related enzymes observed in sesame induce resistance against the pathogen infection in the host. T. harzianum coupled with inoculation of A. sesami enhance the remarkable induction of defence enzyme such as peroxidase (POX, polyphenol oxidase (PPO, phenylalanine ammonia lyase (PAL and also the phenolic content compared with the control. The enzyme activity increased from 48 h of sampling and peaked at 72 h and then decreased after 72 h. In greenhouse and field experiments, soil treatment with a powder formulation of T. harzianum two weeks before planting or at the time of planting reduced significantly the incidence of diseases on both the wild and cultivar Thilarani.The results demonstrate that T. harzianumcan be successfully applied as a biological control against Alternaria leaf spot disease in sesame.

  3. Studies on Exo-Chitinase Production from Trichoderma asperellum UTP-16 and Its Characterization.

    Science.gov (United States)

    Kumar, D Praveen; Singh, Rajesh Kumar; Anupama, P D; Solanki, Manoj Kumar; Kumar, Sudheer; Srivastava, Alok K; Singhal, Pradeep K; Arora, Dilip K

    2012-09-01

    The growth conditions for chitinase production by Trichoderma asperellum UTP-16 in solid state fermentation was optimized using response surface methodology based on central composite design. The chitinase production was optimized, using one-factor at a time approach, with six independent variables (temperature, pH, NaCl, incubation period, nitrogen and carbon sources) and 3.31 Units per gram dry substrate (U gds(-1)) exo-chitinase yield was obtained. A 21.15% increase was recorded in chitinase activity (4.01 U gds(-1)) through surface response methodology, indicates that it is a powerful and rapid tool for optimization of physical and nutritional variables. Further, efficiency of crude enzyme was evaluated against phytopathogenic Fusarium spp. and a mycelial growth inhibition up to 3.5-6.5 mm was achieved in well diffusion assay. These results could be supplemented as basic information for the development of enzyme based formulation of T. asperellum UTP-16 and its use as a biocontrol agent. PMID:23997329

  4. Histopathological studies of sclerotia of phytopathogenic fungi parasitized by a GFP transformed Trichoderma virens antagonistic strain.

    Science.gov (United States)

    Sarrocco, Sabrina; Mikkelsen, Lisbeth; Vergara, Mariarosaria; Jensen, Dan Funck; Lübeck, Mette; Vannacci, Giovanni

    2006-02-01

    The gfp gene from the jellyfish Aequorea victoria, coding for the Green Fluorescent Protein (GFP), was used as a reporter gene to transform a Trichoderma virens strain I10, characterized as having a promising biocontrol activity against a large number of phytopathogenic fungi. On the basis of molecular and biological results, a stable GFP transformant was selected for further experiments. In order to evaluate the effects of GFP transformation on mycoparasitic ability of T. virens I10, sclerotia of Sclerotium rolfsii, Sclerotinia sclerotiorum and S. minor were inoculated with the T. virens strain I10 GFP transformant or the wild type strain. Statistical analysis of percentages of decayed sclerotia showed that the transformation of the antagonistic isolate with the GFP reporter gene did not modify mycoparasitic activity against sclerotia. Sclerotium colonization was followed by fluorescent microscopy revealing intracellular growth of the antagonist in the cortex (S. rolfsii) and inter-cellular growth in the medulla (S. rolfsii, and S. sclerotiorum). The uniformly distributed mycelium of T. virens just beneath the rind of sclerotia of both S. rolfsii and S. sclerotiorum suggests that the sclerotia became infected at numerous randomly distributed locations without any preferential point of entry. PMID:16388938

  5. Trichoderma harzianum T-22 Induces Systemic Resistance in Tomato Infected by Cucumber mosaic virus

    Science.gov (United States)

    Vitti, Antonella; Pellegrini, Elisa; Nali, Cristina; Lovelli, Stella; Sofo, Adriano; Valerio, Maria; Scopa, Antonio; Nuzzaci, Maria

    2016-01-01

    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 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. PMID:27777581

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1987-09-01

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

  7. 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)

    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)

  8. Real-time RT-PCR expression analysis of chitinase and endoglucanase genes in the three-way interaction between the biocontrol strain Clonostachys rosea IK726, Botrytis cinera and strawberry

    DEFF Research Database (Denmark)

    Mamarabadi, Mojtaba; Jensen, Birgit; Jensen, Søren Dan Funck;

    2008-01-01

    Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two endoglucan......Clonostachys rosea is a well-known biocontrol agent against Botrytis cinerea, the causal agent of gray mold in strawberry. The activity of cell wall-degrading enzymes might play a significant role for successful biocontrol by C. rosea. The expression pattern of four chitinases, and two...... endoglucanase genes from C. rosea strain IK726 was analyzed using real-time RT-PCR in vitro and in strawberry leaves during interaction with B. cinerea. Specific primers were designed for ß-tubulin genes from C. rosea and B. cinerea, respectively, and a gene encoding a DNA-binding protein (DBP) from strawberry......, allowing in situ activity assessment of each fungus in vitro and during their interaction on strawberry leaves. Growth of B. cinerea was inhibited in all pathogen-antagonist interactions while the activity of IK726 was slightly increased. In all in vitro interactions, four of the six genes were upregulated...

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

    NARCIS (Netherlands)

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

    2015-01-01

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

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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

  12. 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.

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

    Science.gov (United States)

    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. PMID:25836973

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

    Directory of Open Access Journals (Sweden)

    Aline Crucello

    Full Text Available 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.

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

    Science.gov (United States)

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

    2014-11-01

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

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

    OpenAIRE

    Haryuni -

    2013-01-01

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

  17. Heterologous expression of the glucose oxidase gene in Trichoderma atroviride leads enhanced ability to attack phytopathogenic fungi and induction of plant systemic disease resistance

    Institute of Scientific and Technical Information of China (English)

    Robert L Mach; Brunner Kurt; Matteo Lorito; Susanne Zeilinger; Rosalia Ciliento; Sheridan Woo

    2004-01-01

    @@ A transgenic strain of Trichoderma atroviride that expresses the Aspergillus niger glucose oxidase gene goxA under a homologous pathogen-inducible promoter (nag1) has been constructed, with the aim of increasing the ability of this biocontrol agent (BCA) to attack phytopathogenic fungi and enhance plant systemic disease resistance. The sporulation and growth rate of the transgenic progenies were similar to the wild-type strain Pl. goxA expression occurred immediately after contact with the plant pathogen,and the glucose oxidase formed was secreted extracellularly. The transformed strain SJ3 4, containing 12-14 copies of the transgene, produced significantly less N-acetyl-glucosaminidase and endochitinase then wild type. However, the ability of its culture filtrate to inhibit the germination of Botrytis cinerea spores was increased by about 3-fold. In comparison to P1, the transgenic strain more quickly overgrew and lysed in vitro the pathogens Rhizoctonia solani and Pythium ultimum.

  18. Potential biocontrol agents for biofouling on artificial structures.

    Science.gov (United States)

    Atalah, Javier; Newcombe, Emma M; Hopkins, Grant A; Forrest, Barrie M

    2014-09-01

    The accumulation of biofouling on coastal structures can lead to operational impacts and may harbour problematic organisms, including non-indigenous species. Benthic predators and grazers that can supress biofouling, and which are able to be artificially enhanced, have potential value as augmentative biocontrol agents. The ability of New Zealand native invertebrates to control biofouling on marina pontoons and wharf piles was tested. Caging experiments evaluated the ability of biocontrol to mitigate established biofouling, and to prevent fouling accumulation on defouled surfaces. On pontoons, the gastropods Haliotis iris and Cookia sulcata reduced established biofouling cover by >55% and largely prevented the accumulation of new biofouling over three months. On wharf piles C. sulcata removed 65% of biofouling biomass and reduced its cover by 73%. C. sulcata also had better retention and survival rates than other agents. Augmentative biocontrol has the potential to be an effective method to mitigate biofouling on marine structures.

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

    Science.gov (United States)

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

    2015-09-01

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

  20. Biocontrol agents-mediated suppression of oxalic acid induced cell death during Sclerotinia sclerotiorum-pea interaction.

    Science.gov (United States)

    Jain, Akansha; Singh, Akanksha; Singh, Surendra; Sarma, Birinchi Kumar; Singh, Harikesh Bahadur

    2015-05-01

    Oxalic acid (OA) is an important pathogenic factor during early Sclerotinia sclerotiorum-host interaction and might work by reducing hydrogen peroxide production (H2 O2 ). In the present investigation, oxalic acid-induced cell death in pea was studied. Pea plants treated with biocontrol agents (BCAs) viz., Pseudomonas aeruginosa PJHU15, Bacillus subtilis BHHU100, and Trichoderma harzianum TNHU27 either singly and/or in consortium acted on S. sclerotiorum indirectly by enabling plants to inhibit the OA-mediated suppression of oxidative burst via induction of H2 O2 . Our results showed that BCA treated plants upon treatment with culture filtrate of the pathogen, conferred the resistance via. significantly decreasing relative cell death of pea against S. sclerotiorum compared to control plants without BCA treatment but treated with the culture filtrate of the pathogen. The results obtained from the present study indicate that the microbes especially in consortia play significant role in protection against S. sclerotiorum by modulating oxidative burst and partially enhancing tolerance by increasing the H2 O2 generation, which is otherwise suppressed by OA produced by the pathogen.

  1. Controle do mofo-branco com palhada e Trichoderma harzianum 1306 em soja White mold control with mulch and Trichoderma harzianum 1306 on soybean

    Directory of Open Access Journals (Sweden)

    Claudia Adriana Görgen

    2009-12-01

    Full Text Available O objetivo deste trabalho foi avaliar os efeitos da cobertura vegetal com palhada de Brachiaria ruziziensis, da aplicação de Trichoderma harzianum 1306 e da interação entre esses métodos na redução da densidade de inóculo de Sclerotinia sclerotiorum, na incidência de mofo-branco e na produtividade da soja. As avaliações foram realizadas entre 2006 e 2008 em Jataí, GO, em lavoura comercial, com média de 136,08 escleródios do patógeno m-2. Em março e outubro de 2006, foram aplicados 0, 0,5, 1 e 1,5 L ha-1 do inóculo de T. harzianum 1306 (2x10(9 esporos viáveis mL-1 em parcelas cultivadas com e sem B. ruziziensis. Após dessecação da braquiária com glifosato (2,5 L ha-1 e plantio da soja 'M-Soy 6101' sobre 10,1 Mg ha-1 de palhada, verificou-se redução de 98% de apotécios. Nos tratamentos com palhada e 0,5 e 1 L ha-1 de T. harzianum, verificou-se 72,1 e 84,1% de escleródios parasitados a mais, respectivamente. Em 2006/2007, o rendimento da soja 'M-Soy 6101' foi inversamente proporcional à população de apotécios por metro quadrado, porém 7,6% menor nos tratamentos com palha. Na safra 2007/2008, a incidência da doença em soja 'M-Soy 7908' foi menor nas parcelas com palhada e 0,5 e 1 L ha-1 de T. harzianum 1306, aplicados apenas no ano anterior. O biocontrole com T. harzianum 1306 em campo e uso da palhada de B. ruziziensis é eficiente e viável para o controle do mofo-branco da soja em áreas de Cerrado.The objective of this work was to evaluate the effects of crop cover, using Brachiaria ruziziensis mulch, Trichoderma harzianum 1306 spray treatment and of the interaction between these methods in reducing the density of Sclerotinia sclerotiorum inocula, in white mold incidence and in soybean yield. The evaluations were done from 2006 to 2008 in Jataí (Goiás State, Brazil, in a commercial crop with an average of 136.08 sclerotia m-2. In March and October 2006, 0, 0.5, 1 and 1.5 L ha-1 of T. harzianum 1306 (2x10

  2. Improved delivery of biocontrol Pseudomonas and their antifungal metabolites using alginate polymers.

    Science.gov (United States)

    Russo, A; Moënne-Loccoz, Y; Fedi, S; Higgins, P; Fenton, A; Dowling, D N; O'Regan, M; O'Gara, F

    1996-02-01

    Alginate polymer was evaluated as a carrier for seed inoculation with a genetically modified strain Pseudomonas fluorescens F113LacZY, which protects sugar-beet against Pythium-mediated damping-off. F113LacZY survived in alginate beads at 5 log10 CFU/bead or higher counts for 8 weeks of storage, regardless of the conditions of incubation. In plant inoculation experiments, colonisation of the growing area of the root by F113LacZY, derived from alginate beads placed in the soil next to the seed or from an alginate coating around the seeds, was improved compared with application of just free cells of the strain. F113LacZY trapped in alginate beads was an effective producer of antifungal phloroglucinols as indicated by direct HPLC quantification of phloroglucinols and in vitro inhibition of both the indicator bacterium Bacillus subtilis A1 and the pathogenic fungus Pythium ultimum. Alginate polymer represents a promising carrier for the delivery of biocontrol inoculants for root colonisation and production of antifungal metabolites. PMID:8867631

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

    Science.gov (United States)

    Druzhinina, I S; Kubicek, C P

    2016-01-01

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

  4. Metarhizium anisopliae and Trichoderma viride control colonies of Atta cephalotes in field better than a chemical insecticide

    Directory of Open Access Journals (Sweden)

    Elkin López Arismendy

    2011-12-01

    Full Text Available The leaf-cutting ant Atta cephalotes is an economically important pest in agriculture. These ants use the material they cut to cultivate a fungus from which they fed. In this study, baits prepared with the entomopathogenic fungus Metarhizium anisopliae strain M-137, Trichoderma viride strain T-26, antagonist of the symbiont fungus of A. cephaloies, and a combination of both fungi were applied in order to control A. cephalotes colonies under field conditions. In addition, the insecticide activity of the baits was compared to a chemical product, Pirimifos Metil, which was applied with an air pump. The ants did not detect the fungal agents contained in the baits, and introduced then into their nests without awakening defensive behaviors. The mortality of the bait-treated nests was higher than 80% while the treatment with Pirimifos Metil was effective in only 60% of the nests. Additionally, a week after the application of these treatments, changes in the insects' behavior were observed, reflected mainly in the absence of foraging activity. In conclusion, M. anisopliae was effective in controlling A. cephalotes colonies, and superior to the chemical product Pirimifos Metil. 

  5. Fungicidal compounds from a marine Ascidian-associated fungus Trichoderma harzianum

    Digital Repository Service at National Institute of Oceanography (India)

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

    -infesting fungal pathogen Penicillium griseofulvum. Bioassay guided chemical investigation of the ethyl acetate fraction yielded five compounds (1-5); two alpha-pyrone derivatives, a phenol, a sesquiterpene and an unsaturated hydroxy fatty acid as methylester...

  6. Extending the Use of Bioplastic Granules for the Application of Trichoderma Biocontrol Isolates in Flori/Horticulture and Turfgrass

    Science.gov (United States)

    Bioplastic materials are gaining increasing interest in a variety of different industrial and domestic applications. Beside its usage as mulching films and plant clips in horticulture, no other agricultural applications have been proposed. In 2009 we demonstrated that granules made of the bioplastic...

  7. Mycotoxigenic Fusarium and Deoxynivalenol Production Repress Chitinase Gene Expression in the Biocontrol Agent Trichoderma atroviride P1

    OpenAIRE

    Lutz, Matthias P.; Feichtinger, Georg; Défago, Geneviève; Duffy, Brion

    2003-01-01

    Mycotoxin contamination associated with head blight of wheat and other grains caused by Fusarium culmorum and F. graminearum is a chronic threat to crop, human, and animal health throughout the world. One of the most important toxins in terms of human exposure is deoxynivalenol (DON) (formerly called vomitoxin), an inhibitor of protein synthesis with a broad spectrum of toxigenicity against animals. Certain Fusarium toxins have additional antimicrobial activity, and the phytotoxin fusaric aci...

  8. Effects of Pesticides and Fungicides on Mycelium Growth of Trichoderma harzianum%杀虫剂和杀菌剂对哈茨木霉菌丝生长的影响

    Institute of Scientific and Technical Information of China (English)

    郎剑锋; 孔凡彬; 陆宁海; 杨蕊; 张燕林

    2011-01-01

    通过常规真菌生物量测定的方法,检测了新乡地区7种常见杀虫剂和3种杀菌剂对哈茨木霉(Trichoderma harzianum)菌丝生长的影响.结果表明,十种药剂对哈茨木霉的菌丝生长均有一定的抑制作用,且随着药剂浓度的增加抑制作用越明显,不同的药剂对哈茨木霉的毒力不同.当十种药剂在施用田间推荐剂量时,蛾蛉速杀、链霉素、惊喜一号和吡虫啉对木霉菌丝生长影响不大,可以和哈茨木霉混合使用.%Through use the conventional measurement method for fungus biomass to detect the effects of seven common pesticides and three fungicides in Xinxiang region on mycelium growth of Trichoderma harzianum. The results showed that the seven pesticides and the three fungicides all had a certain inhibitory effect on mycelium growth of Trichoderma harzianum, and the inhibitory effect increased with increasing concentration of pesticides or fungicides, but their toxicity on Trichoderma harzianum had differences. When applying the seven pesticides and the three fungicides with recommend dosage to the field, the 26% of Cyhalothrin + Phoxim EC, streptomycin, acetamiprid WP and 10% of Imidacloprid WP had few effects on mycelium growth of Trichoderma harzianum, so they could mix-uso with Trichoderma harzianum.

  9. Vision and development in Trichoderma atroviride

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

  10. Control of Botrytis cinerea in Eucalyptus globulus Mini-Cuttings Using Clonostachys and Trichoderma Strains Control de Botrytis cinerea en miniestacas de Eucalyptus globulus Utilizando Cepas de Clonostachys y Trichoderma

    Directory of Open Access Journals (Sweden)

    Salomé Zaldúa

    2010-12-01

    Full Text Available Botrytis cinerea Pers. ex Fr. causes the disease known as gray mold in more than 200 hosts. It is one of the most important pathogens in Chilean forest nurseries and Eucalyptus globulus Labill. is one of the most susceptible species, especially in vegetative reproduction systems. Clonostachys and Trichoderma strains were selected as potential biocontrol agents of gray mold in previous research by the authors. The objective of this study was to evaluate the effectiveness of antagonistic fungi to control B. cinerea in E. globulus mini-cuttings. Five fungi strains were tested and applied weekly, two Clonostachys and three Trichoderma (5 x 10(6 conidia mL-1. In addition, comparison treatments were also used: absolute control (water and fungicide application. The experiment was carried out under operational conditions to produce E. globulus mini-cuttings. The Clonostachys UDC-A10 and UDC-A11 strains reduce mini-cutting mortality caused by B. cinerea in 54 and 71%, respectively, and with effects similar to those achieved by fungicides. Clonostachys UDC-A11 reduces the disease progression rate with the same statistical results as fungicides. A negative effect of applying fungicides on rooting of the surviving mini-cuttings was also confirmed. These results demonstrate the effectiveness of Clonostachys as a control agent against gray mold disease in E. globulus mini-cuttings.Botrytis cinerea Pers. ex Fr. ocasiona la enfermedad conocida como moho gris en más de 200 hospederos. En Chile es uno de los patógenos más importantes en viveros forestales, siendo Eucalyptus globulus Labill. una de las especies más susceptibles, especialmente en los sistemas de reproducción vegetativa. En investigaciones previas, realizadas por los autores, se seleccionaron cepas de Clonostachys y Trichoderma como potenciales agentes de biocontrol del moho gris. El objetivo fue evaluar la eficacia de hongos antagonistas en el control de B. cinerea en mini-estacas de E

  11. Challenges and opportunities for the commercialization of postharvest biocontrol

    Science.gov (United States)

    The past twenty years has seen the field of postharvest biocontrol evolve into a sophisticated science with global research programs worldwide, numerous yearly publications, patented technologies, and the development of new commercial products. The use of these products, however, still remains limi...

  12. Screening of biocontrol agents for control of foliar diseases

    NARCIS (Netherlands)

    Köhl, J.

    2009-01-01

    Candidate antagonists for the development of biocontrol agents have to fulfill many criteria. The criterion often investigated first in detail is the antagonistic potential of candidates against the target pathogen. However, candidates must also have high ecological competence, must be suitable for

  13. Grouper as a natural biocontrol of invasive lionfish.

    Science.gov (United States)

    Mumby, Peter J; Harborne, Alastair R; Brumbaugh, Daniel R

    2011-01-01

    Lionfish (Pterois volitans/miles) have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean. PMID:21731769

  14. Grouper as a natural biocontrol of invasive lionfish.

    Directory of Open Access Journals (Sweden)

    Peter J Mumby

    Full Text Available Lionfish (Pterois volitans/miles have invaded the majority of the Caribbean region within five years. As voracious predators of native fishes with a broad habitat distribution, lionfish are poised to cause an unprecedented disruption to coral reef diversity and function. Controls of lionfish densities within its native range are poorly understood, but they have been recorded in the stomachs of large-bodied Caribbean groupers. Whether grouper predation of lionfish is sufficient to act as a biocontrol of the invasive species is unknown, but pest biocontrol by predatory fishes has been reported in other ecosystems. Groupers were surveyed along a chain of Bahamian reefs, including one of the region's most successful marine reserves which supports the top one percentile of Caribbean grouper biomass. Lionfish biomass exhibited a 7-fold and non-linear reduction in relation to the biomass of grouper. While Caribbean grouper appear to be a biocontrol of invasive lionfish, the overexploitation of their populations by fishers, means that their median biomass on Caribbean reefs is an order of magnitude less than in our study. Thus, chronic overfishing will probably prevent natural biocontrol of lionfishes in the Caribbean.

  15. Trichoderma longibrachiatum Evx1 is a fungal biocatalyst suitable for the remediation of soils contaminated with diesel fuel and polycyclic aromatic hydrocarbons.

    Science.gov (United States)

    Andreolli, Marco; Lampis, Silvia; Brignoli, Pierlorenzo; Vallini, Giovanni

    2016-05-01

    Trichoderma sp. strain Evx1 was isolated from a semi-deciduous forest soil in Southern Italy. It decolorizes polynuclear organic dyes and tolerates high concentrations of phenanthrene, anthracene, fluoranthene, and pyrene. The ability of this ascomycete fungus to degrade polycyclic aromatic hydrocarbons was verified in vitro and confirmed by its strong phenoloxidase activity in the presence of gallic acid. Phylogenetic characterization of Trichoderma sp. Evx1 positioned this strain within the species Trichoderma longibrachiatum. The potential use of this species for the bioremediation of contaminated environmental matrices was tested by inoculating diesel-spiked soil with a dense mycelial suspension. The biodegradation percentage of the C12-40 hydrocarbon fraction in the inoculated soil rose to 54.2 ± 1.6 %, much higher than that in non-inoculated soil or soil managed solely by a combination of watering and aeration. The survival and persistence of T. longibrachiatum Evx1 throughout the bioremediation trial was monitored by PCR-DGGE analysis. The fungal strain was still present in the soil 30 days after bioaugmentation. These findings indicate that T. longibrachiatum Evx1 may be a suitable inoculum in bioremediation protocols for the reclamation of soils contaminated by complex mixtures of hydrocarbons. PMID:26832871

  16. Exopolysaccharide from Trichoderma pseudokoningii induces macrophage activation.

    Science.gov (United States)

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

    2016-09-20

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

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

    Science.gov (United States)

    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. PMID:26747625

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

    Science.gov (United States)

    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.

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

    OpenAIRE

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

    2015-01-01

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

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

    Science.gov (United States)

    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. PMID:26378961

  1. Bioaccumulation of mycotoxins by shellfish: contamination of mussels by metabolites of a Trichoderma koningii strain isolated in the marine environment.

    Science.gov (United States)

    Sallenave, C; Pouchus, Y F; Bardouil, M; Lassus, P; Roquebert, M F; Verbist, J F

    1999-01-01

    To determine whether toxic metabolites produced by fungi could cause shellfish toxicities, mussels were contaminated in laboratory conditions by sterile filtrates of a liquid culture of a strain of the fungus Trichoderma koningii previously isolated from a shellfish, the cockle (Cerastoderma edule). Mussels were kept in aerated natural seawater and fed with a culture of the microalga Isochrysis galbana, to which a filtrate of liquid fungal culture was added. Mussels were exposed to contamination for 7 days at 16 or 20 degrees C and extractions were then performed and their activity tested on blowfly larvae. The same toxicity was found in the fungal filtrate and the shellfish, indicating bioaccumulation. The digestive gland was the most toxic part of the mussel, confirming contamination by filtration. Treated mussels produced a mucus which appeared to be a means of eliminating toxic metabolites. PMID:9920482

  2. 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

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

    Directory of Open Access Journals (Sweden)

    Febrilia Nur ‘Aini

    2015-11-01

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

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

    Directory of Open Access Journals (Sweden)

    Denton Jai A

    2011-11-01

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

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

    Science.gov (United States)

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

    2015-04-15

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

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

    Directory of Open Access Journals (Sweden)

    Hudson Teixeira

    2012-12-01

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

  7. Is the High Cu Tolerance of Trichoderma atroviride Isolated from the Cu-Polluted Sediment Due to Adaptation? An In Vitro Toxicological Study

    International Nuclear Information System (INIS)

    The tolerance of Cu by Trichoderma atroviride, a tolerant fungus isolated from the drainage surface sediment of the Serdang Industrial Area was investigated under in vitro conditions. Only this fungus species can tolerate up to 600 mg/ L of Cu on solid medium Potato Dextrose Agar based on the isolation of the most tolerant fungus from the polluted sediment. Toxicity test performed on T. atroviride, showed a maximum tolerance at 300 mg/L of Cu concentration when grown in liquid medium Potato Dextrose Broth (PDB). The EC50 value of the isolate was 287.73 mg/ L of Cu concentration in PDB. The Cu concentration in the drainage surface sediment, where the T. atroviride was isolated from, was 347.64 μg/ g while the geochemical distributions of the non-resistant and resistant fractions of Cu were 99.6 and 0.4 %, respectively. The sediment data indicated that the drainage had greatly received anthropogenic Cu from the nearby industries which are involved in the manufacturing of plastics and electronic products. The present findings indicate that the high Cu tolerance showed by T. atroviride could be due to the well adaptation of the fungus to the Cu polluted sediment. Therefore, T. atroviride could be a potential bioremediator of Cu pollution in the freshwater ecosystem. (author)

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

    Directory of Open Access Journals (Sweden)

    Febri Doni

    2014-05-01

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

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

    Directory of Open Access Journals (Sweden)

    M.R. Reis

    2013-06-01

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

  10. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Directory of Open Access Journals (Sweden)

    Suchada Chantrapromma

    2014-04-01

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

  11. Trichodermaerin: a diterpene lactone from Trichoderma asperellum

    Science.gov (United States)

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

    2014-01-01

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

  12. In-Vitro Efficacy of Trichoderma viride Against Sclerotium rolfsii and Macrophomina phaseolina

    Directory of Open Access Journals (Sweden)

    Khirood DOLEY

    2012-11-01

    Full Text Available The fungal pathogen causes serious widespread losses to agricultural crops worldwide. Therefore, economy of countries may worsen especially of developing countries. In addition, harmful chemical pesticides which are being used today for increasing crop production creates very serious health hazardous problems to human beings and ecosystem as a whole. The antagonistic potential of Trichoderma species which has been long known to control various soil-borne fungal pathogens in biological way may be utilized. The faster growth rates with which it competes with fungal pathogen mainly brings upon their antagonistic characteristics. An investigation was carried out in laboratory condition towards biological efficacy of T. viride on potato dextrose agar (PDA medium for the bio-control of soil-borne plant pathogens Sclerotium rolfsii and Macrophomina phaseolina in in-vitro condition. The dual culture technique was followed in which T. viride showed significant antifungal activities towards both the pathogens. T. viride significantly inhibited the mycelial radial growth of S. rolfsii by 75% and M. phaseolina by 71.42%. The results showed variable mycelial growth rate for all fungal isolates which was determined after 6 days of incubation in which T. viride showed minimum of 4.00 days to completely cover the petri-plates and S. rolfsii showed 4.33 days whereas M. phaseolina showed 6.33 days. Thus, T. viride showed encouraging results regarding their biopesticidal and biofungicidal potential against plant pathogens which may be endorsed to substitute harmful chemical supplements that exists in modern day agricultural practices.

  13. Colonization pattern of the biocontrol strain Pseudomonas chlororaphis MA 342 on barley seeds visualized by using green fluorescent protein.

    Science.gov (United States)

    Tombolini, R; van der Gaag, D J; Gerhardson, B; Jansson, J K

    1999-08-01

    Pseudomonas chlororaphis MA 342 is a potent biocontrol agent that can be used against several seed-borne diseases of cereal crops, including net blotch of barley caused by the fungus Drechslera teres. In this study, strain MA 342 was tagged with the gfp gene (encoding the green fluorescent protein) in order to study the fate of cells after seed inoculation. The gfp-tagged strain, MA 342G2, had the same biocontrol efficacy as the wild type when it was applied at high cell concentrations to seeds but was less effective at lower cell concentrations. By comparing cell counts determined by microscopy to the number of CFU, we found that the number of culturable cells was significantly lower than the total number of bacteria on seeds which were inoculated and dried for 20 h. Confocal microscopy and epifluorescence stereomicroscopy were used to determine the pattern of MA 342G2 colonization and cell aggregation on barley seeds. Immediately after inoculation of seeds, bacteria were found mainly under the seed glume, and there was no particular aggregation pattern. However, after the seeds were sown, irregularly distributed areas of bacterial aggregation were found, which reflected epiphytic colonization of glume cells. There was a trend towards bacterial aggregation near the embryo but never within the embryo. Bacterial aggregates were regularly found in the groove of each seed formed by the base of the coleoptile and the scutellum. Based on these results, we suggest that MA 342 colocalizes with the pathogen D. teres, which facilitates the action of the fungistatic compound(s) produced by this strain.

  14. Isolation, antimicrobial activity, and metabolites of fungus Cladosporium sp. associated with red alga Porphyra yezoensis.

    Science.gov (United States)

    Ding, Ling; Qin, Song; Li, Fuchao; Chi, Xiaoyuan; Laatsch, Hartmut

    2008-03-01

    Cladosporium sp. isolate N5 was isolated as a dominant fungus from the healthy conchocelis of Porphyra yezoensis. In the re-infection test, it did not cause any pathogenic symptoms in the alga. Twenty-one cultural conditions were chosen to test its antimicrobial activity in order to obtain the best condition for large-scale fermentation. Phenylacetic acid, p-hydroxyphenylethyl alcohol, and L-beta-phenyllactic acid were isolated from the crude extract as strong antimicrobial compounds and they are the first reported secondary metabolites for the genus Cladosporium. In addition, the Cladosporium sp. produced the reported Porphyra yezoensis growth regulators phenylacetic acid and p-hydroxyphenylacetic acid. No cytotoxicity was found in the brine shrimp lethality test, which indicated that the environmental-friendly Cladosporium sp. could be used as a potential biocontrol agent to protect the alga from pathogens.

  15. Fermentation and microencapsulation of the nematophagous fungus Hirsutella rhossiliensis in a novel type of hollow beads.

    Science.gov (United States)

    Patel, Anant V; Jakobs-Schönwandt, Desiree; Rose, Thomas; Vorlop, Klaus-Dieter

    2011-03-01

    In this work, fermentation and formulation aspects of the nematophagous fungus Hirsutella rhossiliensis BBA were investigated. When incubated in 2% (w/w) glucose and 0.5% (w/w) yeast extract medium in a 1-L Erlenmeyer flask without baffles, heavy pellet formation was observed. Only 40% of the mycelium had a size less than 500 μm. When a flask with three baffles was used, the portion of mycelium fermentation results were obtained at 0.4 vvm and 400 rpm stirrer speed with 90% mycelium gluten and 0.5% (w/w) yeast extract could be replaced with 3% (w/w) autoclaved baker's yeast which was never used as capsule additive before. Radial growth of mycelium out of dried hollow beads containing 1% (w/w) biomass and 3% (w/w) baker's yeast was faster than for alginate beads containing equivalent amounts of biomass and yeast indicating a higher bio-control potential. PMID:21181150

  16. Phages of Listeria offer novel tools for diagnostics and biocontrol

    Directory of Open Access Journals (Sweden)

    Martin J Loessner

    2014-04-01

    Full Text Available Historically, bacteriophages infecting their hosts have perhaps been best known and even notorious for being a nuisance in dairy-fermentation processes. However, with the rapid progress in molecular microbiology and microbial ecology, a new dawn has risen for phages. This review will provide an overview on possible uses and applications of Listeria phages, including phage-typing, reporter phage for bacterial diagnostics, and use of phage as biocontrol agents for food safety. The use of phage-encoded enzymes such as endolysins for the detection and as antimicrobial will also be addressed. Desirable properties of candidate phages for biocontrol will be discussed. While emphasizing the enormous future potential for applications, we will also consider some of the intrinsic limitations dictated by both phage and bacterial ecology.

  17. Novel routes for improving biocontrol activity of Bacillus based bioinoculants

    Directory of Open Access Journals (Sweden)

    Liming eWu

    2015-12-01

    Full Text Available Biocontrol formulations prepared from plant-growth-promoting bacteria are increasingly applied in sustainable agriculture. Especially inoculants prepared from endospore-forming Bacillus strains have been proven as efficient and environmental-friendly alternative to chemical pesticides due to their long shelf life, which is comparable with that of agrochemicals. However, these formulations of the first generation are sometimes hampered in their action and do not fulfill in each case the expectations of the appliers. In this review we use the well-known plant-associated Bacillus amyloliquefaciens type strain FZB42 as example for the successful application of different techniques offered today by comparative, evolutionary and functional genomics, site-directed mutagenesis and strain construction including marker removal, for paving the way for preparing a novel generation of biocontrol agents.

  18. 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

  19. Cotton Seedling Preemergence Damping-Off Incited by Rhizopus oryzae and Pythium spp. and Its Biological Control with Trichoderma spp.

    Science.gov (United States)

    Howell, C R

    2002-02-01

    ABSTRACT Planting the cotton cv. Sure-Grow 747 in cotton seedling disease plots during the 2001 growing season resulted in high levels of preemergence damping-off among the seedlings. Four cotton pathogens, Pythium aphanidermatum, P. ultimum, an unidentified Pythium sp., and Rhizopus oryzae, were isolated from diseased seed embryos and seedlings. Disease incited by the Pythium spp. could be controlled by seed treatment with Metalaxyl, but disease incited by R. oryzae could not. Seed treatment with Metalaxyl in naturally infested field soil was only partially effective; therefore, symptoms in 47% of the diseased seedlings could be attributed to R. oryzae. Susceptibility to disease appeared to be related to release in the spermosphere, by the germinating seeds, of compounds that stimulate pathogen propagule germination, because exudates from seed of the suscept Sure-Grow 747 and extracts from wheat bran induced pathogen germination and growth, whereas exudates from resistant cv. Stoneville 213 did not. However, even Stoneville 213 became susceptible when infested soil was amended with wheat bran. Seed treatment with preparations of Trichoderma virens parent, mutant, and hybrid strains gave effective biological control of preemergence damping-off. Disease control was attributable to metabolism by the biocontrol agent of pathogen germination stimulants released by the seed, because amendment of pathogen-infested soil with the propagule germination stimulants in wheat bran negated the protective effect of the seed treatment. PMID:18943091

  20. Environmental impact assessment of genetically modified biocontrol agents

    International Nuclear Information System (INIS)

    This review summarises the theoretical basis of risk analysis, and the political and social implications of introducing new biotechnology products in agricultural environments. The main factors to be considered under the present European regulation in the environmental impact assessment of genetically modified biocontrol agents are briefly discussed. Finally, an alternative risk assessment paradigm is proposed for genetically modified microorganisms, which shall consider the intrinsic properties of each antagonist, rather than the method used for generating it

  1. Biocontrol: Bacillus penetrans and Related Parasites of Nematodes

    OpenAIRE

    Sayre, R. M.

    1980-01-01

    Bacillus penetrans Mankau, 1975, previously described as Duboscqia penetrans Thorne 1940, is a candidate agent for biocontrol of nematodes. This review considers the life stages of this bacterium: vegetative growth phase, colony fragmentation, sporogenesis, soil phase, spore attachment, and penetration into larvae of root-knot nematodes. The morphology of the microthallus colonies and the unusual external features of the spore are discussed. Taxonomic affinities with the actinomycetes, partic...

  2. Genome Sequence of the Biocontrol Strain Pseudomonas fluorescens F113

    OpenAIRE

    Redondo-Nieto, M.; M. Barret; Morrisey, J; Germaine, K.; Martínez-Granero, F.; Barahona, E.; Navazo, A.; Sánchez-Contreras, M.; Moynihan, J.; Giddens, S.; Coppoolse, E.; Muriel, C.; Stiekema, W.; Rainey, P; Dowling, D

    2012-01-01

    Pseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) that has biocontrol activity against fungal plant pathogens and is a model for rhizosphere colonization. Here, we present its complete genome sequence, which shows that besides a core genome very similar to those of other strains sequenced within this species, F113 possesses a wide array of genes encoding specialized functions for thriving in the rhizosphere and interacting with eukaryotic organisms.

  3. Biocontrol of Botrytis cinerea by successful introduction of Pantoea ananatis in the grapevine phyllosphere

    Directory of Open Access Journals (Sweden)

    Gasser F

    2012-12-01

    Full Text Available Florian Gasser,1 Massimiliano Cardinale,1 Barbara Schildberger,2 Gabriele Berg11Institute of Environmental Biotechnology, Graz University of Technology, Graz, Austria; 2Höhere Bundesanstalt und Bundesamt für Wein-und Obstbau, Klosterneuburg, AustriaBackground and aims: The fungus Botrytis cinerea is a common problem in viticulture and leads to serious losses in both yield and quality. The objective was to study the potential of the antagonist Pantoea ananatis BLBT1-08 for controlling this disease.Methods: Pathogen suppression by Pantoea treatments was investigated in different field trials and in detached leaf assays. The mode of action was studied by confocal laser scanning microscopy of treated grape leaves and by in vitro assays.Results: The introduction of P. ananatis BLBT1-08 in a 3-year field trial resulted in statistically significant reduction of disease symptoms. However, B. cinerea abundance, measured by quantitative real-time polymerase chain reaction of a B. cinerea specific gene, was not reduced when compared to non-treated, symptom-free leaves. A DsRed fluorescent protein labeled BLBT1-08 strain showed a high phyllosphere competence and competition on the leaf surface, but did not colonize the inner parts of plant tissue. Germination of B. cinerea was not inhibited by BLBT1-08 on the leaf, but mycelial growth and symptoms were suppressed without direct pathogen-antagonist contact. The antimicrobial activity was amino acid and temperature dependent.Conclusion: P. ananatis BLBT1-08 is a competitive and promising biocontrol agent for the control of B. cinerea and is highly effective at reducing disease incidence.Keywords: biological control, sustainable viticulture, antagonism

  4. 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.

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

    Science.gov (United States)

    Zhu, Z X; Zhuang, W Y

    2015-06-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-02

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

  7. Dual utility of a novel, copper enhanced laccase from Trichoderma aureoviridae.

    Science.gov (United States)

    Khambhaty, Yasmin; Ananth, Swetha; Sreeram, Kalarical Janardhanan; Rao, Jonnalagadda Raghava; Nair, Balachandran Unni

    2015-11-01

    Ever since the ability of laccase to oxidize non-phenolic lignin models was described, the oxidative degradation reactions catalyzed by laccase have been widely studied for paper pulp production or detoxification of aromatic pollutants. The viability of developing eco-friendly, laccase aided industrial processes has been explored. Here, we report the isolation and screening of fungi to explore their lignolytic ability on solid media using various substrates as indicators. The promising fungus was cultivated in submerged and solid state conditions. The crude enzyme obtained yielded elevated activity at 75°C and pH 9.0. Addition of CuSO4 increased the activity by almost 25% proving that Cu(2+) catalytically enhances the action of laccases. Decolorization studies were carried out using industrial dye, Remazol Brilliant Blue R (CI 61200) on solid and liquid medium. Visual decolorization was observed within 2 days of inoculation on solid media whereas, liquid medium incorporated with varying concentrations of dye solution showed a final level of decolorization of up to 76%. Bamboo degradation studies revealed a decrease in lignin content by 51 and 43% within a month. To the best of our knowledge, this study for the first time reports that Trichoderma aureoviridae can produce lignolytic enzyme and degrade lignin. PMID:26231326

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

    Science.gov (United States)

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

    2016-02-01

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

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

    Science.gov (United States)

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

    2014-09-01

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

  10. The neutral metallopeptidase NMP1 of Trichoderma guizhouense is required for mycotrophy and self-defence.

    Science.gov (United States)

    Zhang, Jian; Bayram Akcapinar, Gunseli; Atanasova, Lea; Rahimi, Mohammad Javad; Przylucka, Agnieszka; Yang, Dongqing; Kubicek, Christian P; Zhang, Ruifu; Shen, Qirong; Druzhinina, Irina S

    2016-02-01

    Trichoderma guizhouense NJAU 4742 (Harzianum clade) can suppress the causative agent of banana wild disease Fusarium oxysporum f. sp. cubense 4 (Foc4). To identify genes involved in this trait, we used T-DNA insertional mutagenesis and isolated one mutant that was unable to overgrow Foc4 and had reduced antifungal ability. Using the high-efficiency thermal asymmetric interlaced-PCR, the T-DNA was located in the terminator of a neutral metalloprotease gene (encoding a MEROPS family M35 protease), which was named nmp1. The antifungal activity of the mutant was recovered by retransformation with wild-type nmp1 gene. The purified NMP1 (overexpressed in Pichia pastoris) did not inhibit the growth and germination of other fungi in vitro. Its addition, however, partly recovered the antifungal activity of the mutant strain against some fungi. The expression of nmp1 is induced by the presence of fungi and by dead fungal biomass, but the time-course of transcript accumulation following the physical contact depends on mode of interaction: it increases in cases of long-lasting parasitism and decreases if the prey fungus is dead shortly after or even before the contact (predation). We thus conclude that NMP1 protein of T. guizhouense has major importance for mycotrophic interactions and defence against other fungi. PMID:26118314

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

    Science.gov (United States)

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

    2016-02-01

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

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

    Directory of Open Access Journals (Sweden)

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

    2010-01-01

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

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

    Science.gov (United States)

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

    1993-02-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Institute of Scientific and Technical Information of China (English)

    张广志; 杨合同; 张新建; 李纪顺; 陈凯; 黄玉杰

    2013-01-01

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

  16. 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.

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

    Science.gov (United States)

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

    2016-08-01

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

  18. Study on Biological Control Of Rhizoctonia solani via Trichoderma

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

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

  19. The relationship of biofilm production to biocontrol activity of Burkholderia pyrrocinia FP62

    Science.gov (United States)

    Foliar biocontrol agent (BCA) efficacy is often inconsistent due to poor colonization and survival on plant surfaces. Burkholderia pyrrocinia FP62, a superior leaf colonist and BCA of Botrytis cinerea, forms unsaturated biofilms on plant surfaces. To determine the relationship between biocontrol act...

  20. Yeast microflora of nectarines and their potential for biocontrol of brown rot

    Science.gov (United States)

    Resident fruit microflora has been the source of biocontrol agents for the control of postharvest decays of fruits and the active ingredient in commercialized biocontrol products. With the exception of grapes and apples, information on the resident microflora of other fruits is only fragmentary; ho...

  1. Efficacy of Pseudomonas fluorescens for biocontrol of Escherichia coli 0157:H7 on spinach

    Science.gov (United States)

    Control of foodborne pathogens on leafy green vegetables is crucial for consumer food safety. Biocontrol microbes may inhibit or suppress foodborne pathogen growth and enhance existing post-harvest intervention controls. The efficacy of Pseudomonas fluorescens for biocontrol of Escherichia coli O157...

  2. Mutagenesis of Trichoderma Viride by Ultraviolet and Plasma

    Institute of Scientific and Technical Information of China (English)

    姚日生; 李曼曼; 邓胜松; 胡华佳; 王淮; 李凤和

    2012-01-01

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

  3. Mutagenesis of Trichoderma Viride by Ultraviolet and Plasma

    Science.gov (United States)

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

    2012-04-01

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

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

    Science.gov (United States)

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

    2015-04-15

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

  5. Potencialidade antagonística detectada em alguns procariotas agentes de biocontrole de enfermidades de plantas Antagonistic potentiality in prokaryotic biocontrol agents for plant diseases

    Directory of Open Access Journals (Sweden)

    Victor Rafael Barra

    2008-06-01

    Full Text Available Ao longo de vários anos, no Laboratório de Bacteriologia de Plantas e Controle Biológico (LBPCB do Departamento de Fitopatologia da Universidade Federal de Viçosa têm-se, sistematicamente, isolado, de rizosfera, rizoplano e filoplano de plantas cultivadas de importância econômica, milhares de procariotas os quais são testados, um a um, como agentes de biocontrole de enfermidades. A grande maioria deles, como esperado, não exibe qualquer potencialidade como agente de biocontrole e é descartada. Alguns poucos são selecionados, através de demorados ensaios de biocontrole experimental tanto em casa de vegetação como em campo. Neste trabalho, mostra-se que muitos dos organismos em estudo, mesmo havendo sido selecionados, principalmente, por induzirem resistência sistêmica na planta alvo, expressam constitutivamente mecanismos de antagonismo microbiano que parecem fazer parte de seu perfil enquanto agentes de biocontrole.Along several years, at the Laboratory of Plant Bacteriology and Biological Control of the Department of Plant Pathology of the Federal University of Viçosa, thousands of prokaryote isolates have been obtained from the rhizosphere, rhizoplane and phylloplane of economically important plants. Every obtained isolate is tested as biocontrol agents. As expected, the majority does not show any biocontrol potentiality and is discarded. Few are selected through time-consuming experimental biocontrol trials in greenhouse and field. Even being selected mainly for their ability to induce resistance in target plants, many of them constitutively express mechanisms of antagonism against pathogens that seem to be part of their performance while biocontrol agents.

  6. Isolation of Stem rot Disease Causing Organism of Brinjal and their in-vitro Inhibition with Fungicides and Bio-control Agents

    Directory of Open Access Journals (Sweden)

    Shaily Javeria

    2014-09-01

    Full Text Available Different strains of Sclerotinia sclerotiorum were isolated from the diseased samples collected from different hosts and locations. Among the 14 isolates, 12 isolates colonies covered the entire Petri plates within 96 hours but, two isolates from fababean and yellow mustard showed slow colony growth within 96 hours. All isolates produced sclerotia which were varied in number, but the fenugreek isolate produced maximum (43 number of sclerotia and lambs quarter isolate produced minimum number of sclerotia (12 on PDA medium. To examine inhibitory effect of fungicide on the mycelial growth of the pathogen, 9 fungicides were tested in vitro against Sclerotinia sclerotiorum, of those carbendazim, carboxin, topsin-M and carbendazim+ mancozeb (SAAF were found most effective and inhibited the mycelial growth of pathogen up to 100 per cent at 0.05%, 0.1%, and 0.2% concentration. The effect of different bioagents viz., Trichoderma harzianum, T. viride, T. koningii, T. atroviride, T. longibraciatum, Aspergillus niger, Chaetomium globosome and Penicillium notatum in inhibiting the growth of Sclerotinia sclerotiorum was studied through “Dual Culture Technique”. The data showed that among the eight biocontrol agent six were fond effective. The maximum inhibition was found by T. harzianum causing 70.82% inhibition of mycelial growth of the pathogen S. sclerotiorum.

  7. Studies on the management of root-knot nematode, Meloidogyne incognita-wilt fungus, Fusarium oxysporum disease complex of green gram, Vigna radiata cv ML-1108

    Institute of Scientific and Technical Information of China (English)

    HASEEB Akhtar; SHARMA Anita; SHUKLA Prabhat Kumar

    2005-01-01

    Studies were conducted under pot conditions to determine the comparative efficacy of carbofuran at 1 mg a.i./kg soil,bavistin at 1 mg a.i./kg soil, neem (Azadirachta indica) seed powder at 50 mg/kg soil, green mould (Trichoderma harzianum) at 50.0 ml/kg soil, rhizobacteria (Pseudomonas fluorescens) at 50.0 ml/kg soil against root-knot nematode, Meloidogyne incognita-wilt fungus, Fusarium oxysporum disease complex on green gram, Vigna radiata cv ML-1108. All the treatments significantly improved the growth of the plants as compared to untreated inoculated plants. Analysis of data showed that carbofuran and A. indica seed powder increased plant growth and yield significantly more in comparison to bavistin and P.fluorescens. Carbofuran was highly effective against nematode, bavistin against fungus, A. indica seed powder against both the pathogens and both the bioagents were moderately effective against both the pathogens.

  8. 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...

  9. Chitosan enhances parasitism of Meloidogyne javanica eggs by the nematophagous fungus Pochonia chlamydosporia.

    Science.gov (United States)

    Escudero, Nuria; Ferreira, Sebastião R; Lopez-Moya, Federico; Naranjo-Ortiz, Miguel A; Marin-Ortiz, Ana I; Thornton, Christopher R; Lopez-Llorca, Luis V

    2016-04-01

    Pochonia chlamydosporia (Pc), a nematophagous fungus and root endophyte, uses appressoria and extracellular enzymes, principally proteases, to infect the eggs of plant parasitic nematodes (PPN). Unlike other fungi, Pc is resistant to chitosan, a deacetylated form of chitin, used in agriculture as a biopesticide to control plant pathogens. In the present work, we show that chitosan increases Meloidogyne javanica egg parasitism by P. chlamydosporia. Using antibodies specific to the Pc enzymes VCP1 (a subtilisin), and SCP1 (a serine carboxypeptidase), we demonstrate chitosan elicitation of the fungal proteases during the parasitic process. Chitosan increases VCP1 immuno-labelling in the cell wall of Pc conidia, hyphal tips of germinating spores, and in appressoria on infected M. javanica eggs. These results support the role of proteases in egg parasitism by the fungus and their activation by chitosan. Phylogenetic analysis of the Pc genome reveals a large diversity of subtilisins (S8) and serine carboxypeptidases (S10). The VCP1 group in the S8 tree shows evidence of gene duplication indicating recent adaptations to nutrient sources. Our results demonstrate that chitosan enhances Pc infectivity of nematode eggs through increased proteolytic activities and appressoria formation and might be used to improve the efficacy of M. javanica biocontrol. PMID:27020158

  10. Saccharification of Lignocelluloses by Carbohydrate Active Enzymes of the White Rot Fungus Dichomitus squalens.

    Directory of Open Access Journals (Sweden)

    Johanna Rytioja

    Full Text Available White rot fungus Dichomitus squalens is an efficient lignocellulose degrading basidiomycete and a promising source for new plant cell wall polysaccharides depolymerizing enzymes. In this work, we focused on cellobiohydrolases (CBHs of D. squalens. The native CBHI fraction of the fungus, consisting three isoenzymes, was purified and it maintained the activity for 60 min at 50°C, and was stable in acidic pH. Due to the lack of enzyme activity assay for detecting only CBHII activity, CBHII of D. squalens was produced recombinantly in an industrially important ascomycete host, Trichoderma reesei. CBH enzymes of D. squalens showed potential in hydrolysis of complex lignocellulose substrates sugar beet pulp and wheat bran, and microcrystalline cellulose, Avicel. Recombinant CBHII (rCel6A of D. squalens hydrolysed all the studied plant biomasses. Compared to individual activities, synergistic effect between rCel6A and native CBHI fraction of D. squalens was significant in the hydrolysis of Avicel. Furthermore, the addition of laccase to the mixture of CBHI fraction and rCel6A significantly enhanced the amount of released reducing sugars from sugar beet pulp. Especially, synergy between individual enzymes is a crucial factor in the tailor-made enzyme mixtures needed for hydrolysis of different plant biomass feedstocks. Our data supports the importance of oxidoreductases in improved enzyme cocktails for lignocellulose saccharification.

  11. Saccharification of Lignocelluloses by Carbohydrate Active Enzymes of the White Rot Fungus Dichomitus squalens.

    Science.gov (United States)

    Rytioja, Johanna; Hildén, Kristiina; Mäkinen, Susanna; Vehmaanperä, Jari; Hatakka, Annele; Mäkelä, Miia R

    2015-01-01

    White rot fungus Dichomitus squalens is an efficient lignocellulose degrading basidiomycete and a promising source for new plant cell wall polysaccharides depolymerizing enzymes. In this work, we focused on cellobiohydrolases (CBHs) of D. squalens. The native CBHI fraction of the fungus, consisting three isoenzymes, was purified and it maintained the activity for 60 min at 50°C, and was stable in acidic pH. Due to the lack of enzyme activity assay for detecting only CBHII activity, CBHII of D. squalens was produced recombinantly in an industrially important ascomycete host, Trichoderma reesei. CBH enzymes of D. squalens showed potential in hydrolysis of complex lignocellulose substrates sugar beet pulp and wheat bran, and microcrystalline cellulose, Avicel. Recombinant CBHII (rCel6A) of D. squalens hydrolysed all the studied plant biomasses. Compared to individual activities, synergistic effect between rCel6A and native CBHI fraction of D. squalens was significant in the hydrolysis of Avicel. Furthermore, the addition of laccase to the mixture of CBHI fraction and rCel6A significantly enhanced the amount of released reducing sugars from sugar beet pulp. Especially, synergy between individual enzymes is a crucial factor in the tailor-made enzyme mixtures needed for hydrolysis of different plant biomass feedstocks. Our data supports the importance of oxidoreductases in improved enzyme cocktails for lignocellulose saccharification. PMID:26660105

  12. Chaetochromones A and B, two new polyketides from the fungus Chaetomium indicum (CBS.860.68).

    Science.gov (United States)

    Lu, Keyang; Zhang, Yisheng; Li, Li; Wang, Xuewei; Ding, Gang

    2013-01-01

    Chaetochromones A (1) and B (2), two novel polyketides, were isolated from the crude extract of fungus Chaetomium indicum (CBS.860.68) together with three known analogues PI-3(3), PI-4 (4) and SB236050 (5). The structures of these compounds were determined by HRESI-MS and NMR experiments. Chaetochromones A (1) and B (2) are a member of the polyketides family, which might originate from a similar biogenetic pathway as the known compounds PI-3 (3), PI-4 (4) and SB236050 (5). The biological activities of these secondary metabolites were evaluated against eight plant pathogens, including Alternaria alternata, Ilyonectria radicicola, Trichoderma viride pers, Aspergillus niger, Fusarium verticillioide, Irpex lacteus (Fr.), Poria placenta (Fr.) Cooke and Coriolus versicolor (L.) Quél. Compound 1 displayed moderate inhibitory rate (>60%) against the brown rot fungus Poria placenta (Fr.) Cooke, which causes significant wood decay. In addition, the cytotoxic activities against three cancer cell lines A549, MDA-MB-231, PANC-1 were also tested, without any inhibitory activities being detected. PMID:24013408

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

    DEFF Research Database (Denmark)

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

    2001-01-01

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

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

    Science.gov (United States)

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

    2014-02-01

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

  15. Biocontrol of the Brown-banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae, with Entomopathogenic Fungus, Metharhizium anisopliae

    Directory of Open Access Journals (Sweden)

    Mona Sharififard

    2016-01-01

    Full Text Available Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formula­tions of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longi­palpa F. under laboratory and field conditions.Methods: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml-1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs us­ing different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer.Results: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough path­ogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in- water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sun­flower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses.Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising al­ternative for cockroach control.  

  16. Phosphorus uptake of an arbuscular mycorrhizal fungus is not effected by the biocontrol bacterium ¤Burkholderia cepacia¤

    DEFF Research Database (Denmark)

    Ravnskov, S.; Larsen, J.; Jakobsen, I.

    2002-01-01

    intraradices (BEG87) were studied in root-free soil compartments separated from a rooting compartment by a fine nylon-mesh. B. cepacia had no effect on AM fungal biomass and energy reserves measured using the signature fatty acid 16:1omega5 from phospholipid fatty acids (PLFAs) and neutral lipid fatty acids...

  17. Introducing the term 'Biocontrol Plants' for integrated pest management

    Directory of Open Access Journals (Sweden)

    Pia Parolin

    2014-02-01

    Full Text Available Studies of interactions between crops, additional plants, pests and beneficial organisms already exist as well as studies of natural enemy preference, dispersal, and abundance. However, these studies focus on tri-trophic interactions from an "arthropod" point of view. We think that in order to optimize crop protection methods we need to understand the effects that plant structures have on the various arthropods and on subsequent tri-trophic interactions. Although studies and reviews describing the role of secondary plants in Integrated Pest Management (IPM exist, to date a general term which encompasses all plants added to a cropping system with the aim of enhancing IPM strategies has yet to be formulated. Therefore, we suggest a new term, "biocontrol plants", which we define as plants that are intentionally added to a crop system with the aim of enhancing crop productivity through pest attraction and/or pest regulation; a term that will promote the use of biocontrol services, and can ultimately lead to an increase in the sustainability of cropping systems.

  18. Social-insect fungus farming

    DEFF Research Database (Denmark)

    Aanen, Duur Kornelis; Boomsma, Jacobus Jan

    2006-01-01

    basidiomycete genus Termitomyces - whereas the ants are associated with a larger diversity of fungal lineages (all basidiomycetes). The ants and termites forage for plant material to provision their fungus gardens. Their crops convert this carbon-rich plant material into nitrogen-rich fungal biomass to provide...... the farming insects with most of their food ( Figure 1 ). No secondary reversals to the ancestral life style are known in either group, which suggests that the transitions to farming were as drastically innovative and irreversible as when humans made this step about 10,000 years ago....

  19. [Cutaneous mold fungus granuloma from Ulocladium chartarum].

    Science.gov (United States)

    Altmeyer, P; Schon, K

    1981-01-01

    Cutaneous granulomas due to the mold fungus Ulocladium chartarum (Preuss) are described in a 58 year old woman. This fungus is usually harmless for mammalian. It is thought that a consisting immunosuppression (Brill-Symmer's disease, therapy with corticosteroids) was a priming condition for the infection. The route of infection in this patient described is unknown. PMID:7194869

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

    Science.gov (United States)

    Brückner, H; Graf, H; Bokel, M

    1984-11-15

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

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

    Science.gov (United States)

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

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

  2. Potential of the salt-tolerant laccase-producing strain Trichoderma viride Pers. NFCCI-2745 from an estuary in the bioremediation of phenol-polluted environments.

    Science.gov (United States)

    Divya, L M; Prasanth, G K; Sadasivan, C

    2014-06-01

    Industrialization causes the generation of phenolic pollutants in the environment. The ability of laccases to oxidize phenolic compounds and reduce molecular oxygen to water has led to intensive studies on these enzymes. Although salt-tolerant fungi are potential sources of enzymes for industrial applications, they have been inadequately explored for laccase production. This study describes the isolation of a salt- and phenol-tolerant strain of Trichoderma sp. with the ability to produce laccase, and thus with the potential for industrial applications. The coconut husk retting ground in the estuaries of Kerala, India, a saline environment highly polluted with phenolic compounds, was selected for isolating the fungus. Enhanced laccase production was observed at 5-10 ppt salinity. The organism could grow even at 30 ppt salinity with reduced biomass production and laccase secretion. The optimum concentration of different phenolic compounds for enhanced laccase secretion ranged between 20 and 80 mg L(-1) . As the concentration of phenolic compounds increased beyond 200 mg L(-1) , the enzyme activity decreased and was completely inhibited at 800 mg L(-1) . The tolerance of Trichoderma viride Pers. NFCCI-2745 to salinity and various phenolic compounds can be utilized in the bioremediation of highly saline and phenolic compound-rich industrial effluents.

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

    Science.gov (United States)

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

    2008-03-01

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

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

    OpenAIRE

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

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Haryuni -

    2013-09-01

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

  6. 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...

  7. Caracterización molecular y agronómica de aislados de Trichoderma spp nativos del noreste de México

    Directory of Open Access Journals (Sweden)

    José Luis Hernández Mendoza

    2012-06-01

    Full Text Available Normal 0 21 false false false ES-CO X-NONE X-NONE MicrosoftInternetExplorer4 Título en ingles: Molecular and agronomic characterization of Trichoderma spp natives of northeastern Mexico  Resumen Trichoderma sp es un hongo frecuentemente usado en actividades agrícolas, pues actúa como antagonista de diversas especies de hongos fitopatógenos. En este estudio se realizó el aislamiento de cuatro cepas de Trichoderma sp nativas del noreste de México, las cuales fueron identificadas a nivel molecular mediante la secuenciación del ITS 1. Además se evaluó su capacidad antagonista en contra los hongos fitopatógenos Macrophomina phaseolina y Fusarium oxysporum, que afectan severamente cultivos de sorgo, maíz y fríjol en el noreste de México. La identificación se realizó de acuerdo al grado de concordancia con secuencias reportadas y corresponden a las especies T. hammatum (HK701;  T. koningiopsis (HK702; T. asperellum (HK703 y Trichoderma sp (HK704. Por otra parte, las pruebas de antagonismo muestran que los aislados HK701, HK703 y HK704 inhiben por competencia el crecimiento de M. phaseolina y F. oxysporum, mientras que HK702 tiene la capacidad para hiperparasitar dichos fitopatógenos. Finalmente, se evaluó la promoción de crecimiento de T. asperellum HK703, en maíz (Pionner 30P49®, usando para ello concentraciones de tratamiento de 1x10e2 hasta 1x10e6 esp/mL. En estos ensayos se midió la producción de biomasa. Los resultados muestran que en concentraciones intermedias se tiene el mayor incremento en altura de plantas y mayor producción de peso seco en follaje y raíz. Entre los parámetros antes mencionados existen diferencias significativas. Palabras clave: control biológico, antagonismo, biofertilizantes, Zea maiz, Sorghum bicolor. Abstract Trichoderma sp is a fungus often used in agricultural activities, because it acts as an antagonist of several species of plant pathogenic fungi. In this study four strains of Trichoderma

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

    Science.gov (United States)

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

    2013-08-01

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

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

    Directory of Open Access Journals (Sweden)

    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

  10. Risk assessment: progress of quarantine biocontrol research on Chinese Tallow, Melaleuca, and Downy Rose Myrtle

    Science.gov (United States)

    Risk assessments of two biocontrol candidates for Chinese tallow, Triadica sebifera (Euphoriales: Euphorbiaceae), and one for Melaleuca, Melaleuca quinquenervia (Myrtales: Myrtaceae), were conducted during 2009 and continuing into 2010 by USDA scientists located at the Florida Department of Agricul...

  11. Differential effects of organic compounds on cucumber damping-off and biocontrol activity of antagonistic bacteria

    DEFF Research Database (Denmark)

    Li, Bin; Ravnskov, Sabine; Guanlin, X.;

    2011-01-01

    The influence of the organic compounds tryptic soy broth, cellulose, glucose and chitosan on cucumber damping-off caused by Pythium aphanidermatum and biocontrol efficacy of the biocontrol agents (BCAs) Paenibacillus macerans and P. polymyxa were examined in a seedling emergence bioassay. Results...... showed that the organic compounds differentially affected both pathogen and BCAs. Tryptic soy broth, glucose and chitosan increased Pythium damping-off of cucumber, compared to the control treatment without organic compounds, whereas cellulose had no effect. Both Paenibacillus species had biocontrol...... effects against Pythium damping-off compared with the corresponding treatments with P. aphanidermatum alone, but the biocontrol efficacy depended on the type of organic compounds added. Both BCAs counteracted damping-off in treatments with TSB and chitosan. However, P. polymyxa counteracted damping...

  12. Peptaibiomics: microheterogeneity, dynamics, and sequences of trichobrachins, peptaibiotics from Trichoderma parceramosum Bissett (T. longibrachiatum Rifai).

    Science.gov (United States)

    Krause, Corina; Kirschbaum, Jochen; Brückner, Hans

    2007-06-01

    From the culture broth of the filamentous fungus Trichoderma parceramosum, strain CBS 936.69, a mixture of polypeptide antibiotics (pepaibiotics), named trichobrachin (TB), was isolated. Three major groups designated TB I, TB II, and TB III could be separated and isolated by preparative TLC on silica gel. Individual peptides of these three groups were sequenced by on-line LC/ESI-MS(n). The mixture of N-acetylated peptides comprises ten 19-residue peptides with a free C-terminal Gln residue (TB I peptides), two 18-residue peptides with a free C-terminal Gln residue (TB II 1 and 2), seven 20-residue peptides with a C-terminal amide-bound phenylalaninol (TB II 3-10), and 34 eleven-residue peptides with either a C-terminal leucinol or isoleucinol or valinol (TB III 1-34). Monitoring production and degradation of peptaibiotics in a pilot experiment revealed that the biosynthesis of TB II and TB III peptides starts two days after the beginning of fermentation. After five days of fermentation, the concentration of TB II decreased, whereas the amount of TB I increased. This observation unequivocally demonstrates that those two 18-residue TB I and TB II peptides with the free carboxy terminus result from enzymatic C-terminal degradation of the 20-residue TB II peptides. In analogy to the technical terms proteome and proteomics, the terms peptaibiome and peptaibiomics have recently been proposed for the entirety and dynamics of the Aib-containing peptides (comprehensively named peptaibiotics). Consequently, the entire peptaibiome of T. parceramosum grown under submerse conditions in shake-flasks for five days comprises at least 54 peptides differing in main-chain length and microheterogeneity, i.e., exchange of amino acids and the C-terminal 1,2-amino alcohol. PMID:17589878

  13. 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.

  14. Endophytic Bacteria as Biocontrol Agents of Tomato Bacterial Wilt Disease

    Directory of Open Access Journals (Sweden)

    Arika Purnawati

    2014-01-01

    Full Text Available Ralstonia solanacearum is a tomato pathogen causing wilt disease which can lose crop production up to 100%. This problem also has economical impact to Indonesia, though controlling this pathogen has been donebefore. Biological control using endophytic bacteria is one of alternative control methods to support agriculture sustainability. The objective of these experiments are isolating endophytic bacteria taken from healthy tomato stems and root as well as investigating patogens as biocontrol agents of tomato bacterial wilt disease. Among 10 isolates found, only Ps1 and Ps8 can inhibit R. solanacearum based on antagonistic test in vitro using seed coat method and 4-7 mm inhibition zone forming. In vivo test was applied in green house in 30 days old of tomatoes usingendophytic bacteria by theroot soak technique. The result shows Ps1 and Ps8 within 15-16 days incubation period have suppressed 8.07%-9.19% pathogen attack.

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

    OpenAIRE

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

    2014-01-01

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

  16. Biocontrol of larval mosquitoes by Acilius sulcatus (Coleoptera: Dytiscidae

    Directory of Open Access Journals (Sweden)

    Banerjee Siddhartha S

    2008-10-01

    Full Text Available Abstract Background Problems associated with resistant mosquitoes and the effects on non-target species by chemicals, evoke a reason to find alternative methods to control mosquitoes, like the use of natural predators. In this regard, aquatic coleopterans have been explored less compared to other insect predators. In the present study, an evaluation of the role of the larvae of Acilius sulcatus Linnaeus 1758 (Coleoptera: Dytiscidae as predator of mosquito immatures was made in the laboratory. Its efficacy under field condition was also determined to emphasize its potential as bio-control agent of mosquitoes. Methods In the laboratory, the predation potential of the larvae of A. sulcatus was assessed using the larvae of Culex quinquefasciatus Say 1823 (Diptera: Culicidae as prey at varying predator and prey densities and available space. Under field conditions, the effectiveness of the larvae of A. sulcatus was evaluated through augmentative release in ten cemented tanks hosting immatures of different mosquito species at varying density. The dip density changes in the mosquito immatures were used as indicator for the effectiveness of A. sulcatus larvae. Results A single larva of A. sulcatus consumed on an average 34 IV instar larvae of Cx. quinquefasciatus in a 24 h period. It was observed that feeding rate of A. sulcatus did not differ between the light-on (6 a.m. – 6 p.m., and dark (6 p.m. – 6 a.m. phases, but decreased with the volume of water i.e., space availability. The prey consumption of the larvae of A. sulcatus differed significantly (P A. sulcatus larvae, while with the withdrawal, a significant increase (p A. sulcatus in regulating mosquito immatures. In the control tanks, mean larval density did not differ (p > 0.05 throughout the study period. Conclusion the larvae of the dytiscid beetle A. sulcatus proved to be an efficient predator of mosquito immatures and may be useful in biocontrol of medically important mosquitoes.

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

    Science.gov (United States)

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

    2013-11-01

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

  18. Biocontrol of fouling pests: Effect of diversity, identity and density of control agents.

    Science.gov (United States)

    Atalah, Javier; Newcombe, Emma M; Zaiko, Anastasija

    2016-04-01

    Augmentative biocontrol, using native natural enemies, has been suggested as a promising tool to control marine biofouling pests on artificial structures. However, there are still important knowledge gaps to be addressed before biocontrol can be considered as a management tool. In a field experiment on floating marine structures we examined intra- and interspecific consumer interactions among biocontrol agents on different surface orientations. We tested the effect of identity, density and diversity of three invertebrates (the 11-arm seastar Coscinasterias muricata, the sea urchin Evechinus chloroticus and the gastropod Cook's turban Cookia sulcata) to reduce established biofouling and to prevent fouling growth on defouled surfaces. High densities of biocontrol agents were not more effective at fouling control (cover and biomass) than low densities. Nor did multi-species treatments function more effectively than mono-specific ones. However, biocontrol agent identity was important, with the 11-arm seastar and Cook's turban being the most effective at fouling reduction and prevention, respectively. Surface orientation had a strong effect on the effectiveness of control agents, with the best results obtained on vertical compared to diagonal and underside surfaces. This study confirmed the potential of biocontrol as a management tool for marine pest, indicating that identity is more important than richness and density of control agents. It also highlighted the limitations of this approach on diagonal and underside surfaces, where control agents have limited retention ability. PMID:26845376

  19. Microsatellite primers for fungus-growing ants

    DEFF Research Database (Denmark)

    Villesen, Palle; Gertsch, P J; Boomsma, JJ

    2002-01-01

    We isolated five polymorphic microsatellite loci from a library of two thousand recombinant clones of two fungus-growing ant species, Cyphomyrmex longiscapus and Trachymyrmex cf. zeteki. Amplification and heterozygosity were tested in five species of higher attine ants using both the newly...... developed primers and earlier published primers that were developed for fungus-growing ants. A total of 20 variable microsatellite loci, developed for six different species of fungus-growing ants, are now available for studying the population genetics and colony kin-structure of these ants....

  20. Microsatellite Primers for Fungus-Growing Ants

    DEFF Research Database (Denmark)

    Villesen Fredsted, Palle; Gertsch, Pia J.; Boomsma, Jacobus Jan (Koos)

    2002-01-01

    We isolated five polymorphic microsatellite loci from a library of two thousand recombinant clones of two fungus-growing ant species, Cyphomyrmex longiscapus and Trachymyrmex cf. zeteki. Amplification and heterozygosity were tested in five species of higher attine ants using both the newly...... developed primers and earlier published primers that were developed for fungus-growing ants. A total of 20 variable microsatellite loci, developed for six different species of fungus-growing ants, are now available for studying the population genetics and colony kin-structure of these ants....

  1. 40 CFR 180.1293 - Trichoderma gamsii strain ICC 080; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

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

  2. 40 CFR 180.1294 - Trichoderma asperellum strain ICC 012; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

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

  3. 40 CFR 180.1201 - Trichoderma harzianum strain T-39; exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

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

  4. In Vitro Evaluation Of Selected Plant Extracts As Biocontrol Agents Against Black Mold Aspergillus Niger Van Tieghem Of Onion Bulbs Allium Cepa L.

    Directory of Open Access Journals (Sweden)

    Saifeldin A. F. El-Nagerabi

    2015-08-01

    Full Text Available Black mold disease caused by Aspergillus niger V. Tiegh. is the most devastating infection occurs in onions Allium cepa L. under field and store conditions. The use of biocontrol agents is ecofriendly approach for controlling seedborne and soilborne diseases compared to the use of toxic synthetic fungicides. This study has been designed to assess the contamination levels of onion seeds with A. niger and its effect on seed germination and to evaluate the in vitro antifungal activity of Prunus mahaleb seeds Commiphora myrrha resin 0.5 1.0 1.5 2.0 g100 ml Syzygium aromaticum dry buds clove and Panax ginseng roots extracts 0.5 1.0 2.0 2.5 g100 ml against black mold of onion bulbs. The fungus is seedborne pathogen which significantly contaminated onion seeds 89-100 and reduced seed germination 39-83. The extracts of clove caused 43-96 inhibition in spore germination followed by mahaleb 37-96 myrrha 33-88 and ginseng 34-87. The highest concentration of these extracts 3.0 did not affect seed germination but significantly reduced seed contamination by A. niger up to 84 80 71 and 65 for Syzygium aromaticum Syzygium aromaticum Panax ginseng and Prunus mahaleb respectively. The extracts apparently inhibited the fungal growth and mold development on stored onion bulbs which indicates the antifungal property of these extracts against A. niger. Therefore they can be recommended as effective biocontrol agents to reduce seed contamination and enhance the storability of onion bulbs. Thus the use of healthy and certified seeds for onion production is a priority.

  5. The Pochonia chlamydosporia Serine Protease Gene vcp1 Is Subject to Regulation by Carbon, Nitrogen and pH: Implications for Nematode Biocontrol

    Science.gov (United States)

    Ward, Elaine; Kerry, Brian R.; Manzanilla-López, Rosa H.; Mutua, Gerald; Devonshire, Jean; Kimenju, John; Hirsch, Penny R.

    2012-01-01

    The alkaline serine protease VCP1 of the fungus Pochonia chlamydosporia belongs to a family of subtilisin-like enzymes that are involved in infection of nematode and insect hosts. It is involved early in the infection process, removing the outer proteinaceous vitelline membrane of nematode eggs. Little is known about the regulation of this gene, even though an understanding of how nutrients and other factors affect its expression is critical for ensuring its efficacy as a biocontrol agent. This paper provides new information on the regulation of vcp1 expression. Sequence analysis of the upstream regulatory region of this gene in 30 isolates revealed that it was highly conserved and contained sequence motifs characteristic of genes that are subject to carbon, nitrogen and pH-regulation. Expression studies, monitoring enzyme activity and mRNA, confirmed that these factors affect VCP1 production. As expected, glucose reduced VCP1 expression and for a few hours so did ammonium chloride. Surprisingly, however, by 24 h VCP1 levels were increased in the presence of ammonium chloride for most isolates. Ambient pH also regulated VCP1 expression, with most isolates producing more VCP1 under alkaline conditions. There were some differences in the response of one isolate with a distinctive upstream sequence including a variant regulatory-motif profile. Cryo-scanning electron microscopy studies indicated that the presence of nematode eggs stimulates VCP1 production by P. chlamydosporia, but only where the two are in close contact. Overall, the results indicate that readily-metabolisable carbon sources and unfavourable pH in the rhizosphere/egg-mass environment may compromise nematode parasitism by P. chlamydosporia. However, contrary to previous indications using other nematophagous and entomopathogenic fungi, ammonium nitrate (e.g. from fertilizers) may enhance biocontrol potential in some circumstances. PMID:22558192

  6. The Pochonia chlamydosporia serine protease gene vcp1 is subject to regulation by carbon, nitrogen and pH: implications for nematode biocontrol.

    Directory of Open Access Journals (Sweden)

    Elaine Ward

    Full Text Available The alkaline serine protease VCP1 of the fungus Pochonia chlamydosporia belongs to a family of subtilisin-like enzymes that are involved in infection of nematode and insect hosts. It is involved early in the infection process, removing the outer proteinaceous vitelline membrane of nematode eggs. Little is known about the regulation of this gene, even though an understanding of how nutrients and other factors affect its expression is critical for ensuring its efficacy as a biocontrol agent. This paper provides new information on the regulation of vcp1 expression. Sequence analysis of the upstream regulatory region of this gene in 30 isolates revealed that it was highly conserved and contained sequence motifs characteristic of genes that are subject to carbon, nitrogen and pH-regulation. Expression studies, monitoring enzyme activity and mRNA, confirmed that these factors affect VCP1 production. As expected, glucose reduced VCP1 expression and for a few hours so did ammonium chloride. Surprisingly, however, by 24 h VCP1 levels were increased in the presence of ammonium chloride for most isolates. Ambient pH also regulated VCP1 expression, with most isolates producing more VCP1 under alkaline conditions. There were some differences in the response of one isolate with a distinctive upstream sequence including a variant regulatory-motif profile. Cryo-scanning electron microscopy studies indicated that the presence of nematode eggs stimulates VCP1 production by P. chlamydosporia, but only where the two are in close contact. Overall, the results indicate that readily-metabolisable carbon sources and unfavourable pH in the rhizosphere/egg-mass environment may compromise nematode parasitism by P. chlamydosporia. However, contrary to previous indications using other nematophagous and entomopathogenic fungi, ammonium nitrate (e.g. from fertilizers may enhance biocontrol potential in some circumstances.

  7. In Vitro Antifungal Activity of Strains of Trichoderma harzianum

    OpenAIRE

    KÜÇÜK, Çiğdem; Merih KIVANÇ

    2004-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Zayame Vegette Pinto

    2014-06-01

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

  9. In vitro sensitivity of antagonistic Trichoderma atroviride to herbicides

    OpenAIRE

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

    2014-01-01

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

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

    Science.gov (United States)

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

    2016-04-01

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

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

    Science.gov (United States)

    Bailey, M J; Tähtiharju, J

    2003-08-01

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

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

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    MARTINS-CORDER M.P.

    1998-01-01

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

  13. Metabolites from marine fungus Aspergillus sp.

    Digital Repository Service at National Institute of Oceanography (India)

    Wahidullah, S.; Rajmanickam, R.; DeSouza, L.

    Chemical examination of a methanolic extract of the marine fungus, Aspergillus sp., isolated from marine grass environment, yielded a steroid, ergosterol peroxide (1), and a mixture of known glyceride esters (2,3) of unsaturated fatty acids...

  14. A new macrocyclic trichochecene from soil fungus

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    From fermentation broth of soil fungus 254-2 obtained from Yunnan province,a new macrocylic trichochecene was isolated.The structure was determined on the basis of spectroscopic evidences especially the 2-D NMR spectra.

  15. Entomology: A Bee Farming a Fungus.

    Science.gov (United States)

    Oldroyd, Benjamin P; Aanen, Duur K

    2015-11-16

    Farming is done not only by humans, but also by some ant, beetle and termite species. With the discovery of a stingless bee farming a fungus that provides benefits to its larvae, bees can be added to this list.

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

    Science.gov (United States)

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

    2014-08-01

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

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

    Science.gov (United States)

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

    2013-06-15

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

  18. 21 CFR 184.1250 - Cellulase enzyme preparation derived from Trichoderma longibrachiatum.

    Science.gov (United States)

    2010-04-01

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

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

    OpenAIRE

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

    2013-01-01

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

  20. Survival of Spores of Trichoderma longibrachiatum in Space: data from the Space Experiment SPORES on EXPOSE-R

    Science.gov (United States)

    Neuberger, Katja; Lux-Endrich, Astrid; Panitz, Corinna

    2015-01-01

    In the space experiment `Spores in artificial meteorites' (SPORES), spores of the fungus Trichoderma longibrachiatum were exposed to low-Earth orbit for nearly 2 years on board the EXPOSE-R facility outside of the International Space Station. The environmental conditions tested in space were: space vacuum at 10-7-10-4 Pa or argon atmosphere at 105 Pa as inert gas atmosphere, solar extraterrestrial ultraviolet (UV) radiation at λ > 110 nm or λ > 200 nm with fluences up to 5.8 × 108 J m-2, cosmic radiation of a total dose range from 225 to 320 mGy, and temperature fluctuations from -25 to +50°C, applied isolated or in combination. Comparable control experiments were performed on ground. After retrieval, viability of spores was analysed by two methods: (i) ethidium bromide staining and (ii) test of germination capability. About 30% of the spores in vacuum survived the space travel, if shielded against insolation. However, in most cases no significant decrease was observed for spores exposed in addition to the full spectrum of solar UV irradiation. As the spores were exposed in clusters, the outer layers of spores may have shielded the inner part. The results give some information about the likelihood of lithopanspermia, the natural transfer of micro-organisms between planets. In addition to the parameters of outer space, sojourn time in space seems to be one of the limiting parameters.

  1. Enhanced resistance to Botrytis cinerea and Rhizoctonia solani in transgenic broccoli with a Trichoderma viride endochitinase gene

    Institute of Scientific and Technical Information of China (English)

    YU Ya; REN Shu-xin; GUO Yang-dong; ZHANG Lei; LIAN Wei-ran; XU Feng-feng; LI Shuang-tao; XIANG Juan; ZHANG Guo-zhen; HU Zan-min; ZHAO Bing

    2015-01-01

    A endochitinase gene (Tch ) from the fungus Trichoderma viride was introduced into broccoli (Brassica oleracea var. italica) by Agrobacterium-mediated transformation. Sixty-eight putative transformants were obtained and the presence of the Tch gene was conifrmed by both PCR and Southern blot analysis. RT-PCR analysis showed an accumulation of the transcript encoding the endochitinase protein in the transgenic plants. Using real-time quantitative PCR, the expression proifling of endochitinase gene was analyzed. Primary transformants and selfed progeny were examined for expression of the endo-chitinase using a lfuorometric assay and for their resistance to the pathogenic fungi Botrytis cinerea and Rhizoctonia solani. The endochitinase activities in T0 in vitro plants, T0 mature plants and T1 mature plants were correlated with leaf lesions, and the transgenic line T618 had high endochitinse activities of 102.68, 114.53 and 120.27 nmol L–1 MU min–1 mg–1 protein in the three kinds of plants, respectively. The endochitinase activity showed a positive correlation with the resistance to the pathogens. Most transgenic T0 broccoli had increased resistance to the pathogens of B. cinerea and R. solani in leaf assays and this resistance was conifrmed to be inheritable. These ifndings suggested that expression of the Tch gene from T. viride could enhance resistance to pathogenic fungi in Brassica species.

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

    Directory of Open Access Journals (Sweden)

    Christa Ivanova

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

  3. Pathogenic and enzyme activities of the entomopathogenic fungus Tolypocladium cylindrosporum (Ascomycota: Hypocreales) from Tierra del Fuego, Argentina.

    Science.gov (United States)

    Scorsetti, Ana C; Elíades, Lorena A; Stenglein, Sebastián A; Cabello, Marta N; Pelizza, Sebastián A; Saparrat, Mario C N

    2012-06-01

    temperatures of 4 degreeC or 12 degreeC. In accordance with the results on colony diameters, the fungus revealed an ability to degrade casein, chitin derivatives, Tween 20, and urea as evidenced by the appearance of a halo around the fungal colony. Because of its origin and temperature tolerance, this Argentine strain has great potential for use as a biocontrol agent for insect pest control in cold and temperate environments. PMID:23894949

  4. Biodegradation of carbamazepine and clarithromycin by Trichoderma harzianum and Pleurotus ostreatus investigated by liquid chromatography - high-resolution tandem mass spectrometry (FTICR MS-IRMPD).

    Science.gov (United States)

    Buchicchio, Alessandro; Bianco, Giuliana; Sofo, Adriano; Masi, Salvatore; Caniani, Donatella

    2016-07-01

    In this study, the capability of pharmaceutical biodegradation of fungus Trichoderma harzianum was evaluated through the comparison with the well-known biodegradation capability of white-rot fungus Pleurotus ostreatus. The study was performed in aqueous phase under aerobic conditions, using two of the most frequently detected drugs in water bodies: carbamazepine and clarithromycin, with concentrations commonly found in treated wastewater (4μg/l and 0.03μg/l respectively). For the first time, we demonstrated that T. harzianum is able to remove carbamazepine and clarithromycin. The analyses were performed by reversed-phase liquid chromatography/mass spectrometry, using high-resolution Fourier-transform ion cyclotron resonance mass spectrometry upon electrospray ionization in positive ion mode. The high selectivity and mass accuracy provided by high-resolution mass spectrometry, allowed us to identify some unknown metabolites. On the basis of our study, the major metabolites detected in liquid culture treated by T. harzianum were: 14-hydroxy-descladinosyl- and descladinosyl-clarithromycin, which are pharmacologically inactive products not dangerous for the environment. PMID:27039063

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

    Directory of Open Access Journals (Sweden)

    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.

  6. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere

    Directory of Open Access Journals (Sweden)

    Hyun Gi Kong

    2016-04-01

    Full Text Available Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  7. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere.

    Science.gov (United States)

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-04-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer. PMID:27147933

  8. Attachment capability of antagonistic yeast Rhodotorula glutinis to Botrytis cinerea contributes to biocontrol efficacy

    Directory of Open Access Journals (Sweden)

    Boqiang eLi

    2016-05-01

    Full Text Available Rhodotorula glutinis as an antagonism show good biocontrol performance against various postharvest diseases in fruits. In the present study, strong attachment capability of R. glutinis to spores and hyphae of Botrytis cinerea was observed. Further analysis showed that certain protein components on the yeast cell surface played critical role during the interaction between R. glutinis and B. cinerea. The components mainly distributed at the poles of yeast cells and might contain glycosylation modification, as tunicamycin treated yeast cells lost attachment capability to B. cinerea. To investigate contributions of attachment capability of R. glutinis to its biocontrol efficacy, yeast cells were mutagenized with 3% methane-sulfonic acid ethyl ester (EMS, and a mutant CE4 with stable non-attaching phenotype was obtained. No significant difference was found on colony, cell morphology, reproductive ability and capsule formation between the mutant and wild type. However, there was a distinct difference in India ink positive staining patterns between the two strains. Moreover, wild type strain of R. glutinis showed better performance on inhibiting spore germination and mycelial growth of B. cinerea than CE4 strain when yeast cells and B. cinerea were co-cultured in vitro. In biocontrol assay, both wild type and CE4 strains showed significant biocontrol efficacy against gray mould caused by B. cinerea in apple fruit, whereas, control effect of CE4 strain was lower than that of wild type. Our findings provided new evidences that attachment capability of R. glutinis to B. cinerea contributed to its biocontrol efficacy.

  9. Surfactin triggers biofilm formation of Bacillus subtilis in melon phylloplane and contributes to the biocontrol activity.

    Science.gov (United States)

    Zeriouh, Houda; de Vicente, Antonio; Pérez-García, Alejandro; Romero, Diego

    2014-07-01

    The biocontrol activity of many Bacillus species has been traditionally related to the direct antagonism of pathogens. In previous works, we reported that B. subtilis strain UMAF6614 was an efficient biocontrol agent that produced bacillomycin, fengycin and surfactin lipopeptides. Bacillomycins and fengycins were shown to have antagonistic activity towards fungal and bacterial pathogens of cucurbits; however, the functionality of surfactin remained unclear. In this study, the role of surfactin in the biocontrol activity of this strain was investigated. We observed that a deficiency in surfactin production led to a partial reduction of disease suppression by this biocontrol agent, which coincided with a defect in biofilm formation and the colonization of the melon phylloplane. These effects were due to a dramatic reduction in the production of exopolysaccharide and the TasA protein, which are the two major components of the extracellular matrix. We propose that the biocontrol activity of this strain is the result of the coordinated action of the three families of lipopeptides. B. subtilis UMAF6614 produces surfactin to trigger biofilm formation on melon phylloplane, which ensures the long-term persistence and the adequate secretion of suppressive lipopeptides, bacillomycins and fengycins, which efficiently target pathogens.

  10. Draft Genome Sequence of a Biocontrol Rhizobacterium, Chryseobacterium kwangjuense Strain KJ1R5, Isolated from Pepper (Capsicum annuum).

    Science.gov (United States)

    Jeong, Jin-Ju; Park, Hongjae; Park, Byeong Hyeok; Mannaa, Mohamed; Sang, Mee Kyung; Choi, In-Geol; Kim, Ki Deok

    2016-01-01

    Strain KJ1R5 of the rhizobacterium ITALIC! Chryseobacterium kwangjuenseis an effective biocontrol agent against Phytophthora blight of pepper caused by a destructive soilborne oomycete, ITALIC! Phytophthora capsici Here, we present the draft genome sequence of strain KJ1R5, which contains genes related to biocontrol, plant growth promotion, and environmental stress adaptation. PMID:27103726

  11. Evaluation of recycled bioplastic pellets and a sprayable formulation for application of an Aspergillus flavus biocontrol strain

    Science.gov (United States)

    Biocontrol of Aspergillus flavus using inoculated bioplastic granules has been proven to be effective under laboratory and field conditions. In the present study, the use of low-density pellets from recycled bioplastic as a biocontrol strain carrier was evaluated. Applying recycled bioplastic pell...

  12. Optimization of solid fermentation of cellulase from Trichoderma koningii

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

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

    Science.gov (United States)

    Abd El-Baki, G K; Mostafa, Doaa

    2014-12-01

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

  14. Trichoderma strains- Silybum marianum hairy root cultures interactions

    Directory of Open Access Journals (Sweden)

    T. Hasanloo

    2015-03-01

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

  15. Proteomic mapping of secreted proteins of Trichoderma spp.

    Institute of Scientific and Technical Information of China (English)

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

    2004-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Min Li

    2010-08-01

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

  17. Endophytic fungus strain 28 isolated from Houttuynia cordata possesses wide-spectrum antifungal activity.

    Science.gov (United States)

    Pan, Feng; Liu, Zheng-Qiong; Chen, Que; Xu, Ying-Wen; Hou, Kai; Wu, Wei

    2016-01-01

    The aim of this paper is to identify and investigate an endophytic fungus (strain 28) that was isolated from Houttuynia cordata Thunb, a famous and widely-used Traditional Chinese Medicine. Based on morphological methods and a phylogenetic analysis of ITS sequences, this strain was identified as Chaetomium globosum. An antifungal activity bioassay demonstrated that the crude ethyl acetate (EtOAc) extracts of strain 28 had a wide antifungal spectrum and strong antimicrobial activity, particularly against Exserohilum turcicum (Pass.) Leonard et Suggs, Botrytis cinerea persoon and Botrytis cinerea Pers. ex Fr. Furthermore, the fermentation conditions, extraction method and the heat stability of antifungal substances from strain 28 were also studied. The results showed that optimal antifungal activity can be obtained with the following parameters: using potato dextrose broth (PDB) as the base culture medium, fermentation for 4-8d (initial pH: 7.5), followed by extraction with EtOAc. The extract was stable at temperatures up to 80°C. This is the first report on the isolation of endophytic C. globosum from H. cordata to identify potential alternative biocontrol agents that could provide new opportunities for practical applications involving H. cordata. PMID:26991297

  18. Effect of Trichoderma harzianum T4 on Bacterial Community in Watermelon (Citrullus lanatus) Rhizosphere Soil%生防菌哈茨木霉Trichoderma harzianum T4对西瓜根围土壤细菌群落的影响

    Institute of Scientific and Technical Information of China (English)

    夏飞; 张于; 旭热; 王伟

    2013-01-01

      采用平板培养、末端限制性片段长度多态性(terminal restriction fragment length polymorphism, T-RFLP)以及变性梯度凝胶电泳(denatured gradient gel electrophoresis, DGGE)的方法相结合探讨生防菌哈茨木霉 Trichoderma harzianum T4对大棚西瓜根围土壤细菌群落及氨氧化细菌群落的影响,为其在田间应用的生态安全性的评估提供支撑。末端限制性片段长度多态性以及变性梯度凝胶电泳的结果均表明哈茨木霉 T4施入田间约四周内对根围土壤细菌群落产生明显的影响,随后这种扰动现象逐渐减小。对 DGGE中受影响条带的测序结果表明,生防菌 T4促进了假单胞菌 Pseudomonas,芽孢杆菌 Bacillus,苍白杆菌Ochrobactrum 以及中慢生根瘤菌 Mesorhizobium 等细菌类群的生长,对短杆菌 Brevibacterium,克雷白氏肺炎杆菌 Klebsiella pneumoniae,根瘤菌 Rhizobium sp 等表现出抑制作用。生防菌 T4对根围土壤中氨氧化细菌群落并没有产生明显的影响。可见,生防菌木霉 T4引入初期对根围土壤中细菌群落产生明显的扰动,但这种干扰是短暂的,并没有对根围土壤细菌群落形成持续的影响。%The effects of biocontrol strain Trichoderma harzianum T4 on bacterial and ammonia-oxidizing bacteria (AOB) communities in watermelon (Citrullus lanatus) rhizosphere soil were studied using plate colony calculation, T-RFLP and DGGE method, in order to provide a theoretical basis and technique for assessing the microbial ecology risk of biocontrol agents application. Both T-RFLP and DGGE method demonstrated that T. harzianum T4 had short-term influence on rhizosphere soil bacterial communities which lasted about four weeks. Biocontrol strain T. harzianum T4 increased population of some bacteria, such as Pseudomonas, Bacillus, Ochrobactrum and Mesorhizobium. Meanwhile population of other bacteria such as Brevibacterium and Klebsiella pneumoniae were

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

    Institute of Scientific and Technical Information of China (English)

    李广记; 陈捷; 刘铜; 刘力行

    2010-01-01

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

  20. Sequences of metanicins, 20-residue peptaibols from the ascomycetous fungus CBS 597.80.

    Science.gov (United States)

    Kimonyo, Anastase; Brückner, Hans

    2013-05-01

    Four linear 20-residue peptaibols, named metanicins (MTCs) A-D, were isolated from submerged cultures of the ascomycetous fungus CBS 597.80. Structure elucidation was performed by a combination of fast-atom-bombardment mass spectrometry (FAB-MS), electrospray ionization MS, Edman degradation of isolated fragments, and amino acid analysis by ion-exchange and gas chromatography, and enantioselective HPLC. The sequences of MTC A(B) are (amino acid exchange in B and C in parentheses): Ac-Aib-Ala-Aib-Ala-Aib-Ala-Gln-Aib-Val-Aib-Gly-Leu-Aib-Pro-Val-Aib-Aib(D-Iva)-Gln-Gln-Pheol and of MTC C(D) Ac-Aib-Ala-Aib-Ala-Aib-Ala-Gln-Aib-Val-Aib-Gly-Leu-Aib-Pro-Val-Aib-Aib(D-Iva)-Gln-Gln-Pheol (Ac, acetyl; Aib, α-aminoisobutyric acid; Iva, isovaline; Pheol, L-phenylalaninol). The peptides are related, and some of the sequences are identical, to other 20-residue peptaibols isolated from Trichoderma species. MTCs show moderate activities against Micrococcus luteus, Enterococcus faecalis, and Staphylococcus aureus, and very low activities against Bacillus subtilis. The producer has originally been identified and deposited as Metarhizium anisopliae var. anisopliae CBS 597.80. Although this identification has been withdrawn by Centralbureau voor Schimmelcultures (CBS) in the meantime, the accession number will be retained - independently from any taxonomic revisions. PMID:23681727