Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Growth of Schizolobium parahyba var. amazonicum seedlings by strains of Trichoderma spp. under boron rates

Brazil has many important native species, which includes Schizolobium parahyba var. amazonicum (Huber ex Ducke) Barneby, however, the growth of this species is inhibited in soils with low boron contents, or excess boron, which causes phytotoxicity. Applications of strains of Trichoderma spp. increase the plants' tolerance to abiotic stresses, including tolerance to low or excess levels of nutrients such as boron (B). Thus, the objective of this work was to evaluate plant development and the effect of strains of Trichoderma spp. in Schizolobium parahyba var. amazonicum seedlings grown under different boron rates. A randomized block experimental design was used, with a 4×5 factorial scheme, consisting of 4 strains of Trichoderma spp. and 5 B rates (0, 0.5, 1.0, 1.5, and 2.0 mg dm-3), with seven replications, in greenhouse. Plant height, stem diameter, fresh and dry mass, leaf, stem and total were evaluated at 120 days after emergence. The stains of Trichoderma spp. and B rates presented significant interaction for all evaluated variables, decreasing phytotoxicity. The strain IBLF006WP (T. harzianum) showed a higher capacity of increasing the plants' tolerance to boron, followed by URM5911 (T. asperellum). However, the beneficial effect of increasing this tolerance with the application of these strains is only feasible for soils with high contents of this micronutrient in the soil.

Methods of plant growth-promoting fungi application to enhance the growth and yield of wheat var. Ibaa99

Aims@#The main aim of the study was to evaluate some methods of application of Aspergillus niger AD1 and Trichoderma hamatum T-113 for enhancing the growth and yield of wheat var. Ibaa99 in pots and field conditions.@*Methodology and results@#Plant growth-promoting fungi (PGPF) loaded with peat moss were used at a rate of 100, 150 and 200 mL pot-1 or m-2 in filed soil; seed treatment (coating) with fungi suspension 19 × 107, soil treatment and combination of all the three methods was employed in the study. Wheat seeds were sown in pots and field plots during 2018-2019, and data regarding various growth and yield attributes were recorded. In both pot and field trials, the results revealed that the best treatments for the desired plant growth and yield attributes were peat moss 150 mL alone or in combination with soil and seed treatments. The soil physicochemical parameters were also improved after inoculation with selected fungal isolates in different application methods compared with un-inoculated control treatment in both pot and field conditions.@*Conclusion, significance and impact of study@#The PGPF play a vital role represented phytoremediation, phytostimulation and bio-fertilization. The isolates of PGPF, which were applied with peat moss at 150 mL to the pot and in the field alone or combined with seed treatment and soil application, were significantly the best effective method for improving wheat attributes.

Performance of a selected Trichoderma strain as plant pathogen inhibitor and biofertilizer

Aims@#The application of beneficial microbes is a suitable alternative to synthetic pesticides and fertilizers for agriculture. This study was aimed to evaluate the potential of a selected Trichoderma strain as a biocontrol agent against Rhizoctonia sp. and as a biofertilizer to improve paddy growth.@*Methodology and results@#Four Bipolaris strains were identified via DNA barcoding as the cause of brown spot disease, whereas two Rhizoctonia strains were similarly identified as the cause of sheath blight disease in Brunei Darussalam. Eight Trichoderma strains were initially screened using confrontation assay and were found to substantially inhibit the growth of Rhizoctonia sp. Hybrid rice named BDR5 was treated with Trichoderma sp. UBDFM01 and/or Rhizoctonia sp. It was found that the selected strain showed the potential as a biofertilizer by significantly increasing the vigour index I, chlorophyll a, chlorophyll b, total chlorophyll and dry shoot weight of the rice plants. The pathogen negatively affected the plants by significantly reducing the vigour index II, chlorophyll a, chlorophyll a/b ratio, total chlorophyll, and total weight of grains. Trichoderma strain showed the potential as a biocontrol agent by significantly diminishing the negative effects of the pathogen on the chlorophyll a, chlorophyll a/b ratio and total chlorophyll.@*Conclusion, significance and impact of study@#This study highlights the potential of Trichoderma sp. UBDFM01 as a biocontrol agent against Rhizoctonia sp. and also as a biofertilizer for rice plants. In addition, this study is the first to provide DNA-based evidence of Bipolaris sp. and Rhizoctonia sp. as the fungi that caused rice diseases in Brunei Darussalam.

Suppression and management of Meloidogyne incognita in soil using Trichoderma harzianum NFCF160 and Trichoderma virens Isf-77

Aims@#Meloidogyne incognita adversely affects numerous crop plants worldwide. Therefore, the modern world has been moving towards biocontrol methods to prevent nematode attacks. This study was aimed to (i) investigate the potential use of Trichoderma harzianum NFCF160 and T. virens Isf-77 in managing M. incognita in soil and (ii) identify trapping mechanisms employed by both Trichoderma strains to suppress M. incognita.@*Methodology and results@#Three weeks old, Basella alba L. plants were subjected to five different treatments. The above and below ground growth parameters and the galling indices of these plants were measured every four weeks for three sampling times. Trapping mechanisms employed by Trichoderma strains were examined following plate assays. Plants treated with T. harzianum NFCF160 and T. virens Isf-77 had significantly higher values for the total number of leaves (34 ± 2.84) and (27 ± 2.61), fresh weight of the shoot (81 ± 9.51 g) and (91 ± 9.70 g), dry weight of the shoot (71 ± 5.24 g) and (62 ± 5.81 g), respectively eight weeks after inoculation of M. incognita. Significantly low galling indices (2 and 2) were recorded in B. alba treated with Trichoderma strains. Both Trichoderma strains exhibited various nematode-trapping mechanisms, such as non-constricting rings and adhesive spores.@*Conclusion, significance and impact of study@#This investigation highlighted the potential of both Trichoderma strains as biocontrol agents to control M. incognita effect in sustainable agriculture.

Biological control of Meloidogyne javanica in bean plants by Hohenbuehelia spp. and Trichoderma koningiopsis

Nematode control strategies are limited and alternative control is demanded. The main aim was to evaluate control potential of Hohenbuehelia spp. (Hohenbuehelia mastrucata, Hohenbuehelia barbatula, Hohenbuehelia bullulifera, Hohenbuehelia portegna, Hohenbuehelia petaloides and Hohenbuehelia paraguayensis) and Trichoderma koningiopsis against Meloidogyne javanica in bean plants in greenhouse. The parameters evaluated were the number of galls and egg masses which were evaluated in three root regions: basal, intermediate and tip; nematode reproduction factor; vegetal growth parameters (plants height on V1, V2, V4, and R5 growth stages, root length, dry weight of root and aerial part dry weight); Treatments were composed by individual interactions of each fungal isolate with 4,000 eggs of M. javanica, having a total control (bean) and a partial control (bean + nematode) arranged in completely randomized design with four repetitions. Data were tested by Scott­Knott (5%). For vegetal growth parameters, there was only significance for dry weight of roots. All species were able to control nematodes.

Antagonistic microorganisms and nitrogen fertilization in control of tomato southern blight

The present study assessed the efficacy of formulated biocontrol agents and nitrogen fertilization on southern blight control. Antagonism test in vitro was performed to assess the inhibitory activity of Bacillus methylotrophicus and Trichoderma asperellum against the growth of Sclerotium rolfsii. Tomato seedlings were transplanted into the substrate added with ammonium nitrate doses and inoculated with the formulated biocontrol agents Ônix (B. methylotrophicus) or Quality (T. asperellum). Subsequently, seedlings were inoculated with S. rolfsii. Plant mortality, shoot and root weight were assessed 11 days after the last inoculation. Agents had effective inhibitory activity against S. rolfsii; thus, they could reduce southern blight severity when combined with ammonium nitrate. However, plant mortality was not reduced by them.(AU)

Isolation of culturable mycota from Argentinean soils exposed or not-exposed to pesticides and determination of glyphosate tolerance of fungal species in media supplied with the herbicide / Aislamiento de la microbiota cultivable de suelos argentinos expuestos y no expuestos a plaguicidas y determinación de la tolerancia a glifosato de especies fúngicas en medios suplementados con el herbicida

Abstract The current agricultural system has led to the development of glyphosate (GP)-resistant weeds, causing an increase in GP doses and applications. Native mycota of pesticide-contaminated sites are the major source of pesticide-degrading microorganisms. The aims of the present study were to isolate the GP-tolerant culturable mycota in two soils with different pesticide exposure from Córdoba, Argentina, and to evaluate the growth parameters in native fungal isolates in the presence of GP and the effective dose that caused 50% growth reduction. The results showed that the genera Fusarium, Aspergillus, Mucor, Penicil-lium and Sterilia were the prevalent fungi isolated from soils both exposed and not-exposed to pesticides. The highest value (>100 mM) of effective concentration of herbicide that caused 50% growth inhibition (EC50), was found for Trichoderma isolates. Sterilia spp. had EC50 val-ues of 100mM, while Aspergillus spp. and Mucor had EC50 values between 50 and 100mM. The growth rate evaluation varied according to the isolates and GP concentrations. The data showed that all Aspergillus spp., Trichoderma spp., Mucor and three Sterilia spp. had the best growth performance in media supplied with GP after a variable acclimation period. This study provides valuable data for further studies that would allow to know the metabolic capacity of these fungal species that can be potential candidates for GP removal from contaminated environments.

Resumen El sistema agrícola actual condujo ai desarrollo de malezas resistentes a glifos-ato (GP), lo que deriva en el aumento de las dosis y el número de aplicaciones requeridas. La microbiota nativa de sitios contaminados con plaguicidas constituye la fuente principal de microorganismos capaces de degradar este herbicida. Los objetivos del presente estudio fueron aislar la microbiota cultivable tolerante a GP a partir de dos suelos agrícolas con diferente grado de exposición a plaguicidas, ambos de la provincia de Córdoba, Argentina, y evaluar en los hongos nativos aislados los parámetros de crecimiento en presencia de GP y la dosis efectiva que produce un 50% de reducción del crecimiento (EC50). Los géneros Fusarium, Aspergillus, Mucor, Penicillium y Sterilia fueron los de mayor prevalencia, tanto en suelos expuestos como no expuestos a plaguicidas. La mayor EC50 hallada (> 100 mM) correspondió a dos aislamientos de Trichoderma. En dos aislamientos de Sterilia, los valores de EC50 fueron de 100 mM, mientras que en los dos de Aspergillus y en uno de Mucor fueron de entre 50 y 100 mM. La velocidad de crecimiento varió según la concentración de GP empleada y el aislamiento considerado. Todos los aislamientos de Aspergillus, Trichoderma y Mucor, y tres aislamientos de Sterilia mostraron el mejor rendimiento en cuanto al crecimiento en los medios con GP después de un período de aclimatación variable. Este trabajo proporciona datos preliminares valiosos referidos a la capacidad metabólica de estos aislados y su potencial para remover GP en ambientes contaminados.

Monitoramento de fungos anemófilos no ambiente de uma biblioteca no Município de São José do Rio Preto - Sp, Brasil / Anemophilic fungi monitoring in a library environment in the city of São Jose do Rio Preto, Brazil

Os fungos desempenham vários papéis que impactam a humanidade de diversas maneiras. Suas características metabólicas são importantes na biotecnologia, porém, tais microrganismos podem desencadear alguns problemas de saúde pública e até mesmo serem letais. Objetivo: detectar a presença de fungos no acervo de uma biblioteca no município de São José do Rio Preto. Metodologia: foram coletadas quarenta amostras nas superfícies inanimadas (livros, estantes, documentos, mapas, artigos e revistas) das principais salas da biblioteca com o auxílio de swabs umedecidos em solução salina estéril, posteriormente encaminhados ao laboratório de Biomedicina da Universidade Paulista ­ UNIP. As amostras foram semeadas em meio de cultura ágar Sabouraud Dextrose (SDA), tendo adicionado cloranfenicol e incubadas a 30 °C. Foi realizada a colônia gigante em todas as cepas crescidas em SDA para a realização da técnica de microcultivo para a identificação dos fungos, de acordo com o Manual de Detecção e Identificação dos Fungos de Importância Médica da Agência Nacional de Vigilância Sanitária. Resultados: Houve positividade em trinta e uma amostras (78%) e em quatro delas foi observado mais de um tipo de colônia (13%). Das vinte e duas superfícies de livros analisadas, foram isolados e identificados: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. e Nigrospora sp. Nas oito superfícies de estantes: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. e Scopulariopsis sp. e, nos dez documentos: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. e Trichoderma sp. Conclusão: Os fungos encontrados estão amplamente distribuídos no ambiente como solo e ar e, por diversos fatores, instalam-se em locais como bibliotecas. Em condições favoráveis, podem infectar o homem e causar perdas patrimoniais para os acervos.

Fungi play many roles that impact humankind in different ways. Their metabolic characteristics are important in biotechnology; however, these microorganisms can trigger some public health problems or may even be lethal. Objective: detect the presence of fungi in the collection of a public library in the city of São José do Rio Preto, Brazil. Methods: a total of forty samples were collected from inanimate surfaces (books, shelves, documents, maps, articles and magazines) located in the main rooms of the library with swabs soaked in sterile saline solution and sent to the Universidade Paulista ­ UNIP laboratories. The samples were plated in Sabouraud Dextrose Agar (SDA) supplemented with chloramphenicol and incubated at 30 °C. The colonies that grew in SDA were isolated in Potato Dextrose Agar for performing the slide culture technique for the identification of the fungi, performed according to the Manual of Detection and Identification of Fungi of Medical Importance from the Brazilian Health Surveillance Agency (ANVISA). Results: Thirty-one samples (78%) were positive, and in four of them more than one fungus genus was observed (13%). From the twenty-two book surfaces analyzed, the following fungi were isolated and identified: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. and Nigrospora sp. On the eight shelves: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. and Scopulariopsis sp. The ten documents analyzed presented the following fungi: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. and Trichoderma sp.. Conclusion: These fungi are widely distributed in the environment such as in the soil and air, and due to several factors, they colonize public places, such as libraries. In favorable conditions, they may infect humans and cause diseases.

Actividad antimicrobiana del aceite esencial de hojas de Psidium cattleianum Afzel. ex Sabine / Antimicrobial activity of essential oil from Psidium cattleianum Afzel. ex Sabine leaves

La búsqueda de fuentes naturales para controlar los microorganismos es de interés en la producción de alimentos. Este estudio evaluó la composición química y la actividad antimicrobiana del aceite esencial de las hojas de Psidium cattleianum. El aceite esencial se extrajo por hidrodestilación y se identificó por GC-MS. La clase predominante de compuestos fueron los sesquiterpenos (47,6%) y los principales fueron trans-ß-cariofileno (14,7%), 1,8-cineol (11,7%) y É£-muuroleno (5,6%). Actividad antimicrobiana se realizó mediante la técnica de microdilución contra ocho hongos y ocho bacterias. Concentración inhibitoria mínima varió de 0,17 a 11,25 mg mL-1 para hongos y de 1,40 a 16,87 mg mL-1 para bacterias. Principales actividades fueron contra hongos Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730) y Trichoderma viride (IAM 5061), y bacterias Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate) y Staphylococcus aureus (ATCC 6538) con potencial para prevenir enfermedades transmitidas por alimentos.

The search for natural sources to control microorganisms is of interest in food production. This study evaluated the chemical composition and antimicrobial activity of the essential oil from Psidium cattleianum leaves. The essential oil was extracted by hydrodistillation, and identified by GC-MS. The predominant class of compounds was sesquiterpenes (47.6%) and the major compounds were trans-ß-caryophyllene (14.7%), 1,8-cineole (11.7%) and É£-muurolene (5.6%). The antimicrobial activity was carried out by microdillution technique against eight fungi and eight bacteria. The minimum inhibitory concentration ranged from 0.17 to 11.25 mg mL-1 for fungi, and from 1.40 to 16.87 mg mL-1 for bacteria. The highest activities were against fungi Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730), and Trichoderma viride (IAM 5061), and bacteria Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate), and Staphylococcus aureus (ATCC 6538) with potential to prevent foodborne diseases.

Effect of chemical and biological seed treatments on common bean seeds inoculated with Fusarium oxysporum f. sp. phaseoli / Efeito de tratamentos químicos e biológicos de sementes em sementes de feijoeiro inoculadas com Fusarium oxysporum f. sp. phaseoli

Currently, the common bean (Phaseolus vulgaris L.) productivity has been reduced by the fungus Fusarium oxysporum f. sp. phaseoli (Fop), the causative agent of Fusarium wilt. Considering the integrated management of diseases, the objectives of the present work were to verify the compatibility between chemical and biological fungicides for Fusarium oxysporum Schlecht. f. sp. phaseoli Kendrick & Snyder (Fop) control in common bean seeds. In laboratory, the effects of the treatments were evaluated by sanity, germination, seedling total length and seedling dry matter tests. In greenhouse conditions, the emergence speed rate, the percentage of emergence and the rate of pathogen transmission through the pathogen infestation in a substrate to plants were evaluated. Common bean seeds BRS Estilo were artificially inoculated with Fop isolate (IAC 11.299-1). In the seeds' treatment, the chemical fungicides fludyoxonyl, flutriafol, methyl tiofanate, and biological products of Trichoderma sp. (isolates SF04, GF 422 and strain 1306), separately and mixed, were used. Treatments that promoted the best pathogen control in seeds were the combination of methyl tiophanate with biological products. Both flutriafol and GF 422 isolated and in mixed treatments affected the seeds' physiological quality. The protective effect of the products was noted in the transmission test, whose Fop incidence was from 5 to 40% in the hypocotyl and from 5 to 30% in common bean roots.(AU)

Atualmente, a produtividade do feijão comum (Phaseolus vulgaris L.) foi reduzida pelo fungo Fusarium oxysporum f. sp. ­phaseoli (Fop), o agente causador da murcha de Fusarium. Considerando o manejo integrado de doenças, os objetivos do presente trabalho foram verificar a compatibilidade entre fungicidas químicos e biológicos para Fusarium oxysporum Schlecht. f. sp. phaseoli Kendrick & Snyder (Fop) em sementes de feijão. Em laboratório, foram avaliados os efeitos dos tratamentos pelos testes de sanidade, germinação, comprimento total de plântulas e matéria seca de plântulas. Em uma estufa, foram avaliadas a velocidade de emergência, a porcentagem de emergência e a taxa de transmissão de patógenos através da infestação de patógenos no substrato às plantas. As sementes de feijão comum BRS Estilo foram inoculadas artificialmente com isolado Fop (IAC 11.299-1). No tratamento das sementes, foram utilizados os fungicidas químicos fludioxoxil, flutriafol, tiofanato metílico e produtos biológicos de Trichoderma sp. (isolados SF04, GF 422 e cepa 1306), separadamente e misturados. Os tratamentos que promoveram o melhor controle de patógenos nas sementes foram a combinação de tiofanato de metila com produtos biológicos. Tanto o flutriafol quanto o GF 422 isolados e em tratamentos com misturas afetaram a qualidade fisiológica das sementes. O efeito protetor dos produtos foi observado no teste de transmissão, cuja incidência de Fop foi de 5 a 40% no hipocótilo e de 5 a 30% nas raízes do feijão comum.(AU)

Action of Trichodermil 1306 in the control of Fusarium oxysporum f. sp. lycopersici in tomato crops / Ação do Trichodermil 1306 no controle de Fusarium oxysporum f. sp. lycopersici na cultura do tomateiro

Fusarium wilt, caused by Fusarium oxysporum f. sp. lycopersici, is one of the most important diseases of tomato crop in Brazil. Several alternatives to fungicides have been evaluated in search for products that control this disease and have low environmental impact and toxicity to living beings. In this context, the use of Trichoderma spp. is an alternative to agrochemicals, since they are bioregulators and antagonists of several phytopathogens. This work aimed to evaluate the antagonistic and mycoparasitory action of the Trichodermil 1306 biofungicide against F. oxysporum, and its action in the control of fusarium wilt in tomato seeds and seedlings. The biofungicide was used at concentrations of 0, 50, 100, and 150% of the dose recommended for application of 20 L with a backpack sprayer. The experiment was conducted in a completely randomized design with five replications. Data were submitted to analysis of variance and mean compared by Tukey's test at 5% probability. The 100% Trichodermil concentration was the most efficient for treating tomato seeds. Although this concentration did not show differences in relation to the lowest dose used (50%) in the tomato seed antagonism, mycoparasitism and F. oxysporum incidence tests were recommended because that the lowest concentration resulted in 18% lower root protrusion than that observed with 100% dosage. The in vitro antagonistic and mycoparasitory action of this biological fungicide indicates its potential in the control of fusarium wilt in tomato crops.(AU)

A murcha-de-fusário, causada por Fusarium oxysporum f. sp. lycopersici, é uma das mais importantes doenças da cultura do tomate no Brasil. Várias alternativas aos fungicidas têm sido avaliadas na busca de produtos que controlem a doença e tenham baixo impacto ambiental e toxicidade aos seres vivos. Nesse contexto, o uso de Trichoderma spp. é uma alternativa a agrotóxicos, pois são biorreguladores e antagonistas de vários fitopatógenos. O objetivo do trabalho foi avaliar a ação antagonista e micoparasitória do biofungicida Trichodermil 1306 em F. oxysporum, e no controle da murcha-de-fusário em sementes e mudas de tomate. O biofungicida foi utilizado nas concentrações de 0, 50, 100 e 150% da dose recomendada para aplicação com pulverizador costal de 20 L. O experimento foi conduzido em delineamento inteiramente casualizado, com cinco repetições. Os dados foram submetidos à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A concentração de Trichodermil 100% foi a mais eficaz para o tratamento de sementes de tomateiro. Embora essa concentração não tenha se diferenciado da menor dosagem utilizada (50%) para os testes de antagonismo, micoparasitismo e incidência de F. oxysporum em mudas de tomate, sua recomendação se justifica pelo fato da menor concentração ter propiciado protrusão radicular 18% menor do que a observada na concentração de 100%. A ação antagonista e micoparasitória in vitro do fungicida biológico testado no presente estudo indica potencialidade de uso para o controle de murcha-de-fusário em tomateiro.(AU)

Effect of Pleurotus ostreatus (Jacq) and Trichoderma harzianum (Rifai) on Meloidogyne incognita (Kofoid & White) in tomato (Solanum lycopersicum Mill)

Two isolations of fungi from a bank of microorganisms in the Biological Sciences Laboratory at Escuela Superior Politécnica de Chimborazo were tested on the galling caused by Meloidogyne incognita in tomato seedlings grown in pots with substrate infested with a suspension of nematodes, with approximately 2000 juvenile stages (J2) from root galls of plants infested with M. incognita, taken from the Nematology laboratory of the Ecuadorian Agricultural Quality Assurance Agency (AGROCALIDAD). Pleurotus ostreatus was a fungus with nematicidal characteristics through production of toxins; while Trichoderma harzianum is a widely known fungus, although it is a plant growth promoter rather than a nematicide. The two fungi were formulated in wheat straw and rabbit manure. A complete randomized design (CRD) with three replications was used, with a chemical control (Fenamiphos) and an absolute control. Five grams of each formulation was applied per plant before the transplant. The number of galls in the roots, plant height, stem diameter, number of leaves and fresh and dry weight of the aerial part and roots of 180 tomato plants grown in greenhouse were evaluated at 60 days after transplant. The results showed that the two fungi reduced the number of galls and made it possible to obtain dry weights of the aerial and radicular part very close to the chemical control (10.09 and 3.39 g) with 8.68; 8.04; 2.96 and 3.25 g respectively. Besides Trichoderma harzianum proved to be a good promoter of root growth, therefore, the use of these bioformulates is a promising measure for the control of this phytonematode

Multiple Effect of Different Plant Growth Promoting Microorganisms on Beans (Phaseolus vulgaris L. ) Crop

Abstract We evaluated the effect of combined Rhizobium tropici, Trichoderma asperellum and plant growth-promoting rhizobacteria (PGPR) in beans crop. The hypothesis that strains of T. asperullum, R. tropici and PGPR combined could improve growth, biomass accumulation and beans yield was tested under greenhouse and field conditions. The treatments consisted of control, mineral nitrogen application and inoculation, isolated and associated with the following microorganisms: Rhizobium tropici, Bacillus subtilis, Trichoderma asperellum and Burkholderia sp. 10N6. Results were evaluated by shoot dry weight (SDW) and root dry weight (RDW), number of nodules and yield components. In greenhouse environment all the microorganisms behaved similarly, and the treatments inoculated with Burkholderia sp. 10N6 (IBu) and R. tropici (IR) stood out regarding the production components. In field conditions the treatments IR and IRTBa presented the highest values of SDW and RDW. Our results suggest that inoculation with R. tropici, T. asperellum and PGPR may promote beans growth and bring benefits to shoot and root accumulation, increase the number of nodules as well as improve yield components, contributing to a sustainable agriculture.

Enhance of Cellulase Production and Biomass Degradation by Transformation of the Trichoderma reesei RUT-C30∆ zface1 Strain

Abstract The second-generation bioethanol employs lignocellulosic materials degraded by microbial cellulases in their production. The fungus Trichoderma reesei is one of the main microorganisms producing cellulases, and its genetic modification can lead to the optimization in obtaining hydrolytic enzymes. This work carried out the deletion of the sequence that encodes the zinc finger motif of the transcription factor ACE1 (cellulase expression repressor I) of the fungus T. reesei RUT-C30. The transformation of the RUT-C30 lineage was confirmed by amplification of the 989 bp fragment relative to the selection marker, and by the absence of the zinc finger region amplification in mutants, named T. reesei RUT-C30Δzface1. The production of cellulases by mutants was compared to RUT-C30 and measured with substrates carboxymethylcellulose (CMC), microcrystalline cellulose (Avicel®) and Whatman filter paper (PF). The results demonstrated that RUT-C30Δzface1 has cellulolytic activity increased 3.2-fold in Avicel and 2.1-fold in CMC and PF. The mutants presented 1.4-fold higher sugar released in the hydrolysis of the biomass assays. These results suggest that the partial deletion of ace1 gene is an important strategy in achieving bioethanol production on an industrial scale at a competitive price in the fuel market.

Effect of Trichoderma spp. on Growth Promotion and Antioxidative Activity of Pepper Seedlings

Abstract One of the main challenges in pepper production is to enhance seed germination energy and germination, and to grow healthy nursery plants with strong root system. Trichoderma species colonize roots as they grow and provide season-long benefits to plants, which is why Trichoderma species are widely used as plant growth promoter agents and promoters of plant defence mechanisms. This study evaluated the effectiveness of seed biopriming with Trichoderma isolates for growth promotion of pepper plants in early stage and their effects on seedling physiology. Nine out of ten Trichoderma isolates positively affected root weight of pepper seedlings, while three out of ten positively affected shoot weight. Root and shoot lengths were mainly unaffected. Germination energy was positively affected by five isolates with up to 40% increase compared to the control, while germination was significantly enhanced by two isolates with up to 22% increase. Considering seedling physiology, two different strain-dependent modes of actions were expressed. Promising Trichoderma isolates induced formation and accumulation of reactive oxygen species (ROS) which acted as signal molecules that increased germination energy and germination. Positive correlation was found between pyrogallol peroxidase, superoxide dismutase, catalase activity and germination in plants treated with these isolates.

Antagonistic activity of volatile metabolites from Trichoderma asperellum / 生物工程学报

Trichoderma spp. is a kind of filamentous fungi with important biocontrol value. Twelve strains of Trichoderma spp. were isolated from the soils of different types of crops in Shaoxing, Zhejiang and Foshan, Guangdong. The antagonistic resistance to Fusarium oxysporum was compared by plate confrontation test. The further analysis of volatile secondary metabolites for two strains were carried out using HS-SPME-GC-MS analysis. The results showed that T. asperellum ZJSX5003 and GDFS1009 had fast growth ability, and the inhibition effects on F. oxysporum were 73% and 74% respectively. Six identical volatile metabolites were detected as follows 2-Methyl-1-propanol, 3-Methyl-1-butanol, 3-Methyl-3-buten-1-ol, Acetyl methyl carbinol, Butane-2,3-diol and 6-n-pentyl-2H-pyran-2-one (6-PAP). Among them, 6-PAP was validated to have a higher inhibitory effect on F. oxysporum in vitro. This study will provide basis for the development of biocontrol agents with metabolites of Trichoderma, such as 6-PAP.

Combined effect of pseudomonas sp. and trichoderma aureoviride on lettuce growth promotion / Efeito combinado de Pseudomonas sp. e Trichoderma aureoviride na promoção do crescimento de alface

Plant growth promotion by microorganisms may be a viable alternative to increase lettuce production through pathogens control and nutrients absorption increase. Trichoderma and Pseudomonasgenus are examples of widely studied microorganisms with the capacity to promote plant growth. However, there are still gaps regarding the action of the combined effect of these two microorganisms. Therefore, the objective of this study was to evaluate the combined effect of Pseudomonas sp. UAGF14 and Trichoderma aureoviride URM5158 on the development of lettuce plants. The experimental design was completely randomized, with five treatments: CONT (control), CM (soil with organic fertilization), CMB (soil withorganic fertilization and Pseudomonas sp.), CMF (soil with organic fertilization and T. aureoviride), and CMFB (soil with organic fertilization, Pseudomonas sp. and T. aureoviride), with ten repetitions. At 30, 40 and 60 days after sowing, the following parameters were analyzed: plant and canopy height and number of leaves. At 60 days after emergence, shoot dry matter, leaf area, root dry matter, root length and chlorophyll were analyzed. Catalase, peroxidase and polyphenol oxidase enzymatic activity were determined. The CMFB treatment had the highest means of lettuce growth promotion, confirming the synergistic effect of the combination of the two microorganism types, as it increased height, canopy, shoot and root dry matter, and chlorophyll levels compared to CONT, although did not differ from CM in some variables. Enzymatic activity was also influenced by the action of these microorganisms combined, evidencing by polyphenol oxidase increase. The CMFB or CM were efficient in promoting lettuce growth, showing positive response to the plant morphological and physiological characteristics. However, few responses were observed in lettuce plant growth in the first cycle evaluated after 60 days, compared CM and CMFB treatments, but both treatments showed superiority in lettuce plant growth submitted to CONT treatment. Therefore, further studies are needed to estimate the long-term effects of combined effect of Pseudomonas sp. UAGF14 and T. aureoviride URM5158 on crop productivity in field conditions.

A promoção do crescimento das plantas por micro-organismos pode ser uma alternativa viável para aumentar a produção de alface através de controle de patógenos e aumento da absorção de nutrientes. O gênero fúngico Trichoderma e o gênero bacteriano Pseudomonas são exemplos de micro-organismos amplamente estudados com capacidade para promover o crescimento da planta. No entanto, ainda existem lacunas quanto à ação do efeito combinado desses dois micro-organismos. Portanto, o objetivo deste estudo foi avaliar o efeito combinado de Pseudomonas UAGF14 e Trichoderma aureoviride URM5158 sobre o desenvolvimento de plantas de alface. O delineamento experimental foi completamente casualizados, com cinco tratamentos: CONT (controle, sem fertilização orgânica), CM (solo com fertilização orgânica), CMB (solo com fertilização orgânicae Pseudomonas sp.), CMF (solo com fertilização orgânica e T. aureoviride) e CMFB (solo com fertilização orgânica, Pseudomonas sp. e T. aureoviride), com dez repetições. Aos 30, 40 e 60 dias após a semeadura, foram analisados os seguintes parâmetros: altura da planta e dossel e número de folhas. Aos 60 dias após a emergência, a matéria seca da parte aérea, a área foliar, a massa seca das raízes, o comprimento radicular e a clorofila foram analisados. Catalase, peroxidase e atividade enzimática da polifenol oxidase foram determinadas. O CMFB apresentou o maior crescimento de alface, confirmando o efeito benéfico da combinação dos dois tipos de micro-organismos com a planta, na medida em que aumentou a altura, o dossel, a matéria seca da parte aérea e da raiz, e os níveis de clorofila em relação ao CONT, embora não tenha diferido do CM em algumas variáveis. As atividades enzimáticas também foram influenciadas pela ação desses micro-organismos combinados, evidenciada pelo aumento de polifenol oxidase. O CMFB ou CM foram eficientes na promoção do crescimento da alface, mostrando respostas positivas às características morfológicas e fisiológicas. Entretanto, poucas respostas foram observadas no crescimento da alface noprimeiro ciclo da planta avaliado depois de 60 dias, comparando os tratamentos CM e CMFB, mas ambos tratamentos mostraram superioridade em relação ao crescimento das plantas de alface submetidas ao tratamento controle. Por isso, são necessários futuros estudos para estimar à longo prazo o efeito combinado de Pseudomonas sp. UAGF14 e Trichoderma aureoviride URM5158 na produção de cultura em condições de campo.

Trichoderma species diversity in rhizosphere soils and potential antagonism with Fusarium oxysporum / Diversidade de espécies de trichoderma em solos de rizosfera e potencial antagonismo com fusarium oxysporum

Neste estudo foram isoladas seis espécies de trichoderma isoladas de solos rizosféricos de arrozais, bananeiras, dendezeiros, seringueiras, hortaliças e pastagens. As espécies são t. harzianum, t. viride, t. koningii, t. asperellum, and t. parareesei. O estudo morfológico como pigmentação, crescimento de colônias e estudos anatômicos como aparências de conidiação, tamanho de conídios, padrão de ramificação dos conidióforos, formas de phialides, ausência ou presença de clamidósporos foram realizados para identificar as espécies de trichoderma. As espécies de trichoderma harzianum foram abundantes no solo enquanto as de t. viren foram a segunda mais frequente no solo. Todas as espécies apresentaram atividade antagônica contra o fusarium oxysporum. Enquanto t. parareesei apresentou a maior atividade antagônica de 91,10% contra f.oxysporum, relatado como melhor agente antagonista para fitopatógeno.

In this study, six species of trichoderma isolated from rhizospheric soils of rice fields, banana trees, oil palm trees, rubber trees, vegetables and pastures were isolated. The species are t. harzianum, t. viride, t. koningii, t. asperellum, and t. I will stop. Morphological studies such as pigmentation, colony growth and anatomical studies such as appearance of conidia, size of conidia, branching pattern of conidiophores, forms of phialides, absence or presence of chlamydospores were performed to identify the species of trichoderma. The species of trichoderma harzianum were abundant in the soil while those of t. viren were the second most frequent in the soil. All species showed antagonistic activity against fusarium oxysporum. While t. parareesei presented the greatest antagonistic activity of 91.10% against f.oxysporum, reported as the best antagonistic agent for phytopathogen.