Impact of Fusarium spp. on different maize commercial hybrids: disease evaluation and mycotoxin contamination.

Maize is one of the most important crops cultivated worldwide, whose production can be affected by the presence of several pathogens. Fusarium verticillioides and Fusarium graminearum are the most predominant pathogens affecting maize ears. However, few studies have been focused on studying the interaction between both pathogens in field conditions. For this reason, the aim of the present work was to evaluate the interaction between F. graminearum and F. verticillioides in different genotypes of maize under field conditions. Field experiments were carried out during two growing seasons in Azul, Argentina, including 12 commercial hybrids of maize, which were inoculated with F. graminearum, F. verticillioides, and a mixture of both pathogens. Phenotypic traits (plant height, plant diameter, tiller and cob number, and radiation interception), disease evaluation, and mycotoxin contamination were analyzed. The results showed significant differences between genotypes in disease severity (DS) for both years. In general terms, higher values of DS were reported in 2020 (21.70% ± 0.40) than in 2021 (16.50% ± 0.20). Different climatic conditions registered along the assay, especially precipitations and relative humidity, could be responsible for the differences observed over the years. Moreover, no significant correlations were found regarding DS and mycotoxin contamination for each genotype. For these reasons, an automatic correspondence between DS and mycotoxin contamination could lead to wrong agronomic decisions. The present study points out novel information regarding plant-pathogen interaction (maize-F. verticillioides/F. graminearum) under field conditions that could be useful for future maize breeding programmes.

Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L) almacenados en silos bolsa en Argentina / Extrinsic and intrinsic factors associated with mycotoxigenic fungi populations of maize grains (Zea mays L.) stored in silobags in Argentina

Con el objeto de caracterizar las poblaciones fúngicas, en particular las especies potencialmente micotoxigénicas, que pueden contaminar los granos de maíz almacenados en silos bolsa con un contenido de humedad superior al recomendado como seguro, se evaluaron 270 muestras extraídas al inicio, a los 90 días y al final de un período de almacenamiento de 5 meses. En dichas muestras se cuantificó e identificó la biota fúngica y se determinó la contaminación con fumonisinas y aflatoxinas. Asimismo, se evaluó el efecto de factores extrínsecos (ambiente), intrínsecos (granos) y tecnológicos (ubicación de los granos en el perfil del silo bolsa) sobre las poblaciones totales y micotoxigénicas. El pH de los granos y el nivel de O2 se redujeron significativamente a los 5 meses, mientras que la concentración de CO2 se incrementó en igual período. Los recuentos totales de la micobiota fueron significativamente mayores en los granos ubicados en el estrato superior del silo bolsa. Se identificaron especies micotoxigénicas de Fusarium, Aspergillus, Penicillium y Eurotium. La frecuencia de aislamiento de Fusarium verticillioides se redujo al final del almacenamiento y Aspergillus flavus solo se aisló en el inicio del almacenamiento. Los recuentos de Penicillium spp. y Eurotium spp. se incrementaron al final del almacenamiento. El 100 % de las muestras presentaron contaminación con fumonisinas, con niveles máximos de 5,707 mg/kg, mientras que las aflatoxinas contaminaron el 40 % de las muestras con niveles máximos de 0,0008 mg/kg. Las condiciones ambientales y de sustrato generadas durante el almacenamiento produjeron cambios en la composición de las poblaciones fúngicas y limitaron el desarrollo de hongos micotoxigénicos y la producción de micotoxinas.

In order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100 %) with maximum levels of 5.707 mg/kg whereas aflatoxin contaminated only 40 % with maximum levels of 0.0008 mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production.

[Extrinsic and intrinsic factors associated with mycotoxigenic fungi populations of maize grains (Zea mays L.) stored in silobags in Argentina]. / Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L.) almacenados en silos bolsa en Argentina.

In order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100%) with maximum levels of 5.707mg/kg whereas aflatoxin contaminated only 40% with maximum levels of 0.0008mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production.

[Physical and chemical methods for eliminating propagules of indigenous mycorrhizal fungi from soil samples]. / Métodos físicos y químicos para la eliminación de propágulos de hongos formadores de micorrizas nativos de muestras de suelo.

The objective of this work was to evaluate methods to eliminate or reduce the number of indigenous arbuscular mycorrhizal fungi (AMF) from soil samples without affecting their edaphic or microbiological properties. At an early trial we evaluated moist heat (autoclaving), dry heat (oven), sodium hypochlorite (NaClO) and formaldehyde at a range of 100.0-3.3µl/g and 16.7-3.3µl/g respectively. There was no germination in plants of ryegrass (Lolium multiflorum Lam.) sown on substrates receiving NaClO (100.0-33.3µl/g), whereas autoclaving significantly increased the available soil phosphorous content. Both treatments failed to eradicate AMF colonization at 9 weeks; therefore, they were discarded. In a second trial, oven and formaldehyde (10.0µl/g) treatments were analyzed to assess the effects of seed decontamination and AMF reinoculation. Both procedures were effective in reducing or eliminating indigenous AMF at a range of soil P availability of 12-29mg/kg. However, the time between soil treatment and AMF multiplication and safety requirements were greater in the case of formaldehyde application.

Métodos físicos y químicos para la eliminación de propágulos de hongos formadores de micorrizas nativos de muestras de suelo / Physical and chemical methods for eliminating propagules of indigenous mycorrhizal fungi from soil samples

El objetivo de este trabajo fue evaluar métodos para eliminar hongos nativos formadores de micorrizas arbusculares (HMA) o reducir su número en muestras de suelo, sin afectar sus propiedades edáficas y microbiológicas. Se estudió la aplicación de calor húmedo (autoclave), de calor seco (estufa), de hipoclorito de sodio (NaClO) y de formaldehído, en concentraciones entre 100,0 y 3,3 µl/g y 16,7 y 3,3 µl/g, respectivamente. Las semillas de raigrás (Lolium multiflorum Lam.) sembradas en sustratos que recibieron NaClO (100,0-33,3 µl/g) no germinaron y el autoclave incrementó el contenido de fósforo en el sustrato. Estos tratamientos no eliminaron la micorrización por HMA y ambos fueron descartados. En un segundo ensayo se analizaron los tratamientos estufa y formaldehído (10,0 µl/g), asociados o no a la descontaminación de las semillas y a la reinoculación con HMA. Ambos procedimientos redujeron o eliminaron la micorrización por HMA nativos en suelos con 12 a 29 mg/kg de fósforo y permitieron la multiplicación de inóculos de HMA. El tiempo de ventilación de las muestras y los requisitos de seguridad fueron mayores con la aplicación de formaldehído

The objective of this work was to evaluate methods to eliminate or reduce the number of indigenous arbuscular mycorrhizal fungi (AMF) from soil samples without affecting their edaphic or microbiological properties. At an early trial we evaluated moist heat (autoclaving), dry heat (oven), sodium hypochlorite (NaClO) and formaldehyde at a range of 100.0-3.3 µl/g and 16.7-3.3 µl/g respectively. There was no germination in plants of ryegrass (Lolium multiflorum Lam.) sown on substrates receiving NaClO (100.0-33.3 ul/g), whereas autoclaving significantly increased the available soil phosphorous content. Both treatments failed to eradicate AMF colonization at 9 weeks; therefore, they were discarded. In a second trial, oven and formaldehyde (10.0 µl/g) treatments were analyzed to assess the effects of seed decontamination and AMF reinoculation. Both procedures were effective in reducing or eliminating indigenous AMF at a range of soil P availability of 12-29 mg/kg. However, the time between soil treatment and AMF multiplication and safety requirements were greater in the case of formaldehyde application

Biodegradation of phenol in static cultures by Penicillium chrysogenum ERK1: catalytic abilities and residual phytotoxicity.

A phenol-degrading fungus was isolated from crop soils. Molecular characterization (using internal transcribed spacer, translation elongation factor and beta-tubulin gene sequences) and biochemical characterization allowed to identify the fungal strain as Penicillium chrysogenum Thom ERK1. Phenol degradation was tested at 25 degrees C under resting mycelium conditions at 6, 30, 60, 200, 350 and 400 mg/l of phenol as the only source of carbon and energy. The time required for complete phenol degradation increased at different initial phenol concentrations. Maximum specific degradation rate (0.89978 mg of phenol/day/mg of dry weight) was obtained at 200 mg/l. Biomass yield decreased at initial phenol concentrations above 60 mg/l. Catechol was identified as an intermediate metabolite by HPLC analysis and catechol dioxygenase activity was detected in plate assays, suggesting that phenol metabolism could occur via ortho fission of catechol. Wheat seeds were used as phytotoxicity indicators of phenol degradation products. It was found that these products were not phytotoxic for wheat but highly phytotoxic for phenol. The high specific degradation rates obtained under resting mycelium conditions are considered relevant for practical applications of this fungus in soil decontamination processes.

Biodegradation of phenol in static cultures by Penicillium chrysogenum ERK1: catalytic abilities and residual phototoxicity / Biodegradación de fenol en cultivos estáticos por Penicillium chrysogenum ERK1: habilidades catalíticas y fitotoxicidad residual

A phenol-degrading fungus was isolated from crop soils. Molecular characterization (using internal transcribed spacer, translation elongation factor and beta-tubulin gene sequences) and biochemical characterization allowed to identify the fungal strain as Penicillium chrysogenum Thorn ERK1. Phenol degradation was tested at 25 °C under resting mycelium conditions at 6, 30, 60, 200, 350 and 400 mg/l of phenol as the only source of carbon and energy. The time required for complete phenol degradation increased at different initial phenol concentrations. Maximum specific degradation rate (0.89978 mg of phenol/day/mg of dry weight) was obtained at 200 mg/l. Biomass yield decreased at initial phenol concentrations above 60 mg/l. Catechol was identified as an intermediate metabolite by HPLC analysis and catechol dioxygenase activity was detected in plate assays, suggesting that phenol metabolism could occur via ortho fission of catechol. Wheat seeds were used as phototoxicity indicators of phenol degradation products. It was found that these products were not phytotoxic for wheat but highly phytotoxic for phenol. The high specific degradation rates obtained under resting mycelium conditions are considered relevant for practical applications of this fungus in soil decontamination processes.

Un aislamiento fúngico capaz de degradar fenol como única fuente de carbono y energía fue aislado de suelos agrícolas. La caracterización molecular (basada en el empleo de secuencias de espaciadores de transcriptos internos, de factores de la elongación de la traducción y del gen de la beta-tubulina) y la caracterización bioquímica permitieron identificar a esta cepa como Penicillium chrysogenum Thom ERK1. Se estudió la degradación de fenol a 25 °C en cultivos estáticos con 6, 30, 60, 200, 350 y 400 mg/l de fenol inicial. El tiempo requerido para completar la degradación de fenol aumentó al elevarse las concentraciones iniciales de dicho compuesto. La máxima tasa de degradación específica (0,89978 mg de fenol/día/mg de peso seco) se obtuvo con 200 mg/l. El rendimiento en biomasa disminuyó con concentraciones Iniciales de fenol mayores de 60 mg/l. Se identificó al catecol como intermediarlo metabolico por HPLC y se observó actividad de catecol dioxigenasa en placa, lo que sugiere que el metabolismo de degradación del fenol ocurre vía orto fisión del catecol. Se utilizaron semillas de trigo como indicadores de fitotoxicidad de los productos de degradación. Estos productos no fueron fitotóxicos para trigo, mientras que el fenol mostró una alta fitotoxicidad. La alta tasa de degradación específica obtenida en condiciones estáticas resulta de gran interés para la aplicación de este hongo en procesos de descontaminación de suelos.

Surface mycobiota on Argentinean dry fermented sausages.

The diversity of the mycobiota community occurring on the surface of fermented dry sausages and in the environment of the processing plants was studied. The manufacturing plants (five in total) were located in three different areas of southern Buenos Aires province (Argentina). Samples were collected from two types of fermented sausages (short and long ripening time) at two times of the year (winter and summer) and at different types of plants (artisanal and industrial). A total of 342 samples were examined and 822 isolates belonging to six genera and 16 fungal species were identified. In most cases, Penicillium was the genus most frequently isolated. Penicillium nalgiovense, P. nordicum, P. solitum and P. chrysogenum were the most abundant species. The characteristics of the plants and the season were the influencing factors on the composition of the mycobiota. Under conditions of high humidity and in ripening rooms with reduced ventilation, Mucor racemosus was most prevalent. Eurotium amstelodami and Aspergillus spp were detected mainly during the summer when the temperature was higher. A high number of species isolated from the surface of the sausage was also isolated from air samples. Geotrichum candidum, Alternaria infectoria and P. glabrum were found only in air samples. P. chrysogenum, P. nalgiovense and P. nordicum showed different levels of antibacterial activity in sensitive bioassay with Micrococcus luteus.