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Cordyceps militaris is a fungus with numerous therapeutic properties that has gained worldwide popularity due to its potential health benefits. The fruiting body of this mushroom is highly expensive and takes a longer time to produce, making mycelial a sustainable and cost-effective alternative. The study investigates and optimizes cultural and nutritional conditions to maximize mycelial biomass. The initial optimization was done by the conventional single-factor approach, followed by Plackett-Burman design to screen the most significant variables, with yeast extract, temperature, and glucose being the most significant, contributing 11.58%, 49.74%, and 27.98%, respectively, in mycelial biomass production. These variables were then optimized using response surface methodology (RSM) based on central composite design (CCD). The study observed that temperature and glucose had the highest impact on mycelial biomass, with p-values of 0.0128 and 0.0191, respectively. Under the optimized conditions, temperature 20 °C, glucose 2.5% (w/v), and yeast extract 0.8% (w/v), the maximal yield of mycelial biomass reached 547 ± 2.09 mg/100 mL, which was 1.95-fold higher than the yield in the basal medium. These findings suggest that optimizing the cultural and nutritional conditions can enhance mycelial biomass production of Cordyceps militaris, offering a sustainable and cost-effective source of this valuable fungus.

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The use of agrochemicals has caused environmental problems and toxicity to humans, so natural alternatives for disease control during harvest and postharvest have been evaluated. The aim of this study was to evaluate cinnamon essential oil, neem oil, and black sapote fruit extract for in vitro inhibition of fungi isolated from chayote fruit. The extracts were applied at 300, 350, and 400 ppm in Petri dishes and the mycelial growth of Fusarium oxysporum, Fusarium solani, Goetrichum sp., and Phytophthora capsici was evaluated for 7 days, and the percentage of mycelial growth inhibition per day was calculated. Cinnamon oil showed a fungicidal effect at all concentrations. Neem oil at 400 ppm showed a 42.3% reduction in the growth of F. solani and 27.8% reduction in the growth of F. oxysporum, while at 350 ppm it inhibited the mycelial growth of Phytophthora capsici by 53.3% and of Goetrichum sp. by 20.9%; finally, the black sapote extract at 400 ppm inhibited 21.9-28.6% of the growth of all fungi. The growth of postharvest fungi on chayote fruit could be prevented or reduced by applying the plant extracts evaluated at adequate concentrations.

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Monilinia fructicola (Wint.) Honey is a plant pathogenic fungus that infects stone fruits such as peach, nectarine and plum, which are high demand cultivars found in Brazil. This pathogen may remain latent in the host, showing no apparent signs of disease, and consequently may spread to different countries. The aim of this study was to evaluate the activity of hydroalcoholic extract (HydE) obtained from Lactarius deliciosus (L.) Sf. Gray a mushroom, against M. fructicola phytopathogenic-induced mycelial growth. In addition, the purpose of this study was to examine phytotoxicity attributed to HydE using Brassica oleracea seeds, as well as cytotoxic analysis of this extract on cells of mouse BALB/c monocyte macrophage cell line (J774A.1 cell line) (ATCC TIB-67). The L. deliciosus HydE inhibited fungal growth and reduced phytopathogen mycelial development at a concentration of 1.25 mg/ml. Our results demonstrated that the extract exhibited phytotoxicity as evidenced by (1) interference on germination percentage and rate index, (2) decreased root and initial growth measures, and (3) lower fresh weight of seedlings but no cytotoxicity in Vero cell lines. Data suggest that the use of the L. deliciosus extracts may be beneficial for fungal control without any apparent adverse actions on mouse BALB/c monocyte macrophage cell line (J774A.1 cell line) viability.

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This paper aims to investigate, for the first time, physicochemical characteristics and antioxidant capacity of Psidium myrtoides ripe and unripe fruit. In addition, essential oil was extracted from fresh leaves (PM-EO) collected in dry and rainy seasons to investigate the influence of seasonality and, after that, to evaluate its antifungal activity on mycelial growth of Colletotrichum gloeosporioides. Both GC-FID and GC-MS analyses revealed that major components determined in PM-EO were ß-caryophyllene (20.0% and 32.9%) and limonene (10.4% and 19.8%) in rainy and dry seasons, respectively. High antioxidant capacity was displayed by unripe fruit (IC50 = 3.57 mg/mL) and ripe ones (IC50 = 3.68 mg/mL). Both showed high total phenolic and tannin contents. In sum, PM-EO showed satisfactory antifungal activity, since its inhibitory action on mycelial growth of C. gloeosporioides was above 70% in the dry season, while it exhibited weak activity in the rainy season.

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RESUMEN La cepa mexicana CP-145 de Ganoderma lucidum debido a la importancia medicinal que ha presentado últimamente, la presente investigación tuvo como objetivo evaluar el efecto de la temperatura y medio de cultivo sobre el crecimiento micelial óptimo en diferentes rangos de pH. Los tratamientos correspondieron en la utilización del medio de cultivo papa dextrosa agar (PDA) y extracto de malta agar (EMA), con dos niveles de temperatura (25 y 28 °C) y seis rangos de pH (4.0, 4.5, 5.0, 5.5, 6.0 y 6.5). El diseño experimental fue completamente al azar con medidas repetidas a través del tiempo, analizados con el paquete REPEATED MEASURE y el efecto tiempo con PROC MIXED de SAS. Como resultado se obtuvieron que el efecto de la temperatura y medios de cultivo en los diferentes rangos de pH, presentaron diferencias significativas (P ≤ 0.05). El crecimiento micelial óptimo de la cepa mexicana de G. lucidum fue en el medio de cultivo EMA en los rangos de pH de 4.0 y 4.5 con 8.3 y 8.2 cm respectivamente. De igual forma, en los rangos de pH 4.0 y 4.5 se obtuvieron los crecimientos miceliales óptimos a temperatura de 25 °C con 8.1 y 8.0 cm respectivamente. El cual concluyó esta investigación que el crecimiento micelial óptimo de la cepa mexicana fueron a pH 4.0 y 4.5, temperatura de 25 °C y medio de cultivo EMA.

ABSTRACT The Mexican strain CP-145 of Ganoderma lucidum due to the medicinal importance it has presented lately, the present investigation had as objective to evaluate the effect of temperature and culture medium on the optimal mycelial growth in different pH ranges. The treatments corresponded to the use of potato dextrose agar (PDA) and malt extract agar (EMA), with two temperature levels (25 and 28 °C) and six pH ranges (4.0, 4.5, 5.0, 5.5, 5.5, 6.0 and 6.5). The experimental design was completely randomised with repeated measures over time, analysed with the REPEATED MEASURE package and the time effect with PROC MIXED of SAS. As a result, the effect of temperature and culture media in the different pH ranges showed significant differences (P ≤ 0.05). The optimal mycelial growth of the Mexican strain of G. lucidum was in the EMA culture medium in the pH ranges of 4.0 and 4.5 with 8.3 and 8.2 cm respectively. Similarly, in the pH ranges 4.0 and 4.5 the optimum mycelial growth was obtained at 25 °C with 8.1 and 8.0 cm respectively. This research concluded that the optimal mycelial growth of the Mexican strain was at pH 4.0 and 4.5, temperature of 25 °C and EMA culture medium.

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The development of greener nano-constructs with noteworthy biological activity is of supreme interest, as a robust choice to minimize the extensive use of synthetic drugs. Essential oils (EOs) and their constituents offer medicinal potentialities because of their extensive biological activity, including the inhibition of fungi species. However, their application as natural antifungal agents are limited due to their volatility, low stability, and restricted administration routes. Nanotechnology is receiving particular attention to overcome the drawbacks of EOs such as volatility, degradation, and high sensitivity to environmental/external factors. For the aforementioned reasons, nanoencapsulation of bioactive compounds, for instance, EOs, facilitates protection and controlled-release attributes. Nanoliposomes are bilayer vesicles, at nanoscale, composed of phospholipids, and can encapsulate hydrophilic and hydrophobic compounds. Considering the above critiques, herein, we report the in-house fabrication and nano-size characterization of bioactive oregano essential oil (Origanum vulgare L.) (OEO) molecules loaded with small unilamellar vesicles (SUV) nanoliposomes. The study was focused on three main points: (1) multi-compositional fabrication nanoliposomes using a thin film hydration-sonication method; (2) nano-size characterization using various analytical and imaging techniques; and (3) antifungal efficacy of as-developed OEO nanoliposomes against Trichophyton rubrum (T. rubrum) by performing the mycelial growth inhibition test (MGI). The mean size of the nanoliposomes was around 77.46 ± 0.66 nm and 110.4 ± 0.98 nm, polydispersity index (PdI) of 0.413 ± 0.015, zeta potential values up to -36.94 ± 0.36 mV were obtained by dynamic light scattering (DLS). and spherical morphology was confirmed by scanning electron microscopy (SEM). The presence of OEO into nanoliposomes was displayed by attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy. Entrapment efficiency values of 79.55 ± 6.9% were achieved for OEO nanoliposomes. In vitro antifungal activity of nanoliposomes tested against T. rubrum strains revealed that OEO nanoliposomes exhibited the highest MGI, 81.66 ± 0.86%, at a concentration of 1.5 µL/mL compared to the rest of the formulations. In summary, this work showed that bioactive OEO molecules with loaded nanoliposomes could be used as natural antifungal agents for therapeutical purposes against T. rubrum.

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The rubber tree (Hevea brasiliensis) is a tropical perennial crop for the primary source of natural rubber. Colletotrichum gloeosporioides from Hevea brasiliensis (C. gloeosporioides Hb) and Colletotrichum acutatum from Hevea brasiliensis (C. acutatum Hb) are the causal agents of rubber tree anthracnose and lead to serious loss of natural rubber production. Inoculation tests showed that C. gloeosporioides Hb possessed higher pathogenicity than C. acutatum Hb to the rubber tree. Genomic analysis revealed that an unknown gene, named CgNPG1 (a Novel Pathogenic Gene 1), was presented in the genome of C. gloeosporioides Hb but not identified in C. acutatum Hb. CgNPG1 was predicted to encode a small secretory protein without any conserved domain. To investigate the functions of CgNPG1 in C. gloeosporioides Hb and in C. acutatum Hb, the gene deletion and overexpression mutants were generated. The phenotype analysis showed that deletion of CgNPG1 led to changed conidia morphology, decreased mycelial growth, conidiation, conidia germination rate, appressorium formation rate, and pathogenicity of C. gloeosporioides Hb to the rubber tree. Meanwhile, heterogeneous expression of CgNPG1 in C. acutatum Hb significantly changed the conidia morphology and improved the mycelial growth rate, conidiation, conidia germination rate, appressorium formation rate, and the pathogenicity of C. acutatum Hb to the rubber tree. Consistently, CgNPG1 increased the expression level of CaCRZ1 and CaCMK1 in C. acutatum Hb. These data suggested that CgNPG1 contributed to mycelial growth, conidiation, the development of invasive structures, and the pathogenicity of Colletotrichum to the rubber tree, which might be related to the modulation of CaCRZ1 and mitogen-activated protein kinase CMK1. Our results provided new insight into CgNPG1 in regulating growth and pathogenicity of the Colletotrichum spp.

RESUMO

Abstract Biochar (BCH) is a solid product, rich in carbon, obtained by heating biomass under controlled conditions of oxygenation, in a process known as pyrolysis. Its benefits are associated with improvements to the physical, chemical and biological properties of soil. Furthermore, BCH can affect the growth of phytopathogenic microorganisms. Despite research advances in this area, there is still a lack of information on the effect of BCH concentration on different soilborne microorganisms. This work evaluated the direct effect of sewage sludge biochar (SSB) on the in vitro growth of different soilborne fungi. Eight phytopathogens [Fusarium oxysporum, F. oxysporum f. sp. lycopersici (CEN 1456), Macrophomina phaseolina (429), Sclerotinia sclerotiorum, S. sclerotiorum (CEN 1147), Sclerotium rolfsii (CEN 216), Sclerotium cepivorum (CEN 1357), Rhizoctonia solani] were evaluated. Additionally, a biological control agent [Trichoderma afroharzianum (T-22)] was also studied. Microorganisms were subjected to growth in PDA (Potato-Dextrose-Agar) culture medium enriched with SSB. Biochars, produced at 300 and 500 ºC, were applied at different doses: 0.0 (control), 0.2, 0.4, 0.6, 0.8 and 1.0 (g of SSB/100 ml of PDA). Biochars showed inhibitory effects on the mycelial growth of the different fungi evaluated. It was observed that there is a certain specificity of biochar concentration that must be evaluated for the control of phytopathogens. In most cases, the 0.4 and 0.6% concentrations had a greater inhibitory effect on phytopathogens and did not affect the biological control agent. Therefore, SSB proved to be a promising product for the control of different soilborne phytopathogens.

RESUMO

In this study, five new recently described Monosporascus species, M. brasiliensis, M. caatinguensis, M. mossoroensis, M. nordestinus, and M. semiaridus, which were found on weeds collected from cucurbit cultivation fields in northeastern Brazil, are characterized regarding mycelial growth at different pH levels and salinity (NaCl) concentrations, their pathogenicity to selected cucurbit species, and their sensitivity to fungicides with different modes of action. Our results reveal great variability among the representative isolates of each Monosporascus spp. All of them showed a wide range of tolerance to different pH levels, and NaCl significantly reduced their in vitro mycelial growth, although no concentration was able to inhibit them completely. In pathogenicity tests, all seedlings of cucurbits evaluated, melon, watermelon, cucumber, and pumpkin, were susceptible to the five Monosporascus spp. in greenhouse experiments using artificial inoculation of roots. Moreover, all Monosporascus spp. were highly susceptible to the fungicides fludioxonil and fluazinam. Our findings provide relevant information about the response of these new Monosporascus spp. to environmental factors, plant genotypes and fungicides.

RESUMO

The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed. (AU)

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The high diversity of the genus Geastrum and the difficulty of obtaining mycelial cultures impairs the study of the ecophysiology and the exploration of the biotechnological potential of the taxon. In this study, different culture media were tested to obtain mycelial cultures for G. lloydianum and G. subiculosum collected in the Brazilian Amazon. Data on spore germination, and isolation of monokaryotic cultures and in vitro sexual reproduction are presented, as well as a brief morphological description of the cultures obtained. For both species, Potato Dextrose Agar (PDA) was the most promising of the tested culture media. The highest growth in agar culture ever recorded for this genus is reported (4.9 mm per week for G. lloydianum and 7.5 mm for G. subiculosum). In the PDA culture medium, spores germinated after 35-40 days of incubation and the isolation of monokaryotic cultures of the two species, as well as in vitro sexual crosses, were successfully performed.(AU)

A alta diversidade do gênero Geastrum e a dificuldade de obtenção de culturas miceliais prejudicam o estudo ecofisiológico e a exploração do potencial biotecnológico do táxon. Nesse estudo, foram testados diferentes meios de cultivo, visando a obtenção de culturas miceliais para G. lloydianum e G. subiculosum coletadas na Amazônia brasileira. A germinação dos esporos, o isolamento das culturas monocarióticas e o cruzamento sexual in vitro são apresentados, além de uma breve descrição morfológica das culturas obtidas. O meio de cultura Batata Dextrose Ágar (BDA) foi o mais promissor dentre os meios de cultura testados no cultivo das duas espécies. Reportamos o maior crescimento em cultura de ágar já registrado para esse gênero (4,9 mm por semana para G. lloydianum e 7,5 mm por semana para G. subiculosum). Nesse meio de cultivo, os esporos germinaram após 35-40 dias de incubação e o isolamento de culturas monocarióticas das duas espécies, assim com os cruzamentos sexuais in vitro, foram realizados com sucesso.(AU)

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Frente a necessidade de preservação e a manutenção de materiais biológicos, dentre eles fungos com potencial para controle biológico, para o desenvolvimento biotecnológico e científico, que vêm ganhando destaque no cenário mundial. Sendo necessário a adequação de métodos de preservação que além de garantia a sobrevivência destes microrganismos permitam a conservação de suas características morfológicas, fisiológicas e genéticas, no entanto, não existe um método ideal ou universal para a conservação de materiais biológicos. Diante desta necessidade o presente trabalho teve como objetivo de avaliar a eficácia e viabilidade de três métodos de preservação de isolados do fungo Phomadimorpha (repicagens periódicas, Castellani e fragmentos de papel-filtro), em dois períodos de avaliação, seis e doze meses após o armazenamento. Estudou-se a eficácia e viabilidade, através do crescimento micelial do fungo em meio de cultivo contendo batata-dextrose-ágar. Houve variabilidade entre os métodos de preservação do isolado do fungo P. dimorpha para o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial, nos dois períodos de avaliação, após seis e doze meses de armazenamento. O método de preservação em fragmento de papel filtro mostrou-se como o mais eficaz na preservação do isolado do fungo P. dimorpha nos dois períodos de avaliação, após seis e doze meses de armazenamento, sendo ideal para obter o maior o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial.

Facing the need for preservation and maintenance of biological materials, among them fungi with potential for biological control, for biotechnological and scientific development, which are gaining prominence in the world scenario. It is necessary to adapt preservation methods that besides guaranteeing the survival of these microorganisms allow the conservation of their morphological, physiological and genetic characteristics, however, there is no ideal or universal method for the conservation of biological materials. In view of this need, the present work had the objective of evaluating the efficacy and feasibility of three methods for the preservation of Phoma dimorpha (periodic transfer, Castellani and filter paper fragments) isolates in two evaluation periods, six and twelve months after the storage. Efficacy and viability were studied by mycelial growth of the fungus in a culture medium containing potato-dextrose-agar. There was variability between the preservation methods of the P. dimorpha fungus isolate for mycelial growth, method efficacy and mycelial growth rate index, in the two evaluation periods, after six and twelve months of storage. The filter paper fragment preservation method was the most effective in reserving the P. dimorpha fungus isolate in the two evaluation periods, after six and twelve months of storage, being ideal to obtain the highest mycelial growth, efficacy of the method and mycelial growth rate index.

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Frente a necessidade de preservação e a manutenção de materiais biológicos, dentre eles fungos com potencial para controle biológico, para o desenvolvimento biotecnológico e científico, que vêm ganhando destaque no cenário mundial. Sendo necessário a adequação de métodos de preservação que além de garantia a sobrevivência destes microrganismos permitam a conservação de suas características morfológicas, fisiológicas e genéticas, no entanto, não existe um método ideal ou universal para a conservação de materiais biológicos. Diante desta necessidade o presente trabalho teve como objetivo de avaliar a eficácia e viabilidade de três métodos de preservação de isolados do fungo Phomadimorpha (repicagens periódicas, Castellani e fragmentos de papel-filtro), em dois períodos de avaliação, seis e doze meses após o armazenamento. Estudou-se a eficácia e viabilidade, através do crescimento micelial do fungo em meio de cultivo contendo batata-dextrose-ágar. Houve variabilidade entre os métodos de preservação do isolado do fungo P. dimorpha para o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial, nos dois períodos de avaliação, após seis e doze meses de armazenamento. O método de preservação em fragmento de papel filtro mostrou-se como o mais eficaz na preservação do isolado do fungo P. dimorpha nos dois períodos de avaliação, após seis e doze meses de armazenamento, sendo ideal para obter o maior o crescimento micelial, eficácia do método e índice de velocidade do crescimento micelial.(AU)

Facing the need for preservation and maintenance of biological materials, among them fungi with potential for biological control, for biotechnological and scientific development, which are gaining prominence in the world scenario. It is necessary to adapt preservation methods that besides guaranteeing the survival of these microorganisms allow the conservation of their morphological, physiological and genetic characteristics, however, there is no ideal or universal method for the conservation of biological materials. In view of this need, the present work had the objective of evaluating the efficacy and feasibility of three methods for the preservation of Phoma dimorpha (periodic transfer, Castellani and filter paper fragments) isolates in two evaluation periods, six and twelve months after the storage. Efficacy and viability were studied by mycelial growth of the fungus in a culture medium containing potato-dextrose-agar. There was variability between the preservation methods of the P. dimorpha fungus isolate for mycelial growth, method efficacy and mycelial growth rate index, in the two evaluation periods, after six and twelve months of storage. The filter paper fragment preservation method was the most effective in reserving the P. dimorpha fungus isolate in the two evaluation periods, after six and twelve months of storage, being ideal to obtain the highest mycelial growth, efficacy of the method and mycelial growth rate index.(AU)

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Botrytis cinerea is a worldwide spread fungus that causes the grey mold disease, which is considered the most important factor in postharvest losses in fresh fruit crops. Consequently, the control of gray mold is a matter of current and relevant interest for agricultural industries. In this work, a series of phenylpropanoids derived from eugenol were synthesized and characterized. Their effects on the mycelial growth of a virulent and multi-resistant isolate of B. cinerea (PN2) have been evaluated and IC50 values for the most active compounds range between 31⁻95 ppm. The antifungal activity exhibited by these compounds is strongly related to their chemical structure, i.e., increasing activity has been obtained by isomerization of the double bond or introduction of a nitro group on the aromatic ring. Based on the relationship between the fungicide activities and chemical structure, a mechanism of action is proposed. Finally, the activity of these compounds is higher than that reported for the commercial fungicide BC-1000 that is currently employed to combat this disease. Thus, our results suggest that these compounds are potential candidates to be used in the design of new and effective control with inspired natural compounds of this pathogen.

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A uva é uma fruta amplamente aceita pelos consumidores, além de ser produzida em larga escala no Brasil. Entretanto é uma cultura muito suscetível a perdas em pré-colheita ocasionadas por doenças e, entre elas, pode-se citar a podridão cinzenta do cacho, cujo agente causal é o fungo Botrytis cinerea. A busca por formas alternativas para o controle desse patógeno pode minimizar o uso de agrotóxicos, garantindo um alimento mais seguro. Objetivou-se avaliar o potencial antifúngico in vitro do extrato alcoólico de própolis, álcool de cereais e óleo essencial de manjericão (Ocimum basilicum), autoclavados ou não, sobre o crescimento micelial de Botrytis cinerea, ao longo do tempo de exposição. Os tratamentos avaliados foram: extrato alcoólico de própolis e álcool de cereais, ambos nas concentrações 0, 2,5, 6,25, 12,5%; e óleo essencial de manjericão, nas concentrações 0, 0,25, 0,50, 1%. Para a obtenção do crescimento micelial foram realizadas medidas em sentidos diametralmente opostos, com um paquímetro digital, durante 0, 24, 48, 72, 96, 120 e 144 h de exposição, para o cálculo da taxa de inibição do crescimento micelial (ICM, %). O experimento foi realizado em duplicata, sendo os tratamentos dispostos em esquema fatorial 4x7 (concentrações x tempos) e os dados foram analisados por comparação de médias (teste de Tukey, p ≤ 0,05), desdobrando a interação, quando significativa. Todos os tratamentos contendo óleo essencial de manjericão autoclavado e não autoclavado foram eficientes na inibição do crescimento micelial de B. cinerea. A autoclavagem do extrato alcoólico de própolis proporciona maior eficácia para o controle do patógeno do que a não autoclavagem. Para o álcool de cereais, a autoclavagem interfere na eficiência de controle de B. cinerea.

Grape is a fruit widely accepted by consumers, besides being produced in large scale in Brazil. However, it is a crop very susceptible to losses in pre-harvest and, among the main diseases responsible for these losses, we can mention the gray rot of the bunch, whose causal agent is the fungus Botrytis cinerea. The search for alternative ways to control this pathogen can minimize the use of agrochemicals, ensuring a safer food. The aim of this study was to evaluate the in vitro antifungal potential of propolis alcoholic extract, cereal alcohol and basil oil (Ocimum basilicum), autoclaved or not, on the mycelial growth of Botrytis cinerea, during the exposure time. The treatments evaluated were: alcoholic extract of propolis and cereal alcohol, both in concentrations 0.0, 2.5, 6.25, 12.5%; and basil oil, at concentrations 0.25, 0.50, 1.00%. In order to obtain the mycelial growth, measurements were carried out in diametrically opposite directions, with a digital caliper, during 0, 24, 48, 72, 96, 120 and 144 hours of exposure, to calculate the rate of inhibition of mycelial growth (IMG, %). The experiment was performed in duplicate, with the treatments arranged in a factorial scheme 4x7 (concentrations x times) and the data were analyzed by means comparison (Tukey's test, p ≤ 0.05), unfolding the interaction, when significant. All treatments containing autoclaved and non-autoclaved basil essential oil were efficient in inhibiting the mycelial growth of B. cinerea. The autoclaving of the propolis alcoholic extract provides greater efficacy for the control of the pathogen than non-autoclaving. For cereal alcohol, autoclaving interferes with the control efficiency of B. cinerea.

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RESUMO

A uva é uma fruta amplamente aceita pelos consumidores, além de ser produzida em larga escala no Brasil. Entretanto é uma cultura muito suscetível a perdas em pré-colheita ocasionadas por doenças e, entre elas, pode-se citar a podridão cinzenta do cacho, cujo agente causal é o fungo Botrytis cinerea. A busca por formas alternativas para o controle desse patógeno pode minimizar o uso de agrotóxicos, garantindo um alimento mais seguro. Objetivou-se avaliar o potencial antifúngico in vitro do extrato alcoólico de própolis, álcool de cereais e óleo essencial de manjericão (Ocimum basilicum), autoclavados ou não, sobre o crescimento micelial de Botrytis cinerea, ao longo do tempo de exposição. Os tratamentos avaliados foram: extrato alcoólico de própolis e álcool de cereais, ambos nas concentrações 0, 2,5, 6,25, 12,5%; e óleo essencial de manjericão, nas concentrações 0, 0,25, 0,50, 1%. Para a obtenção do crescimento micelial foram realizadas medidas em sentidos diametralmente opostos, com um paquímetro digital, durante 0, 24, 48, 72, 96, 120 e 144 h de exposição, para o cálculo da taxa de inibição do crescimento micelial (ICM, %). O experimento foi realizado em duplicata, sendo os tratamentos dispostos em esquema fatorial 4x7 (concentrações x tempos) e os dados foram analisados por comparação de médias (teste de Tukey, p ≤ 0,05), desdobrando a interação, quando significativa. Todos os tratamentos contendo óleo essencial de manjericão autoclavado e não autoclavado foram eficientes na inibição do crescimento micelial de B. cinerea. A autoclavagem do extrato alcoólico de própolis proporciona maior eficácia para o controle do patógeno do que a não autoclavagem. Para o álcool de cereais, a autoclavagem interfere na eficiência de controle de B. cinerea.(AU)

Grape is a fruit widely accepted by consumers, besides being produced in large scale in Brazil. However, it is a crop very susceptible to losses in pre-harvest and, among the main diseases responsible for these losses, we can mention the gray rot of the bunch, whose causal agent is the fungus Botrytis cinerea. The search for alternative ways to control this pathogen can minimize the use of agrochemicals, ensuring a safer food. The aim of this study was to evaluate the in vitro antifungal potential of propolis alcoholic extract, cereal alcohol and basil oil (Ocimum basilicum), autoclaved or not, on the mycelial growth of Botrytis cinerea, during the exposure time. The treatments evaluated were: alcoholic extract of propolis and cereal alcohol, both in concentrations 0.0, 2.5, 6.25, 12.5%; and basil oil, at concentrations 0.25, 0.50, 1.00%. In order to obtain the mycelial growth, measurements were carried out in diametrically opposite directions, with a digital caliper, during 0, 24, 48, 72, 96, 120 and 144 hours of exposure, to calculate the rate of inhibition of mycelial growth (IMG, %). The experiment was performed in duplicate, with the treatments arranged in a factorial scheme 4x7 (concentrations x times) and the data were analyzed by means comparison (Tukey's test, p ≤ 0.05), unfolding the interaction, when significant. All treatments containing autoclaved and non-autoclaved basil essential oil were efficient in inhibiting the mycelial growth of B. cinerea. The autoclaving of the propolis alcoholic extract provides greater efficacy for the control of the pathogen than non-autoclaving. For cereal alcohol, autoclaving interferes with the control efficiency of B. cinerea.(AU)

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RESUMO

Abstract Antibiosis is the mechanism by which certain microorganisms respond to the presence of others, secreting compounds or metabolites capable of inhibiting or impeding their development. The crude extract of Trichoderma contains a mixture of secondary compounds, which may show antibiotic effect, and has been used for the prospect of this fungus for biological control and other industrial purposes. Faced with the increasing demand of agriculture for ecologically compatible alternatives for the management of diseases, this work aimed to investigate the spectrum of action of Non-Volatile Metabolites (NVMs) of Trichoderma isolates against different plant pathogenic fungi. The antagonistic potential of NVMs was evaluated through the incorporation method of the filtered liquid extract in PDA medium. The assays showed that all the NVMs produced inhibited the fungus Sclerotinia sclerotiorum similarly. On the other hand, strains CEN1245 and CEN1274, both belonging to the species Trichoderma brevicompactum, showed broad spectrum against Sclerotium rolfsii, Colletotrichum gloesporioides, Verticillium dahliae, Fusarium oxysporum and Cylindrocladium sp. The present study describes isolates producing non-volatile metabolites with broad spectrum of antifungal action, as well as pathogen-specific. The Trichoderma spp. NVMs obtained from different soil samples cultivated with vegetables, cassava and maize were efficient in inhibiting plant pathogenic fungi belonging to other patossystems, such as forest or fruit, which could increase their potential application in biological control of plant diseases. In addition, these antagonistic fungi should be studied in greater depth for the identification of bioactive molecules of industrial interest or in commercial formulations of products for biological control of plant pathogens.

Resumo Antibiose é um mecanismo pelo qual certos microrganismos respondem à presença de outros, secretando compostos ou metabólitos capazes de inibir ou impedir o seu desenvolvimento. O extrato bruto de Trichoderma contém uma mistura de compostos secundários e tem sido utilizado na prospecção deste fungo para o controle biológico e demais fins industriais. Diante da crescente demanda da agricultura por alternativas ecologicamente compatíveis para o manejo de doenças, este trabalho teve como objetivo investigar o espectro de ação de Metabólitos Não Voláteis (MNVs), produzidos por isolados de Trichoderma, contra diferentes fungos fitopatogênicos. O potencial antagônico dos MNVs foi avaliado através do método de incorporação do extrato líquido filtrado em meio BDA. Os ensaios mostraram que todos os MNVs produzidos inibiram de forma semelhante o fungo Sclerotinia sclerotiorum. Por outro lado, os isolados CEN1245 e CEN1274, ambos Trichoderma brevicompactum, mostraram um amplo espectro de ação, atuando contra Sclerotium rolfsii, Colletotrichum gloesporioides, Verticillium dahliae, Fusarium oxysporum e Cylindrocladium sp. O presente estudo descreve isolados que produziram metabólitos não voláteis com amplo espectro de ação antifúngico, assim como patógeno-específico. Os MNVs de Trichoderma spp. obtidos de diferentes amostras de solo cultivadas com vegetais, mandioca e milho, foram eficientes na inibição de fungos fitopatogênicos pertencentes a outros patossistemas, como os de espécies florestais e frutíferas, o que poderia aumentar sua potencial aplicação no controle de doenças de plantas. Adicionalmente, estes fungos antagonistas devem ser mais bem estudados para identificação de moléculas bioativas de interesse industrial ou formulação de produtos para o controle biológico de patógenos de plantas.

RESUMO

Three non-ionic surfactants: Tween20, Tween80 and Breakthru (®) were screened for their effects on spore germination and mycelial growth rates and for their influence on three isolates of Beauveria bassiana spore germination at various temperatures. Tween20 and Tween80 were compatible with all the B. bassiana isolates in the germination studies, but inhibited germination at higher surfactant concentrations, irrespective of the conidial concentrations . Breakthru (®) had an inhibitory effect on germination even at the lowest concentration of 0.1% on all the B. bassiana isolates. The effects of the surfactants on spore germination did not correspond with their effects on colony growth. Conidial viability within the same formulation declined significantly with increases in temperature, irrespective of the surfactant. The optimal temperature for conidial germination of B. bassiana isolates was approximately 25 °C with an upper limit at 30 °C. Isolate 7320 was identified as the least affected by the different surfactants. This isolate was able to germinate rapidly in a broad temperature range of 25-30 °C after 24 h, this characteristic being an essential factor in controlling house fly populations in poultry houses.

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RESUMO

Three non-ionic surfactants: Tween20, Tween80 and Breakthru® were screened for their effects on spore germination and mycelial growth rates and for their influence on three isolates of Beauveria bassiana spore germination at various temperatures. Tween20 and Tween80 were compatible with all the B. bassiana isolates in the germination studies, but inhibited germination at higher surfactant concentrations, irrespective of the conidial concentrations. Breakthru® had an inhibitory effect on germination even at the lowest concentration of 0.1% on all the B. bassiana isolates. The effects of the surfactants on spore germination did not correspond with their effects on colony growth. Conidial viability within the same formulation declined significantly with increases in temperature, irrespective of the surfactant. The optimal temperature for conidial germination of B. bassiana isolates was approximately 25 °C with an upper limit at 30 °C. Isolate 7320 was identified as the least affected by the different surfactants. This isolate was able to germinate rapidly in a broad temperature range of 25–30 °C after 24 h, this characteristic being an essential factor in controlling house fly populations in poultry houses.

Assuntos

RESUMO

Three non-ionic surfactants: Tween20, Tween80 and Breakthru® were screened for their effects on spore germination and mycelial growth rates and for their influence on three isolates of Beauveria bassiana spore germination at various temperatures. Tween20 and Tween80 were compatible with all the B. bassiana isolates in the germination studies, but inhibited germination at higher surfactant concentrations, irrespective of the conidial concentrations. Breakthru® had an inhibitory effect on germination even at the lowest concentration of 0.1% on all the B. bassiana isolates. The effects of the surfactants on spore germination did not correspond with their effects on colony growth. Conidial viability within the same formulation declined significantly with increases in temperature, irrespective of the surfactant. The optimal temperature for conidial germination of B. bassiana isolates was approximately 25 °C with an upper limit at 30 °C. Isolate 7320 was identified as the least affected by the different surfactants. This isolate was able to germinate rapidly in a broad temperature range of 25–30 °C after 24 h, this characteristic being an essential factor in controlling house fly populations in poultry houses.(AU)

Assuntos