Evaluation of PSP1 biostimulant on Fusarium graminearum-wheat pathosystem: impact on disease parameters, grain yield, and grain quality.

BACKGROUND: Plant defense elicitors are valuable tools in sustainable agriculture, providing an environmentally friendly and effective means of enhancing plant defense and promoting plant health. Fusarium head blight (FHB) is one of the most important fungal diseases of cereal crops worldwide. The PSP1 is a novel biopesticide formulated based on an elicitor, the extracellular protein AsES, from the fungus Sarocladium strictum. The present work aimed to evaluate the effectiveness of PSP1 in controlling FHB under field conditions. Experiments were conducted during three consecutive growing seasons (2019, 2020, and 2021). Three biostimulant treatments were tested in different physiological stages (from late tillering to heading stage), and FHB inoculations were performed at anthesis. Disease parameters, seed parameters, grain yield, and grain quality parameters were evaluated. RESULTS: Depending on the year and the genotype, reductions in disease incidence (up to 11%) and disease severity (up to 5%) were reported, although these differences could not be attributed to the use of the PSP1 biostimulant. Occasional improvements in seed parameters and grain quality were observed, suggesting that early treatments could work better than late treatments, probably due to early activation/priming of defense response mechanisms. However, more studies are deemed necessary. CONCLUSION: The use of PSP1 biostimulant in commercial wheat crops could be a biological alternative or complement to traditional chemical fungicides to manage FHB. The reduced environmental impact and the potential benefits in grain yield and quality are other reasons that can generate new adherents of this technology in worldwide agriculture systems in the coming years. © 2024 Society of Chemical Industry.

Whole-genome single nucleotide polymorphism analysis for typing the pandemic pathogen Fusarium graminearum sensu stricto.

Recent improvements in microbiology and molecular epidemiology were largely stimulated by whole- genome sequencing (WGS), which provides an unprecedented resolution in discriminating highly related genetic backgrounds. WGS is becoming the method of choice in epidemiology of fungal diseases, but its application is still in a pioneer stage, mainly due to the limited number of available genomes. Fungal pathogens often belong to complexes composed of numerous cryptic species. Detecting cryptic diversity is fundamental to understand the dynamics and the evolutionary relationships underlying disease outbreaks. In this study, we explore the value of whole-genome SNP analyses in identification of the pandemic pathogen Fusarium graminearum sensu stricto (F.g.). This species is responsible for cereal diseases and negatively impacts grain production worldwide. The fungus belongs to the monophyletic fungal complex referred to as F. graminearum species complex including at least sixteen cryptic species, a few among them may be involved in cereal diseases in certain agricultural areas. We analyzed WGS data from a collection of 99 F.g. strains and 33 strains representing all known cryptic species belonging to the FGSC complex. As a first step, we performed a phylogenomic analysis to reveal species-specific clustering. A RAxML maximum likelihood tree grouped all analyzed strains of F.g. into a single clade, supporting the clustering-based identification approach. Although, phylogenetic reconstructions are essential in detecting cryptic species, a phylogenomic tree does not fulfill the criteria for rapid and cost-effective approach for identification of fungi, due to the time-consuming nature of the analysis. As an alternative, analysis of WGS information by mapping sequence data from individual strains against reference genomes may provide useful markers for the rapid identification of fungi. We provide a robust framework for typing F.g. through the web-based PhaME workflow available at EDGE bioinformatics. The method was validated through multiple comparisons of assembly genomes to F.g. reference strain PH-1. We showed that the difference between intra- and interspecies variability was at least two times higher than intraspecific variation facilitating successful typing of F.g. This is the first study which employs WGS data for typing plant pathogenic fusaria.

Fusarium graminearum Species Complex: A Bibliographic Analysis and Web-Accessible Database for Global Mapping of Species and Trichothecene Toxin Chemotypes.

Fusarium graminearum is ranked among the five most destructive fungal pathogens that affect agroecosystems. It causes floral diseases in small grain cereals including wheat, barley, and oats, as well as maize and rice. We conducted a systematic review of peer-reviewed studies reporting species within the F. graminearum species complex (FGSC) and created two main data tables. The first contained summarized data from the articles including bibliographic, geographic, methodological (ID methods), host of origin and species, while the second data table contains information about the described strains such as publication, isolate code(s), host/substrate, year of isolation, geographical coordinates, species and trichothecene genotype. Analyses of the bibliographic data obtained from 123 publications from 2000 to 2021 by 498 unique authors and published in 40 journals are summarized. We describe the frequency of species and chemotypes for 16,274 strains for which geographical information was available, either provided as raw data or extracted from the publications, and sampled across six continents and 32 countries. The database and interactive interface are publicly available, allowing for searches, summarization, and mapping of strains according to several criteria including article, country, host, species and trichothecene genotype. The database will be updated as new articles are published and should be useful for guiding future surveys and exploring factors associated with species distribution such as climate and land use. Authors are encouraged to submit data at the strain level to the database, which is accessible at https://fgsc.netlify.app.

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.

Diversity of Mobile Genetic Elements in the Mitogenomes of Closely Related Fusarium culmorum and F. graminearum sensu stricto Strains and Its Implication for Diagnostic Purposes.

Much of the mitogenome variation observed in fungal lineages seems driven by mobile genetic elements (MGEs), which have invaded their genomes throughout evolution. The variation in the distribution and nucleotide diversity of these elements appears to be the main distinction between different fungal taxa, making them promising candidates for diagnostic purposes. Fungi of the genus Fusarium display a high variation in MGE content, from MGE-poor (Fusarium oxysporum and Fusarium fujikuroi species complex) to MGE-rich mitogenomes found in the important cereal pathogens F. culmorum and F. graminearum sensu stricto. In this study, we investigated the MGE variation in these latter two species by mitogenome analysis of geographically diverse strains. In addition, a smaller set of F. cerealis and F. pseudograminearum strains was included for comparison. Forty-seven introns harboring from 0 to 3 endonucleases (HEGs) were identified in the standard set of mitochondrial protein-coding genes. Most of them belonged to the group I intron family and harbored either LAGLIDADG or GIY-YIG HEGs. Among a total of 53 HEGs, 27 were shared by all fungal strains. Most of the optional HEGs were irregularly distributed among fungal strains/species indicating ancestral mosaicism in MGEs. However, among optional MGEs, one exhibited species-specific conservation in F. culmorum. While in F. graminearum s.s. MGE patterns in cox3 and in the intergenic spacer between cox2 and nad4L may facilitate the identification of this species. Thus, our results demonstrate distinctive traits of mitogenomes for diagnostic purposes of Fusaria.

Effects of Fusarium graminearum and Fusarium poae on disease parameters, grain quality and mycotoxins contamination in bread wheat (Part I).

BACKGROUND: Wheat is the most important winter crop in the world, being affected by the presence of fungal, mainly those belonging to the Fusarium genus. Fusarium head blight (FHB) is a serious disease that causes important economic damage and quantitative/qualitative losses, with Fusarium graminearum and Fusarium poae being two of the most isolated species worldwide. The present study aimed to evaluate the interaction between F. graminearum and F. poae and the effects on disease parameters, grain quality and mycotoxin contamination on five wheat genotypes under field conditions during three growing seasons. RESULTS: Statistical differences between Fusarium treatments were found for disease parameters, grain quality and mycotoxin contamination during the 2014/2015 growing season. High values of incidence (58.00 ± 8.00%), severity (6.28 ± 1.51%) and FHB index (4.72 ± 1.35) were observed for F. graminearum + F. poae treatment. Regarding grain quality, the results showed that the degradation of different protein fractions depends on each Fusarium species: glutenins were degraded preferably by F. graminearum (-70.82%), gliadins were degraded preferably by F. poae (-29.42%), whereas both protein fractions were degraded when both Fusarium species were present (-60.91% and -16.51%, respectively). Significant differences were observed for mycotoxin contamination between genotypes, with Proteo being the most affected (DON = 12.01 ± 3.67 µg g-1 ). In addition, we report that 3-ADON predominated over 15-ADON in the three seasons evaluated. CONCLUSION: Variations in plant-pathogen interaction (Fusarium-wheat pathosystem) should be considered at least in years with favorable climatic conditions for FHB development, as a result of the potential impact of this disease on grain quality and mycotoxin contamination. © 2019 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.

Development of PSP1, a Biostimulant Based on the Elicitor AsES for Disease Management in Monocot and Dicot Crops.

In this work, we present a novel biostimulant for sustainable crop disease management, PSP1, based on the plant defense-elicitor AsES, an extracellular protease produced by the strawberry fungal pathogen Acremonium strictum. Fungal fermentation conditions and downstream processing were determined to maximize extracellular protein production, product stability and a high plant defense-eliciting activity, as monitored by anthracnose resistance in supernatant-treated strawberry plants subsequently infected with a virulent strain of Colletotrichum acutatum. Fermentation batches were shown to reduce anthracnose development by 30-60% as compared to infected non-treated plants. Product formulation was shown to be stable for 6 months when stored at temperatures up to 45°C and toxicological tests showed that PSP1 was harmless to beneficial organisms and non-toxic to mammalian species at concentrations 50 times higher than those used in plant experiments. Furthermore, disease protection studies using dilutions of PSP1 indicated that there is a minimum threshold protease activity needed to induce pathogen defense in strawberry and that this induction effect is dose-independent. A significant characteristic of PSP1 is its broad-range protection against different diseases in various crop species. In soybean, PSP1 reduced the symptomatology by 70% of Corynespora cassiicola, etiological agent of the target spot. This protection effect was similar to the commercial inducer BION 500 WG based on BTH, and both products were shown to induce an oxidative burst and up-regulated PR1-gene expression in soybean. Furthermore, a double PSP1-treatment on greenhouse-grown sugarcane plants provided protection against bacterial red stripe disease caused by Acidovorax avenae and a double foliar application of PSP1 on field-grown wheat plants significantly increased resistance against Fusarium graminearum, causal agent of head blight disease, manifested mainly in an increased seed germination rate. In summary, these disease protection studies demonstrated an effective control against both bacterial and fungal pathogens in both monocot and dicot crop species, which together with its low production cost, effectiveness at low concentrations, long shelf-life, tolerance to high temperatures, harmlessness to non-target organisms and simple handling and application, make PSP1 a very promising candidate for effective and sustainable disease management in many crop species.

Compatibility of chemical insecticides and entomopathogenic fungi for control of soybean defoliating pest, Rachiplusia nu / Compatibilidad de insecticidas químicos y hongos entomopatógenos para el control de Rachiplusia nu, plaga defoliadora de la soja

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) is one of the major lepidopteran pests defoliating soybeans (Glycine max Merrill) in Argentina. The combined use of chemical insecticides and entomopathogenic fungi is a promising pest-control option to minimize adverse chemical effects. In this work, we evaluated the interactions between five insecticides-two being considered biorational-and five fungal entomopathogenic strains under laboratory conditions in order to determine the possible usefulness of combinations of these agents against R. nu. The insecticides were tested for compatibility at four doses by in vitro bioassay and for the lethality of R. nu by inoculations at three doses. Fungal strains were applied at 1 x 10(8), 1 x 10(6), and 1 x 10(4) conidia/ml. The combinations of those insecticides with Beauveria bassiana (LPSc 1067, LPSc 1082, LPSc 1098), Metarhizium anisopliae (LPSc 907), and Metarhizium robertsii (LPSc 963) caused higher R. nu-larval mortalities than any of the individual agents alone. We observed significant differences in the in vitro conidial viability, vegetative growth, and conidia production of the five strains of entomopathogenic fungi exposed to different doses of the chemical insecticides. The combination gamma-cyhalothrin-LPSc-1067 caused the highest percent mortality of R. nu larvae, with synergism occurring between the two agents at 50% and 25% of the maximum field doses.

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) es una de las principales plagas de lepidópteros defoliadores del cultivo de soja (Glycine max Merrill) en Argentina. El uso combinado de insecticidas químicos y hongos entomopatógenos es una opción de control de plagas prometedora para minimizar los efectos químicos adversos. En este trabajo se evaluaron las interacciones entre 5 insecticidas -2 de ellos considerados biorracionales-- y 5 cepas fúngicas entomopatógenas en condiciones de laboratorio, para determinar la posible utilidad de combinaciones de estos agentes frente a R. nu. Se evaluó la compatibilidad de los insecticidas a 4 dosis mediante bioensayos in vitro y la letalidad de aquellos sobre R. nu mediante inoculaciones a 3 dosis. Las cepas fúngicas se evaluaron a concentraciones de 1 x 10(8), 1 x 10(6) y 1 x 10(4) conidios/ml. Las combinaciones de estos insecticidas con Beauveria bassiana (LPSc1067, LPSc 1082, LPSc 1098), Metarhizium anisopliae (LPSc 907) y Metarhizium robertsii (LPSc 963) causaron una mayor mortalidad de larvas de R. nu que cualquiera de los agentes individuales. Asimismo, se observaron diferencias significativas en la viabilidad de los conidios in vitro, el crecimiento vegetativo y la producción de conidios de las 5 cepas de hongos entomopatógenos expuestos a diferentes dosis de los insecticidas químicos. La combinación gamma-cialotrina-LPSc-1067 causó el mayor porcentaje de mortalidad de larvas de R. nu, con un efecto de sinergismo entre los 2 agentes al 50 y el 25% de las dosis de campo recomendadas por el fabricante.

Natural Contamination with Mycotoxins Produced by Fusarium graminearum and Fusarium poae in Malting Barley in Argentina.

Two of the most common species of toxin-producing Fusarium contaminating small cereal grains are Fusarium graminearum and F. poae; with both elaborating diverse toxins, especially deoxynivalenol (DON) and nivalenol (NIV), respectively. The objective of our work during the 2012-2014 growing seasons was to screen crops for the most commonly isolated Fusarium species and to quantify DON and NIV toxins in natural malting-barley samples from different producing areas of Argentina. We identified 1180 Fusarium isolates in the 119 samples analyzed, with 51.2% being F. graminearum, 26.2% F. poae and 22.6% other species. We found high concentrations of mycotoxins, at maximum values of 12 µg/g of DON and 7.71 µg/g of NIV. Of the samples, 23% exhibited DON at an average of 2.36 µg/g, with 44% exceeding the maximum limits (average of 5.24 µg/g); 29% contained NIV at an average of 2.36 µg/g; 7% contained both DON and NIV; and 55% were without DON or NIV. Finally, we report the mycotoxin contamination of the grain samples produced by F. graminearum and F. poae, those being the most frequent Fusarium species present. We identified the main Fusarium species affecting natural malting-barley grains in Argentina and documented the presence of many samples with elevated concentrations of DON and NIV. To our knowledge, the investigation reported here was the first to quantify the contamination by Fusarium and its toxins in natural samples of malting barley in Argentina.

Compatibility of chemical insecticides and entomopathogenic fungi for control of soybean defoliating pest, Rachiplusia nu.

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) is one of the major lepidopteran pests defoliating soybeans (Glycine max Merrill) in Argentina. The combined use of chemical insecticides and entomopathogenic fungi is a promising pest-control option to minimize adverse chemical effects. In this work, we evaluated the interactions between five insecticides-two being considered biorational-and five fungal entomopathogenic strains under laboratory conditions in order to determine the possible usefulness of combinations of these agents against R. nu. The insecticides were tested for compatibility at four doses by in vitro bioassay and for the lethality of R. nu by inoculations at three doses. Fungal strains were applied at 1×108, 1×106, and 1×104conidia/ml. The combinations of those insecticides with Beauveria bassiana (LPSc 1067, LPSc 1082, LPSc 1098), Metarhizium anisopliae (LPSc 907), and Metarhizium robertsii (LPSc 963) caused higher R. nu-larval mortalities than any of the individual agents alone. We observed significant differences in the in vitro conidial viability, vegetative growth, and conidia production of the five strains of entomopathogenic fungi exposed to different doses of the chemical insecticides. The combination gamma-cyhalothrin-LPSc-1067 caused the highest percent mortality of R. nu larvae, with synergism occurring between the two agents at 50% and 25% of the maximum field doses.

Resistance of Fusarium poae in Arabidopsis leaves requires mainly functional JA and ET signaling pathways.

Fusarium poae has been considered as a minor species among those that cause the FHB disease but in recent years several researchers have documented a high frequency of occurrence in several crops. We evaluated the ability of F. poae to produce symptoms in A. thaliana leaves. Moreover, we analyzed the defense of A. thaliana against F. poae using SA, JA, and ET mutants and we monitored the expression level of genes involved in the main signaling pathways related to plant defense. Symptoms were observed in the inoculated leaves demonstrating the ability of F. poae to infect A. thaliana leaves. Moreover, the npr1-1 mutants presented low symptoms compared to Col-0, etr2-1, and coi1-1 and that the coi1-1 mutant was the most susceptible genotypes followed by etr2-1 genotypes. The RT-PCR revealed that PDF1.2, CHI/PR3, and ERF1, three important JA-ET responsive genes and NPR1 and PR1, which are regulated by SA signaling, were expressed upon F. poae inoculation. Our results suggest that JA and ET could play a key role in Arabidopsis leaves defense against F. poae representing the first evaluation of the response of the main A. thaliana phytohormones involved in plant defense in the presence of F. poae.

Characterization of small-spored Alternaria from Argentinean crops through a polyphasic approach.

Small-spored Alternaria have been isolated from a wide variety of food crops, causing both economic losses and human health risk due to the metabolites produced. Their taxonomy has been discussed widely, but no scientific consensus has been established in this field to date. Argentina is a major exporter of agricultural products, so it is essential to thoroughly understand the physiological behaviour of this pathogen in a food safety context. Thus, the objective of this work was to characterize small-spored Alternaria spp. obtained from tomato fruits, pepper fruits, wheat grains and blueberries from Argentina by a polyphasic approach involving metabolomic and phylogenetic analyses based on molecular and morphological characters. Morphological analysis divided the population studied into three groups; A. arborescens sp.-grp., A. tenuissima sp.-grp., and A. alternata sp.-grp. However, when these characters were simultaneously analysed with molecular data, no clearly separated groups were obtained. Haplotype network and phylogenetic analysis (both Bayesian and maximum parsimony) of a conserved region yielded the same result, suggesting that all isolates belong to the same species. Furthermore, no correlation could be established between morphological species-groups and a metabolite or group of metabolites synthesized. Thus, the whole set of analyses carried out in the present work supports the hypothesis that these small-spored Alternaria isolates from food belong to the same species. Identification at species level through classical morphology or modern molecular techniques does not seem to be a useful tool to predict toxicological risk in food matrices. The detection of any small-spored Alternaria from Section Alternaria (D.P. Lawr., Gannibal, Peever & B.M. Pryor 2013) in food implies a potential toxicological risk.

ToxGen: an improved reference database for the identification of type B-trichothecene genotypes in Fusarium.

Type B trichothecenes, which pose a serious hazard to consumer health, occur worldwide in grains. These mycotoxins are produced mainly by three different trichothecene genotypes/chemotypes: 3ADON (3-acetyldeoxynivalenol), 15ADON (15-acetyldeoxynivalenol) and NIV (nivalenol), named after these three major mycotoxin compounds. Correct identification of these genotypes is elementary for all studies relating to population surveys, fungal ecology and mycotoxicology. Trichothecene producers exhibit enormous strain-dependent chemical diversity, which may result in variation in levels of the genotype's determining toxin and in the production of low to high amounts of atypical compounds. New high-throughput DNA-sequencing technologies promise to boost the diagnostics of mycotoxin genotypes. However, this requires a reference database containing a satisfactory taxonomic sampling of sequences showing high correlation to actually produced chemotypes. We believe that one of the most pressing current challenges of such a database is the linking of molecular identification with chemical diversity of the strains, as well as other metadata. In this study, we use the Tri12 gene involved in mycotoxin biosynthesis for identification of Tri genotypes through sequence comparison. Tri12 sequences from a range of geographically diverse fungal strains comprising 22 Fusarium species were stored in the ToxGen database, which covers descriptive and up-to-date annotations such as indication on Tri genotype and chemotype of the strains, chemical diversity, information on trichothecene-inducing host, substrate or media, geographical locality, and most recent taxonomic affiliations. The present initiative bridges the gap between the demands of comprehensive studies on trichothecene producers and the existing nucleotide sequence databases, which lack toxicological and other auxiliary data. We invite researchers working in the fields of fungal taxonomy, epidemiology and mycotoxicology to join the freely available annotation effort.

Depicting the Discrepancy between Tri Genotype and Chemotype on the Basis of Strain CBS 139514 from a Field Population of F. graminearum Sensu Stricto from Argentina.

Recent studies on a field population of F. graminearum sensu stricto from Argentina revealed an atypical panel of strains identified through PCR genotyping as 15ADON genotypes, but producing high levels of 3ADON. Based on representative strain CBS 139514, we asked if the discrepancy between the trichothecene genotype and chemotype might result from an inter-chemotype recombination of the chemotype-determining genes. To answer this, we sequenced the complete core Tri gene cluster (around 30,200 bp) from this strain and compared its sequence to sequence data of typical type B trichothecene genotypes/chemotypes. Sequence alignment showed that CBS 139514 has an identical sequence within the entire core Tri cluster to the 15ADON genotype. The revealed discrepancy underlines the need for using both molecular and chemical methods for reliable characterization of toxigenic strains of Fusarium.

Endophytic fungi from selected varieties of soybean (Glycine max L. Merr.) and corn (Zea mays L.) grown in an agricultural area of Argentina / Hongos endófitos aislados de cultivares de soja (Glycine max L. Merr) y maíz (Zea mays L.) presentes en áreas agrícolas argentinas

Endophytic fungi are ubiquitous and live within host plants without causing any noticeable symptoms of disease. Little is known about the diversity and function of fungal endophytes in plants, particularly in economically important species. The aim of this study was to determine the identity and diversity of endophytic fungi in leaves, stems and roots of soybean and corn plants and to determine their infection frequencies. Plants were collected in six areas of the provinces of Buenos Aires and Entre Ríos (Argentina) two areas were selected for sampling corn and four for soybean. Leaf, stem and root samples were surface-sterilized, cut into 1 cm² pieces using a sterile scalpel and aseptically transferred to plates containing potato dextrose agar plus antibiotics. The species were identified using both morphological and molecular data. Fungal endophyte colonization in soybean plants was influenced by tissue type and varieties whereas in corn plants only by tissue type. A greater number of endophytes were isolated from stem tissues than from leaves and root tissues in both species of plants. The most frequently isolated species in all soybean cultivars was Fusarium graminearum and the least isolated one was Scopulariopsis brevicaulis. Furthermore, the most frequently isolated species in corn plants was Aspergillus terreus whereas the least isolated one was Aspergillus flavus. These results could be relevant in the search for endophytic fungi isolates that could be of interest in the control of agricultural pests.

Los hongos endófitos son ubicuos y se encuentran en el interior de los tejidos de las plantas de manera asintomática. Se sabe muy poco acerca de la diversidad y la función de estos hongos, particularmente en especies de importancia económica. El objetivo de este trabajo fue determinar la diversidad y la frecuencia de colonización de hongos endófitos en raíces, tallos y hojas de 2 variedades de maíz y de 4 variedades de soja; las muestras se tomaron de 6 áreas diferentes ubicadas en las provincias de Buenos Aires y Entre Ríos (Argentina). Con un bisturí estéril se obtuvieron porciones de 1 cm² de raíz, tallo y hoja, que fueron colocados en placas con agar papa dextrosa más antibiótico. Las especies de hongos fueron identificadas a partir de características morfológicas y moleculares. La colonización de hongos endófitos en soja estuvo influenciada por la variedad y por el tipo de tejido, en tanto que en el maíz solo hubo influencia del tipo de tejido. El mayor número de endófitos se encontró en los tallos de ambas especies. El aislamiento más frecuente en todas las variedades de soja fue Fusarium graminearum y el menos frecuente Scopulariopsis brevicaulis. En ambas variedades de maíz la especie con mayor frecuencia de aislamiento fue Aspergillus terreus y la de menor fue Aspergillus flavus. Estos resultados son relevantes para la búsqueda de especies de hongos endófitos que podrían ser de interés en el control de plagas agrícolas.

Endophytic fungi from selected varieties of soybean (Glycine max L. Merr.) and corn (Zea mays L.) grown in an agricultural area of Argentina.

Endophytic fungi are ubiquitous and live within host plants without causing any noticeable symptoms of disease. Little is known about the diversity and function of fungal endophytes in plants, particularly in economically important species. The aim of this study was to determine the identity and diversity of endophytic fungi in leaves, stems and roots of soybean and corn plants and to determine their infection frequencies. Plants were collected in six areas of the provinces of Buenos Aires and Entre Ríos (Argentina) two areas were selected for sampling corn and four for soybean. Leaf, stem and root samples were surface-sterilized, cut into 1cm(2) pieces using a sterile scalpel and aseptically transferred to plates containing potato dextrose agar plus antibiotics. The species were identified using both morphological and molecular data. Fungal endophyte colonization in soybean plants was influenced by tissue type and varieties whereas in corn plants only by tissue type. A greater number of endophytes were isolated from stem tissues than from leaves and root tissues in both species of plants. The most frequently isolated species in all soybean cultivars was Fusarium graminearum and the least isolated one was Scopulariopsis brevicaulis. Furthermore, the most frequently isolated species in corn plants was Aspergillus terreus whereas the least isolated one was Aspergillus flavus. These results could be relevant in the search for endophytic fungi isolates that could be of interest in the control of agricultural pests.

Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis.

Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience.

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)

Primer registro de Talaromyces udagawae relacionado con cuerpos humanos en descomposición, en suelo de la Argentina / First record of Talaromyces udagawae in soil related to decomposing human remains in Argentina

The morphologic features of Talaromyces udagawae Stolk and Samson are here described and illustrated. This teleomorphic Ascomycota fungus was isolated from soil obtained in Buenos Aires province (Argentina) from beneath a human cadaver in an advanced state of decomposition. After washing and serial dilution of the soil along with moist-chamber techniques for fungal cultivation, T. udagawae formed very restricted colonies of bright yellow color on different growth media with 8-ascospored asci. The ascospores were ellipsoidal and ornamented. The anamorphic state was not observed. Molecular-genetic techniques identified the species. The present record is the first of the species in Argentina, pointing it as a tool to identify soils where cadaver decomposition occurs.

Se describen e ilustran las características morfológicas de Talaromyces udagawae Stolk y Samson. Se aisló el estado teleomórfico de este hongo Ascomycota de suelo obtenido en la provincia de Buenos Aires (Argentina), por debajo de un cadáver humano en avanzado estado de descomposición. Las muestras de suelo fueron analizadas mediante lavado, dilución seriada y cámaras húmedas, técnicas ampliamente usadas para el estudio de hongos de suelo. T. udagawae formó colonias muy restringidas de color amarillo brillante en diferentes medios de cultivo, con ascos con 8 ascosporas. Las ascosporas eran elipsoidales y ornamentadas. No fue hallado el estado anamórfico. La especie también fue identificada mediante técnicas moleculares. El presente registro es el primero de la especie en la Argentina y el único que la postula como herramienta para identificar suelos donde ocurre una descomposición cadavérica.