Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature / Fornecendo inóculo para estudos de Didymella bryoniae: o efeito do espectro de luz e armazenamento em baixa temperatura

Abstract The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus' survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores' size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.

Resumo A esporulação de Didymella bryoniae in vitro é de grande importância para estudos que requerem inóculo puro e em grandes quantidades. Assim, os objetivos deste estudo foram identificar a melhor condição para esporulação de D. bryoniae combinando diferentes espectros de luz (luz UV-A ou UV-B, luz branca e escuro contínuo) com distintos meios de cultura (PDA, V8, ML e PDAB) e, avaliar a sobrevivência do fungo armazenado a -20°C ao longo do tempo. As amostras de fungo só esporularam quando submetidas ao tratamento com luz UV-B, independentemente do meio de cultura. Maior aparecimento de esporos do tipo conídio foi observado no meio PDAB, e a menor produção ocorreu no meio ML. Estruturas reprodutivas, como peritécios e picnídeos, foram observadas em todos os meios de cultura. No entanto, houve uma variação considerável na quantidade de cada estrutura entre os diferentes meios de cultura. Os meios ML e V8 apresentaram maior número de peritécios e os meios PDA e PDAB apresentaram maior proporção de picnídeos em relação aos peritécios. A duração do armazenamento a -20°C não afetou o crescimento micelial ou a taxa de crescimento micelial. Em conclusão, a luz UV-B é essencial para a esporulação de D. bryoniae in vitro. Além disso, a composição do meio de cultura influencia o tipo de estrutura fúngica produzida, bem como o tamanho e a quantidade dos esporos. O congelamento a -20°C é uma técnica eficiente que pode ser usada para armazenar D. bryoniae por pelo menos cinco meses sem perda de viabilidade

Diversity of culturable endophytic fungi vary through time in Manihot esculenta Crantz / A diversidade de fungos endofíticos cultiváveis ​​varia ao longo do tempo em Manihot esculenta Crantz

Abstract Endophytic fungi are a ubiquituos group that colonize all plant species on earth. Studies comparing the location of endophytic fungi within the leaves and the sampling time in Manihot esculenta Crantz (cassava) are limited. In this study, mature leaves of M. esculenta from Panama were collected in order to compare the cultivable diversity of endophytic fungi and to determine their distribution within the leaves. A total of one hundred sixty endophytes belonging to 97 species representing 13 genera and 8 morphospecies determined as mycelia sterilia that containing 63 isolates were isolated. Cladosporium, Nigrospora, Periconia, and mycelia sterilia 1 and 3 were the most predominant isolated endophytes. We detected that endophytes varied across the sampling time, but not amongst locations within leaves. The endophytes composition across sampling and the location of endophytes within leaf was similar, except for Periconia and mycelia sterilia 3 and 7. The data generated in this study contribute to the knowledge on the biodiversity of endophytic fungi in Panama, and establish the bases for future research focused on understanding the function of endophytes in M. esculenta crops.

Resumo Os fungos endofíticos são um grupo ubiquituo que colonizam todas as espécies de plantas na terra. Os estudos que comparam a localização dos fungos endofíticos dentro das folhas de Manihot esculenta Crantz (mandioca) e o tempo de amostragem são muito escassos. Neste estudo, folhas maduras de M. esculenta foram coletadas do Panamá com a finalidade de comparar a diversidade cultivável de endófitos e determinar sua distribuição dentro das folhas. Um total de 170 endófitos foram isolados de 97 espécies que representam 13 gêneros e 8 morfoespécies determinadas como micélios esterilizados contendo 63 isolados. Os fungos Cladosporium, Nigrospora, Periconia e mycelia sterilia 1 e 3 foram os isolados mais predominantes. Também detectamos que os endófitos variaram ao longo do tempo de amostragem, mas não entre os locais dentro das folhas. A composição de endófitos na amostragem e localização de endófitos dentro da folha foi semelhante, exceto para Periconia e mycelia sterilia 3 e 7. Os dados gerados neste estudo contribuem para o conhecimento da biodiversidade de fungos endofíticos no Panamá e estabelecem as bases para pesquisas sobre o entendimento da função de endófitos em culturas de M. esculenta.

Community structure of endophytic fungi in roots and leaves of Fagopyrum mill and Avena sativa in a Chinese northern cold region

In order to explore the endophytic fungi of Fagopyrum Mill and Avena sativa, Illumina Miseq high-throughput sequencing was used to analyze the community structure and diversity of endophytic fungi in leaves and roots of buckwheat and oat at the mature stage. The results of community structure showed that there were 205 operational taxonomic units (OTUs) in buckwheat roots and 181 OTUs in buckwheat leaves based on 97% sequence similarity level. There were 152 OTUs and 127 OTUs in the root and the leaf of oat, respectively. At the phylum level, Ascomycota and Basidiomycota were the dominant endophytic fungi in buckwheat roots and leaves, while Ascomycota was the dominant endophytic fungus in oat roots and leaves. Alpha diversity analysis showed that the Ace index, Chao index and Shannon index of buckwheat roots were higher than that of buckwheat leaves, and the three indices of oat roots were also higher than that of oat leaves, indicating that the richness and diversity of endophytic fungi community in roots were higher than that in leaves. Biomarkers were found by significant difference analysis in buckwheat and oat. The endophytic functional groups of buckwheat and oat were mainly distributed in Pathotroph and Saprotroph. The results of this study laid a foundation for fully exploiting the dominant endophytic fungal resources of buckwheat and oat and further developing microbial fertilizers.

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.

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.

Gene clone and functional identification of sterol glycosyltransferases from Paris polyphylla var. yunnanensis / 中国中药杂志

In this study, the authors cloned a glycosyltransferase gene PpUGT2 from Paris polyphylla var. yunnanensis with the ORF length of 1 773 bp and encoding 590 amino acids. The phylogenetic tree revealed that PpUGT2 belonged to the UGT80A subfamily and was named as UGT80A49 by the UDP-glycosyltransferase(UGT) Nomenclature Committee. The expression vector pET28a-PpUGT2 was constructed, and enzyme catalytic reaction in vitro was conducted via inducing protein expression and extraction. With UDP-glucose as sugar donor and diosgenin and pennogenin as substrates, the protein was found with the ability to catalyze the C-3 hydroxyl β-glycosylation of diosgenin and pennogenin. To further explore its catalytic characteristic, 15 substrates including steroids and triterpenes were selected and PpUGT2 showed its activity towards the C-17 position of sterol testosterone with UDP-glucose as sugar donor. Homology modelling and molecule docking of PpUGT2 with substrates predicted the key residues interacting with ligands. The re-levant residues of PpUGT2-ligand binding model were scanned to calculate the corresponding mutants, and the optimized mutants were obtained according to the changes in binding affinity of the ligand with protein and the surrounding residues within 5.0 Å of ligands, which had reference value for design of the mutants. This study laid a foundation for further exploring the biosynthetic pathway of polyphyllin as well as the structure of sterol glycosyltransferases.

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.

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.

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.

Utilization of used fungus-growing materials of Gastrodia elata / 中国中药杂志

This study aims to explore the resource utilization of used fungus-growing materials produced in the cultivation of Gastrodia elata. To be specific, based on the production practice, this study investigated the recycling mechanism of used fungus-growing materials of G. elata by Phallus inpudicus. To screen edible fungi with wide adaptability, this study examined the allelopathic effects of Armillaria mellea secretions on P. impudicus and 6 kinds of large edible fungi and the activities of enzymes related to degradation of the used fungus-growing materials of G. elata. The results showed that P. impudicus can effectively degrade cellulose, hemicellulose, and lignin in used fungus-growing materials of G. elata. The cellulase activity of A. mellea was significantly higher than that of P. impudicus, and the activities of lignin peroxidase, polyphenol oxidase, and xylanase of P. impudicus were significantly higher than those of A. mellea, which was the important reason why A. mellea and P. impudicus used different parts and components of the used fungus-growing materials to absorb carbon sources and develop ecological niche differences. The growth of P. impudicus was significantly inhibited on the used fungus-growing materials of G. elata. The secretions of A. mellea had allelopathic effects on P. impudicus and other edible fungi, and the allelopathic effects were related to the concentration of allelopathy substances. The screening result showed that the growth and development of L. edodes and A. auricular were not significantly affected by 30% of A. mellea liquid, indicating that they had high resistance to the allelopathy of A. mellea. The results showed that the activities of extracellular lignin peroxidase, polyphenol oxidase, and xylanase of the two edible fungi were similar to those of P. impudicus, and the cellulase activity was higher than that of P. impudicus. This experiment can be further verified by small-scale production tests.

Cloning, expression and activity analysis of cutinase from Sclerotinia sclerotiorum / 生物工程学报

Cutinase can degrade aliphatic and aromatic polyesters, as well as polyethylene terephthalate. Lack of commercially available cutinase calls for development of cost-effective production of efficient cutinase. In this study, eight cutinase genes were cloned from Sclerotinia sclerotiorum. The most active gene SsCut-52 was obtained by PCR combined with RT-PCR, expressed in Escherichia coli BL21 and purified by Ni-NTA affinity chromatography to study its characteristics and pathogenicity. Sscut-52 had a total length of 768 bp and 17 signal peptides at the N terminals. Phylogenetic analysis showed that its amino acid sequence had the highest homology with Botrytis keratinase cutinase and was closely related to Rutstroemia cutinase. Sscut-52 was highly expressed during the process of infecting plants by Sclerotinia sclerotiorum. Moreover, the expression level of Sscut-52 was higher than those of other cutinase genes in the process of sclerotia formation from mycelium. The heterologously expressed cutinase existed in the form of inclusion body. The renatured SsCut-52 was active at pH 4.0-10.0, and mostly active at pH 6.0, with a specific activity of 3.45 U/mg achieved. The optimum temperature of SsCut-52 was 20-30 ℃, and less than 60% of the activity could be retained at temperatures higher than 50 ℃. Plant leaf infection showed that SsCut-52 may promote the infection of Banlangen leaves by Sclerotinia sclerotiorum.

Four new diphenyl ether derivatives from a mangrove endophytic fungus Epicoccum sorghinum / 中国天然药物

Four new diphenyl ethers, named epicoccethers K-N (1-4), were purified from the fermentation medium of a fungus Epicoccum sorghinum derived from Myoporum bontioides, and identified through HR-ESI-MS and NMR spectral analysis. Except that compound 1 showed moderate antifungal activity against Penicillium italicum and Fusarium graminearum, the other three compounds showed stronger activity against them than triadimefon. All of them showed moderate or weak antibacterial activity towards Staphylococcus aureus and Escherichia coli with O6 and O78 serotypes except that 3 was inactive to E. coli O6.

Presence of hyphae in chromoblastomycosis examinations: an enigma to be solved

Abstract The detection of muriform cells in direct mycological or anatomopathological examination is considered pathognomonic for chromoblastomycosis. The morphological aspect that these fungal structures acquire were called "Borelli spiders", when associated with hyphae. Reports of this association have been described for decades, initially related to more pathogenic agents of this mycosis. More recent studies have shown aspects related to the host's immunity that participate in this process, as well as an association with a worse disease prognosis. The present study discloses the findings of complementary examinations with the presence of "Borelli's spiders" in a patient diagnosed with chromoblastomycosis.

Sensitivity of hyperparasitic fungi to alternative products for use in the control of papaya black spot / Sensibilidade de fungos hiperparasitas a produtos alternativos para uso no controle da pinta preta do mamoeiro

Abstract The use of more than one control technique can maximize the reduction of the damages caused by the fungus Asperisporium caricae causal agent of the black spot in the papaya crop. The objective of this work was to evaluate the sensitivity of the fungi Hansfordia pulvinata and Acremonium spp. to alternative products with potential for use in the control of the black-spotted ptarmigan. Three isolates of Acremonium spp. (A-598, A-602 and A-617) and an isolate of H. pulvinata (H-611) were grown in BDA medium containing Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separately. The Amistar 500WG ® fungicide was used as a positive control and pure BDA as a negative control. The toxicity of the tested products was determined based on the values of the biological index, derived from the means of mycelial growth, sporulation and germination of conidia, in each experimental unit. In this way it was possible to select the products classified as compatible for all isolates, and to test them in vivo. In the greenhouse, only the isolates and isolates with selected products, were applied in papaya plants with foliar symptoms of black-spotted. The incidence of leaves with hyperparasites and the percentage of black-painted lesions colonized by the tested isolates were evaluated. Thus, the H-611 isolate proved to be compatible with most of the alternative products tested, except with Hortifos®. Bion® and Matrix® products were compatible with all tested isolates and could be used in conjunction with Acremonium spp. and H. pulvinata to control the papaya black spot, since these products did not present toxicity on the hyperparasitic fungi.

Resumo O uso de mais de uma técnica de controle pode maximizar a redução dos danos causados pelo fungo Asperisporium caricae agente causal da pinta preta na cultura do mamoeiro. O objetivo deste trabalho foi avaliar a sensibilidade dos fungos Hansfordia pulvinata e Acremonium spp. a produtos alternativos com potencial para uso no controle da pinta-preta. Três isolados de Acremonium spp. (A-598, A-602 e A-617) e um isolado de H. pulvinata (H-611) foram cultivados em meio BDA contendo Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separadamente. Fungicida Amistar 500WG® foi utilizado como controle positivo e BDA puro, como controle negativo. A toxicidade dos produtos testados foi determinada com base nos valores do índice biológico, provenientes das médias do crescimento micelial, esporulação e germinação de conídios, em cada unidade experimental. Dessa forma foi possível selecionar os produtos classificados como compatíveis para todos os isolados, e testá-los in vivo. Em casa de vegetação, somente os isolados e os isolados com produtos selecionados, foram aplicados em plantas de mamoeiro com sintomas foliares de pinta-preta. Avaliaram-se a incidência de folhas com hiperparasitas e a porcentagem de lesões de pinta-preta colonizadas pelos isolados testados. Assim o isolado H-611 mostrou-se compatível com a maioria dos produtos alternativos testados, exceto com Hortifos®. Os produtos Bion® e Matriz® apresentaram compatibilidade com todos os isolados testados, podendo ser utilizados em conjunto com o Acremonium spp. e H. pulvinata para controlar a pinta-preta do mamoeiro, uma vez que estes produtos não apresentaram toxidade sobre os fungos hiperparasitas.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; never-theless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Efficacy of a commercial fungal formulation containing Duddingtonia flagrans (Bioverm®) for controlling bovine gastrointestinal nematodes / Eficácia de uma formulação comercial contendo Duddingtonia flagrans (Bioverm®) no controle de nematódeos gastrintestinais de bovinos

Abstract Bioverm® (Duddingtonia flagrans) is a fungal formulation indicated for controlling gastrointestinal nematodes in ruminants and horses, which has recently been authorized for commercialization in Brazil. The objective was to determine the efficiency of Bioverm® against larvae of gastrointestinal nematodes after passage through the gastrointestinal tract of cattle. Twelve animals were used, divided into two groups. In the treated group, a single dose of 1 g of Bioverm® per 10 kg of live weight (containing 105 chlamydospores of D. flagrans) was provided for each animal. Fecal samples were obtained from the animals in each group at 12, 24, 36, 48, 60 and 72 hours after administration. In assay A, 2 g of feces were added to Petri dishes containing 2% agar-water medium. In assay B, coprocultures were performed. In both assays, the peak of larval predation occurred within 48 hours after administration of Bioverm®. In assay A, a significant larval reduction (P < 0.05) was seen at 48 h (88.2%). In assay B, significant reductions (P < 0.05) were seen at 36 h (43.7%) and 48 h (82.3%). Bioverm® showed high predatory capacity after passage through the gastrointestinal tract of cattle and was effective for controlling gastrointestinal nematodes.

Resumo O Bioverm® (Duddingtonia flagrans) é uma formulação fúngica indicada para o controle de nematódeos gastrintestinais de ruminantes e equídeos, recentemente autorizado para a comercialização no Brasil. Objetivou-se determinar a eficiência do Bioverm® contra larvas de nematódeos gastrintestinais após a passagem pelo trato gastrintestinal de bovinos. Foram utilizados doze bovinos divididos em dois grupos. No grupo tratado, foi fornecida, por animal, a dose única de 1g (105 clamidósporos de D. flagrans) do Bioverm® para cada 10 kg de peso vivo. Foram obtidas amostras fecais dos animais de cada grupo a partir de 12, 24, 36, 48, 60 e 72 horas após a administração. No ensaio A, 2g de fezes foram adicionadas em placas de Petri contendo meio ágar-água 2%. No ensaio B, foram realizadas coproculturas. Em ambos os ensaios, o pico de predação larval ocorreu em 48 horas após a administração do Bioverm®. No ensaio A, houve redução larval significativa (P<0,05) em 48h (88,2%). No ensaio B, as reduções significativas (P<0,05) ocorreram em 36h (43,7%) e 48h (82,3%). O Bioverm® apresentou elevada capacidade predatória após a passagem pelo trato gastrintestinal de bovinos, sendo eficaz no controle dos nematódeos gastrintestinais.

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.

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.

Neoscytalidium dimidiatum un raro agente de onicomicosis no dermatofítica en Chile / Neoscytalidium dimidiatuma rare agent ofnon dermatophyticonychomycosisin Chile

El término de onicomicosis se emplea para describir las infecciones de las uñas causadas por diferentes grupos taxonómicos fúngicos ya sea filamentosos como levaduriformes. A pesar de que estas patologías son causadas en los vertebrados principalmente por integrantes de la Familia Artrodermatáceae (Onygenales), la micología médica aplicó para ellos la terminología más específica de dermatofitosis, por ser un grupo ecológico de mayor importancia y presencia clínica. Las dermatomicosis de piel y fanéreos, representan un conjunto de infecciones producidas por especies fúngicas distribuidas en ambientes diversos, capaces de crecer a temperaturas de 37° y que actúan usualmente como patógenos oportunistas cuando existe generalmente un factor predisponente en el huésped. Se destaca la colonización en una uña de los pies en un hombre de 49 años por Neoscytalidium dimidiatum(Penz.) Crous & Slippers, un reconocido fitopatógeno de rápido crecimiento, común en zonas tropicales y subtropicales, que presentó la capacidad de invadir tejidos queratinizados con un aspecto clínico indistinguible de los causadas por dermatofitos. Por la rara presencia de este hongo en nuestra zona geográfica (provincia de Valparaíso, Chile), se aportan los principales datos morfofisiológicos,taxonómicos y moleculares utilizados en su diagnóstico. (AU)

The term onychomycosis is used to describe nail infections caused by different fungal taxonomic groups, either filamentous or yeast. Despite the fact that these pathologies are caused in vertebrates mainly by members of the Artrodermatáceae Family (Onygenales), medical mycology applied the more specific terminology of dermatophytosis for them, as it is an ecological group of greater importance and clinical presence. Skin and pharynx dermatomycosis represent a set of infections produced by fungal species distributed in diverse environments, capable of growing at temperatures of 37° and that usually act as opportunistic pathogens when there is a predisposing factor in the host. The colonization on a toenail in a 49-year-old man by Nesoscytalidium dimidiatumis highlighted (Penz.) Crous & Slippers, a recognized fast-growing phytopathogen, common in tropical and subtropical areas, which presented the ability to invade keratinized tissues with a clinical appearance indistinguishable from those caused by dermatophytes. Due to the rare presence of this fungus in our geographical area (Valparaíso province, Chile), the main morphophysiological, taxonomic and molecular data used in its identificationare provided. (AU)