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Number of results: 14
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Abstract

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

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Authors and Affiliations

Agnieszka Wolna-Maruwka
Tomasz Piechota
Alicja Niewiadomska
Jacek Dach
Magdalena Szczech
Małgorzata Jędryczka
Agnieszka A. Pilarska
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Abstract

In view of the ecological hazards of chemicals, pot experiments were conducted to determine the efficacy of Trichoderma sp. against Macrophomina phaseolina. Greenhouse evolution of the interaction between M. phaseolina isolates and Trichoderma sp. isolates revealed a very highly significant (p = 0.0000). M. phaseolina isolate x antagonist isolate interaction for all the following parameters: preemergence damping-off, postemergence damping-off, survival, plant height, and dry weight. This interaction implies that a single isolate of antagonist can be highly effective againstan isolate of M. phaseolina, but may have only minimal effectsonotherisolatesof M. phaseolina. Therefore, isolates of antagonist should be tested against as many isolates of M. phaseolina as possible, as this will improve the chance of identifying antagonist isolates effective against several isolates of M. phaseolina.

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Authors and Affiliations

Aly A. Aly
Mohamed A. Abdel-Sattar
Moawad R. Omar
Kamel A. Abd-Elsalam
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Abstract

Potato (Solanum tuberosum L.), an important food crop in the world, is susceptible to many fungal pathogens including Alternaria solani and Fusarium oxysporum causing Fusarium wilt and early blight diseases. Mycoparasitic fungi like Trichoderma encode chitinases, cell wall degrading enzymes, with high antifungal activity against a wide range of phytopathogenic fungi. In this study, a binary vector harboring endochitinase gene of ~1,000 bp was constructed and used to transform potato nodes through Agrobacterium-mediated transformation. Out of several primary transformants, two transgenic potato lines were verified for transgene insertion and integration by Southern blot. In a pot experiment for Fusarium resistance, the transgenic potato lines didn’t show any symptoms of disease, instead they remained healthy post infection. The transgenic potato lines exhibited 1.5 fold higher mRNA expression of endochitinase at 7 days as compared to 0 day post fungus inoculation. It was evident that the mRNA expression decreased over days of inoculation but was still higher than at 0 day and remained stable upto 30 days post inoculation. Similarly, for A. solani infection assay, the mRNA expression of the endochitinase gene was 3 fold higher 7 days post inoculation compared to expression at 0 day. Although the expression decreased by1.2 fold during subsequent days post infection, it remained stable for 30 days, suggesting that protection in transgenic potato plants against fungal pathogens was achieved through an increase in endochitinase transcript.

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Authors and Affiliations

Neelam Fatima
Bushra Tabassum
Iqra Yousaf
Momina Malik
Anwar Khan
Imtiaz Ahmad Sajid
Muhammad Tariq
Nida Toufiq
Saman Riaz
Idrees Ahmad Nasir
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Abstract

In this study Trichoderma harzianum strain A was isolated from the rhizosphere of an argan tree in southern Morocco. Trichoderma harzianum strain A had previously demonstrated a high antagonistic potential in vitro by direct confrontation and in vivo on wheat plants in pots under greenhouse conditions against Zymoseptoria tritici, the agent of septoria leaf blotch. In this study, the activity of filtrates prepared from the liquid culture of T. harzianum A alone and from the confrontation medium with two Z. tritici strains [G1-1 (durum wheat) and A5-1 (soft wheat)] on the inhibition of Z. tritici pycnidiospore germination was studied by nephelometry. The results of the antibiosis assay revealed that filtrate 0 (A in confrontation with G1-1) and F3 (A against A5-1) showed 95% of G1-1 and A5-1 pycnidiospore inhibition at 9/10 dilution of the undiluted filtrates after 4 days of incubation. To understand and explain the antifungal activity of these filtrates, the extraction and identification of secondary molecules of peptaibiotic nature secreted by T. harzianum A in the three studied filtrates were performed. According to the results of high-performance liquid chromatography-mass spectrometry (HPLC-MS) analyses, 38 peptaibiotic molecules reported in the literature for their antifungal activity were identified in the different extracts at high concentrations (high peak intensities). These molecules are divided into nine groups, namely: Trichocryptin, Trichobrevin, Triochocryptin, Hypocompactin, Hyporodicin, Trichocompactin, Alamethicine, Trichoferin, and Trichokonin. It was also shown that the presence of the pathogen induces the production of peptaibols by the antagonistic strain of Trichoderma.
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Authors and Affiliations

Ilham Barakat
1
ORCID: ORCID
Noureddine Chtaina
1
Taha El Kamli
2
Philippe Grappin
3
Mohammed El Guilli
4
Brahim Ezzahiri
1

  1. Department of Production, Protection, and Biotechnology, Hassan II Institute of Agronomic and Veterinary Medicine, Rabat, Morocco
  2. Department of Veterinary, Hassan II Institute of Agronomic and Veterinary Medicine, Rabat, Morocco
  3. FungiSem, IRHS, INRA-University of Angers-Agrocampus-Ouest, Angers, France
  4. Laboratory of Phytopathology and Post-Harvest Quality, Regional Centre for Agronomic Research, Kénitra, Morocco
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Abstract

The objective of this study was to evaluate the antagonistic activity of Trichoderma spp. against wild pathogen Fusarium oxysporum F.28.1A, which causes wilt disease on sesame. Twenty-six isolates of Trichoderma spp. isolated from soil samples were tested to control F. oxysporum F.28.1A. Prescreening showed that five isolates were T-02B1, T-18B2, T-20B1, T-28B1, and T-29A1, based on the lowest values of colony radius of F. oxysporum F.28.1A. The selected isolates were identified by their ITS region as T. yunnanense T-02B1, T. lentiforme T-18B2, T. asperellum T-20B1, T. hamatum T-28B1, and T. hamatum T-29A1, with similarities around 96–100%. The isolates selected were able to produce enzymes including chitinase, exo-β-1,3-glucanase, and endo-β-1,3-glucanase at levels of 0.34–0.44, 0.017–0.034, and 0.032–0.121 UI · ml –1, respectively, which were considered to be a mechanism to prevent the growth of F. oxysporum F.28.1A. The isolates tested were applied in soil pots to prevent damage from F. oxysporum F.28.1A as a following experiment. The greenhouse experiment was arranged in a completely randomized design with 10 treatments, including a negative control, application of only F. oxysporum F.28.1A, application of both F. oxysporum F.28.1A and fungicide chemicals, application of both F. oxysporum F.28.1A and Trichoderma spp. DHCT, application of T. yunnanense T-02B1, application of T. lentiforme T-18B2, application of T. asperellum T-20B1, application of T. hamatum T-28B1, application of T. hamatum T-29A1 and a mixture of the five selected isolates of Trichoderma spp. with their total population equal to that in individual strain application. The results showed that the five mixed isolates of Trichoderma had a synergistic effect on the reduction of the disease’s prevalence by 35% compared to the negative control treatment.
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Authors and Affiliations

Nguyen Quoc Khuong
1
Cao Thi Thuy Trang
1
Do Thi Xuan
2
Le Thanh Quang
2
Tran Ngoc Huu
1
Ly Ngoc Thanh Xuan
3
Jun-Ichi Sakagami
4
Le Vinh Thuc
1

  1. Faculty of Crop Science, College of Agriculture, Can Tho University, Viet Nam
  2. Biotechnology Research and Development Institute, Can Tho University, Viet Nam
  3. Experimental and Practical Area, An Giang University – Vietnam National University, Ho Chi Minh City, Viet Nam
  4. Tropical Crop Science Laboratory, Faculty of Agriculture, Kagoshima University, Japan
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Abstract

In this work, response surface optimization strategy was employed to enhance the biodegradation process of fresh palm oil mill effluent (POME) by Aspergillus niger and Trichoderma virens. A central composite design (CCD) combined with response surface methodology (RSM) were employed to study the effects of three independent variables: inoculum size (%), agitation rate (rpm) and temperature (°C) on the biodegradation processes and production of biosolids enriched with fungal biomass protein. The results achieved using A. niger were compared to those obtained using T. virens. The optimal conditions for the biodegradation processes in terms of total suspended solids (TSS), turbidity, chemical oxygen demand (COD), specific resistance to filtration (SRF) and production of biosolids enriched with fungal biomass protein in fresh POME treated with A. niger and T. virens have been predicted by multiple response optimization and verified experimentally at 19% (v/v) inoculum size, 100 rpm, 30.2°C and 5% (v/v) inoculum size, 100 rpm, 33.3°C respectively. As disclosed by ANOVA and response surface plots, the effects of inoculum size and agitation rate on fresh POME treatment process by both fungal strains were significant.

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Authors and Affiliations

Noorbaizura Jalaludin
Roshanida A. Rahman
Firdausi Razali
Hind F.A Barghash
Siti Sabrina Mohd Sukri
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Abstract

Ability of five strains of Trichoderma pseudokoningii (antagonists) to suppress radial growth of Fusarium verticillioides (Sacc.) Nirenberg (= Fusarium moniliforme Sheldon) was examined in vitro These were T. pseudokoningii strai n1 (IMI 380933), strain 2 (IMI 380937), strain 3 (IMI 3809 39), strain 4 (IMI 380940) a nd strain 5 (IMI 380941). Each strain was paired with pathogen by inoculating at opposite ends of 9 cm petri plates using three pairing methods. Gradings were assigned to varied growth inhibition of pathogen by antagonists and analysed using GLM procedure (SAS). Growth suppression of F. verticillioides by all strains of T. pseudokoningii was significantly different (R2 = 0.98, p = 0.05) from control in all pairing methods. It differed significantly (p > 0.0003) among the strains in all pairing methods. Growth suppression also differed significantly among (p>0.0001) and within (p > 0.018) pairing methods. Growth suppression was best when antagonists were inoculated before pathogen. Suppression mechanisms include mycoparasitism and competition for space and nutrients. T. pseudokoningii strains 3 and 4 had the best (p = 0.05) growth suppression of F. verticillioides and could be used as biocontrol agents for endophytic F. verticillioides in maize plant. This experiment was conducted in the search for resedent microorganisms that might be capable of checking F. verticillioides within maize plant by competitive exclusion in subsequent experiment.

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Authors and Affiliations

Ayodele Adegboyega Sobowale
Kitty Francies Cardwell
Adegboyega Christopher Odebode
Ranajit Bandyopadhyay
Segun Gbolagade Jonathan
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Abstract

Six isolates of Trichoderma spp. (belonging to species; Trichoderma harzianum and T. longibrachiatum) were applied as seed or soil treatments to suppress damping-off of seedlings of ten cotton cultivars under greenhouse conditions. In most cases, cultivar x isolate interaction was a highly significant (p < 0.01) source of variation in the tested seedling growth parameters: incidence of disease, seedling height, and seedling dry weight. This interaction implies that a single isolate of Trichoderma can be highly effective in controlling the disease on a cotton cultivar but may have minimal efficiency in controlling the disease on another cultivar. It was also found that, in most cases, cultivar x isolate x application method was a highly significant source of variation (p < 0.01) in the tested growth parameters. Cotton cultivars showed differences in the disease reaction to the biocontrol agents. In the experiments evaluating the Trichoderma antagonists and their effect on seedling disease, a highly significant (p < 0.01) experimental treatment interaction was found. This interaction suggests that the outcome of cultivar x isolate interaction is markedly affected by the application method. Thus, the application method should be chosen to maximize the outcome of this interaction. The degree of the control of seedling disease in cotton differed according to the isolates of antagonists, the application method and cultivars.

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Authors and Affiliations

Asran-Amal Abdel-Mongy
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Abstract

Fusarium avenaceum, F. oxysporum, Alternaria alternata, Botrytis cinerea, Rhizoctonia solani, Mucor sp., Aspergillus niger, Penicillium spp. were the most often isolated species from fiber seeds. The antagonistic action of Trichoderma lignorum T 13–82 in relation to seed contaminants was evaluated. The use of Trichodermin-BL, based on that antagonist, applied as pre-sowing seed treatment and on growing plants decrease diseases incidence. The application of Trichodermin-BL improved a set of biometrical and physiological parameters during crop vegetation, increased the yield and raised flax fiber quality.

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Authors and Affiliations

Lyudmila Pristchepa
Dmitry Voitka
Evgeniya Kasperovich
Natalya Stepanova
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Abstract

This study was carried out for the estimation of polyphenols (TP) and induction of oxidative enzymes polyphenol oxidase (PPO) and peroxidase (POD) in sunflower plants through seed immersion in agrochemicals of salicylic acid (SA) and water soluble chitosan (CH) in addition to a conidial suspension of Trichoderma harzianum and then analysis of plant content of carbohydrates and protein. The highest level of PPO 253.3 U ꞏ min –1 was detected in 50 ppm SA for 6 h. Next was T. harzianum when catalyzed PPO with 193.67 U ꞏ min –1. Peroxidase was substantially catalyzed in accordance with the increment of inducers. Sunflower roots induced TP with up to 4.88 mg ꞏ g –1 in plants treated with SA at 50 ppm for 6 h and then declined with an increasing SA dose. The total carbohydrate content in leaves of 320 mg ꞏ 100 g –1 was found in treatments of CH at 50 ppm for 6 h. In roots, a carbohydrate content of 500 mg ꞏ 100 g –1 was observed using CH 75 ppm for 6 h. Trichoderma harzianum remarkably increased proteins in leaves and roots by up to 25% compared to 16.9% in the control. These results suggest that inducing the plants’ own defense mechanism by applying salicylic acid and chitosan and bio-control of T. harzianum may offer alternative methods for controlling charcoal rot of sunflower due to the creation of defensive enzymes and could support plant vigor by enhancement of its protein and carbohydrate content.
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Authors and Affiliations

Khadeeja Ahmed Sido
1
Wazeer Ali Hassan
1
ORCID: ORCID

  1. Plant Protection Department, College of Agricultural Engineering Sciences, University of Duhok, Kurdistan Region, Duhok, Iraq
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Abstract

Clethodim herbicide (Cle) and three Trichoderma strains (Tri) were applied either alone or in combination (Cle + Tri) for controlling weeds, root knot nematodes (Meloidogyne arenaria) and Rhizoctonia root rot disease (Rhizoctonia solani) as well as for evaluating their effects on total microbial count in the rhizosphere and the number of Rhizobium nodules on roots in two faba bean cultivars cultivated in naturally heavily infested fields. The evaluated characters were very similar for the two tested cultivars (Nubariya 1 and Sakha 3). Treatment with Cle alone highly reduced the fresh and dry matter of tested weeds (Amaranthus viridis, Cynodon dactylon and Cenchrus ciliaris), followed by Cle + Tri and Tri alone. Cle + Tri highly reduced nematode parameters viz. numbers of J2 in soil or roots, females, eggs, galls and egg-masses when compared with each treatment alone. Tri alone caused a great decrease in Rhizoctonia root rot infection, followed by Cle + Tri and Cle alone. Total microbial count and Rhizobium nodules were affected only with Cle treatment. Plant growth parameters (shoot length, shoot fresh and dry weight and numbers of branches and leaves) and yield parameters (fresh pod and dry weight, seed number per pod, seed weight and ash pod weight of plant) were greatly improved for Cle + Tri treatments when compared with either Tri or Cle alone.

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Authors and Affiliations

Mahmoud A.T. El-Dabaa
Hassan Abd-El-Khair
ORCID: ORCID
Wafaa M.A. El-Nagdi
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Abstract

A representative group of hydrophilic fungi from the genus Trichoderma isolated from lignocellulose composts with varying degrees of maturity was analyzed for their ability to biodegrade a harmful anthraquinone dye, i.e. Remazol Brilliant Blue R (RBBR). In RBBR-containing post-culture liquids, there were determined the degree of RBBR decolorization, horseradish peroxidase-like, superoxide dismutase-like, and xylanase activities, and the concentrations of low-molecular phenolic compounds. The study results demonstrated that Trichoderma asperellum, T. harzianum, and T. lixii strains isolated from compost containing larger amounts of easily available lignocellulose fractions, i.e. grasses, exhibit higher RBBR decolorization effi ciency ranging from 0.3 to 62% than T. citrinoviride strains isolated from compost II, which contained greater quantities of hardly degradable lignocellulose. The decolorization of remazol blue R by the investigated Trichoderma strains intensified signifi cantly with the increase in peroxidase activity and it was correlated with a decline in the content of low-molecular phenolic compounds. The dynamics of changes in the horseradish peroxidase-like, superoxide dismutase, and xylanase activities in the aqueous post-culture liquids of the investigated fungal strains depended largely on the duration of the culture. Given their ability to adapt to water environments, e.g. wastewater, and to decolorize and detoxify the RBBR anthraquinone dye, Trichoderma fungi can be used for bioremediation of such environments.

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Authors and Affiliations

Justyna Bohacz
1

  1. University of Life Sciences in Lublin, Faculty of Agrobioengineering, Department of Environmental Microbiology, Poland
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Abstract

This work aimed to evaluate the potential of Trichoderma asperellum organic extract and its emulsion to control cocoa black pod disease caused by Phytophthora megakarya. Organic extract was obtained after fermentation of T. asperellum and its emulsion prepared by emulsification. The in vitro antimicrobial assays of organic extract and its emulsion were evaluated and the in situ tests were carried out on detached cocoa pods. T. asperellum inhibited the mycelia growth of P. megakarya at the rates of 52% and 100%, respectively, on dual culture and the cellophane plate. This antagonist produced lytic enzymes such as cellulase, amylase, lipase and protease. The organic extract contained alkaloid, flavonoid and phenol compounds. The emulsion obtained was stable. At 100 μg · ml -1, the extract and its formulation completely inhibited the mycelial growth of P. megakarya. Similarly, when infected detached cocoa pods were sprayed with extract or emulsion, there was a significant reduction of necrosis both for healing and prevention with the latter being the most efficient. For the preventive tests, the total inhibition was recorded at 3000 μg · ml -1 and 1000 μg · ml -1, respectively, with crude organic extract and its emulsion· For curative tests, total inhibition was obtained at 4000 μg · ml -1 and 3000 μg · ml -1, respectively, for preventive and curative tests. There was a significant and positive correlation between the content of biochemical markers and the reduction of necrosis on cocoa pods after treatment with the extract or its formulation. T. asperellum organic extract emulsion could be used as an alternative in the bio- protection of cocoa black pods disease.
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Authors and Affiliations

Yves Olivier Youassi Youassi
1
Henri Tibo Ambata Ambata
1
Mylaure Benisa Jiogue
1
Fabiola Jeannette Dikongue Ndondoni
1
Moïse Ntah A Ayong
1
Marie Amperes Bedine Boat
2
Severin Tchameni Nguemezi
1
ORCID: ORCID
Modeste Lambert Sameza
1

  1. Department of Biochemistry, Faculty of Science, University of Douala, Douala, Cameroon
  2. Department of Agriculture, Faculty of Agronomy and Agronomic Sciences, University of Dschang, Dschang, Cameroon
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Abstract

Many species of Trichoderma produce secondary metabolites such as volatile organic compounds (VOCs) that reduce plant diseases and promote their growth. In this work we evaluated the antagonistic effects of VOCs released by eight strains of two Trichoderma species against Pyrenophora teres Drechsler, the causal agent of barley net blotch. Antagonism was estimated based on the percentage of mycelial growth inhibition according to the confronted cultures method. VOCs extraction and identification were performed by gas chromatography and mass spectrometry, through different methodologies for VOCs emitted by antagonists and pathogens alone or when confronted. VOCs produced by all Trichoderma strains inhibited mycelial growth of the pathogen in a range of 3 to 32%, showing weak and unpigmented mycelia with vacuolization. In addition, P. teres stimulated the release of VOCs by both Trichoderma species. The major groups of VOCs detected were sesquiterpenes, followed by diterpenes, terpenoids and eight-carbon compounds. This is the first report about characterization of volatiles emitted by Trichoderma in the presence of P. teres.
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Authors and Affiliations

Paulina Moya
Juan Roberto Girotti
Andrea Vanesa Toledo
Sisterna Marina Noemi

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