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Maize (Zea mays L.), a major crop in Argentina and a staple food around the world, is affected by the emergence and re-emergence of foliar diseases. Agrochemicals are the main control strategy nowadays; however, they can cause resistance in insects and microbial pathogens and have negative effects on the environment and human health. An emerging alternative is the use of living organisms, i.e. microbial biocontrol agents, to suppress plant pathogen populations. This is a risk-free approach when the organisms acting as biocontrol agents come from the same ecosystem as the foliar pathogens they are meant to antagonize. Some epiphytic microorganisms may form biofilm by becoming aggregated and attached to a surface, as is the case of spore-forming bacteria from the genus Bacillus. Their ability to sporulate and their tolerance to long storage periods make them a frequently used biocontrol agent. Moreover, the biofilm that they create protects them against different abiotic and biotic factors and helps them to acquire nutrients, which ensures their survival on the plants they protect. This review analyzes the interactions that the phyllosphere-inhabiting Bacillus genus establishes with its environment through biofilm, and how this lifestyle could serve to design effective biological control strategies.

The objective of this study was to evaluate different doses of butylated hydroxytoleuene (BHT) against the confused flour beetle, Tribolium confusum Jacquelin du Val (Coleptera: Tenebrionidae). The effects on feeding behavior, growth, food consumption and total protein content of insects treated with different doses (1.48, 3.18, 6.36 and 9.54 mg g–1) of BHT were evaluated. Antifeedant activity was observed only at 1.48 mg g–1 in the choice assay, with a feeding deterrence index (FDI) of 0.41; this concentration also deterred feeding by 41.6%. All treatments showed feeding suppression index (FSI) and feeding inhibition index (FII) as

The present study evaluates the possible modes of action of antagonistic bacteria and their tolerance to UV radiation, temperature and osmotic stress. The partial 16S-23S rRNA gene sequencing of eight antagonistic bacteria had a high match with three bacterial genera: Curtobacterium, Pantoea and Bacillus. In this study, the three Bacillus isolates showed the most relevant production of enzymes, volatile organic compounds and antibiosis against Exserohilum turcicum. Respect to UV radiation and temperature Pantoea and Bacillus isolates were more tolerant; whereas the eight isolates were tolerant to osmotic stress in varying degree. The three Bacillus isolates have the greatest potential as biocontrol agents for foliar diseases in maize. The antagonistic action could be explained through different modes of action such as enzymes, volatile organic compounds and/or direct antibiosis by other secondary metabolites. Bacillus isolates tolerance to environmental stresses including UV radiation, temperature and osmotic stress is relevant for survival and persistence on the leaf surface. This work provides new information about the mode of action of antagonistic bacteria with proven efficacy against maize leaf pathogens. In addition, it provides information about the tolerance of antagonistic bacteria against different stress conditions. The data of the present study could contribute to the development of a successful foliar biofungicide. Fil: Sartori, Melina Victoria. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Bonacci, Martin Miguel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Barra, Paula Sabina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Fessia, Alumine Soledad. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Etcheverry, Miriam Graciela. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Nesci, Andrea Verónica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina

A third-generation enzymatic biosensor was developed to quantify sterigmatocystin (STEH). It was based on a glassy carbon electrode modified with a composite of the soybean peroxidase enzyme (SPE) and chemically reduced graphene oxide. The optimal conditions to construct the biosensor were obtained through an experimental design based on the response surfaces methodology. The experiments were performed in 0.1 mol L−1 phosphate buffer solution, pH 5. Amperometric measurements were carried out at − 0.09 V vs Ag/AgCl (3 mol L−1 NaCl). The biosensor showed a lineal response in the concentration range from 6.9 × 10−9 to 5.0 × 10−7 mol L−1. The limit of detection was 2.3 × 10−9 mol L−1 for a signal: noise ratio of 3: 1. Values of the apparent Michaellis-Menten constant, K M app , obtained by using both Lineweaver-Burk and Eadi-Hofstee methods were (1.5 ± 0.2) × 10−6 and (1.2 ± 0.2) × 10−6 mol L−1, respectively. STEH was analyzed in corn samples spiked with STEH, with an average recovery of 96.5%. The biosensor was also used to determine STEH in corn samples inoculated with the Aspergillus flavus fungus, which is an aflatoxins producer. Considering that STEH is a precursor of aflatoxin B1 (AFB1) in its biological transformation, its decrease over time was related to the production of AFB1. The STEH concentration determined using the biosensor was in very good agreement with that determined by HPLC.

The aims of this study were to determine the occurrence of Fusarium graminearum species complex (FGSC) on soybean pods, seeds and roots, including rhizoplane, during the period of soybean crop in rotation with wheat and to evaluate the FGSC dynamics on wheat and soybean residues during two soybean growing seasons in rotation with wheat, particularly F. graminearum sensu stricto (FGss).Soybean roots, pods and seeds were analysed during 2012/13 and 2013/14 seasons. The morphological identification of FGSC and mycotoxin analysis was done. Crop residues were taken in both soybean season in wheat rotation and FGss were quantificated by real-time PCR. The results showed that Fusarium species, mainly FGSC, survive in a soybean crop in rotation with wheat. Isolation frequency of these species was higher on soybean pods than on seeds at R6 stage. Deoxynivalenol contamination on soybean seeds was higher in the 2013/14 season in comparison with the 2012/13 season. Low isolation levels of Fusarium species and species that did not belong to FGSC were observed in soybean root, whereas in rhizoplane a higher level was observed. Fusarium species inoculum on residues remained stable during crop succession and the FGSC were recovered from both wheat and soybean residues. Real time PCR data showed a higher DNA concentration of FGss in wheat residues in the first developmental stages of soybean plants, being the levels more significant during 2012/13 season. With regard to soybean residues collected during the wheat growing stages, an increase in DNA from anthesis until wheat harvest was observed.In a no-till production system, the populations of FGSC can colonize wheat and soybean residues to become an inoculum source.This study provides new data on the occurrence of FGSC populations in soybean plant and FGss on residues in soybean-wheat rotation, a cultural practice commonly used in in Argentina.

Exserohilum turcicum and Puccinia sorghi cause foliar fungal diseases that affect maize crop in Argentina. These diseases, northern leaf blight and common rust respectively, are presented each year with different levels of severity affecting significantly the yield in susceptible hybrids. Disease control usually consists in the use of resistant cultivars and chemical control. Biological control as a preventive method is a viable alternative to evaluate. The aims of this study were to evaluate the natural incidence of both foliar diseases in maize after application of two antagonists, to determine the survival of the antagonists in the maize phyllosphere and to evaluate the effect of inoculation on grain yield at harvest. Plants treated with both biological control agents showed significant reductions in the incidence of both foliar diseases. In northern leaf blight the reduction was higher than 50% during 40 days in plants treated with Bacillus spp. Moreover, grain yield was significantly higher as compared to control treatments.

Northern corn leaf blight (NCLB) caused by Exserohilum turcicum is an endemic foliar disease in the Argentinean maize production area. Before applying a control method, it is essential to focus on factors that lead to the survival and conservation of E. turcicum inoculum. However, there is little information about the life cycle of this emerging pathogen in our country. The objective of the present work was to analyse the growth ability of three E. turcicum isolates under different type and conditions of maize, wheat and soybean residues, matric potential, temperature and their interactions. Statistical analysis demonstrated a significant effect of three factors on growth rate: residues-based media, matric potential and temperature. Among them the major effect was produced by matric potential followed by temperature. Although there were no significant differences in the growth rate among different residues types, the range of conditions in which growth occurred was different. According to these results, soybean residues would allow a better survival of E. turcicum inoculum under no-tillage system and providing an inoculum source for maize infections in subsequent year. Significance and Impact of the Study: Exserohilum turcicum is an emerging pathogen in Argentina that has caused significant economic losses in different maize growing areas. However, at present there is a lack of information about the life cycle of this fungal pathogen. The present study shows the influence of abiotic factors such as temperature and water potential on the growth of E. turcicum on different crop residues used in rotations with maize under no-till system cultivation. According to our results, soybean residues allowed the pathogen growth in a wider range of conditions compared to wheat and maize residue, providing an inoculum source for maize infections in subsequent season. Fil: Montemarani, Analía Mariana. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Sartori, Melina Victoria. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Nesci, Andrea Verónica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Etcheverry, Miriam Graciela. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Instituto de Investigación en Micología y Micotoxicología. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación en Micología y Micotoxicología; Argentina

Fusarium graminearum (Schwabe) is reported as the main causal agent of Fusarium head blight in Argentina. The disease causes great losses in humid and semi-humid regions of the world, reducing grain yield and quality. During 2012/13 harvest season, a severe epidemic occurred in Argentina. The aims of this work were to determine the F. graminearum incidence and deoxynivalenol accumulation in wheat grain and flour samples obtained from two of the main wheat growing regions from Argentina. Levels of the pathogen and deoxynivalenol content were correlated in heads, grains and flour. Out of 69 wheat grain samples, 55 (79.7%) showed deoxynivalenol levels between 0.4 and 8.5 μg/kg. Fusarium graminearum was the main species isolated, the isolation frequency ranged from 30 to 52% of the total grains analyzed. Correlations were observed between deoxynivalenol content, % of F. graminearum infection, presence of the pathogen in heads, grain and flour. Fil: Palazzini, Juan Manuel. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Fumero, María Verónica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Yerkovich, Nadia. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cuniberti, M.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Córdoba. Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Microbial bioremediation emerged some decades ago as an eco-friendly technology to restore polluted sites. Traditionally, the search for microorganisms suitable for bioremediation has been based on the selection of isolated strains able to remove a specific type of pollutant. However, this strategy has now become obsolete, since co-pollution is a global reality. Thus, current studies attempt to find bacterial cultures capable of coping with a mixture of organic and inorganic compounds. In this sense, the bacterial consortium SFC 500-1 has demonstrated efficiency for Cr(VI) and phenol removal, both of which are found in many industrial wastewaters. In the present study, the ability of SFC 500-1 for simultaneous removal was improved through its entrapment in a Ca-alginate matrix. This strategy led to an increased removal of Cr(VI), which was partially reduced to Cr(III). Immobilised cells were able to tolerate and degrade phenol up to 1,500 mg/l at high rates, forming catechol and cis,cis-muconate as oxidation intermediates. Successful removal potential through 5 cycles of reuse, as well as after long-term storage, was another important advantage of the immobilised consortium. These characteristics make SFC 500-1 an interesting system for potential application in the biotreatment of co-polluted effluents. Fil: Ontañon, Ornella Mailén. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: González, Paola Solange. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Agostini, Elizabeth. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The aims of this study were, to analyze in vitro phosphate solubilization activity of six native peanut bacteria and to determine the effect of single and mixed inoculation of these bacteria on peanut and maize plants. Ability to produce organic acids and cofactor PQQ, to solubilize FePO4 and AlPO4 and phosphatase activity were analyzed. Also, the ability to solubilize phosphate under abiotic stress and in the presence of pesticides of the selected bacteria was determined. The effect of single and mixed bacterial inocula was analyzed on seed germination, maize plant growth and in a crop rotation plant assay with peanut and maize. The six strains produced gluconic acid and five released cofactor PQQ into the medium. All bacteria showed ability to solubilize phosphate from FePO4 and AlPO4 and phosphatase activity. The ability of the bacteria to solubilize tricalcium phosphate under abiotic stress and in presence of pesticides indicated encouraging results. Bacterial inoculation on peanut and maize increased seed germination, plant́s growth and P content. Phosphate solubilizing bacteria used in this study showed efficient phosphate mineralizing and solubilization ability and would be potential P-biofertilizers for peanut and maize. Fil: Anzuay, María Soledad. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Ruiz Ciancio, Maria Gabriela. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina Fil: Ludueña, Liliana Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina Fil: Angelini, Jorge Guillermo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina Fil: Barros, Germán Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina Fil: Pastor, Nicolás Alejandro. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina Fil: Taurian, Tania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquimicas y Naturales. Departamento de Ciencias Naturales. Cátedra de Biologia General; Argentina

The aim of this study was to analyze if cofactor pyrroquinoline quinone from Serratia sp. S119 is involved in the inorganic phosphate solubilization mechanism and in its ability to promote the plant growth. Site directed mutagenesis was performed to obtain a pqqE- minus mutant of strain Serratia sp. S119. The phosphate solubilization ability, gluconate and PQQ production of the mutant Serratia sp. RSL (pqqE-) was analyzed. Mutant RSL (pqqE-) showed significant decrease in P soluble and gluconic acid levels produced and undetectable levels of PQQ cofactor compared with wild-type strain. Complementation with synthetic PQQ cofactor restored P solubilization and gluconate production reaching the levels produced by wild-type strain. PqqE gene sequence indicated that it is highly conserved within Serratia strains and its product shows conserved motifs found in other PqqE proteins of several bacteria. The effect of the inoculation of the PQQ- mutant on peanut and maize plants was evaluated in pot assays. Plants growth parameters showed no differences among the different treatments indicating that PQQ from Serratia sp. S119 is not involved in the growth promotion of these plants. PQQ cofactor is essential for phosphate solubilization ability of Serratia sp. S119 but is not required for growth promotion of peanut and maize plants., Facultad de Ciencias Exactas, Centro de Investigación y Desarrollo en Fermentaciones Industriales

Mycotoxins including aflatoxins, deoxynivalenol, fumonisins and ochratoxin A are among the main fungal secondary metabolites detected as natural contaminants in South America in different commodities such as peanuts (aflatoxins), cereals (deoxynivalenol and fumonisins) or grapes (ochratoxin A). Different strategies including crop rotation, tillage practices, fungicide application and planting less susceptible cultivars are used in order to reduce the impact of these mycotoxins in both food and feed chains. The development of fungicide resistance in many fungal pathogens as well as rising of public concern on the risks associated with pesticide use led to the search for alternative environmentally friendly methods. Biological control of plant pathogens and toxigenic fungi offers an alternative that can complement chemical control in the frame of an integrated pest management to reduce the impact of mycotoxins in the food and feed chains. The advances made in Argentina on reducing the impact of toxigenic fungi and mycotoxins in peanut, grapes and cereals using the biocontrol strategy are summarised. Native bacteria, yeasts and filamentous fungi have been selected to evaluate them as potential biocontrol agents. Field trials showed that Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were effective at reducing deoxynivalenol accumulation in wheat. The application of Clonostachys rosea isolates on wheat stubble reduced Fusarium colonisation on the stubble. Bacillus amyloliquefaciens and Microbacterium oleovorans showed good activity to control both Fusarium verticillioides growth and the accumulation of fumonisins at pre-harvest stage in maize. Control of toxigenic Aspergillus flavus and aflatoxin accumulation in peanuts was achieved using a native atoxigenic Aspergillus flavus strain based on competitive exclusion of the toxigenic strains. Kluyveromyces thermotolerans strains were used as biocontrol agents to reduce the impact of Aspergillus section Nigri and ochratoxin A accumulation in grapes. Fil: Chulze, Sofia Noemi. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina Fil: Palazzini, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Torres, Adriana Mabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Barros, Germán Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología. Cátedra de Micología; Argentina Fil: Ponsone, Maria Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; Argentina. Instituto Nacional de Tecnología Agropecuaria; Argentina Fil: Geisen, R.. Max Rubner-Institut; Alemania Fil: Schmidt Heydt, M.. Max Rubner-Institut; Alemania Fil: Köhl, J.. Wageningen UR. Plant Research International, Wageningen; Países Bajos

Soybean (Glycine max L.) is one of the main crops in Argentina. Most of the studies of pathogenicity in the Fusarium graminearum complex have focused on strains isolated from wheat and maize, and there is little information on strains isolated from soybean. Our objective in the present study was to compare the pathogenicity among soybean isolates of different phylogenetic species within the Fusarium graminearum complex on soybean seedlings under controlled conditions. Six strains representing three different phylogenetic species (F. graminearum, F. meridionale and F. cortaderiae) were identified by partial sequencing of the Translation Elongation Factor -1α gene (TEF-1) and evaluated for pathogenicity. All six strains reduced emergence, mainly by causing pre-emergence damping-off, seedling height and root dry weight and produced abnormal seedlings. The mean disease severity averaged across all isolates was approximately 3.0 in a 0–4 rating scale where 0 = healthy seedling and 4 = dead seedling. Significant differences in pathogenicity were observed among F. graminearum, F. meridionale and F. cortaderiae. These results are consistent with the hypothesis that different phylogenetic species within the Fusarium graminearum complex isolated from soybean are pathogenic under controlled conditions to soybean seedlings in Argentina. The present study demonstrates for the first time the pathogenic effect of F. meridionale on soybean in Argentina. Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Alaniz Zanon, Maria Silvina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Chiotta, María Laura. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Reynoso, Maria Marta. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina Fil: Scandiani, María Mercedes. Laboratorio Agrícola Río Paraná, San Pedro; Argentina Fil: Chulze, Sofia Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina

Biocontrol by competitive exclusion has been developed as the most promising means of controlling aflatoxins in peanuts. A 2-year study was carried out to determine the efficacy of an Aspergillus flavus strain as biocontrol agent to reduce aflatoxin production in peanuts under field conditions in Argentina. The competitive strain used was a nontoxigenic A. flavus (AFCHG2) naturally occurring in peanut from Cordoba, Argentina. The inoculum was produced through solid-state fermentation on long grain rice and applied at rate of 50 kg inoculum/ha. The incidence of the released strain within the A. flavus communities in soil and peanuts was determined using the shift in the ratio toxigenic:nontoxigenic and VCG analysis. During the 2009/2010 growing season, treatments produced significant reductions in the incidence of toxigenic isolates of A. flavus/Aspergillus parasiticus in soil and peanuts. However, no preharvest aflatoxin contamination was observed. In the 2010/2011 growing season, plants were exposed to late season drought conditions that were optimal for aflatoxin contamination. Significant reductions in aflatoxin levels averaging 71% were detected in treated plots with different inoculation treatments. The results suggest that using the strategy of competitive exclusion A. flavus AFCHG2 can be applied to reduce aflatoxin contamination in Argentinean peanuts.

Fusarium equiseti and Fusarium semitectum represent the most abundant species in the Fusarium complex isolated from flowers, soybean pods and seeds in Argentina. The aim of the present study was to assess the production of major type A and type B trichothecenes (diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin, nivalenol, deoxynivalenol) and zearalenone by 40 F. equiseti and 22 F. semitectum isolates on rice culture. Mycotoxins were determined by HPLC with fluorescence detection after derivatisation with 1-anthronylnitrile for type A trichothecenes (i.e. diacetoxyscirpenol, neosolaniol, T-2 toxin and HT-2 toxin), by HPLC with UV detection for type B trichothecenes (i.e. nivalenol and deoxynivalenol), and by TLC for zearalenone. A total of 22 of 40 F. equiseti isolates produced diacetoxyscirpenol, nivalenol and ZEA alone or in combination, whereas only two of 20 F. semitectum isolates were nivalenol and ZEA producers. Both Fusarium species did not produce any deoxynivalenol, neosolaniol, T-2 toxin and HT-2 toxin. The variable retention in toxigenicity displayed by both fungal species suggests that these species have a saprophytic lifestyle in the soybean agroecosystem in Argentina.

The natural occurrence of alternariol (AOH) and alternariol monomethyl ether (AME) in soya beans harvested in Argentina was evaluated. Both toxins were simultaneously detected by using HPLC analysis coupled with a solid phase extraction column clean-up. Characteristics of this in-house method such as accuracy, precision and detection and quantification limits were defined by means of recovery test with spiked soya bean samples. Out of 50 soya bean samples, 60% showed contamination with the mycotoxins analyzed; among them, 16% were only contaminated with AOH and 14% just with AME. Fifteen of the positive samples showed co-occurrence of both mycotoxins analyzed. AOH was detected in concentrations ranging from 25 to 211 ng/g, whereas AME was found in concentrations ranging from 62 to 1,153 ng/g. Although a limited number of samples were evaluated, this is the first report on the natural occurrence of Alternaria toxins in soya beans and is relevant from the point of view of animal public health.

BACKGROUND:Fusariumgraminearum andF.verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans. RESULTS: F.graminearum grew better when was incubated at 15, 20 and 15 – 20 ◦ C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µ gg −1 , respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 ◦ Cf or zearalenone and 20 ◦ C for deoxynivalenol. F.verticillioides grew better at 25 ◦ C in culture medium and at 15/20 ◦ C and 15/25 ◦ C on soybean seeds. Fumonisin B1 was produced only in culture medium, and the maximum level (7.38 µ gg −1 ) was found at 15 ◦ C after 7 days of incubation. CONCLUSION: When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions. c � 2012 Society of Chemical Industry

Soybean (Glycine max L.), the main source of protein throughout the world, is used both as a food and a feedstuff. Currently, limited information about the occurrence of Fusarium species and mycotoxins in soybean grain and by-products is available. The aims of the present study were: (1) to identify toxigenic Fusarium species associated with soybean during crop reproductive stages; (2) to determine the occurrence of deoxynivalenol (DON) and nivalenol (NIV) in soybean seeds; (3) to determine the genotype and chemotype of selected Fg complex strains using molecular and chemical analysis, respectively; and (4) to characterize the strains using AFLP(s) markers. One soybean field located at Córdoba Province, Argentina, was monitored and samples of soybean tissue were harvested at three reproductive stages: flowering (R2), full seed (R6) and full maturity (R8). A total of 389 Fusarium strains F. equiseti (40%) was the most frequently species recovered followed by F. semitectum (27%) and F. graminearum (Fg) (11%). From the 40 soybean samples analysed, only two presented detectable DON levels. Based on DON occurrence on soybean seeds at ripening stages, the toxigenic ability of Fg complex strains isolated from soybean seeds, pods and flowers were analysed. The trichothecene genotype was determined by a multiplex PCR using primers based on Tri3, Tri5 and Tri7 toxin genes and then the chemotype was verified by chemical analysis. Most Fg complex strains showed 15-ADON genotype and five strains presented a DON/NIV; these also produced both toxins under in vitro culture. Neither the NIV nor the 3-ADON genotypes were detected among the members of the population evaluated. All the 15-ADON genotype strains were characterized as F. graminearum sensu stricto (lineage 7), while the strains presented a DON/NIV genotype were characterized as F. meridionale (lineage 2). The present study contributes new information on the occurrence of Fusarium species and trichothecenes toxins on soybean at the pre-harvest stages. Also, this is the first report on the chemotype, genotype and lineages among Fg complex isolated from soybean.

In the last years Conyza bonariensis has become an important weed and control is difficult with the use of current technology in Argentinean pampas region. The increasing prevalence of herbicide-resistant weed species, public concern related to pesticide use and the introduction of government policies for pesticide reduction, is driving the search for alternative methods to chemical control. The aims of the present study were to detect fungal diseases associated with C. bonariensis, to identify fungal isolates from the symptomatic leaves and to confirm through Koch’s postulates the isolates pathogenicity. Mycological analysis of symptomatic leaves showed the presence of twelve genera of filamentous fungi. Among 116 isolates, Colletotrichum spp. was the most prevalent genus followed by Nigrospora spp. and Septoria spp. In the pathogenicity assays, 22 out of 116 isolates were able to comply with the Koch’s postulates. The pathogenic isolates were included into three genera Alternaria spp., Colletotrichum spp. and Septoria spp. This study provides the first report that demonstrates pathogenicity of fungal isolates on C. bonariensis in Argentina and represents the first step in a future biocontrol program.

The objective of this study was to determine the effect of water activity (a(w); 0.995, 0.98, 0.96, 0.94, 0.92, and 0.90), temperature (5, 18, 25, and 30 degrees C), incubation time (7 to 35 days), and their interactions on mycelial growth and alternariol (AOH) and alternariol monomethyl ether (AME) production. Two Alternaria alternata strains isolated from soybeans in Argentina were grown on 2% soybean extract agar. Maximum growth rates were obtained at the highest a(w) (0.995) and 25 degrees C, with growth decreasing as the water availability of the medium was reduced. Maximum amount of AOH was produced at 0.98 a(w) and 25 degrees C for both strains. Maximum AME production was obtained for both strains at 30 degrees C but different a(w) values, 0.92 and 0.94, for the strains RC 21 and RC 39, respectively. The concentrations of both toxins varied considerably depending on the a(w) and temperature interactions assayed. The two metabolites were produced from 5 to 30 degrees C and at a(w) values of 0.92 to 0.995. Although at 5 and 18 degrees C little mycotoxin was produced at a(w) lower than 0.94. Two-dimensional profiles of a(w) by temperature interactions were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soybeans. All the conditions of a(w) and temperature that resulted in maximum production of both toxins are those found during soybean development in the field. Thus, field conditions are likely to be conducive to optimum A. alternata growth and toxin production.

Aims: To determine the effects of water activity (aW; 0·995–0·90), temperature (5, 18, 25 and 30°C), time of incubation (7–35 days) and their interactions on tenuazonic acid (TA) production on 2% soybean-based agar by two Alternaria alternata strains isolated from soybean in Argentina. Methods and Results: TA production by two isolates of A. alternata was examined under interacting conditions of aW, temperature and time of incubation on 2% soybean-based agar. Maximum TA production was obtained for both strains at 0·98 aW, but at 30 and 25°C for the strains for RC 21and RC 39, respectively. The toxin concentration varied considerably depending on aW, temperature, incubation time and strain interactions. TA was produced over the temperature range from 5 to 30°C and aW range from 0·92 to 0·995, however at 5 and 18°C little TA was produced at aW below 0·94. Contour maps were developed from these data to identify areas where conditions indicate a significant risk for TA accumulation. Conclusions: The optimum and marginal conditions for TA production by A. alternata on soybean-based agar were identified. The results indicated that TA production by A. alternata is favoured by different temperatures in different strains. Significance and Impact of the Study: Data obtained provide very useful information for predicting the possible risk factors for TA contamination of soybean as the aW and temperature range used in this study simulate those occurring during grain ripening. The knowledge of TA production under marginal or sub-optimal temperature and aW conditions for growth are relevant as improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality.

A Bt maize hybrid and its non-transgenic counterpart harvested during 2002/2003 and 2003/2004 harvest seasons from different locations within the maize-growing area in Argentina were compared for fungal and mycotoxin contamination. Fusarium species were the most prevalent on both genotypes with an isolation frequency >60% across all locations. The percentage of infection was lower in Bt maize than in non-Bt maize (P1, B2 and B3) in the Bt and the non-Bt hybrid were significantly different (P

BACKGROUND: Accumulation of patulin in apples and pears and subsequently in their by-products is caused mostly by Penicillium expansum which causes blue mould. Fruit pH and other parameters are sometimes characteristic of a certain variety and thus the use of particular varieties might affect patulin content in the final products. The aim of this study was to evaluate the influence of fruit variety and pH on patulin accumulation. Patulin accumulation in both apple and pear juices at different pH and also in different apple varieties (Golden, Gala and Fuji) and pear varieties (Blanquilla and Conference) was assessed. RESULTS: The pH of juices significantly influenced patulin accumulation, especially in apple juice in which the highest amounts of patulin were detected at pH 3.5. In fruits, the pH values of the substrate were determinant only under cold storage. Thus, Golden apples, which presented a lower pH, accumulated more patulin at 1 ◦ C. However, this trend was not observed at other temperatures in which varieties with higher amounts of organic acids (Golden and Fuji apples) accumulated most patulin. In the pear varieties, significant differences in pH did not lead to significant differences in patulin accumulation. CONCLUSION: Although pH influences patulin accumulation, other factors such as organic acid content may play an important role. Concerning the apple and pear varieties we studied, it seems that rather than variety, other parameters like pH or acidic content may be more important. These parameters vary significantly depending on the degree of ripeness of the fruit. Gala apple, which is used in Spanish juice production, was the only variety that accumulated dramatically higher amounts of patulin.  2008 Society of Chemical Industry

A rapid and accurate method of quantifying deoxynivalenol (DON) and nivalenol (NIV) in soybean and soy flour is described. The samples were extracted with acetonitrile:water (84:16, v/v) and cleaned through a solidphase extraction (SPE) column. The mycotoxins were separated, detected and quantified by reversed-phase high performance liquid chromatography (HPLC) with UV detection (220 nm) using water:methanol (88:12, v/v) as mobile phase. Characteristics of this in-house method such as accuracy, precision and detection and quantification limits were defined by means of a recovery test with spiked soybean and soy flour samples. The detection limit (LOD) was 0.1 µg/g for DON and 0.2 µg/g for NIV, based on a signal-noise ratio 3:1. Quantification limit (LOQ) was established as three times the detection limit.

Aims: The objectives of this study were: (i) to evaluate genetic relatedness among Aspergillus section Flavi strains isolated from soil and peanut seeds in Argentina; (ii) to determine if AFLP molecular markers could be useful to identify isolates up to species level, and to correlate these markers with the isolates’ toxigenic potentials and/or vegetative compatibility group (VCG) affiliations. Methods and Results: Amplified fragment length polymorphism (AFLPs) analysis was applied to compare 82 isolates of Aspergillus section Flavi. Cluster analysis showed a clear separation of A. flavus and A. parasiticus, and comparison of fingerprints revealed several specific markers for each group of isolates. AFLP analysis indicates that no genotypical differences can be established between aflatoxigenic and nonaflatoxigenic producers in both species analysed. In addition, candidate AFLP markers associated with a particular VCG were not found. Conclusions: There was a concordance between morphological identification and separation up to species level using molecular markers. The findings of specific bands for A. flavus and A. parasiticus may be useful for the design of specific PCR primers in order to differentiate these species and detect them in food. Significance and Impact of the Study: The present study provides new data on molecular characterization of Aspergillus section Flavi in Argentina.

The distribution of the soil borne fungal population in maize non-rhizospheric soil in experimental trials were studied during the harvest seasons 2000/2001 and 2001/2002. The effect of different tillage practices was evaluated. Soil samples were collected from three tillage treatments, conventional tillage, reduced tillage and no tillage, with and without grazing. The distribution of fungal propagules in different treatments was compared. Soil samples were collected from the top soil (3 cm) before planting, in the sprong. No tillage without grazing and reduced tillage with grazing had the highest densities of filamentous fungi (5.71 and 5.85 log, respectively). The mycological survey showed the presence of six genera of filamentous fungi. They were Aspergillus , Fusarium , Penicillium , Trichoderma , Cladosporium , Alternaria . The species identified from genus Aspergillus were A. flavus , A. restrictus , A. candidus , A. parasiticus and other Aspergillus spp. The species identified from genus Penicillium were P. pinophylum , P. citreonigrum , P. implicatum , P. purpurogenum , P. minioluteum , P. waksmanii , P. restrictum , P. rugulosum , P. funiculosum and P. variabile . Species from genus Fusarium were identified as F. verticillioides , F. oxysporum , F. proliferatum and others Fusarium species. The isolation frequency of Aspergillus increased in the no tillage and grazing practices treatments. The data indicate that different agricultural practices impact soil inocula of principal maize toxigenic fungi in the production area. To our knowledge, this is the first report on the effect of tillage management practices on fungal populations in preharvest maize agroecosystem in Argentina.

The genetic diversity of Aspergillus flavus populations isolated from the peanut-cropped soils in the peanut-growing region at Cordoba Province was evaluated by analysis of vegetative compatibility group (VCG). VCGs were determined through complementation assays between nitrate-nonutilizing (NNO) mutants. Fifty-six VCGs were identified from 100 isolates. Twenty-five VCGs contained two or more isolates and 31 VCGs contained only a single isolate. In general, there were significant differences among VCGs in aflatoxin and CPA production. One VCG group included a single atoxigenic strain since it was neither aflatoxin nor cyclopiazonic acid producer. This isolate could be useful as a biological control agent, since it was unable to form a stable heterokaryon in the complementation test with the other isolates. Seven A. flavus isolated from soil were atypical because they simultaneously produced aflatoxins B, G and CPA. q 2005 Elsevier Ltd. All rights reserved.

The Aspergillus flavus population was evaluated in the period 1998–2001 in soil samples from the peanut-growing region in Argentina. A total of 369 A flavus isolates were examined for sclerotia, aflatoxin and cyclopiazonic acid production. The L phenotype was isolated in a higher percentage than the S phenotype and represented 59% of the total isolates. Statistical analysis showed significant differences between L, S and non-sclerotial strains with regard to aflatoxin and cyclopiazonic acid production (p < 0.05). The S strains produced higher mycotoxin levels than the L and non-sclerotial strains. About 10% of the S strains had an unusual pattern of mycotoxin production because they simultaneously produce aflatoxins B and G and CPA. The SBG strains isolated in the present study have all morphological and microscopic characteristics of A flavus. These strains are of concern in food safety, as there is a higher probability of aflatoxin contamination in peanuts. Copyright © 2005 Society of Chemical Industry

Aspergillus species from section Flavi were isolated from soil samples in three peanut-growing regions of Cordoba Province, Argentina. The samples were collected during the planting and harvest periods. Both total fungal population and Aspergillus species from section Flavi showed no significant differences between planting and harvest time in two of the regions evaluated. Only in one region were there significant differences in cfu g−1 of total fungal population and Aspergillus species from section Flavi. A flavus was the dominant species isolated in all three localities during the planting and harvest periods. There were significant differences (p < 0.05) in the ratio of toxigenic and atoxigenic strains dependent on the period and the region evaluated. In one region, higher frequencies of toxigenic A flavus and A parasiticus in soil were found and a high contamination level of aflatoxins was detected in peanut seeds. Copyright © 2003 Society of Chemical Industry

Production of aflatoxin B1 and fumonisin B1 in pure and mixed cultures of Aspergillus flavus and Fusarium proliferatum were determined on irradiated maize seeds inoculated with different spore concentrations at 0.97 water activity (aw) and a temperature of 25 °C. The highest levels of aflatoxin B1 were produced by A. flavus at the lowest levels of inoculum (103 spore ml−1). There was no spore concentration influence on fumonisin B1 production after 10, 20 and 35 days of incubation. When A. flavus was co-inoculated with F. proliferatum, aflatoxin B1 production was inhibited. The higher the inocula levels of Fusarium produced, the higher the inhibition and this inhibition increased during the incubation period. Total inhibition was reached at 35 days of incubation. There was no interaction influence on fumonisin B1 production at all inoculum levels assayed. These results suggest that under optimal environmental conditions of substrate, water activity and temperature, the interaction between A. flavus and F. proliferatum could produce inhibition of aflatoxin B1 and stimulation of fumonisin B1. Copyright © 1999 John Wiley & Sons, Ltd.

A study has been carried out in Argentina on samples of corn genotypes from a breeding station as well as in commercially available corn meal. All samples were analyzed for fungal infection and aflatoxin B1.Mycological analysis of corn genotypes showed the presence of three principal genera of filamentous fungi Fusarium (100%), Penicillium (67%) and Aspergillus (60%). In the genus Fusarium three species were identified, F. moniliforme (42%), F. nygamai (56%) andF. proliferatum (1.8%). Eight species ofPenicillium were identified, the predominant species isolated were P. minioluteum, P. funiculosum and P. variabile. In the genus ranked third in isolation frequency, two species were identified, A. flavus and A. parasiticus, the percentage of infection was 78% and 21%, respectively. Only one corn genotype was contaminated with aflatoxin B1 at a level of 5 ppb. The cornmeal samples showed great differences in fungal contamination, the values ranging from 1 × 101 to 7 × 105 cfu g−1. Fusarium (68%), Aspergillus (35%) and Penicillium (21%) were the most frequent genera isolated. Among the genus, Aspergillus, A. parasiticus (38%) was the most frequent species isolated. All the samples of corn meal were negative to aflatoxin B1. These results indicate a low degree of human exposure to aflatoxins in Argentina through the ingestion of maize or corn meal.

The aim of this study was to evaluate whether the interaction of Vicia sativa with a bacterial strain capable of using phenol as sole carbon and energy sources can moderate adverse effects of this pollutant in plant tissues. A bacterial strain identified as Bacillus sp., isolated from a heavily polluted environment, was inoculated at different stages of growth. In root elongation assay, inoculated seeds showed higher values of relative root elongation and germination index than uninoculated ones in the presence of high phenol concentrations. Thus, common vetch–Bacillus sp. association could be important at the first stages of development allowing this plant to grow in highly polluted environments. Besides, phenol removal was largely accelerated in phenol-spiked soils, after 48 h of treatment with uninoculated/inoculated plants rather than by adsorption or biodegradation of the bacterial strain. Peroxidase and ascorbate peroxidase activities increased significantly in uninoculated plants, while superoxide dismutase activity, chlorophyll, malondialdehyde, and H2O2 levels of aerial parts remained unaltered in uninoculated/inoculated plants treated with the pollutant, demonstrating that the efficient response to oxidative damage did not depend on the inoculation. Fil: Ibañez, Sabrina Guadalupe. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Merini, Luciano Jose. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barros, Germán Gustavo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Microbiología e Inmunología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Medina, Maria Ines. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Agostini, Elizabeth. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Fusarium graminearum and F. verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans.F. graminearum grew better when was incubated at 15, 20 and 15-20 °C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µg g(-1), respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 °C for zearalenone and 20 °C for deoxynivalenol. F. verticillioides grew better at 25 °C in culture medium and at 15/20 °C and 15/25 °C on soybean seeds. Fumonisin B(1) was produced only in culture medium, and the maximum level (7.38 µg g(-1)) was found at 15 °C after 7 days of incubation.When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions.

Soybean (Glycine max L. Merr.) is an Asiatic leguminous plant cultivated in many parts of the world for its oil and proteins, which are extensively used in the manufacture of animal and human foodstuffs (FAO, 2004a; Hepperly, 1985). The production reached 47.5 million tons during the 2006/2007 harvest season ranking Argentina third as soybean producer in the world. In Argentina, during the last quarter of the century, soybean production has increased at an unprecedented rate from a cultivated area of 38.000 hectares in 1970 to 16 million hectares today. Around 70% of the soybean harvested is processed, providing 81% and 36% of the world's exported soybean oil and meal, respectively (SAGPyA, 2010). Soybean is often attacked by fungal infections during cultivation, or post-harvest (in transit or in storage), significantly affecting its productivity. Seeds and infected harvest debris are the main sources of primary infections, and the level of seed damage depends on environmental conditions such as high relative humidity, dew, and temperatures above 25 oC. These species can be potential mycotoxin producers. Mycotoxins (from “myco” fungus and toxin) are relatively low-molecular weight, fungal secondary metabolic products that may affect exposed vertebrates such as animals in a variety of ways. Mycotoxins are considered secondary metabolites because they are not necessary for fungal growth and are simply a product of primary metabolic processes. The functions of mycotoxins have not been clearly established, but they are believed to play a role in eliminating other microorganisms competing in the same environment. They are also believed to help parasitic fungi invade host tissues. The amount of toxins needed to produce adverse health effects varies widely among toxins, as well as within each person’s immune system (Brase et al., 2009). Some mycotoxins are carcinogenic, some are vasoactive, and some cause central nervous system damage. The mycotoxins can be acutely or chronically toxic, or both, depending on the kind of toxin, the dose, the health, the age and nutritional status of the exposed individual or animal, and the possible synergistic effects between mycotoxins. The most frequently studied mycotoxins are produced by species of Aspergillus, Penicillium, Fusarium and Alternaria. There is an increasing world consumer demand for high quality and inocuous food and drink products with the lowest possible level of contaminants such as mycotoxins. As a result, the food industry in the developed world demands raw ingredients of the best

The aim of this study was to determine the effects of water activity (a(w)) (0.99-0.90), temperature (15, 25 and 30°C) and their interactions on growth and alternariol (AOH) and alternariol monomethyl ether (AME) production by Alternaria alternata on irradiated soya beans. Maximum growth rates were obtained at 0.980 a(w) and 25°C. Minimum a(w) level for growth was dependent on temperature. Both strains were able to grow at the lowest a(w) assayed (0.90). Maximum amount of AOH was produced at 0.98 a(w) but at different temperatures, 15 and 25°C, for the strains RC 21 and RC 39 respectively. Maximum AME production was obtained at 0.98 a(w) and 30°C for both strains. The concentration range of both toxins varied considerably depending on a(w) and temperature interactions. The two metabolites were produced over the temperature range 15 to 30°C and a(w) range 0.99 to 0.96. The limiting a(w) for detectable mycotoxin production is slightly greater than that for growth. Two-dimensional profiles of a(w)× temperature were developed from these data to identify areas where conditions indicate a significant risk from AOH and AME accumulation on soya bean. Knowledge of AOH and AME production under marginal or sub-optimal temperature and a(w) conditions for growth can be important since improper storage conditions accompanied by elevated temperature and moisture content in the grain can favour further mycotoxin production and lead to reduction in grain quality. This could present a hazard if the grain is used for human consumption or animal feedstuff.

Aflatoxins are produced by Aspergillus flavus and A. parasiticus in oil-rich seed and grain crops and are a serious problem in agriculture, with aflatoxin B1 being the most carcinogenic natural compound known. Sexual reproduction in these species occurs between individuals belonging to different vegetative compatibility groups (VCGs). We examined natural genetic variation in 758 isolates of A. flavus, A. parasiticus and A. minisclerotigenes sampled from single peanut fields in the United States (Georgia), Africa (Benin), Argentina (Córdoba), Australia (Queensland) and India (Karnataka). Analysis of DNA sequence variation across multiple intergenic regions in the aflatoxin gene clusters of A. flavus, A. parasiticus and A. minisclerotigenes revealed significant linkage disequilibrium (LD) organized into distinct blocks that are conserved across different localities, suggesting that genetic recombination is nonrandom and a global occurrence. To assess the contributions of asexual and sexual reproduction to fixation and maintenance of toxin chemotype diversity in populations from each locality/species, we tested the null hypothesis of an equal number of MAT1-1 and MAT1-2 mating-type individuals, which is indicative of a sexually recombining population. All samples were clone-corrected using multi-locus sequence typing which associates closely with VCG. For both A. flavus and A. parasiticus, when the proportions of MAT1-1 and MAT1-2 were significantly different, there was more extensive LD in the aflatoxin cluster and populations were fixed for specific toxin chemotype classes, either the non-aflatoxigenic class in A. flavus or the B1-dominant and G1-dominant classes in A. parasiticus. A mating type ratio close to 1∶1 in A. flavus, A. parasiticus and A. minisclerotigenes was associated with higher recombination rates in the aflatoxin cluster and less pronounced chemotype differences in populations. This work shows that the reproductive nature of the population (more sexual versus more asexual) is predictive of aflatoxin chemotype diversity in these agriculturally important fungi., Author Summary Fungal pathogen populations have mixed proportions of vegetative propagation and sexual reproduction ranging from predominantly clonal to varying levels of sexuality. Aflatoxins are the most potent naturally occurring carcinogens known and aflatoxin-producing Aspergillus flavus and A. parasiticus show extensive genetic and mycotoxin diversity. Population genetic studies and experimental matings in the laboratory have revealed the underlying genetic mechanisms and adaptive processes that create and maintain aflatoxin diversity. These studies provided unequivocal evidence of meiosis, crossing over, and aflatoxin heritability, but whether these processes directly influence genetic diversity in nature with respect to aflatoxin formation is not clear. Here, our work with A. flavus, A. parasiticus and A. minisclerotigenes from fields in different continents shows that populations with higher mean recombination rates exhibit less variability in aflatoxin profiles due to genetic intermixing, whereas populations with lower recombination rates have greater variability in aflatoxin profiles due to increased fixation of specific toxin chemotypes. Therefore, sexuality generates novel toxin chemotypes but tends to equalize toxin differences in populations. Our study highlights how an understanding of variation in mating-type frequency, fertility and recombination in these fungi is crucial for the selection of nontoxigenic biocontrol strains for long-term reduction of aflatoxins in target regions.