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.

Contamination of Fusarium spp. and mycotoxins at different ear physiological stages of maize in Argentina.

Maize is one of the most important crops worldwide, being affected by several fungal species under field conditions. The study of plant-pathogen interaction plays a key role because fungal diseases are responsible for reducing grain yield and quality by increasing mycotoxin production. Thus, the present work aimed to evaluate the interaction of F. graminearum and F. verticillioides and mycotoxin production under field conditions along different physiological stages. During the 2019/2020 and 2020/2021 growing seasons, twelve maize genotypes were inoculated at the flowering stage (silking). Four treatments were applied using one isolate of each species: (i) F. graminearum; (ii) F. verticillioides; (iii) a combined inoculum (F. graminearum + F. verticillioides); (iv) and control treatment. Fungal diversity, disease evaluation, and mycotoxin contamination were evaluated at three different physiological stages: T1 (from R2 to R4), T2 (from R4 to R6), and T3 (at harvest time). A total of 15,907 Fusarium isolates were obtained. The results showed a predominance of F. verticillioides over F. graminearum in both years evaluated, reporting an increase in the occurrence of this species at late stages. Regarding mycotoxin contamination, no evidence was found supporting antagonism or synergism regarding isolates of both species used as inoculum under field conditions. The results reported in the present manuscript point out a major influence of climatic conditions on F. verticillioides predominance over F. graminearum, mainly during the late physiological stages. Furthermore, no clear relationship between mycotoxin concentration and physiological stages was established, suggesting that other grain factors, such as water activity and pH, could modulate mycotoxin production and accumulation under field conditions.

Alternaria in malting barley: Characterization and distribution in relation with climatic conditions and barley cultivars.

Alternaria is one of the main fungal genera affecting the quality of barley grains. In this study, a polyphasic approach was carried out to characterise the Alternaria population infecting different cultivars of barley grains from the major producing regions of Argentina in the 2014 and 2015 seasons. Its relationship with Fusarium and correlations between predominant species, barley cultivars, and climatic conditions in the growing regions were evaluated. Alternaria incidence exceeded that of Fusarium in all the barley samples and was higher in the drier season (21% in 2014 and 42% in 2015 vs. 6% and 4%, respectively). The main Alternaria species-groups identified were present in both growing seasons in similar frequencies (A. tenuissima sp.-grp., 83.4% in 2014 and 81.7% in 2015; A. infectoria sp.-grp., 11.7% in 2014 and 11.3% in 2015). The dominant Alternaria species-group isolated and identified based on morphological characteristics, DNA sequencing, and metabolite profile was A. tenuissima (72.9%), followed by A. infectoria (14.6%). An association between their frequency and field temperature was observed; A. tenuissima sp.-grp. was more frequent in northern localities, where higher temperatures were registered, while the opposite was observed for A. infectoria sp.-grp. A smaller percentage of A. arborescens sp.-grp. (5%), A. alternata sp.-grp. (3.9%) and A. vaccinii (1.4%) were also identified. Both secondary metabolite profiles and phylogenetic analysis were useful to distinguish isolates from Alternaria section Alternaria and section Infectoriae. Regarding metabolite profiles, alternariol was the most frequent compound produced by isolates of the section Alternaria. Infectopyrones and novae-zelandins were produced by most of the isolates from section Infectoriae. The barley cultivars analysed in this study did not show a particular susceptibility regarding the Alternaria population composition, except for Andreia, which presented the highest frequency of contamination with A. tenuissima sp.-grp. The rest of the cultivars, when grown in different regions, showed different proportion of the Alternaria sp.-grps., suggesting that other factors were determinant in their distribution. The results obtained in the present study will be a valuable tool for health authorities to assess the need for regulations on Alternaria mycotoxins, given the high incidence of Alternaria spp. in barley and the diversity of metabolites that might contaminate the grains.

Effects of Fusarium graminearum and Fusarium poae on disease parameters, grain quality and mycotoxin contamination in barley (part II).

BACKGROUND: Barley is one of the most sown crops in the world, with multiple uses such as human consumption, animal feed and for the malting industry. This crop is affected by different diseases, such as Fusarium Head Blight (FHB), that causes losses in yield and quality. In the last years F. graminearum and F. poae were two of the most frequently isolated species in barley grains, so the aim of this study was to evaluate the interaction between these Fusarium species and the effects on disease parameters, grain quality and mycotoxin contamination on five barley genotypes under field conditions. RESULTS: Statistical differences between Fusarium treatments for some parameters depending mainly on the year/genotype were found. The results showed that the germination process was affected by both Fusarium species. As to grain quality and the different hordein fractions, it was observed that F. graminearum affects preferentially D and C-hordeins. Different concentrations of nivalenol, deoxynivalenol and their acetylated derivatives (3-acetyl deoxynivalenol (3-ADON), 15-acetyl deoxynivalenol (15-ADON)) were detected. CONCLUSIONS: In the present work, no evidence of synergism between F. graminearum and F. poae were found regarding disease parameters and mycotoxin contamination. However, at least in the years with favorable climatic conditions to FHB development and depending on the barley genotype, a continuous monitoring is deemed necessary to prevent the negative impact on protein composition and germinative parameters © 2020 Society of Chemical Industry.

Natural occurrence of Alternaria mycotoxins in malting barley grains in the main producing region of Argentina.

BACKGROUND: Barley (Hordeum vulgare L.) is one of the most important cereals worldwide, and its quality is affected by fungal contamination such as species of the genus Alternaria. No information is available about the occurrence of Alternaria mycotoxins in Argentinean barley grains, which is of concern, because they can be transferred into malt and beer. The aim of this study was to analyze the occurrence of alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TeA) in malting barley grains from the main producing region of Argentina during the 2014 and 2015 growing seasons. RESULTS: The most frequent mycotoxin was AOH (64%), which was detected at higher levels (712 µg kg-1 ) compared with other studies, followed by TeA (37%, 1522 µg kg-1 ), while AME was present in five samples in the 2015 growing season only, with a mean of 4876 µg kg-1 . A similar frequency of mycotoxin occurrence was observed in both years (80.8 vs 85.3%), but more diverse contamination was found in 2015, which was characterized by lower accumulated precipitation. Nevertheless, AOH was more frequently found in 2014 than in 2015 (80.8 and 47.1% respectively). A negative correlation between AOH concentration and temperature was observed. The susceptibility of different barley varieties to mycotoxin accumulation varied with the mycotoxin, geographical location and meteorological conditions. CONCLUSION: The results obtained in the present work represent a tool for risk assessment of exposition to these mycotoxins and could be used by food safety authorities to determine the need for their regulation. Furthermore, the establishment of a hazard analysis and critical control point (HACCP) system to minimize fungal and mycotoxin contamination in barley from farm to processing could be apply to ensure food safety. © 2019 Society of Chemical Industry.

Pathogenic ability and saline stress tolerance of two Fusarium isolates from Odontesthes bonariensis eggs / Patogenicidad y tolerancia al estrés salino de dos aislamientos de Fusarium aislados de huevos de Odontesthes bonariensis

Background. Several fungal species represent a potential risk to embryos of Odontesthes bonariensis (Cuvier and Valenciennes, 1835), a euryhaline freshwater fish that lives in the Pampean inland waters and has potential economic relevance. Aims. To identify two fungi isolated from O. bonariensis eggs exposed to saline conditions and to characterize their pathogenicity and tolerance to sodium chloride solutions. Methods. The isolates were identified by morphological features, and a preliminar phylogenetic analysis using sequences of translation elongation factor 1-alpha (EF-1α) and calmodulin (CAM) was performed. Koch's postulates were tested to identify the causative agent of fungal infection. The influence of NaCl on the fungal growth was evaluated in in vitro assays. Results. The isolates LPSC 1001 and 1002 were identified as representatives of the genus Fusarium, and belonging to the Fusarium incarnatum-Fusarium equiseti species complex (FIESC) and the Fusarium solani species complex (FSSC), respectively. Histological observations on eggs exposed in vitro to both isolates in infectivity assays confirmed the ability of the fungal isolates to penetrate to egg's chorionic membrane, leading to the death of embryos. Increasing NaCl concentration in the culture medium reduced the growth of the isolates LPSC 1001 and 1002, being completely inhibited at 160 and 120 g/l NaCl respectively. Conclusions. The isolates LPSC 1001 (FIESC) and 1002 (FSSC) were identified as fungal pathogens to O. bonariensis eggs. The use of NaCl solutions as antifungal treatment was not effective to control the infection with these strains (AU)

Antecedentes. Diversas especies de hongos pueden representar un riesgo importante para los embriones de Odontesthes bonariensis (Cuvier & Valenciennes 1835), un pez de agua dulce eurihalino que vive en las aguas interiores pampeanas y tiene una relevancia económica potencial. Objetivos. Identificar dos especies de hongos de huevos de O. bonariensis expuestos a condiciones salinas y caracterizar su patogenicidad y tolerancia a las soluciones de cloruro de sodio. Métodos. Se identificaron los aislamientos por sus características morfológicas, y se realizó un análisis filogenético preliminar utilizando secuencias de traslación del factor 1 alfa elongación (EF- 1α) y calmodulina (CAM). Se llevaron a cabo los postulados de Koch para identificar el agente causante de la infección fúngica. La influencia de NaCl sobre el crecimiento fúngico se evaluó en ensayos in vitro. Resultados. Se identificaron los aislamientos LPSC 1001 y 1002 como representantes del género Fusarium, y pertenecientes al complejo de especies Fusarium incarnatum-Fusarium equiseti (FIESC) y al complejo de especies de Fusarium solani (FSSC), respectivamente. Las observaciones histológicas en los huevos expuestos in vitro a ambos aislamientos en los ensayos de infectividad confirmaron la capacidad de estos para penetrar en la membrana coriónica, lo que condujo a la muerte de los embriones. El aumento de la concentración de NaCl en el medio de cultivo redujo el crecimiento de los aislamientos LPSC 1001 y 1002, quedando completamente inhibidos a 160 y 120 g/l de NaCl, respectivamente. Conclusiones. Se identificaron los aislamientos LPSC 1001 (FIESC) y 1002 (FSSC) como hongos patógenos para los huevos de O. bonariensis; el uso de soluciones de NaCl como tratamiento antifúngico no resultó eficaz para el control de la infección con estas cepas (AU)

Pathogenic ability and saline stress tolerance of two Fusarium isolates from Odontesthes bonariensis eggs.

BACKGROUND: Several fungal species represent a potential risk to embryos of Odontesthes bonariensis (Cuvier and Valenciennes, 1835), a euryhaline freshwater fish that lives in the Pampean inland waters and has potential economic relevance. AIMS: To identify two fungi isolated from O. bonariensis eggs exposed to saline conditions and to characterize their pathogenicity and tolerance to sodium chloride solutions. METHODS: The isolates were identified by morphological features, and a preliminar phylogenetic analysis using sequences of translation elongation factor 1-alpha (EF-1α) and calmodulin (CAM) was performed. Koch's postulates were tested to identify the causative agent of fungal infection. The influence of NaCl on the fungal growth was evaluated in in vitro assays. RESULTS: The isolates LPSC 1001 and 1002 were identified as representatives of the genus Fusarium, and belonging to the Fusarium incarnatum-Fusarium equiseti species complex (FIESC) and the Fusarium solani species complex (FSSC), respectively. Histological observations on eggs exposed in vitro to both isolates in infectivity assays confirmed the ability of the fungal isolates to penetrate to egg's chorionic membrane, leading to the death of embryos. Increasing NaCl concentration in the culture medium reduced the growth of the isolates LPSC 1001 and 1002, being completely inhibited at 160 and 120g/l NaCl respectively. CONCLUSIONS: The isolates LPSC 1001 (FIESC) and 1002 (FSSC) were identified as fungal pathogens to O. bonariensis eggs. The use of NaCl solutions as antifungal treatment was not effective to control the infection with these strains.

[Fusarium graminearum presence in wheat samples for human consumption]. / Presencia de Fusarium graminearum en muestras de trigo destinado 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.

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

Fusarium poae Pathogenicity and Mycotoxin Accumulation on Selected Wheat and Barley Genotypes at a Single Location in Argentina.

Fusarium poae is a relatively weak pathogen with increasing importance in cereal grains, principally due to its capacity to produce several mycotoxins. In this study, we evaluated the pathogenicity and toxin accumulation of individual F. poae isolates on wheat and barley under natural conditions for 3 years. Analysis of variance demonstrated significant differences for year-genotype, year-isolate, genotype-isolate, and year-genotype-isolate interactions for both incidence and disease severity. Based on contrast analysis, 'Apogee' was more susceptible than the other wheat genotypes, wheat genotypes were more susceptible than barley genotypes, durum wheat genotypes were more susceptible than bread wheat genotypes, and barley genotype 'Scarlett' had greater symptom development per spike than the other barley genotypes. Neither HT-2 nor T-2 toxins were detected in the grain samples. However, high levels of nivalenol were found in both wheat and barley samples. The increased reported isolation of F. poae from wheat and barley and the high capacity of this fungus to produce nivalenol underlie the need for more studies on F. poae-host interactions, especially for barley.

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

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  1Ag/g DON, 26

ranged from 0,5 and 0,99Ag/g, and the remaining 50

had values lower than 0,5Ag/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.

Development of a PCR assay to detect the potential production of nivalenol in Fusarium poae.

Fusarium species can produce mycotoxins, which can contaminate cereal-based food producing adverse effects for human and animal health. In recent years, the importance of Fusarium poae has increased within the Fusarium head blight complex. Fusarium poae is known to produce trichothecenes, especially nivalenol, a potent mycotoxin able to cause a variety of toxic effects. In this study, a specific primer pair was designed based on the tri7 gene to detect potential nivalenol-producing F. poae isolates. A total of 125 F. poae, four F. cerealis, two F. culmorum, one F. langsethiae, one F. sporotrichioides and seven F. graminearum, plus F. austroamericanum, F. meridionale, F. graminearum sensu stricto and F. cortaderiae from the NRRL collection were analysed, and only F. poae isolates gave a positive result for the presence of a 296-bp partial tri7 DNA fragment. Moreover, the primer set was tested from cereal seed samples where F. poae and other Fusarium species with a negative result for the specific reaction ( F. graminearum, F. oxysporum, F. chlamydosporum, F. sporotrichioides, F. equiseti and F. acuminatum) were isolated, and the expected fragment was amplified. We developed a rapid and reliable PCR assay to detect potential nivalenol-producing F. poae isolates.