Reduced Risk of Oat Grain Contamination with Fusarium langsethiae and HT-2 and T-2 Toxins with Increasing Tillage Intensity.

Frequent occurrences of high levels of Fusarium mycotoxins have been recorded in Norwegian oat grain. To elucidate the influence of tillage operations on the development of Fusarium and mycotoxins in oat grain, we conducted tillage trials with continuous oats at two locations in southeast Norway. We have previously presented the content of Fusarium DNA detected in straw residues and air samples from these fields. Grain harvested from ploughed plots had lower levels of Fusarium langsethiae DNA and HT-2 and T-2 toxins (HT2 + T2) compared to grain from harrowed plots. Our results indicate that the risk of F. langsethiae and HT2 + T2 contamination of oats is reduced with increasing tillage intensity. No distinct influence of tillage on the DNA concentration of Fusarium graminearum and Fusarium avenaceum in the harvested grain was observed. In contrast to F. graminearum and F. avenaceum, only limited contents of F. langsethiae DNA were observed in straw residues and air samples. Still, considerable concentrations of F. langsethiae DNA and HT2 + T2 were recorded in oat grain harvested from these fields. We speculate that the life cycle of F. langsethiae differs from those of F. graminearum and F. avenaceum with regard to survival, inoculum production and dispersal.

Different Resistance to DON versus HT2 + T2 Producers in Nordic Oat Varieties.

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.

Weather Patterns Associated with DON Levels in Norwegian Spring Oat Grain: A Functional Data Approach.

Fusarium graminearum is regarded as the main deoxynivalenol (DON) producer in Norwegian oats, and high levels of DON are occasionally recorded in oat grains. Weather conditions in the period around flowering are reported to have a high impact on the development of Fusarium head blight (FHB) and DON in cereal grains. Thus, it would be advantageous if the risk of DON contamination of oat grains could be predicted based on weather data. We conducted a functional data analysis of weather-based time series data linked to DON content in order to identify weather patterns associated with increased DON levels. Since flowering date was not recorded in our dataset, a mathematical model was developed to predict phenological growth stages in Norwegian spring oats. Through functional data analysis, weather patterns associated with DON content in the harvested grain were revealed mainly from about three weeks pre-flowering onwards. Oat fields with elevated DON levels generally had warmer weather around sowing, and lower temperatures and higher relative humidity or rain prior to flowering onwards, compared to fields with low DON levels. Our results are in line with results from similar studies presented for FHB epidemics in wheat. Functional data analysis was found to be a useful tool to reveal weather patterns of importance for DON development in oats.

Microdochium majus and other fungal pathogens associated with reduced gluten quality in wheat grain.

The bread-making quality of wheat depends on the viscoelastic properties of the dough in which gluten proteins play an important role. The quality of gluten proteins is influenced by the genetics of the different wheat varieties and environmental factors. Occasionally, a near complete loss of gluten strength, measured as the maximum resistance towards stretching (Rmax), is observed in grain lots of Norwegian wheat. It is hypothesized that the loss of gluten quality is caused by degradation of gluten proteins by fungal proteases. To identify fungi associated with loss of gluten strength, samples from a selection of wheat grain lots with weak gluten (n = 10, Rmax < 0.3 N) and strong gluten (n = 10, Rmax ≥ 0.6 N) was analyzed for the abundance of fungal operational taxonomic units (OTUs) using DNA metabarcoding of the nuclear ribosomal Internal Transcribed Spacer (ITS) region ITS1. The DNA quantities for a selection of fungal pathogens of wheat, and the total amount of fungal DNA, were analyzed by quantitative PCR (qPCR). The mean level of total fungal DNA was higher in grain samples with weak gluten compared to grain samples with strong gluten. Heightened quantities of DNA from fungi within the Fusarium Head Blight (FHB) complex, i.e. Fusarium avenaceum, Fusarium graminearum, Microdochium majus, and Microdochium nivale, were observed in grain samples with weak gluten compared to those with strong gluten. Microdochium majus was the dominant fungus in the samples with weak gluten. Stepwise regression modeling based on different wheat quality parameters, qPCR data, and the 35 most common OTUs revealed a significant negative association between gluten strength and three OTUs, of which the OTU identified as M. majus was the most abundant. The same analysis also revealed a significant negative relationship between gluten strength and F. avenaceum detected by qPCR, although the DNA levels of this fungus were low compared to those of M. majus. In vitro growth rate studies of a selection of FHB species showed that all the tested isolates were able to grow with gluten as a sole nitrogen source. In addition, proteins secreted by these fungi in liquid cultures were able to hydrolyze gluten substrate proteins in zymograms, confirming their capacity to secrete gluten-degrading proteases. The identification of fungi with potential to influence gluten quality can enable the development of strategies to minimize future problems with gluten strength in food-grade wheat.

Removal of Small Kernels Reduces the Content of Fusarium Mycotoxins in Oat Grain.

Cereal grain contaminated by Fusarium mycotoxins is undesirable in food and feed because of the harmful health effects of the mycotoxins in humans and animals. Reduction of mycotoxin content in grain by cleaning and size sorting has mainly been studied in wheat. We investigated whether the removal of small kernels by size sorting could be a method to reduce the content of mycotoxins in oat grain. Samples from 24 Norwegian mycotoxin-contaminated grain lots (14 from 2015 and 10 from 2018) were sorted by a laboratory sieve (sieve size 2.2 mm) into large and small kernel fractions and, in addition to unsorted grain samples, analyzed with LC-MS-MS for quantification of 10 mycotoxins. By removing the small kernel fraction (on average 15% and 21% of the weight of the samples from the two years, respectively), the mean concentrations of HT-2+T-2 toxins were reduced by 56% (from 745 to 328 µg/kg) in the 2015 samples and by 32% (from 178 to 121 µg/kg) in the 2018 samples. Deoxynivalenol (DON) was reduced by 24% (from 191 to 145 µg/kg) in the 2018 samples, and enniatin B (EnnB) by 44% (from 1059 to 594 µg/kg) in the 2015 samples. Despite low levels, our analyses showed a trend towards reduced content of DON, ADON, NIV, EnnA, EnnA1, EnnB1 and BEA after removing the small kernel fraction in samples from 2015. For several of the mycotoxins, the concentrations were considerably higher in the small kernel fraction compared to unsorted grain. Our results demonstrate that the level of mycotoxins in unprocessed oat grain can be reduced by removing small kernels. We assume that our study is the first report on the effect of size sorting on the content of enniatins (Enns), NIV and BEA in oat grains.

Multimycotoxin and fungal analysis of maize grains from south and southwestern Ethiopia.

The natural occurrence of fungi, mycotoxins and fungal metabolites was investigated in 100 samples of maize grains collected from south and southwestern Ethiopia in 2015. The maize samples were contaminated by Fusarium, Aspergillus and Penicillium species. Using liquid chromatography tandem mass spectrometry 127 secondary metabolites were analysed. Zearalenone was the most prevalent mycotoxin, occurring in about 96% of the samples. Zearalenone sulfate was the second most prevalent, present in 81% of the samples. Fumonisin B1 was detected in 70% of the samples with a mean level of 606 µg kg-1 in positive samples, while FB2, FB3 and FB4 were detected in 62%, 51% and 60% of the maize samples with mean levels of 202, 136 and 85 µg kg-1, respectively. Up to 8% of the samples were contaminated with aflatoxins, with a maximum level of aflatoxin B1 of 513 µg kg-1. Results were higher than earlier reports for maize from Ethiopia.

Complete genome sequence of Luteibacter rhizovicinus strain LJ96T, isolated from the rhizosphere of barley (Hordeum vulgare L.) in Denmark.

We present the complete genome sequence of Luteibacter rhizovicinus type strain LJ96T, a yellow-pigmented gammaproteobacterium isolated from the rhizosphere of barley (Hordeum vulgare) Johansen et al. (2005) , a species with numerous potential applications. The genome sequence was deposited to NCBI GenBank with the accession number CP017480.

Global transcriptome changes in perennial ryegrass during early infection by pink snow mould.

Lack of resistance to pink snow mould (Microdochium nivale) is a major constraint for adaptation of perennial ryegrass (Lolium perenne L.) to continental regions with long-lasting snow cover at higher latitudes. Almost all investigations of genetic variation in resistance have been performed using cold acclimated plants. However, there may be variation in resistance mechanisms that are functioning independently of cold acclimation. In this study our aim was to identify candidate genes involved in such resistance mechanisms. We first characterized variation in resistance to M. nivale among non-acclimated genotypes from the Norwegian cultivar 'Fagerlin' based on relative regrowth and fungal quantification by real-time qPCR. One resistant and one susceptible genotype were selected for transcriptome analysis using paired-end sequencing by Illumina Hiseq 2000. Transcriptome profiles, GO enrichment and KEGG pathway analysis indicate that defense response related genes are differentially expressed between the resistant and the susceptible genotype. A significant up-regulation of defense related genes, as well as genes involved in cell wall cellulose metabolic processes and aryl-alcohol dehydrogenase (NADP+) activity, was observed in the resistant genotype. The candidate genes identified in this study might be potential molecular marker resources for breeding perennial ryegrass cultivars with improved resistance to pink snow mould.

Faces of a changing climate: semi-quantitative multi-mycotoxin analysis of grain grown in exceptional climatic conditions in Norway.

Recent climatological research predicts a significantly wetter climate in Southern Norway as a result of global warming. Thus, the country has already experienced unusually wet summer seasons in the last three years (2010-2012). The aim of this pilot study was to apply an existing multi-analyte LC-MS/MS method for the semi-quantitative determination of 320 fungal and bacterial metabolites in Norwegian cereal grain samples from the 2011 growing season. Such knowledge could provide important information for future survey and research programmes in Norway. The method includes all regulated and well-known mycotoxins such as aflatoxins, trichothecenes, ochratoxin A, fumonisins and zearalenone. In addition, a wide range of less studied compounds are included in the method, e.g., Alternaria toxins, ergot alkaloids and other metabolites produced by fungal species within Fusarium, Penicillium and Aspergillus. Altogether, 46 metabolites, all of fungal origin, were detected in the 76 barley, oats and wheat samples. The analyses confirmed the high prevalence and relatively high concentrations of type-A and -B trichothecenes (e.g., deoxynivalenol up to 7230 µg/kg, HT-2 toxin up to 333 µg/kg). Zearalenone was also among the major mycotoxins detected (maximum concentration 1670 µg/kg). Notably, several other Fusarium metabolites such as culmorin, 2-amino-14,16-dimethyloctadecan-3-ol and avenacein Y were co-occurring. Furthermore, the most prevalent Alternaria toxin was alternariol with a maximum concentration of 449 µg/kg. A number of Penicillium and Aspergillus metabolites were also detected in the samples, e.g., sterigmatocystin in concentrations up to 20 µg/kg.