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

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.

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.

Identification of locally isolated entomopathogenic Fusarium species from the soil of Changa Manga Forest, Pakistan and evaluation of their larvicidal efficacy against Aedes aegypti / Identificação de espécies de Fusarium entomopatogênicas localmente isoladas do solo da Floresta de Changa Manga, Paquistão e avaliação de sua eficácia larvicida contra Aedes aegypti

Dengue fever vectored by the mosquito Aedes aegypti is one of the most rapidly spreading insect-borne diseases. Current reliance of dengue vector control is mostly on chemical insecticides. Growing insecticide resistance in the primary mosquito vector, Aedes aegypti, limits the effectiveness of vector control through chemical insecticides. These chemical insecticides also have negative environmental impacts on animals, plants and human health. Myco-biocontrol agents are naturally occurring organisms and are found to be less damaging to the environment as compared to chemical insecticides. In the present study, entomopathogenic potential of local strains of fungi isolated from soil was assessed for the control of dengue vector. Local fungal isolates presents better alternative to introducing a foreign biocontrol strain, as they may be better adapted to environmental conditions of the area to survive and may have more entomopathogenic efficacy against target organism. Larvicidal efficacy of Fusarium equiseti and Fusarium proliferatum was evaluated against Aedes aegypti. Local strains of F. equiseti (MK371718) and F. proliferatum (MK371715) were isolated from the soil of Changa Manga Forest, Pakistan by using insect bait method. Larvicidal activity of two Fusarium spp. was tested against forth instar larvae of A. aegypti in the laboratory, using concentrations 105, 106, 107 and 108 conidia /ml. LC50 values for F. equiseti after 24h, 48h, 72h and 96h of exposure were recorded as 3.8x108, 2.9x107, 2.0x107, and 7.1x106 conidia /ml respectively while LC50 values for F. proliferatum were recorded as 1.21x108, 9.6x107, 4.2x107, 2.6x107 conidia /ml respectively after 24h, 48h, 72h and 96h of exposure. The results indicate that among two fungal strains F. equiseti was found to be more effective in terms of its larvicidal activity than F. proliferatum against larvae of A. aegypti.

A dengue transmitida pelo mosquito Aedes aegypti é uma das doenças transmitidas por insetos de propagação mais rápida. A dependência atual do controle do vetor da dengue é principalmente de inseticidas químicos. O aumento da resistência a inseticidas no principal vetor do mosquito, Aedes aegypti, limita a eficácia do controle do vetor por meio de inseticidas químicos. Esses inseticidas químicos também têm impactos ambientais negativos sobre os animais, plantas e saúde humana. Os agentes de micobiocontrole são organismos que ocorrem naturalmente e são menos prejudiciais ao meio ambiente em comparação com os inseticidas químicos. No presente estudo, avaliou se o potencial entomopatogênico de cepas locais de fungos isolados do solo para o controle do vetor da dengue. Isolados de fungos locais apresentam melhor alternativa para a introdução de uma cepa de biocontrole estrangeira, pois podem ser mais bem adaptados às condições ambientais da área para sobreviver e podem ter maior eficácia entomopatogênica contra o organismo-alvo. A eficácia larvicida de Fusarium equiseti e Fusarium proliferatum foi avaliada contra Aedes aegypti. Cepas locais de F. equiseti (MK371718) e F. proliferatum (MK371715) foram isoladas do solo de Changa Manga Forest, Paquistão, usando o método de isca para insetos. Atividade larvicida de dois Fusarium spp. foi testado contra larvas de quarto ínstar de A. aegypti em laboratório, nas concentrações 105, 106, 107 e 108 conídios / ml. Os valores de LC50 para F. equiseti após 24 h, 48 h, 72 h e 96 h de exposição foram registrados como 3,8x 108, 2,9x107, 2,0x107 e 7,1x106 conídios / ml, respectivamente, enquanto os valores de LC50 para F. proliferatum foram registrados como 1,21x108, 9,6 x107, 4,2x107, 2,6x107 conídios / ml, respectivamente, após 24 h, 48 h, 72 h e 96 h de exposição. Os resultados indicam que entre duas cepas de fungos F. equiseti se mostrou mais eficaz em termos de atividade [...].

Ceratocystis fimbriata agente causal de la enfermedad de marchitez vascular de Tectona grandis L.f. (teca) en Ecuador / Ceratocystis fimbriata causative agent of vascular wilt disease of Tectona grandis L.f. (teak) in Ecuador

Tectona grandis L.f. es una especie forestal de madera dura que, a pesar de no ser nativa de América, su plasticidad de adaptación ha permitido que en Ecuador se establezcan plantaciones intensivas con fines de exportación. Una compleja enfermedad con características de marchitez vascular está matando miles de árboles de diferentes edades en el país. Se planteó conocer el agente fitopatógeno causante de la patogénesis y muerte de los árboles. Se aislaron los hongos Fusarium sp. y Ceratocystis fimbriata Ellis & Halst. desde árboles enfermos. A nivel de invernadero se plantearon los postulados de Koch, para el efecto 30 plantas de T. grandis de cuatro meses de edad por cada tratamiento, se inocularon con los microrganismos como se describe a continuación: T1= C. fimbriata, T2 = Fusarium sp., T3 = C. fimbriata + Fusarium sp., T4 = agar-agar (control). Se empleó un diseño completo al azar (DCA) y las plantas se evaluaron a los 45 días después de inoculadas. Los tratamientos C. fimbriata, y C. fimbriata + Fusarium sp., mostraron mayores volúmenes aparentes de necrosis, con 1.52 cm3 y 1.93 cm3, y plantas muertas por la infección durante el tiempo de evaluación, mientras que Fusarium sp. mostró baja o nula patogenicidad y comportamiento similar al control, con 0.27 cm3 y 0.16 cm3 respectivamente. Estos resultados sugieren que el agente causal de la enfermedad de marchitez vascular en T. grandis es C. fimbriata y sería el primer reporte del fitopatógeno atacando teca en Ecuador. (AU)

Tectona grandisL.f. it is a hardwood forest species, which despite not being native to America, its adaptive plasticity has allowed intensive plantations to be established for export purposes in Ecuador. A complex disease with characteristics of vascular wilt is killing thousands of trees of different ages in the country. It was proposed to know the phytopathogenic agent causing the pathogenesis and death of the trees. Fusarium sp. andCeratocystis fimbriata Ellis & Halst. were isolated from diseased trees. At the greenhouselevel, Koch's postulates were proposed, for the effect 30 four month old T. grandisplants for each treatment were inoculated with the microorganisms as described below: T1 = C. fimbriata, T2 = Fusariumsp ., T3 = C. fimbriata+ Fusariumsp., T4 = agar-agar (control). A complete randomized design (DCA) was used and the plants were evaluated 45 days after inoculation. The treatments C. fimbriata, and C. fimbriata+ Fusariumsp. showed higher apparent volumes of necrosis, with 1.52 cm3and 1.93 cm3, and plants killed by the infection during the evaluation time, while Fusariumsp. showed low or no pathogenicity and behavior similar to the control, with 0.27 cm3and 0.16 cm3respectively. These results suggest that the causative agent of vascular wilt disease in T. grandisis C. fimbriataand it would be the first report of phytopathogen attacking teak in Ecuador. (AU)

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

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

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

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

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

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

Correlation between pathogenic species and clinical findings, disease severity, and visual outcome in patients with fungal keratitis / Correlação entre espécies patogênicas e achados clínicos, gravidade da doença e desfecho visual em pacientes com ceratite fúngica

ABSTRACT Purpose: To determine whether a correlation exists between pathogenic species and clinical findings, disease severity, and visual outcome in patients with keratitis and fungal growth in microbiological culture. Methods: A retrospective study of patients with fungal growth in the microbiological culture of corneal scrapings. Patients were treated at an ophthalmologic reference center in Southeastern Brazil from January 1992 to October 2015. Results: Medical records of 181 patients (131 males and 50 females) with a mean age of 47 ± 18 years were analyzed. The three most common etiologies were Fusarium sp. (38.7%), Aspergillus sp. (15%), and Candida sp. (13.2%). Among these, Fusarium sp. was the most frequent in patients aged £50 years (p=0.002) and in those with a recent history of a foreign body and/or ocular trauma (p=0.01). Candida sp. was the most frequent etiology in patients aged >50 years (p=0.002), in those with postoperative ocular surgery (p=0.002); in those with a previous ocular pathology (p=0.0007); and in immunodepressed patients (p=0.0004). Conclusion: Fusarium sp. was predominant in patients aged £50 years and those with a recent history of foreign body and/or ocular trauma, whereas Candida sp. was predominant in older adults, in those with a postoperative ocular surgery, in those with a previous ocular pathology, and in immunodepressed patients.

RESUMO Objetivo: Determinar se existe uma correlação entre espécies patogênicas e achados clínicos, gravidade da doença e resultado visual em pacientes com ceratite e crescimento de fungos em cultura microbiológica. Métodos: Estudo retrospectivo de pacientes com crescimento de fungos na cultura microbiológica de raspado de córnea. Os pacientes foram tratados em um centro de referência oftalmológica no Sudeste do Brasil de janeiro de 1992 a outubro de 2015. Resultados: Foram analisados registros médicos de 181 pacientes (131 homens e 50 mulheres) com idade média de 47 ± 18 anos. As três etiologias mais comuns foram Fusarium sp. (38,7%), Aspergillus sp. (15%) e Candida sp. (13,2%). Entre estas, Fusarium sp. foi a mais frequente em pacientes com idade £50 anos (p=0,002) e naqueles com história recente de corpo estranho e/ou trauma ocular (p=0,01). Candida sp. foi a etiologia mais frequente em pacientes com idade >50 anos (p=0,002), naqueles com cirurgia ocular pós-operatória (p=0,002); naqueles com patologia ocular prévia (p=0,0007); e em pacientes imunodeprimidos (p=0,0004). Conclusão: Fusarium sp. foi predominante em pacientes com idade £50 anos e naqueles com história recente de corpo estranho e/ou trauma ocular; enquanto Candida sp. foi predominante em adultos mais velhos, naqueles com cirurgia ocular pós-operatória, naqueles com patologia ocular prévia e em pacientes imunodeprimidos.

Hongos fitopatógenos asociados a la enfermedad de marchitez vascular y muerte regresiva en plantaciones de Tectona grandis L. f. (teca) en el Trópico Húmedo Ecuatoriano / Phytopathogen fungi related to vascular wilt and die back in Tectona grandis L. f (teak) field in Ecuadorian Tropical Rainforest

Plantaciones de teca (Tectona grandis L.f.) en Ecuador están siendo afectadas por una compleja enfermedad de marchitez vascular y muerte regresiva, con características epidémicas, sin que hasta el momento se conozca el o los agentes causales. Se planteó describir la sintomatología de la enfermedad e identificar los hongos fitopatógenos asociados a árboles enfermos en el Trópico Húmedo Ecuatoriano mediante morfofisiología. Se seleccionaron tres plantaciones de 2, 5 y 7 años de edad, en cada una se delimitó tres parcelas de 500 m2. Se realizó la descripción sintomatológica, evaluó la incidencia y severidad de la enfermedad empleando una escala de cinco categorías. Por parcela se diseccionaron tres árboles, cuyos tejidos se llevaron al laboratorio, donde se emplearon tres estrategias metodológicas (cámara húmeda, sandwiches de zanahoria, y medio de cultivo papa-dextrosa-agar; PDA) para estimular la expresión de los fitopatógenos. En árboles enfermos se detectó clorosis, pérdida de turgencia, ápices de crecimiento secos, emisión de brotes epicormicos en el fuste, y marchitez fulminante. Se aisló e identificó Ceratocystis fimbriata Ellis & Halst., y especies de Fusarium de forma consecutiva con las tres estrategias metodológicas empleadas. La incidencia de la enfermedad fue del 16.6%, 15.2%, y 11.6% para las plantaciones de 2, 5 y 7 años, respectivamente. Los árboles enfermos en la plantación de 2 años se encontraron en las escalas 2, 4 y 5, mientras que en plantaciones de 5 y 7 años se ubicaron en las escalas 2, 3 y 5 de progreso de la enfermedad...(AU)

Teak plantations (Tectona grandis L. f.) in Ecuador are being affected by a complex disease of vascular wilt and dieback, with epidemic characteristics, without knowing the causal agent(s) so far. We proposed to describe the symptomatology of the disease and identify phytopathogenic fungi associated with diseased trees in the Ecuadorian Humid Tropic by morphophysiology. Three plantations of 2, 5 and 7 years of age were selected, in each three plots of 500 m2 were delimited. The symptomatologic description was made, evaluated the incidence and severity of the disease using a scale of five categories. By plot, three trees were dissected, whose tissues were taken to the laboratory, where three methodological strategies were used (wet chamber, carrot sandwiches, and potatodextrose-agar culture medium, PDA) to stimulate the expression of phytopathogens. In diseased trees, chlorosis, turgor loss, dry growth apices, emission of epicormic shoots in the stem, and fulminating wilt were detected. It was isolated and identified Ceratocystis fimbriata Ellis & Halst., and Fusarium species. consecutively with the three methodological strategies employed. The incidence of the disease was 16.6%, 15.2%, and 11.6% for plantations of 2, 5 and 7 years, respectively. The sick trees in the plantation of 2 years were found in scales 2, 4 and 5, while in plantations of 5 and 7 years they were located in scales 2, 3 and 5 of progress of the disease. ..(AU)

Evaluación de la capacidad patogénica de Fusarium spp. en modelos vegetal y murino / Evaluation of Fusarium spp. pathogenicity in plant and murine models

El género Fusarium es ampliamente conocido por su capacidad fitopatógena, típicamente asociada al marchitamiento vascular. Sin embargo, se ha reportado como un patógeno oportunista en pacientes inmunocompetentes e inmunocomprometidos, por lo que puede ser considerado como un microorganismo de interés en estudios de patogenicidad en diferentes hospederos. Este trabajo evaluó la capacidad patogénica de aislamientos de Fusarium spp. de diferentes orígenes en hospederos vegetales y en un hospedero animal (modelo murino). Doce aislamientos de Fusarium spp. de origen vegetal, animal superficial, humano superficial y humano sistémico fueron inoculados en plantas de tomate, gulupa y clavel, y en ratones BALB/c, inmunocompetentes e inmunosuprimidos. Las pruebas de patogenicidad en plantas no mostraron todos los síntomas asociados al marchitamiento vascular en los tres modelos vegetales, pero la colonización y la necrosis de los haces vasculares observada en todos los casos, independientemente de la especie de Fusarium y el origen del aislamiento, demostró el potencial infeccioso de Fusarium spp. en las diferentes especies de plantas. Por otro lado, las pruebas de patogenicidad en el modelo murino evidenciaron alteraciones del comportamiento. Asimismo, se observó en el modelo murino que todos los aislamientos infectaron y colonizaron diferentes órganos, independientemente de su origen, de la especie o del estado inmunitario del hospedero, pero solamente cinco (de diferente origen y correspondientes a diferentes especies) generaron mortalidad. En contraste, la prueba de inoculación superficial no evidenció lesiones ni colonización. Los resultados observados indican el potencial papel patogénico de los aislamientos de Fusarium spp. en los diferentes tipos de hospederos. Sin embargo, es necesario profundizar en estudios de factores de patogenicidad que expliquen la capacidad de este género para colonizar múltiples hospederos.

The genus Fusarium is widely recognized for its phytopathogenic capacity. However, it has been reported as an opportunistic pathogen in immunocompetent and immunocompromised patients. Thus, it can be considered a microorganism of interest in pathogenicity studies on different hosts. Therefore, this work evaluated the pathogenicity of Fusarium spp. isolates from different origins in plants and animals (murine hosts). Twelve isolates of Fusarium spp. from plants, animal superficial mycoses, and human superficial and systemic mycoses were inoculated in tomato, passion fruit and carnation plants, and in immunocompetent and immunosuppressed BALB/c mice. Pathogenicity tests in plants did not show all the symptoms associated with vascular wilt in the three plant models; however, colonization and necrosis of the vascular bundles, regardless of the species and origin of the isolates, showed the infective potential of Fusarium spp. in different plant species. Moreover, the pathogenicity tests in the murine model revealed behavioral changes. It was noteworthy that only five isolates (different origin and species) caused mortality. Additionally, it was observed that all isolates infected and colonized different organs, regardless of the species and origin of the isolates or host immune status. In contrast, the superficial inoculation test showed no evidence of epidermal injury or colonization. The observed results in plant and murine models suggest the pathogenic potential of Fusarium spp. isolates in different types of hosts. However, further studies on pathogenicity are needed to confirm the multihost capacity of this genus.

Pathogenicity and transmission of fungi detected on Passiflora alata seeds / Patogenicidade e transmissão de fungos detectados em sementes de Passiflora alata

Passion fruit is usually propagated by seeds because of the ease and lower cost in seedling production. However, the seed is the most efficient agent for the spread of pathogens. The damages from seed-borne diseases occur mainly during the germination stages or at the formation of seedlings in nurseries. Considering the need for knowledge on the pathology of sweet passion fruit seeds, the objective was to evaluate the transmission and pathogenicity of the fungi Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. and Lasiodiplodia theobromae, known as potentially pathogenic to this crop, and isolated from sweet passion fruit seeds. Therefore, tests on seed health, germination and seedling emergence in a sterilized commercial substrate were conducted using seeds from this species, inoculated with those fungal isolates. Leaves, stems and fruit from this plant were also inoculated with the same fungi. Alternaria sp., Fusarium spp. and L. theobromae were identified in seedlings obtained from inoculated seeds, confirming the transmission of these fungi by seeds. L. theobromae was also considered the most harmful fungus to passion fruit crop, as it causes seed rot and other disease symptoms on the leaves, stem and fruit. These findings inferred that healthy seeds of sweet passion fruit are essential for producing seedlings and to prevent the spread of the diseases caused by these fungi to exempt areas.(AU)

O maracujazeiro geralmente é propagado por meio de sementes em virtude da facilidade e do menor custo na produção de mudas. No entanto, a semente é o agente mais eficiente de disseminação de patógenos, sendo que os danos decorrentes das doenças transmitidas por elas ocorrem principalmente durante os estágios de germinação ou na formação de mudas nos viveiros. Considerando a necessidade de informações acerca da patologia de sementes de maracujá-doce nesse contexto, objetivou-se obter informações sobre a transmissão e a patogenicidade dos fungos Alternaria sp., Botrytis fabae, Cladosporium cladosporioides, Fusarium spp. e Lasiodiplodia theobromae, isolados de sementes de maracujá-doce e potencialmente patogênicos à cultura. Para tanto, testes de sanidade, germinação e emergência de plântulas em substrato comercial esterilizado foram conduzidos com sementes dessa espécie, inoculadas com esses isolados. Folhas, colo e frutos dessa planta também foram inoculados com os mesmos fungos. Alternaria sp., Fusarium spp. e L. theobromae foram identificados em plântulas obtidas de sementes inoculadas, confirmando a transmissão por sementes. L. theobromae foi considerado o mais agressivo à cultura do maracujá, por ter causado podridão nas sementes, além de maiores lesões nas folhas, no colo da planta e nos frutos. Dessa forma, infere-se que a obtenção de sementes de maracujá-doce sadias é imprescindível para a produção de mudas, evitando-se assim a disseminação desses patógenos em áreas isentas.(AU)

Seeds of weeds as an alternative host of phytopathogens / Sementes de plantas daninhas como hospedeiras alternativas de fitopatógenos

Weeds compete with agricultural crops for water, light, nutrients and space, besides having an extensive seed bank. However, another aspect to be considered relates to few studies pointing out weeds as hosts of phytopathogenic fungi. Many fungi, the main cause of diseases in plants, are known to use seeds as an efficient means of survival and dispersal. The objective of this work was to evaluate the health of weed seeds and the pathogenicity of fungi associated to plants of agricultural importance. The seeds were collected manually in Cerrado areas located in the municipality of Gurupi, Tocatins, Brazil. The blotter test method was used to evaluate seed health. The incidence of fungi was evaluated through an individual analysis of seeds using a stereoscopic and an optical microscope. The pathogenicity of fungi from weed seeds was evaluated by inoculation in plants of agronomic interest and, when pathogenic, we inoculated them in the host plant of the fungus. Weed seeds have been identified in fungi of the genus Alternaria, Aspergillus, Bipolaris, Cladosporium, Curvularia, Fusarium, Nigrospora, Papularia, Rhizopus and Pythium. The seeds of Acanthospermum australe, Bidens pilosa, Cenchrus echinatus, Digitaria horizontalis, Echinochloa crus-pavonis, Eleusine indica, Ipomoea sp., Pennisetum setosum, Sida rhombifolia, Spermacoce latifolia, Tridax procumbens and Vernonia polyanthes carry and disseminate fungi that, once inoculated, cause infection in plants of agricultural importance, such as Oryza sativa, Phaseolus vulgaris, Vigna unguiculata, Zea mays and Glycine max.(AU)

As plantas daninhas competem com culturas agrícolas por água, luz, nutrientes e espaço, além de possuírem um extenso banco de sementes. Entretanto, outra vertente a ser considerada é quanto aos poucos estudos relacionando plantas daninhas como hospedeiras de fungos fitopatogênicos. É sabido que muitos fungos, principais causadores de doenças em plantas, utilizam as sementes como meio eficiente de sobrevivência e de dispersão. Dessa forma, o trabalho objetivou avaliar a sanidade de sementes de plantas daninhas e a patogenicidade dos fungos associados às plantas de importância agrícola. As sementes foram coletadas manualmente em áreas de cerrado localizadas no município de Gurupi, Tocantins, utilizando o método blotter test para avaliação da sanidade. A incidência dos fungos foi avaliada com auxílio de microscópio estereoscópico e ótico. A patogenicidade dos fungos oriundos das sementes de plantas daninhas foi avaliada por meio da inoculação em plantas de interesse agronômico e, quando patogênico, a inoculação foi na própria planta daninha hospedeira do fungo. Foram identificados os fungos dos gêneros Alternaria, Aspergillus, Bipolaris, Cladosporium, Curvularia, Fusarium, Nigrospora, Papularia, Rhizopus e Pythium. As sementes de Acanthospermum australe, Bidens pilosa, Cenchrus echinatus, Digitaria horizontalis, Echinochloa crus-pavonis, Eleusine indica, Ipomoea sp., Pennisetum setosum, Sida rhombifolia, Spermacoce latifolia, Tridax procumbens e Vernonia polyanthes transportam e disseminam fungos que, uma vez inoculados, causam infecção em plantas de importância agrícola, como Oryza sativa, Phaseolus vulgaris, Vigna unguiculata, Zea mays e Glycine max.(AU)

Patogenicidad de especies de Fusarium asociadas a la pudrición basal del ajo en el centro norte de México / Fusarium species associated with basal rot of garlic in North Central Mexico and its pathogenicity

El ajo en México es uno de los cultivos de hortalizas más rentables, más del 83% de esta superficie es aportada por los estados de Zacatecas, Guanajuato, Sonora, Puebla, Baja California y Aguascalientes. La pudrición basal ocasionada por Fusarium spp. se encuentra ampliamente distribuida a nivel mundial; esta enfermedad se ha convertido en una limitante en zonas productoras de cebolla y ajo, no solo en México, sino también en otros países, En México, se ha informado la presencia de Fusarium oxysporum en plantas en Guanajuato y en semillas de ajo en Aguascalientes. En el estado de Morelos se ha reportado la presencia de Fusarium culmorum en cultivares de cebolla. Asimismo, en Aguascalientes se tienen antecedentes de otras especies como Fusarium proliferatum, Fusarium verticillioides, Fusarium solani y Fusarium acuminatum. Para este trabajo se planteó como objetivo identificar las especies de Fusarium encontradas en los estados de Zacatecas, Guanajuato y Aguascalientes, y evaluar su patogenicidad. Se realizaron recolectas de plantas con síntomas de la enfermedad en los estados antes mencionados. De los muestreos realizados se identificaron las especies F. oxysporum, F. proliferatum, F. verticillioides, F. solani y F. acuminatum; las cepas de Aguascalientes identificadas como AGS1A (F. oxysporum), AGS1B (F. oxysporum) y AGSY-10 (F. acuminatum) fueron las que presentaron bajo condiciones de invernadero un mayor índice de severidad.

Garlic in Mexico is one of the most profitable vegetable crops, grown in almost 5,451 ha; out of which more than 83% are located in Zacatecas, Guanajuato, Sonora, Puebla, Baja California and Aguascalientes. Blossom-end rot caused by Fusarium spp is widely distributed worldwide and has been a limiting factor in onion and garlic production regions, not only in Mexico but also in other countries. The presence of Fusarium oxysporum has been reported in Guanajuato and Aguascalientes. Fusarium culmorum has been reported in onion cultivars of Morelos; and Fusarium proliferatum, Fusarium verticillioides, Fusarium solani and Fusarium acuminatum have been previously reported in Aguascalientes. The goal of this work was identifying the Fusarium species found in Zacatecas, Guanajuato and Aguascalientes, to assess their pathogenicity. Plants with disease symptoms were collected from hereinabove mentioned States. The samples resulted in the identification of: F. oxysporum, F. proliferatum, F. verticillioides, F. solani and F. acuminatum species; out of which Aguascalientes AGS1A (F. oxysporum), AGS1B (F. oxysporum) and AGSY-10 (F. acuminatum) strains showed higher severity under greenhouse conditions.

Detección de fusarium semitectum en vainas de plantas de moringa (moringa oleifera lam) / Detection of fusarium semitectum from pods of moringa plants (moringa oleífera lam)

Se analizaron muestras de Moringa oleifera procedentes de la provincia Habana, Cuba con el objetivo de determinar el agente causal de los daños. Las plantas presentaban afectaciones en las vainas, tallo y follaje iniciadas por insecto seguida por síntomas típicos de hongos fitopatógenos. Las muestras fueron procesadas y se determinó que el agente causal es el hongo fitopatógeno Fusarium semitectum. Este hongo se asocia a Fusicladium sp causando daños severos en la planta. En Cuba se ha aislado fundamentalmente en lotes de semillas de moringa.

It analyzed samples of Moringa oleifera from Habana province, Cuba with the objective of to determinate the causal agent of the damages. The plants shows affectations in pods, stems and foliage initiated for insect straight on typical symptoms of phytopathongens fungi. The samples were preprocessed and it determined that causal agent is phytopathongens fungus Fusarium semitectum. This fungus is associated to Fusicladium sp caused severe damages in the plant. In Cuba it has isolated principally from fractions of moringa seeds.

Construction of cDNA expression library of watermelon for isolation of ClWRKY1 transcription factors gene involved in resistance to Fusarium wilt.

Full-length cDNAs are very important for genome annotation and functional analysis of genes. The number of full-length cDNAs from watermelon remains limited. Here we report first the construction of a full-length enriched cDNA library from Fusarium wilt stressed watermelon (Citrullus lanatus Thunb.) cultivar PI296341 root tissues using the SMART method. The titer of primary cDNA library and amplified library was 2.21 × 106 and 2.0 × 1010 pfu/ml, respectively and the rate of recombinant was above 85%. The size of insert fragment ranged from 0.3 to 2.0 kb. In this study, we first cloned a gene named ClWRKY1, which was 1981 bp long and encoded a protein consisting of 394 amino acids. It contained two characteristic WRKY domains and two zinc finger motifs. Quantitative real-time PCR showed that ClWRKY1 expression levels reached maximum level at 12 h after inoculation with Fusarium oxysporum f. sp. niveum. The full-length cDNA library of watermelon root tissues is not only essential for the cloning of genes which are known, but also an initial key for the screening and cloning of new genes that might be involved in resistance to Fusarium wilt.

Presencia de Fusarium graminearum en muestras de trigo destinado al consumo humano / Fusarium graminearum presence in wheat samples for human consumption

La fusariosis es una de las enfermedades más importantes de los cereales, Fusarium graminearum es su principal agente etiológico. Este hongo posee la capacidad de producir distintos tipos y niveles de toxinas, en especial deoxinivalenol (DON). En la campaña 2012-2013 se dieron condiciones ambientales predisponentes para el desarrollo de esta enfermedad. El objetivo de este trabajo fue evaluar la presencia del hongo y el contenido de DON en 50 muestras de trigo. Los resultados demostraron la presencia de Fusarium graminearum en el 80 % de las muestras analizadas. El 24 % de las muestras presentó valores de DON ≥ 1µg/g, el 26 % varió entre 0,5 y 0,99µg/g, mientras que el 50 % restante mostró valores inferiores a 0,5µg/g. Se observó correlación entre la presencia de Fusarium graminearum y de DON. Es necesario establecer valores límites de DON en granos de trigo destinados al consumo humano

One of the most important diseases in cereal crops is Fusarium head blight, being Fusarium graminearum the main etiological agent. This fungus has the ability to produce a wide spectrum and quantity of toxins, especially deoxynivalenol (DON). During the last crop season (2012-2013) the climatic conditions favored Fusarium colonization. The objective of this work was to determine the presence of this fungus as well as the DON content in 50 wheat grain samples. Our results showed that 80% of the samples were contaminated with Fusarium graminearum. Twenty four percent (24%) of the samples contained ≥ 1µg/g DON, 26% ranged from 0,5 and 0,99µg/g, and the remaining 50% had values lower than 0,5µg/g. Correlation was found between the presence of Fusarium graminearum and DON. It is necessary to establish DON limit values in wheat grains for human consumption

Queratitis micótica: identificación de especies en Valparaíso / Fungal keratitis: identification of species in Valparaíso

Introducción: La queratitits micótica puede ser provocada por una amplia variedad de especies, tanto por hongos filamentosos como levaduras. Objetivo: Conocer las especies involucradas en queratitits micótica durante el decenio 2003-2013 en la ciudad de Valparaíso. Materiales y método: Se revisaron los casos de queratitis micótica diagnosticados durante el periodo 2003-2013 en la cátedra de Micología de la Universidad de Valparaíso. Las muestras fueron tomadas con tórulas estériles, las cuales fueron sembradas en agar sabouraud y una vez obtenido el desarrollo de las especies se traspasaron a medios de cultivo especiales. La identificación se realizó por morfofisiología y en el caso de Candida además por pruebas bioquímicas. Resultados: Se diagnosticaron 10 pacientes con queratitis micótica, de ellos 6 fueron hombres. 8 pacientes tenían el antecedente de lesión traumática corneal. Las especies más frecuentes fueron Aspergillus fumigatus y Fusarium oxysporum. También se aislaron especies como Fusarium dimerumy Exophiala jeanselmei. Conclusión: De los 10 casos de queratitis micótica diagnosticados, la mayoría tenía el antecedente de lesión traumática corneal. Los géneros más aislados fueron Aspergillus y Fusarium.

Introduction: Fungal keratitis can be caused by a wide variety of species, either filamentous as yeast fungi. Objective: To determine the species involved in fungal keratitis during the decade 2003-2013 in the city of Valparaiso. Materials and Methods: The cases of fungal keratitis diagnosed in the depar tment of Mycology, University of Valparaiso, were reviewed for the period 2003-2013. Samples were taken with sterile swabs, which were seeded in Sabouraud agar and after obtaining the development of species, were transferred to special culture media. Identification was performed by morphophysiology and in the case of Candida further biochemical tests. Results: 10 patients were diagnosed with fungal keratitis, of which 6 were men. 8 patients had a history of traumatic corneal injury. The most frequent species were Aspergillus fumigatus and Fusarium oxysporum. Species Fusarium dimerum and Exophiala jeanselmei were also isolated. Conclusion: Of the 10 cases of fungal keratitis diagnosed, most had a history of traumatic corneal injury. The most isolated genus were Aspergillus and Fusarium.

Biochemical markers assisted screening of Fusarium wilt resistant Musa paradisiaca (L.) cv. Puttabale micropropagated clones.

An efficient protocol was standardized for screening of panama wilt resistant Musa paradisiaca cv. Puttabale clones, an endemic cultivar of Karnataka, India. The synergistic effect of 6-benzyleaminopurine (2 to 6 mg/L) and thidiazuron (0.1 to 0.5 mg/L) on MS medium provoked multiple shoot induction from the excised meristem. An average of 30.10 ± 5.95 shoots was produced per propagule at 4 mg/L 6-benzyleaminopurine and 0.3 mg/L thidiazuron concentrations. Elongation of shoots observed on 5 mg/L BAP augmented medium with a mean length of 8.38 ± 0.30 shoots per propagule. For screening of disease resistant clones, multiple shoot buds were mutated with 0.4% ethyl-methane-sulfonate and cultured on MS medium supplemented with Fusarium oxysporum f. sp. cubense (FOC) culture filtrate (5–15%). Two month old co-cultivated secondary hardened plants were used for screening of disease resistance against FOC by the determination of biochemical markers such as total phenol, phenylalanine ammonia lyase, oxidative enzymes like peroxidase, polyphenol oxidase, catalase and PR-proteins like chitinase, β-1-3 glucanase activities. The mutated clones of M. paradisiaca cv. Puttabale cultured on FOC culture filtrate showed significant increase in the levels of biochemical markers as an indicative of acquiring disease resistant characteristics to FOC wilt.

Negative correlation between phospholipase and esterase activity produced by Fusarium isolates

Fusarium species have emerged as one of the more outstanding groups of clinically important filamentous fungi, causing localized and life-threatening invasive infections with high morbidity and mortality. The ability to produce different types of hydrolytic enzymes is thought to be an important virulence mechanism of fungal pathogens and could be associated with the environment of the microorganism. Here, we have measured the production of two distinct lipolytic enzymes, phospholipase and esterase, by sixteen Fusarium isolates recovered from the hospital environment, immunocompromised patients’ blood cultures, foot interdigital space scrapings from immunocompromised patients, and foot interdigital space scrapings from immunocompetent patients (4 isolates each). Fourteen of these 16 isolates were identified asFusarium solani species complex (FSSC) and two were identified as F. oxysporum species complex (FOSC). Some relevant genus characteristics were visualized by light and electron microscopy such as curved and multicelled macroconidia with 3 or 4 septa, microconidia, phialides, and abundant chlamydospores. All Fusarium isolates were able to produce esterase and phospholipase under the experimental conditions. However, a negative correlation was observed between these two enzymes, indicating that a Fusarium isolate with high phospholipase activity has low esterase activity and vice versa. In addition, Fusarium isolated from clinical material produced more phospholipases, while environmental strains produced more esterases. These observations may be correlated with the different types of substrates that these fungi need to degrade during their nutrition processes.

Comparative study of the pathogenicity of seabed isolates of Fusarium equiseti and the effect of the composition of the mineral salt medium and temperature on mycelial growth

The pathogenicity of seven strains of Fusarium equiseti isolated from seabed soil was evaluated on different host plants showing pre and post emergence damage. Radial growth of 27 strains was measured on culture media previously adjusted to different osmotic potentials with either KCl or NaCl (-1.50 to - 144.54 bars) at 15º, 25º and 35º C. Significant differences and interactive effects were observed in the response of mycelia to osmotic potential and temperature.

Micoparasitismo biotrofico de Fusarium oxysporum sobre Cunnighamella sp / Biotrophic mycoparasitism of Fusarium oxysporum on Cunnighamella sp

Se describe un caso de micoparasitismo biotrófico de ocurrencia natural en el suelo, entre las hifas de una cepa de Fusarium oxysporum complex y Cunninghamella sp. Las hifas de F. oxysporum se desarrollaron sobre las células vivas del hospedador, mostrando 2 tipos de efectos parasíticos: uno de enrollamiento y otro de contacto con penetración de las hifas, sin la aparente eliminación del hospedador. Esta situación poco común en la literatura, demuestra las capacidades adaptativas de esta especie al micoparasitismo en grupos filogenéticamente distantes.

This paper describes a case of mycoparasitism naturally occurring, where Fusarium oxysporum parasitizes hyphae of Cunninghamella sp, to show mycoparasitism between the two fungi. This is a case of biotrophic mycoparasitism by contact. The hyphae of F. oxysporum developed closely along the living cells of the host showing mycoparasitic effect, some for a loop, and other contact with penetration of the hyphae. This situation is rare in the literature, demonstrates the adaptive capacities of this species to mycoparasitism in phylogenetically distant groups.

Growth of in vitro Fusarium oxysporum f. sp. niveum in chemically defined media amended with gallic acid

Gallic acid was artificially added to the media to grow Fusarium oxysporum f.sp.niveum to investigate its effect on the pathogenic fungus. Results indicate that gallic acid inhibited the growth of F. oxysporum f.sp.niveum. The colony diameter, the conidia germinating rate and the conidia yield were reduced by 5.7-22.9 percent percent, 35.8-55.6 percent and 38.9-62.2 percent respectively. However, the virulence factors by the fungus were stimulated. The activity of pectinase, proteinase and cellulase increased by 12.3-627.8 percent, 11.8-41.2 percent and 0.5-325.0 percent respectively, while the activity of amylase increased slightly. The results suggest that gallic acid repressed growth but facilitated the relative pathogenicity of invading pathogens.