Antónia Šrobárová

Toxigenic Fusarium species of Liseola section in pre-harvest maize ear rot, and associated mycotoxins in Slovakia

The occurrence of Fusarium species of Liseola section and related toxins was investigated for two years (1996 and 1998) on maize ear rot samples collected in the most important areas for maize growing in Slovakia. The species most frequently isolated was F. verticillioides, followed by F. proliferatum in 1996 and F. subglutinans in 1998. Most of the strains belonged to mating populations A, D, and E of the teleomorph Gibberella fujikuroi. Fusarium graminearum was also frequently recovered in both the years of investigations. Toxin analysis of maize ears showed that most of the samples (21 out of 22) were contaminated with at least one toxin. In particular, the concentration of fumonisin B1, and fumonisin 2 was up to 26.9 and 5.1μgg-1, respectively in 1996, and up to 12.1 and 6.3μgg-1, respectively in 1998. Beauvericin was detected only in one sample in 1996. Seven samples in 1996 were contaminated by fusaproliferin up to 8.2μgg-1, but just traces of the toxin were found in one sample in 1998. All 29 strains of F. verticillioides, two of three strains of F. proliferatum and none of eight F. subglutinans strains isolated from samples produced fumonisin B1 in culture on whole maize kernels (0.1–5646 and 940–1200μugg-1, respectively). Two strains of F. subglutinans and two of F. proliferatum produced beauvericin (up to 65 and 70μgg-1, respectively). Ten strains of F. verticillioides produced beauvericin: 9 strains produced a low amount (up to 3μgg-1), while only one of them produced a high level of toxin (375μgg-1). Fusaproliferin was produced by two F. proliferatum strains (220 and 370μgg-1), by seven F. subglutinans (20–1335μgg-1) and by three F. verticillioides (10–35μgg-1). This is the first report on fusaproliferin production by F. verticillioides, although at low level.

Response of oat cultivars to Fusarium infection with a view to their suitability for food use

Oats as a source of antioxidants and complex polysaccharides are currently an important component in human nutrition. Producing healthy, safe and high-quality grain for this purpose depends upon growing oat cultivars with improved resistance to diseases caused by Fusarium spp. producing mycotoxins. Thirteen cultivars of naked (Ábel, Detvan, Izák and Avenuda) and covered (Zvolen, Auron, Atego, Flämingsstern, Kanton, Viktor, Zlaťák, Euro and Ardo) oats were inoculated with conidial suspensions of F. culmorum isolate in the field at flowering in 2006 and 2007. After harvest, reduction in thousand-kernel weight (R-TKW), reduction in panicle-kernel weight (R-PKW), and deoxynivalenol (DON) content in grain and hulls were determined. The ELISA immunochemical method was employed for the quantitative analyses of DON. Values of yield components (R-TKW; R-PKW) were 35.4% and 31.1% lower in dehulled covered oats than in naked oat cultivars. The DON accumulation was highest in hulls as compared with DON content in kernels of naked and covered oat cultivars. Accumulation of DON in dehulled covered cultivars was 34.4% lower than the average contamination in naked cultivars. When the cultivars were compared, there were positive correlations between R-TKW and R-PKW and also between DON content and R-PKW. With a view to growing oat cultivars for production of cereal foods, it was shown that dehulling of covered oat grain resulted in substantially reduced DON content.

Beauvericin decreases cell viability of wheat.

Recently, beauvericin (BEA) has been recognized as an important toxic compound synthesized by several Fusarium strains, infecting maize, wheat, and rice, worldwide. The effects of BEA on mammalian cells have been studied; however, its effects on the function of host plant cells are largely unknown. The purpose of our work was to assess whether BEA can affect the root and leaf cells of wheat cultivar (cv.) ‘Arina’ seedlings, using a cytotoxicity assay and fluorescence microscopy. Toxigenicity during wheat germination was higher in BEA‐treated wheat seedlings than in non‐treated seedlings (control). Leaf primordial, situated at the base and the tips of treated leaves, were more affected by BEA compared to the control when assayed in medium for cell viability measured by luminescent equipment. BEA‐Treated plant cells secrete adenosine triphosphate (ATP) to the extracellular matrix and invoke more luminescence by luciferase than the non‐treated seedlings. Our results were confirmed by fluorescence microscopy following ‘4′,6‐diamidino‐2‐phenylindole’ (DAPI) staining and by confocal microscopy. In addition, the bioluminescent protein luciferase was observed in the intracellular space indicating presence of ATP. The incidence of nuclear fragmentation increased significantly in cells of seedlings treated with BEA at 40 μM concentration implying that the intracellular phytotoxin BEA plays an important role, possibly as a mediator in cell‐death signalling.

Diversity of the Fusarium species associated with head and seedling blight on wheat in Slovakia

Species associated with Fusarium head blight are depending on the production and edaphic conditions. The differences are found in the representation of various Fusarium spp. in the diseases, which sporadically occur all over the territory of Slovakia, in all agricultural production types. We identified fifteen Fusarium species during ten years of investigation. Most of the mentioned species F. culmorum (W.G. Smith) Sacc., F. graminearum Schwabe, recently F. cerealis (Cooke) Sacc. (crookwellense Burgess, Nelson & Tousson) and F. sambucinum Fuckel in diseased caryopsis are seed transmitted. The significant differences among species and intra species in cultural and pathogenicity assays in vitro and in vivo were correlated. Some of them are able to produce toxic metabolites — deoxynivalenone, which probably play a role in the aggressiveness of the pathogen and promote disease development and pathogen colonization.

Alternaria mycotoxins associated with grape berries in vitro and in situ

The aim of this study was to assess the production of secondary metabolites of Alternaria species isolated from grape berries and determined the occurrence of Alternaria mycotoxins in vitro and in dried berries. Direct morphological examination on different media was used for identification and HPLC/MS method for quantification of Alternaria mycotoxins. We isolated A. alternata and A. tenuissima and randomly selected strains in vitro and in dried berries. In vitro we identified the production of Alternaria metabolites, included mycotoxins: alternariol, alternariol methylether, tenuazonic acid. Beside that one in dry berries were in measurable concentrations: macrosporin A, tentoxin and altenuete (I, II).

Penicillium strains isolated from Slovak grape berries taxonomy assessment by secondary metabolite profile

The secondary metabolite profiles of microfungi of the genus Penicillium isolated from samples of grape berries collected in two different phases during two vegetative seasons in Slovakia is described to assess the taxonomy. Three Slovak vine regions have been selected for this study, based on their climatic differences and national economic importance. Cultures of microfungi isolated from berries were incubated on different selective media for macro and micromorphology identification. The species Penicillium brevicompactum, Penicillium crustosum, Penicillium chrysogenum, Penicillium expansum, Penicillium palitans and Penicillium polonicum were identified according to growth and morphology. The related strains were found to produce a broad spectrum of fungal metabolites, including roquefortine C, chaetoglobosin A, penitrem A, cyclopeptin, cyclopenin, viridicatin, methylviridicatin, verrucofortine, secalonic acid D, cyclopiazonic acid, fumigaclavine and mycophenolic acid. Chemotaxonomy was performed using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Dried grape berries were also analyzed allowing to assess the presence of patulin, roquefortine C and penicillic acid; this last one has been identified in dried berries but not in vitro.

Response to Fusarium culmorum inoculation in barley

In field tests replicated in 2004 and 2005, 32 cultivars of spring barley were assessed for resistance to Fusarium head blight (FHB) by single floret inoculation and spray inoculation with Fusarium culmorum (W. G. Smith) Sacc. It was found that the weather conditions in individual years affect to a large extent the progression of FHB and production of mycotoxin deoxynivalenol (DON). At the same time, in both years the cultivars reacted to F. culmorum infection similarly with respect to areas under disease progress curve (AUDPC) values and content of mycotoxin DON. Spraying inoculation led to stronger infection. The biggest differences in AUDPC values were observed between the cultivars Brise and Celinka, and weak reaction was found in the cultivars Kompakt and Madonna. The cultivars Kompakt and Tolar were most resistant towards FHB. In both monitored years the variety Ludan contained the lowest amounts of mycotoxin DON. Cultivars with high infection and low DON content (r = 0.78) showed weak positive relationship between resistance to FBH and accumulation of DON (concentration 70–200 mg/kg). This is the first information on FHB and in vivo concentrations of DON in certificated barley cultivars in Slovakia.

Berries contamination by microfungi in Slovakia vineyard regions, Impact of climate conditions on microfungi biodiversity

BACKGROUND Weather conditions can selectively promote the growth of particular fungal species, which cause rotting and spoilage of grape berries before harvest. The presence of pathogenic fungi can lead to the development of opportunist microfungi that can produce mycotoxins and cause grapes and wine contamination. OBJECTIVE The variation of climatic conditions allowed to design a pilot study and address relevant questions for risk assessment of climate related mycotoxins production in grapes and in wine. METHODS Microfungi contaminating berries during the vegetative period of year 2008 and 2009 in vineyards regions of Slovakia at the early veraison and at the ripening in harvest time have been identified. RESULTS Spoilage fungi were more abundant in veraison for both years in all the studied geographical regions, with an average temperature of 20°C, humidity between 60 and 80%, and precipitation in the range 6-5mm. Much more strains of toxigenic fungi were found during veraison and ripening in the case of a year temperature variation in the range 17-22.5°C, humidity values ranging from 58 to 80% and precipitations in the range 0.7-8.4mm. CONCLUSIONS The results suggest that there is an effect of the climatic conditions on microfungi biodiversity.

Cultural characteristics and zearalenone production by strains ofFusarium from wheat

Conidia of the speciesFusariumculmorum /W.G.Sm./ Sacc. andFusariumgraminearum Schwabe are characterized by variability in zearalenone production and dimensions depending on the substrate. The sporulation of isolates from some wheat eultivars have been deprived in vivo and in vitro in the first passage, but not their pathogenicity and toxic metabolites production. Nonsporulating strains produced lower quantités of zearalenone than sporulating ones. Liquid filtrates of such nonsporulating strains had a high phytotoxic effect on wheat caryopses. The crystalline toxin DAS /0,25 ug/ml/ had low phytotoxic effect on wheat caryopses.

Zearalenone production in wheat cultivars infected with the fungusFusarium graminearum Schwabe

In wheat plants of the eultivars “Danubia”, “Agra”, “Selekta” and “Jubilejna” the fungusFusariumgraminsarum Schwabe produced toxic metabolite zearalenone/F-2/ which simultaneously influenced the development of plants characterized by a lower germinating capacity, a reduced growth rate and a higher production of side branches. The presence ofFusariumgraminearum was confirmed only in infected plants after plating of organs (root, stem base, stem) and soil on agar medium. The mycotoxin production is dependent on the pathogen development in host plants. The F-2 level progressed from the root into the soil, stem base and stem. The highest F-2 production was identified in cultivar “Selekta” the lowest in cultivar “Danubia”. The highest F-2 level (in all wheat eultivars) was identified in the stem base.