During the last ten years, Norwegian cereal grain industry has experienced large challenges due to Fusarium spp. and Fusarium mycotoxin contamination of small-grained cereals. To prevent severely contaminated grain lots from entering the grain supply chain, it is important to establish surveys for the most prevalent Fusarium spp. and mycotoxins. The objective of our study was to quantify and calculate the associations between Fusarium spp. and mycotoxins prevalent in oats and spring wheat. In a 6-year period from 2004-2009, 178 grain samples of spring wheat and 289 samples of oats were collected from farmers' fields in South East Norway. The grains were analysed for 18 different Fusarium-mycotoxins by liquid chromatography -mass spectrometry. Generally, the median mycotoxin levels were higher than reported in Norwegian studies covering previous years. The DNA content of Fusarium graminearum, Fusarium culmorum, Fusarium langsethiae, Fusarium poae and Fusarium avenaceum were determined by quantitative PCR. We identified F. graminearum as the main deoxynivalenol (DON) producer in oats and spring wheat, and F. langsethiae as the main HT-2 and T-2-toxins producer in oats. No association was observed between quantity of F. graminearum DNA and quantity of F. langsethiae DNA nor for their respective mycotoxins, in oats. F. avenaceum was one of the most prevalent Fusarium species in both oats and spring wheat. The following ranking of Fusarium species was made based on the DNA concentrations of the Fusarium spp. analysed in this survey (from high to low): F. graminearum = F. langsethiae = F. avenaceum > F. poae > F. culmorum (oats); F. graminearum = F. avenaceum > F. culmorum > F. poae = F. langsethiae (spring wheat). Our results are in agreement with recently published data indicating a shift in the relative prevalence of Fusarium species towards more F. graminearum versus F. culmorum in Norwegian oats and spring wheat.
The increased occurrence of Fusarium-mycotoxins in Norwegian cereals over the last decade, is thought to be caused by increased inoculum resulting from more cereal residues at the soil surface as a result of reduced tillage practices. In addition, weather conditions have increasingly promoted inoculum development and infection by Fusarium species. The objective of this work was to elucidate the influence of different tillage regimes (autumn plowing; autumn harrowing; spring plowing; spring harrowing) on the inoculum potential (IP) and dispersal of Fusarium spp. in spring oats. Tillage trials were conducted at two different locations in southeast Norway from 2010 to 2012. Oat residues from the previous year’s crop were collected within a week after sowing for evaluation. IP was calculated as the percentage of residues infested with Fusarium spp. multiplied by the proportion of the soil surface covered with residues. Fusarium avenaceum and F. graminearum were the most common Fusarium species recovered from oat residues. The IP of Fusarium spp. was significantly lower in plowed plots compared to those that were harrowed. Plowing in either the autumn or spring resulted in a low IP. Harrowing in autumn was more effective in reducing IP than the spring harrowing, and IP levels for the spring harrowed treatments were generally higher than all other tillage treatments examined. Surprisingly low levels of F. langsethiae were detected in the residues, although this species is a common pathogen of oat in Norway. The percentage of the residues infested with F. avenaceum, F. graminearum, F. culmorum, and F. langsethiae generally related to the quantity of DNA of the respective Fusarium species determined using quantitative PCR (qPCR). Fusarium dispersal, quantified by qPCR analysis of spore trap samples collected at and after heading, generally corresponded to the IP. Fusarium dispersal was also observed to increase after rainy periods. Our findings are in line with the general understanding that plowing is a means to reduce the IP of Fusarium spp. in cereal fields. The main inoculum source for F. langsethiae remains unclear. Our results will be useful in the development of forecasting tools to calculate the risk of Fusarium in cereals.
High concentrations of the mycotoxin deoxynivalenol (DON), produced by Fusarium graminearum have occurred frequently in Norwegian oats recently. Early prediction of DON levels is important for farmers, authorities and the Cereal Industry. In this study, the main weather factors influencing mycotoxin accumulation were identified and two models to predict the risk of DON in oat grains in Norway were developed: (1) as a warning system for farmers to decide if and when to treat with fungicide, and (2) for authorities and industry to use at harvest to identify potential food safety problems. Oat grain samples from farmers' fields were collected together with weather data (2004)(2005)(2006)(2007)(2008)(2009)(2010)(2011)(2012)(2013).A mathematical model was developed and used to estimate phenology windows of growth stages in oats (tillering, flowering etc.). Weather summarisations were then calculated within these windows, and the Spearman rank correlation factor calculated between DONcontamination in oats at harvest and the weather summarisations for each phenological window. DON contamination was most clearly associated with the weather conditions around flowering and close to harvest. Warm, rainy and humid weather during and around flowering increased the risk of DON accumulation in oats, as did dry periods during germination/seedling growth and tillering. Prior to harvest, warm and humid weather conditions followed by cool and dry conditions were associated with a decreased risk of DON accumulation. A prediction model, including only pre-flowering weather conditions, adequately forecasted risk of DON contamination in oat, and can aid in decisions about fungicide treatments.
Fusarium langsethiae is a recently characterized fungus within the genus Fusarium. It is found as a grain contaminant of small grain cereals such as oats and barley, and to a lesser extent wheat. Fusarium langsethiae is particularly widespread in the Nordic countries and the UK where it poses a serious problem as the main producer of T-2 and HT-2 mycotoxins. The biology of F. langsethiae and its interaction with the plant remains poorly understood, partly hampered by difficulties reproducing a natural level of infection under controlled conditions. The reported study was designed as a series of glasshouse experiments to advance our understanding of F. langsethiae biology by investigating alternative infection routes and its proliferation in oats, Avena sativa. Various methods of seed, soil, and seedling inoculation, boot injection and spray inoculation, were tested. The results clearly show a strong preference of F. langsethiae for the panicle, ruling out alternative infection routes. At relatively low temperatures spray infection, accompanied by prolonged humidity, ensured a thorough establishment of the fungus both at flowering and at early dough stage. Boot injection proved to be a reliable working tool for production of an even and predictable grain infection. Apart from in the panicle, considerable fungal proliferation was only detected in flag leaf nodes, and was a direct consequence of the boot injection method. Fungal presence in the node tissue also correlated with significant stunting of infected shoots. In light of the results the pathogenic and endophytic abilities of F. langsethiae are discussed.
As has been observed in several European countries, the frequency of Fusarium head blight (FHB) caused by members of the Fusarium graminearum species complex (FGSC) has increased in Norwegian cereals in recent years, resulting in elevated levels of deoxynivalenol in cereal grains. The objective of this study was to determine if this increase was associated with changes in FGSC composition within Norway. FGSC isolates collected from wheat, oats and barley in Norway during two periods, mainly 1993-1998 and 2004-2007, were characterized to determine species and trichothecene genotype composition and to assess levels of genetic variation and population structure. In vitro growth rates at different temperatures and aggressiveness in spring wheat were further characterized for a sub-selection of isolates. All Norwegian isolates were identified as F. graminearum. The 3-acetyl-deoxynivalenol (3-ADON) trichothecene type was dominant. However, isolates with the 15-ADON chemotype were detected in Norway for the first time and may represent a recent introduction of this trichothecene type. Bayesian-model based clustering and analyses of genetic differentiation indicated the persistence over the last 20 years of two sympatric and partially admixed populations of F. graminearum in Norway. Significant differences in average in vitro growth rates and aggressiveness were observed between these two populations. Our results demonstrate that the recent increase in prevalence of the FGSC in Norwegian cereals do not correspond to any dramatic changes in FGSC species or trichothecene chemotype composition. However, significant changes in population frequencies were observed among Norwegian F. graminearum.
High concentrations of the mycotoxins HT-2 and T-2 (HT2 + T2), primarily produced by Fusarium langsethiae, have occasionally been detected in Norwegian oat grains. In this study, we identified weather variables influencing accumulation of HT2 + T2 in Norwegian oat grains. Oat grain samples from farmers' fields were collected together with weather data (2004)(2005)(2006)(2007)(2008)(2009)(2010)(2011)(2012)(2013). Spearman rank correlation coefficients were calculated between the HT2 + T2 contamination in oats at harvest and a range of weather summarisations within estimated phenological windows of growth stages in oats (tillering, flowering etc.). Furthermore, we developed a mathematical model to predict the risk of HT2 + T2 in oat grains. Our data show that adequate predictions of the risk of HT2 + T2 in oat grains at harvest can be achieved, based upon weather data observed during the growing season. Humid and cool conditions, in addition to moderate temperatures during booting, were associated with increased HT2 + T2 accumulation in harvested oat grains, whereas warm and humid weather during stem elongation and inflorescence emergence, or cool weather and absence of rain during booting reduced the risk of HT2 + T2 accumulation. Warm and humid weather immediately after flowering increased the risk, while moderate to warm temperatures and absence of rain during dough development, reduced the risk of HT2 + T2 accumulation in oat grains. Our data indicated that HT2 + T2 contamination in oats is influenced by weather conditions both pre-and post-flowering. These findings are in contrast with a previous study examining the risk of deoxynivalenol contamination in oat reporting that toxin accumulation was mostly influenced by weather conditions from flowering onwards.
Over recent decades, the Norwegian cereal industry has had major practical and financial challenges associated with the occurrence of Fusarium head blight (FHB) pathogens and their associated mycotoxins in cereal grains. Deoxynivalenol (DON) is one of the most common Fusarium-mycotoxins in Norwegian oats, however T-2 toxin (T2) and HT-2 toxin (HT2) are also commonly detected. The aim of our study was to rank Nordic spring oat varieties and breeding lines by content of the most commonly occurring Fusarium mycotoxins (DON and HT2 + T2) as well as by the DNA content of their respective producers. We analyzed the content of mycotoxins and DNA of seven fungal species belonging to the FHB disease complex in grains of Nordic oat varieties and breeding lines harvested from oat field trials located in the main cereal cultivating district in South-East Norway in the years 2011–2020. Oat grains harvested from varieties with a high FHB resistance contained on average half the levels of mycotoxins compared with the most susceptible varieties, which implies that choice of variety may indeed impact on mycotoxin risk. The ranking of oat varieties according to HT2 + T2 levels corresponded with the ranking according to the DNA levels of Fusarium langsethiae, but differed from the ranking according to DON and Fusarium graminearum DNA. Separate tests are therefore necessary to determine the resistance towards HT2 + T2 and DON producers in oats. This creates practical challenges for the screening of FHB resistance in oats as today’s screening focuses on resistance to F. graminearum and DON. We identified oat varieties with generally low levels of both mycotoxins and FHB pathogens which should be preferred to mitigate mycotoxin risk in Norwegian oats.
The aim of this study was to evaluate the performance and usefulness of three rapid test kits for analysis of HT-2 and T-2 toxins (HT-2 and T-2), two of the most potent trichothecenes commonly found in European oats. Concentrations of these two toxins combined (HT-2+T-2) were analysed in naturally contaminated oat samples (n=68) using the following test kits: Ridascreen® FAST T-2 Toxin (‘Fast ELISA’), DRAFT Ridascreen® HT-2/T-2 (‘Standard ELISA’, not commercially available), and the lateral flow device ROSA® HT-2-T-2 (‘Rosa LFD’). Mycotoxin analysis by LC-MS/MS was used as a reference method. Rosa LFD offered the best reliability, achieving detection that was stable across toxin levels, whereas detection by both ELISA kits differed significantly among toxin levels (P<0.01). The kits were also evaluated regarding agreement with the reference method (measured as Cohen's kappa) at a HT-2+T-2 concentration of 1000 μg/kg in naturally contaminated oats. Agreement was greatest for Rosa LFD (89.2%), intermediate for Standard ELISA (66.8%), and lowest for Fast ELISA (62.2%). Rosa LFD showed cross-reaction of 100% with both T-2 and HT-2. For the ELISA kits, cross-reactions were 100% with T-2 but below 100% with HT-2. Therefore, to estimate the sum of HT-2 and T-2 in an oat sample, it was necessary to re-calculate the data from both ELISA kits according to the known cross-reaction of each kit with HT-2 and the concentration ratio of HT-2 to T-2 in Norwegian oats. Rosa LFD had the highest correlation with LC-MS/MS (R2=0.94), and the corresponding R2 values for Fast and Standard ELISA were 0.61 and 0.83, respectively. Rosa LFD was well suited for on-site detection. Standard ELISA allows simultaneous testing of several samples that are useful for centralised laboratories.
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