Production, partial purification, and characterization of a glucoamylase from Epicoccum nigrum

Starch processing industries use amylases, accounting for approximately 30% of the world's enzyme market. Previously, an amylase-producing strain of Epicoccum nigrumwas isolated from maize grains. Although E. nigrumamylase production is already reported in the literature, no published data on production optimization or characterization of the produced enzyme exists. The objectives of this work were to improve the amylase production by theE. nigrumPG 16 strain and to purify and characterize the produced enzyme. The E. nigrumPG 16 amylase production best conditions in submerged culture were: inoculum of 4% (v v-1) of a five-days-old stationary culture homogenate, agitation at 100 rpm, 25°C, natural light, 72 hours of incubation, starch as thecarbon source, and an initial medium pH of 7.0. A molecular exclusion chromatography profile has shown the production of only one amylase, which was partially purified with ammonium precipitation and dialysis. The enzyme optima pH and temperature are 6.0 and 50°C, respectively. The partially purified enzyme lostits activity when incubated for 30 min in temperatures above 40°C, presenting a T50of 46.25°C. The KMand Vmaxof the partially purified enzyme are 1.72 mg mL-1of starch and 0.15 mgmin-1of degraded starch, respectively. The ion Ca2+slightly activated the studied enzyme. The ions Cu2+, Zn2+, and Fe3+and the detergents SDS and Tween 80 acted as inhibitors of the studied enzyme. The partially purified enzyme released glucose from p-nitrophenyl α-D-glucopyranoside (p-NPG). Glucose was the enzyme's main product from starch hydrolysis, as evidenced by thin-layer chromatography. The E. nigrumPG 16 studied enzyme is a glucoamylase and represents an alternative for enzymatic starch hydrolysis.(AU)

Endophytic Bacillus subtilis P10 from Prunus cerasifera as a biocontrol agent against tomato Verticillium wilt / Bacillus subtilis P10 endofítico de Prunus cerasifera como agente de biocontrole contra murcha de Verticillium de tomate

Abstract Endophytic bacteria serve key roles in the maintenance of plant health and growth. Few studies to date, however, have explored the antagonistic and plant growth-promoting (PGP) properties of Prunus cerasifera endophytes. To that end, we isolated endophytic bacteria from P. cerasifera tissue samples and used a dual culture plate assay to screen these microbes for antagonistic activity against Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum, and F. moniliforme. Of the 36 strains of isolated bacteria, four (strains P1, P10, P16, and P20) exhibited antagonistic effects against all five model pathogens, and the P10 strain exhibited the strongest antagonistic to five pathogens. This P10 strain was then characterized in-depth via phenotypic assessments, physiological analyses, and 16s rDNA sequencing, revealing it to be a strain of Bacillus subtilis. Application of a P10 cell suspension (1×108 CFU/mL) significantly enhanced the seed germination and seedling growth of tomato in a greenhouse setting. This P10 strain further significantly suppressed tomato Verticillium wilt with much lower disease incidence and disease index scores being observed following P10 treatment relative to untreated plants in pot-based experiments. Tomato plants that had been treated with strain P10 also enhanced defense-related enzymes, peroxidase, superoxide dismutase, and catalase activity upon V. dahliae challenge relative to plants that had not been treated with this endophytic bacterium. The results revealed that the P10 bacterial strain has potential value as a biocontrol agent for use in the prevention of tomato Verticillium wilt.

Resumo As bactérias endofíticas desempenham papel fundamental na manutenção da saúde e do crescimento das plantas. Poucos estudos até o momento, no entanto, exploraram as propriedades antagônicas e promotoras de crescimento de plantas (PGP) de endófitos de Prunus cerasifera. Para esse fim, isolamos bactérias endofíticas de amostras de tecido de P. cerasifera e usamos um ensaio de placa de cultura dupla para rastrear esses micróbios quanto à atividade antagonista contra Verticillium dahliae, Botryosphaeria dothidea, Fusarium oxysporum, F. graminearum e F. moniliforme. Das 36 cepas de bactérias isoladas, quatro (cepas P1, P10, P16 e P20) exibiram efeitos antagônicos contra todos os cinco patógenos modelo, e a cepa P10 exibiu o antagonista mais forte para cinco patógenos. Essa cepa P10 foi então caracterizada em profundidade por meio de avaliações fenotípicas, análises fisiológicas e sequenciamento de rDNA 16s, revelando ser uma cepa de Bacillus subtilis. A aplicação de uma suspensão de células P10 (1 × 108 UFC / mL) aumentou significativamente a germinação das sementes e o crescimento das mudas de tomate em casa de vegetação. Essa cepa P10 suprimiu ainda mais a murcha de Verticillium do tomate com incidência de doença muito menor e pontuações de índice de doença sendo observadas após o tratamento com P10 em relação a plantas não tratadas em experimentos baseados em vasos. As plantas de tomate que foram tratadas com a cepa P10 também aumentaram as enzimas relacionadas à defesa, peroxidase, superóxido dismutase e atividade da catalase após o desafio de V. dahliae em relação às plantas que não foram tratadas com essa bactéria endofítica. Os resultados revelaram que a cepa bacteriana P10 tem valor potencial como agente de biocontrole para uso na prevenção da murcha de Verticillium em tomate.

Production and marketing of Tuber floridanum ­ ecology and gastronomic value of a recently described truffle species

Tuber floridanum is the first truffle species reported in Brazil and little is known about its nutritional properties and ecology. Therefore, this study aimed to (1) determine T. floridanum aroma and nutritional properties and (2) analyze cultivation and fruiting conditions, including potential crop pests and market value. Fruiting bodies of T. floridanum showed a slightly ellipsoid shape and smooth to shallow groovy surface. The mass of ascocarps ranged from 0.13 g to 15.95 g, and the truffle was composed of 71 % carbohydrates, 19 % protein, 5 % ash, and 5 % of fat. Octan-3-one, 1-methoxy-3-methyl-butane, and 1-methoxy-2-methyl-butane determined T. floridanum characteristic aroma. Soil properties were also specific for this truffle, differing from optimal soils where commercial truffles are produced. In addition, the occurrence of earwigs (Anisolabididae, Anisolabidinae) feeding on truffles (26 % of all ascocarps) in pecan orchards may reduce truffle quality and cause financial losses. T. floridanum aroma has a unique composition, completely different from any commercial and non-commercial truffle species analyzed so far. Soil conditions in pecan orchards were also specific for truffle fruiting. Further studies should elucidate other ecological conditions for the co-production of T. floridanum with pecan.(AU)

Filtrates from cultures of endophytic fungi isolated from leaves of Copaifera oblongifolia (Fabaceae) affect germination and seedling development differently / Fungos endofíticos isolados de folhas de Copaifera oblongifolia (Fabaceae) afetam diferentemente a germinação e o desenvolvimento das plântulas

Abstract Interactions between endophytic fungi (EFs) and their host plants range from positive to neutral to negative. The results of such interactions can vary depending on the organ of the infected host plant. EFs isolated from the leaves of some species of plants have potential for use as agents to inhibit seed germination and control invasive plants. The objectives of this study were to identify EFs present in the leaves of Copaifera oblongifolia and to evaluate the role of these fungi in seed germination and seedling development. A total of 11 species of EFs were isolated, which were identified using the internal transcribed spacers (ITS) sequence of the nuclear ribosomal DNA. The isolated species of EFs are generalists and probably are transmitted horizontally. Laboratory tests revealed that filtrates of these fungal isolates differently affect seed germination and seedling development of C. oblongifolia. The species Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum and Phomopsis sp. negatively affected seed germination, with N. parvum standing out for its negative effects, inhibiting seedling germination and survival in 89 and 222%, respectively. In addition, Cochliobolus intermedius negatively affected seedling development. Thus, the combined use of N. parvum and C. intermedius, or products from the metabolism of these microorganisms, in the control of invasive plants deserves attention from future studies.

Resumo As interações entre fungos endofíticos (FEs) e suas plantas hospedeiras variam de positivas, neutras a negativas. Os resultados destas interações podem variar dependendo do órgão da planta hospedeira infectada. FEs isolados de folhas de algumas espécies de plantas têm potencial para serem usados como agentes inibidores da germinação de sementes e no controle de plantas invasoras. Os objetivos deste estudo foram identificar os FEs presentes nas folhas de Copaifera oblongifolia e avaliar o papel destes fungos na germinação das sementes e no desenvolvimento das plântulas. Um total de 11 espécies de FEs foi isolado das folhas de C. oblongifolia e identificado através da sequência dos espaçadores internos transcritos do DNA ribossomal nuclear. As espécies de FEs isoladas são generalistas e provavelmente devem ser transmitidas horizontalmente. Os resultados dos testes de germinação mostraram que filtrados destes isolados fúngicos podem afetar diferentemente a germinação das sementes e o desenvolvimento das plântulas de C. oblongifolia. As espécies Curvularia intermedia, Neofusicoccum parvum, Pseudofusicoccum stromaticum e Phomopsis sp. afetaram negativamente a germinação das sementes de C. oblongifolia. Dentre estas espécies devemos destacar que N. parvum reduziu a germinação e a sobrevivência das plântulas em 89 e 222%, respectivamente. Além disso, Cochiliobolus intermedius afetou negativamente o desenvolvimento das plântulas. Assim, o uso combinado de N. parvum e C. intermedius, ou de produtos do metabolismo destas espécies de fungos, têm potencial para serem usados no manejo de plantas invasoras.

Oxidative stress caused by lead in the lichen Xanthoria parietina

The assessment of the air quality is a major concern to the current time. The monitoring and maintenance of air quality necessarily pass by detecting and estimating the overall air pollution. The use of lichens must be an assessmenttool to be studied. In our work we were interested about the toxicity of lead on the various parameters of stress in the lichen Xanthoria parietina. For this purpose, lichen thalli have been incubated at lead concentrations of 0, 0.5, 1.0, 5.0 and 10.0 mM, for time scale of 0, 24, 48 and 96 hours. The obtained results showed that lead has an action on the various studied parameters, and the intensity of oxidative stress observed in lichens thalli depends on the concentration, and time of exposure. Lead induced a decrease in chlorophyll and protein contents, and an increase in the contents of catalase, hydrogen peroxide and reduced glutathione. Furthermore, the results also showed that high concentrations of lead caused total destruction of reduced glutathione.(AU)

Efficiency of different strains of Trichoderma on the control of Sclerotinia sclerotiorum, Sclerotium rolfsii and Sclerotium cepivorum

The aim of this work was to verify the efficiency of different isolates of Trichoderma spp.on the control of Sclerotiniasclerotiorum, Sclerotiumrolfsii and Sclerotiumcepivorum, and the influence they pose on the conidia production of Trichoderma spp.For mycelial growth, discs with inoculum of phytopathogens were placed on the center of the Petri dishes followed by the addition of two Trichodermasp. discs on the opposite sides of the plate after 24hours. Every 12hoursdata were collected from colonies diameters and used for the analyses of Mycelial Growth Index (MGI) and Area Under the Curve of Mycelial Growth (AUCMG). The analyses were performed by a completely randomized design with two controls, a negative one without Trichodermasp. and one with acommercial strain of Trichodermaharzianum. Spore solution for evaluation of conidia production were made by adding 10 mL of distilled water and scratching the surface of the colonies. For S. cepivorum, all Trichoderma spp. strains reduced both indexes tested. However, while for MGI S. sclerotiorum also presented some reduction on the growth rate, the total area of this fungus was not affected. Sclerotiumrolfsii strains of Trichodermasp. from Lages and Curitibanosshowed an effect on the reduction of AUCMG of this fungus, although none of the Trichodermaaffected the growth rate of this phytopathogen. On the presence of S. sclerotiorumand S. cepivorum, none of the Trichodermaspp. showed any difference on conidia production when compared among themselves, nonetheless we did notice that on the presence of S. cepivorum, the strain from Rio do Sul retained its reproductive ability compared to control. Results obtained from this research can demonstrate the importance of biocontrol agents against different plant pathogens since it might have a specific antagonist-pathogen relation.(AU)

New treatments to disinfect vine cuttings with Phaeomoniella chlamydospora and the reason for the control

Petri disease is a problem for vineyard caused mainly by the fungus Phaeomoniella chlamydospora. Contaminated seedlings are source of inoculum for the disease. Treatment to disinfect vine rootstock cuttings for seedling production is hot water treatment (HWT) by 50 °C for 30 min, but the efficiency is contested. To improve its efficacy, the study aimed to assess the combination of the following methods and the reason for the control: i) exposition of the fungus to five different temperatures in HWT bath for 30 min; ii and iii) exposition of the fungus and also plants infected with P. chlamydospora to different disinfection treatments (biofumigation = soil + cabbage at 40 °C; temperatures of 40 and 23 °C, all in microcosm), in different periods (7, 14 and 21 days), with and without additional HWT (51 °C for 30 min). The results showed that HWT with high temperatures (55­70 °C) for 30 min inactivated the fungus. Biofumigation technique at 40 °C and the temperature solely of 40 °C applied for up to 21 days and combined with HWT (51 °C for 30 min) inhibited mycelial growth and inactivated the fungus in vine plant tissues without compromising the rooting.

Ameliorative effects of morel mushroom (Morchella esculenta) against Cadmium-induced reproductive toxicity in adult male rats / Efeitos benéficos do cogumelo morel (Morchella esculenta) contra a toxicidade reprodutiva induzida por cádmio em ratos machos adultos

Cadmium (Cd) is one of the major toxicants, which affects human health through occupational and environmental exposure. In the current study, we evaluated the protective effects of morel mushrooms against Cd-induced reproductive damages in rats. For this purpose, 30 male rats were divided into 6 groups (n=5/group), the first group served as the control group, second group was treated with an intraperitoneal (i.p) injection of 1 mg/kg/day of Cd. Third and fourth groups were co-treated with 1 mg/kg/day of Cd (i.p) and 10 and 20 mg/kg/day of morel mushroom extract (orally) respectively. The final 2 groups received oral gavage of 10 and 20 mg/kg/day of morel mushroom extract alone. After treatment for 17 days, the animals were euthanized, and testes and epididymis were dissected out. One testis and epididymis of each animal were processed for histology, while the other testis and epididymis were used for daily sperm production (DSP) and comet assay. Our results showed that Cd and morel mushrooms have no effect on animal weight, but Cd significantly decreases the DSP count and damages the heritable DNA which is reversed in co-treatment groups. Similarly, the histopathological results of tests and epididymis show that morel mushrooms control the damage to these tissues. Whereas the morel mushroom extract alone could enhance the production of testosterone. These results conclude that morel mushrooms not only control the damage done by Cd, but it could also be used as a protection mechanism for heritable DNA damage.

O cádmio (Cd) é um dos principais tóxicos, que afeta a saúde humana por meio da exposição ocupacional e ambiental. No presente estudo, avaliamos os efeitos protetores dos cogumelos morel contra os danos reprodutivos induzidos pelo Cd em ratos. Para tanto, 30 ratos machos foram divididos em 6 grupos (n = 5 / grupo); o primeiro grupo serviu de controle, o segundo grupo foi tratado com injeção intraperitoneal (i.p) de 1 mg / kg / dia de Cd. O terceiro e o quarto grupos foram cotratados com 1 mg / kg / dia de Cd (i.p) e 10 e 20 mg / kg / dia de extrato de cogumelo morel (por via oral), respectivamente. Os dois grupos finais receberam gavagem oral de 10 e 20 mg / kg / dia de extrato de cogumelo morel sozinho. Após o tratamento por 17 dias, os animais foram sacrificados e os testículos e o epidídimo foram dissecados. Um testículo e epidídimo de cada animal foram processados para histologia, enquanto o outro testículo e epidídimo foram usados para produção diária de esperma (DSP) e ensaio cometa. Nossos resultados mostraram que os cogumelos Cd e morel não têm efeito sobre o peso do animal, mas o Cd diminui significativamente a contagem de DSP e danifica o DNA hereditário, que é revertido em grupos de cotratamento. Da mesma forma, os resultados histopatológicos dos testículos e do epidídimo mostram que os cogumelos morel controlam os danos a esses tecidos. Considerando que o extrato de cogumelo morel sozinho pode aumentar a produção de testosterona. Esses resultados concluem que os cogumelos morel não apenas controlam os danos causados pelo Cd, mas também podem ser usados como um mecanismo de proteção para danos hereditários ao DNA.

Characterization of the antagonism of the mycorrhizal fungus Waitea circinata against Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens and Sarocladium oryzaerice pathogens / Caracterização do antagonismo do fungo micorrízico Waitea circinata contra os patógenos do arroz Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens e Sarocladium oryzae

Rice production is affected by important pathogens such as Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens and Sarocladium oryzae, and orchid mycorrhizal fungi can contribute to the biocontrol of these diseases. The aim of this study was to characterize the antagonism mechanisms of W. circinata against rice pathogens. The indoleacetic acid (IAA) and lytic enzymes of W. circinata were quantified. The effectiveness of the bioagent and its metabolites was assessed in vitro. Antagonism in vivo against rice blast was also investigated. Waitea circinata produced IAA (2.3 µg ml¹), cellulase and polyphenol oxidase in a qualitative test. Glucanase, chitinase and protease were also produced in culture with cell walls and in co-culture with pathogens to explain antibiosis. W. circinata acts on direct antagonism by antibiosis and volatile metabolites. Mycelial suspension reduced M. oryzae conidial germination, appressorium formation, and rice blast by 61, 82 and 84%, respectively. W. circinata, a unique bioagent, controls four rice pathogens from mechanisms of parasitism and antibiosis. The results could contribute to new formulations containing this fungus, enzymes as well as its volatile metabolites.

A produção de arroz é afetada por patógenos importantes como Magnaporthe oryzae, Cochliobolus miyabeanus, Monographella albescens e Sarocladium oryzae, e fungos micorrízicos de orquídeas podem contribuir para o biocontrole dessas doenças. O objetivo deste estudo foi caracterizar os mecanismos de antagonismo de W. circinata contra patógenos do arroz. O ácido indolacético (AIA) e as enzimas líticas de W. circinata foram quantificadas. A eficácia do bioagente e seus metabólitos foi avaliada in vitro. O antagonismo in vivo contra a brusone do arroz também foi investigado. Waitea circinata produziu AIA (2,3 µg ml¹), celulase e polifenol oxidase em teste qualitativo. Glucanase, quitinase e protease também foram produzidas em cultura com paredes celulares e em co-cultura com patógenos para explicar a antibiose. W. circinata atua no antagonismo direto por antibiose e metabólitos voláteis. A suspensão micelial reduziu a germinação de conídios de M. oryzae, a formação de apressórios e a brusone do arroz em 61, 82 e 84%, respectivamente. W. circinata, um bioagente único, controla quatro patógenos do arroz a partir de mecanismos de parasitismo e antibiose. Os resultados podem contribuir para novas formulações contendo este fungo, enzimas, bem como seus metabólitos voláteis.

Chemical control of dieback and mango malformation in a semiarid region

The São Francisco Sub medium Valley is a great mango exporter in Brazil. In this semiarid region, diseases such as dieback and mango malformation have reduced the production of this crop. In this work the effectiveness of different active ingredients (flutriafol, methyl-thiophanate, copper hydroxide and fluxapyroxad mixed with pyraclostrobin) was assessed for the control of these diseases in a mango orchard. Monthly assessments of disease incidence were carried out. There was a difference among treatments for the two diseases studied, with flutriafol as the most effective fungicide against dieback and, for the malformation, the mixture of fluxapyroxad and pyraclostrobin and methyl-thiophanate showed the best results.

Antifungal activity of homeopathic medicines against the white mold causing agent Sclerotinia sclerotiorum

White mold disease, caused by fungus Sclerotinia sclerotiorum (Lib.) de Bary., is a disease hard to control due to the high amount of sclerotia produced, which guarantees its survival in the soil for years leading to significant yield losses. Alternative techniques to control the pathogen have been researched, including homeopathy. The present work aimed to evaluate the in vitro antifungal effect of homeopathic medicines on S. sclerotiorum mycelial growth. Homeopathic medicines Sulphur, fungal sclerotium Nosode and Calcarea carbonica, in 30CH, 200CH and 1000CH dynamizations were tested. Assays were carried out in a completely randomized design, with four repetitions. Experiments were performed through the addition of homeopathic medicines on the surface of plates containing culture medium, followed by insertion of a disc containing fungus mycelia and incubation. Control treatment received no homeopathic medicine. The mycelial progression was monitored by seven halo diameter measurements during experiment period. All homeopathic medicines tested and their dynamizations were able to inhibit partially the development of the fungus. Calcarea carbonica at the dynamization of 1000 CH showed the best inhibitory effect on S. sclerotiorum, which under its effect produced a mycelial halo 40% smaller than the control treatment.

Antifungal activity of homeopathic medicines against the white mold causing agent Sclerotinia sclerotiorum

White mold disease, caused by fungus Sclerotinia sclerotiorum (Lib.) de Bary., is a disease hard to control due to the high amount of sclerotia produced, which guarantees its survival in the soil for years leading to significant yield losses. Alternative techniques to control the pathogen have been researched, including homeopathy. The present work aimed to evaluate the in vitro antifungal effect of homeopathic medicines on S. sclerotiorum mycelial growth. Homeopathic medicines Sulphur, fungal sclerotium Nosode and Calcarea carbonica, in 30CH, 200CH and 1000CH dynamizations were tested. Assays were carried out in a completely randomized design, with four repetitions. Experiments were performed through the addition of homeopathic medicines on the surface of plates containing culture medium, followed by insertion of a disc containing fungus mycelia and incubation. Control treatment received no homeopathic medicine. The mycelial progression was monitored by seven halo diameter measurements during experiment period. All homeopathic medicines tested and their dynamizations were able to inhibit partially the development of the fungus. Calcarea carbonica at the dynamization of 1000 CH showed the best inhibitory effect on S. sclerotiorum, which under its effect produced a mycelial halo 40% smaller than the control treatment.(AU)

Effect of essential oils on the development of Colletotrichum sp. fungus in fragments of Feijoa sellowiana fruits

The aim of this work was to evaluate the effect of the essential oils of Syzygium aromaticum, Cymbopogon citratus, Eucalyptus citriodora and Rosmarinus officinalis on the mycelial development of the fungus Colletotrichum sp. in fragments of Feijoa sellowiana fruits. The essential oils were incorporated in the PDA (Potato-Dextrose-Agar) medium in the concentrations of 250, 500 and 1000 ppm, 0 ppm (PDA only) (negative control), and fungicide fluazinam 1% (positive control). The area under the mycelial growth curve (AUMGC) and percent inhibition of mycelial growth (PIMG) were calculated. In the second evaluation, fruits fragments bordering the disease symptom were immersed in essential oils aqueous solution of S. aromaticum, C. citratus, and E. citriodora, at the concentration of 5000 ppm, 0 ppm (water only - negative control) and fluazinam 1% (positive control). The immersion times in the treatments were: 2, 4, 8, 12 and 24 hours, with subsequent incubation in Agar-Agar medium at 25°C. This evaluation was performed daily for 15 days, observing the moment of fungal germination through the emission of the mycelium. It was verified from the obtained results that all treatments reduced the fungal growth, and the essential oils of C. citratus and S. aromaticum totally inhibited its growth from the dose 500 and 1000 ppm, respectively. Regarding the test on fruit fragments, the essential oil of S. aromaticum at the immersion times of 12 and 24 hours was effective in inhibiting the fungus until the 15th day of evaluation.

Effect of essential oils on the development of Colletotrichum sp. fungus in fragments of Feijoa sellowiana fruits

The aim of this work was to evaluate the effect of the essential oils of Syzygium aromaticum, Cymbopogon citratus, Eucalyptus citriodora and Rosmarinus officinalis on the mycelial development of the fungus Colletotrichum sp. in fragments of Feijoa sellowiana fruits. The essential oils were incorporated in the PDA (Potato-Dextrose-Agar) medium in the concentrations of 250, 500 and 1000 ppm, 0 ppm (PDA only) (negative control), and fungicide fluazinam 1% (positive control). The area under the mycelial growth curve (AUMGC) and percent inhibition of mycelial growth (PIMG) were calculated. In the second evaluation, fruits fragments bordering the disease symptom were immersed in essential oils aqueous solution of S. aromaticum, C. citratus, and E. citriodora, at the concentration of 5000 ppm, 0 ppm (water only - negative control) and fluazinam 1% (positive control). The immersion times in the treatments were: 2, 4, 8, 12 and 24 hours, with subsequent incubation in Agar-Agar medium at 25°C. This evaluation was performed daily for 15 days, observing the moment of fungal germination through the emission of the mycelium. It was verified from the obtained results that all treatments reduced the fungal growth, and the essential oils of C. citratus and S. aromaticum totally inhibited its growth from the dose 500 and 1000 ppm, respectively. Regarding the test on fruit fragments, the essential oil of S. aromaticum at the immersion times of 12 and 24 hours was effective in inhibiting the fungus until the 15th day of evaluation.(AU)

Management of white mold of soybean using winter cereal straw / Manejo do mofo-branco na cultura da soja pelo uso de cobertura do solo com cereais de inverno

The management of white mold (Sclerotinia sclerotiorum (Lib.) De Bary) has been one of the main production limitations faced by soybean (Glycine max L.) producers. Considering the complex management of this disease and resistance structure of the pathogen, the present study was conducted in the municipalities of Guarapuava and Palmas in Paraná with the objective of managing the white mold of soybean using straws of winter cereals, such as oat, rye, and triticale. Initially, the three winter cereals were cultivated simultaneously in both the study areas. Straw production, plant height, and shoot fresh and dry weight were evaluated. Subsequently, BMX Apollo soybean was cultivated on cereal straws, and the incidence and severity of white mold were evaluated. In the in vitro experiment, 20 sclerotia covered by a layer of cereal (oat, rye, and triticale) straws were added and carpogenic germination was observed only in the control treatment after 98 days. Regardless of the study site, rye presented greater height and fresh weight than the remaining two cereals. Soybean cultivation on winter cereal straw reduced the incidence and severity of white mold. Cultivation on rye straw reduced mold incidence by 77.7% and 76.6% in Palmas and Guarapuava, respectively.(AU)

O manejo do mofo branco (Sclerotinia sclerotiorum) tem sido um dos principais limitantes de produção enfrentados por produtores de soja. Tendo em vista a complexidade do manejo desta doença e da estrutura de resistência do patógeno, o presente trabalho foi conduzido nos municípios de Guarapuava e Palmas - Paraná visando o manejo do mofo branco na cultura da soja utilizando palhadas de cereais de inverno, aveia, centeio e triticale. Inicialmente, cultivou-se os diferentes cereais de inverno, simultaneamente, nas duas áreas de estudo (Guarapuava e Palmas - PR). Avaliou-se a produção de palhada, a altura de plantas, a massa verde e seca da parte aérea. Posteriormente, semeou-se soja BMX Apolo sobre as palhadas de cereais e avaliou-se a incidência e a severidade do mofo branco. No experimento in vitro, adicionou-se 20 escleródios cobertos por uma camada de palhas (aveia, centeio e triticale) e após 98 dias, observou-se germinação carpogênica somente no tratamento testemunha. Independentemente do local de estudo, o centeio destacou-se com maiores altura e massa verde. O cultivo sobre palhada de cereais de inverno reduziu a incidência e severidade de mofo branco com destaque para a palhada de centeio, reduzindo a incidência da doença em 77,7% e 76,6%, em Palmas e Guarapuava, respectivamente.(AU)

Fungicidal potential of essential oils in control of Fusarium spp. and Sclerotinia sclerotiorum / Potencial fungicida de óleos essenciais no controle de Fusarium spp. e Sclerotinia sclerotiorum

The use of highly toxic pesticides to control soil pathogens, such as Fusarium spp. and Sclerotinia sclerotiorum has generated concern, due to the irreversible impacts caused on the environment, in addition to selecting resistant isolates. In this way, essential oils appear as an efficient alternative in control of diseases. Facing the problem of soil pathogens control and high antimicrobial fungicide that essential oils present, this work aimed to evaluate the in vitro fungicidal potential of essential oils in control of Fusarium spp. and S. sclerotiorum. A completely randomized design, factorial scheme 2×4×8 was used, with two isolates (Fusarium spp. and S. sclerotiorum), four essential oils (Aloysia citriodora, Cymbopogon winterianus, Lippia alba and Ocimum americanum), eight essential oil concentrations (0.0; 0.2; 0.4; 0.6; 0.8; 1.0; 1.2 and 1.4 ?L·mL-1), and ten replicates. The essential oils inhibited mycelial growth of the fungi in different concentrations, being their potential justified by the presence of antifungal chemical compounds. Essential oils of A. citriodora, C. winterianus, L. alba and O. americanum present high fungicidal potential, being viable alternatives for formulation of commercial products, boosting the pesticides industry.(AU)

O uso de pesticidas com alta toxicidade para controlar patógenos do solo, como Fusarium spp. e Sclerotinia sclerotiorum, tem gerado preocupação, devido aos impactos irreversíveis causados no meio ambiente, além de selecionar isolados resistentes. Dessa forma, os óleos essenciais surgem como uma alternativa eficiente no controle de doenças. Diante da problemática de controle de patógenos do solo e alto potencial antimicrobiano que os óleos essenciais possuem, este trabalho teve como objetivo avaliar o potencial fungicida de óleos essenciais no controle de Fusarium spp. e S. sclerotiorum, in vitro. Utilizou-se um delineamento inteiramente randomizado, esquema fatorial 2×4×8, com dois isolados (Fusarium spp. e S. sclerotiorum), quatro óleos essenciais (Aloysia citriodora, Cymbopogon winterianus, Lippia alba e Ocimum americanum) e oito concentrações de óleo essencial (0,0; 0,2; 0,4; 0,6; 0,8; 1,0; 1,2 e 1,4 ?L·mL-1), com dez repetições. Os óleos essenciais inibiram o crescimento micelial dos fungos em diferentes concentrações, sendo seu potencial justificado pela presença de compostos químicos antifúngicos. Os óleos essenciais de A. citriodora, C. winterianus, L. alba e O. americanum apresentam alto potencial fungicida, sendo alternativas viáveis para formulação de produtos comerciais, impulsionando a indústria de agrotóxicos.(AU)

Fungicidal potential of essential oils in control of Fusarium spp. and Sclerotinia sclerotiorum / Potencial fungicida de óleos essenciais no controle de Fusarium spp. e Sclerotinia sclerotiorum

The use of highly toxic pesticides to control soil pathogens, such as Fusarium spp. and Sclerotinia sclerotiorum has generated concern, due to the irreversible impacts caused on the environment, in addition to selecting resistant isolates. In this way, essential oils appear as an efficient alternative in control of diseases. Facing the problem of soil pathogens control and high antimicrobial fungicide that essential oils present, this work aimed to evaluate the in vitro fungicidal potential of essential oils in control of Fusarium spp. and S. sclerotiorum. A completely randomized design, factorial scheme 2×4×8 was used, with two isolates (Fusarium spp. and S. sclerotiorum), four essential oils (Aloysia citriodora, Cymbopogon winterianus, Lippia alba and Ocimum americanum), eight essential oil concentrations (0.0; 0.2; 0.4; 0.6; 0.8; 1.0; 1.2 and 1.4 ?L·mL-1), and ten replicates. The essential oils inhibited mycelial growth of the fungi in different concentrations, being their potential justified by the presence of antifungal chemical compounds. Essential oils of A. citriodora, C. winterianus, L. alba and O. americanum present high fungicidal potential, being viable alternatives for formulation of commercial products, boosting the pesticides industry.(AU)

O uso de pesticidas com alta toxicidade para controlar patógenos do solo, como Fusarium spp. e Sclerotinia sclerotiorum, tem gerado preocupação, devido aos impactos irreversíveis causados no meio ambiente, além de selecionar isolados resistentes. Dessa forma, os óleos essenciais surgem como uma alternativa eficiente no controle de doenças. Diante da problemática de controle de patógenos do solo e alto potencial antimicrobiano que os óleos essenciais possuem, este trabalho teve como objetivo avaliar o potencial fungicida de óleos essenciais no controle de Fusarium spp. e S. sclerotiorum, in vitro. Utilizou-se um delineamento inteiramente randomizado, esquema fatorial 2×4×8, com dois isolados (Fusarium spp. e S. sclerotiorum), quatro óleos essenciais (Aloysia citriodora, Cymbopogon winterianus, Lippia alba e Ocimum americanum) e oito concentrações de óleo essencial (0,0; 0,2; 0,4; 0,6; 0,8; 1,0; 1,2 e 1,4 ?L·mL-1), com dez repetições. Os óleos essenciais inibiram o crescimento micelial dos fungos em diferentes concentrações, sendo seu potencial justificado pela presença de compostos químicos antifúngicos. Os óleos essenciais de A. citriodora, C. winterianus, L. alba e O. americanum apresentam alto potencial fungicida, sendo alternativas viáveis para formulação de produtos comerciais, impulsionando a indústria de agrotóxicos.(AU)

Contribution to the knowledge of Camillea (Ascomycota, Graphostromataceae) in the Amazon forest in Pará, Brazil / Contribuição ao conhecimento de Camillea (Ascomycota, Graphostromataceae) na floresta amazônica no Pará, Brasil

Three species of the Ascomycetes genus Camillea were recorded in a fragment of Amazon rainforest in the region of Santarém, Pará state, Brazil. The occurrence of C. leprieurii, C. cyclops and C. bilabiata expand the range of distribution of these species in the state. Camillea leprieurii has previous records in the regions of Marabá, Oriximiná, Itaituba and Novo Progresso, while C. cyclops had been recorded in the west of the state. This is the first record of C. bilabiata for Pará. We provide a morphological description of the specimens and an identification key for Camillea species found in Pará.(AU)

Três espécies de Ascomycetes, gênero Camillea, foram registradas em um fragmento de floresta Amazônica na região de Santarém, Pará, Brasil. A ocorrência de C. leprieurii, C. cyclops e C. bilabiata expande a área de distribuição dessas espécies no estado. Camillea leprieurii foi registrada previamente nas regiões de Marabá, Oriximiná, Itaituba e Novo Progresso, enquanto C. cyclops havia sido registrada apenas no oeste do estado. Este é o primeiro registro de C. bilabiata no Pará. Fornecemos uma descrição morfológica dos espécimes e uma chave de identificação para as espécies de Camillea encontradas no Pará.(AU)

Diversity of bacterial strains in biochar-enhanced Amazon soil and their potential for growth promotion and biological disease control in tomato / Diversidade de isolados bacterianos em solo amazônico enriquecido com biocarvão e seu potencial para a promoção de crescimento e controle biológico de doenças em tomate

The use of bacteria in growth promotion and biological control of plant diseases can minimize environmental contamination caused by the indiscriminate use of pesticides and chemical fertilizers. We aimed to evaluate growth promotion and biological control of Corynespora cassiicola in tomato seedlings mediated by beneficial bacteria isolated from a non-rhizospheric Amazon soil containing different amounts of biochar, and to identify to which groups of bacteria the strains belong. We obtained 200 strains of bacteria from experimental plots containing biochar doses of 0, 40, 80 and 120 t ha-1. Of these, 53 strains were selected by root colonization tests. Based on growth promotion parameters, 25 strains were screened, identified by molecular characterization and evaluated for indoleacetic acid (IAA) production, phosphate solubilization and biological control. The best dose of biochar for colony formation was 40 t ha-1, and a regression model indicated 34 t ha-1 as the optimal dose. The production of IAA was observed in 18 (75%) strains, and two (8%) strains were able to solubilize phosphate. The efficiency in root growth promotion was up to 125%, and the percentage of plant protection ranged from 50 to 59%. Molecular characterization showed that the bacteria used in this study belong to the genera Bacillus and Lysinibacillus.(AU)

O uso de bactérias na promoção do crescimento e no controle biológico de doenças em plantas pode minimizar a contaminação ambiental causada pela aplicação indiscriminada de pesticidas e fertilizantes químicos. Objetivamos avaliar a promoção do crescimento e o controle biológico de Corynespora cassiicola em mudas de tomate mediadas por bactérias benéficas isoladas de solo amazônico não-rizosférico contendo diferentes dosagens de biocarvão, e identificar a quais grupos de bactérias os isolados pertencem. Obtivemos 200 isolados de parcelas experimentais contendo doses de biocarvão de 0, 40, 80 e 120 t ha-1. Destes, 53 foram selecionados por testes de colonização radicular. Com base nos parâmetros de promoção do crescimento, 25 isolados foram selecionados, identificados através de análise molecular e avaliados para produção de ácido indolacético (AIA), solubilização de fosfato e controle biológico. A melhor dose de biocarvão para a formação de colônias foi 40 t ha-1, e um modelo de regressão indicou 34 t ha-1como dose ótima. A produção de AIA foi observada em 18 (75%) isolados e dois (8%) isolados foram capazes de solubilizar fosfato. A eficiência na promoção do crescimento das raízes foi de até 125%, e a porcentagem de proteção das plantas variou de 50,3 a 59,0%. A caracterização molecular indicou que as bactérias utilizadas nesse estudo pertencem aos gêneros Bacillus e Lysinibacillus.(AU)

Endophytic fungi from Brachiaria grasses in Brazil and preliminary screening of Sclerotinia sclerotiorum antagonists

Fungal endophytes of Brachiaria, a nonhost of Sclerotinia sclerotiorum, may harbor species with antagonistic effects against this plant pathogen. The objective of this work was to investigate the diversity of endophytic fungi associated with different Brachiaria species and hybrids and evaluate their potential to inhibit the plant pathogen S. sclerotiorum. Stem samples from 39 Brachiaria spp. plants were collected in pasture fields and experimental areas of three states of Brazil resulting in 74 endophytes isolated. Twenty-eight species were identified by sequences of the Internal Transcribed Spacer (ITS) and 18S rDNA regions. Paraconiothyrium sp. was the most abundant endophyte, accounting for 24 % (14 isolates) of total, and it was isolated from B. ruziziensis, B. decumbens, B. humidicola, and B. brizantha. Phoma sorghina was the second most abundant taxon, followed by Sarocladium strictum, and Plenodomus sp. In vitro analyses showed that Paraconiothyrium sp., Sarocladium kiliense, Acremonium curvulum, Setophoma terrestris, Dissoconium sp., and Cladosporium flabelliforme exhibited antagonistic activity against S. sclerotiorum, with percentages of growth inhibition ranging from 25 to 60 (p < 0.05). Paraconiothyrium sp. BBXE1 (60 %), BBPB4.1 (60 %), BCMT4.1 (54 %), and S. kiliense (54 %) showed the highest values of Antagonism Percentages (AP). Therefore, fungi with inhibitory activity against S. sclerotiorum such as Paraconiothyrium sp. are naturally endophytic in Brachiaria grasses.