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.

Longevity of Plant Pathogens in Dry Agricultural Seeds during 30 Years of Storage.

Plant diseases may survive and be spread by infected seeds. In this study we monitored the longevity of 14 seed-borne pathogens in 9 crop species commonly grown in the Nordic countries, in addition to a sample of sclerotia of Sclerotinia sclerotiorum. The data from the first 30 years of a 100-year seed storage experiment located in a natural -3.5 °C environment (permafrost) in Svalbard, Norway, are presented. To date, the pathogens, tested by traditional seed health testing methods (freezing blotter, agar plates, growing on tests), have survived. Linear regression analyses showed that the seed infection percentages of Drechslera dictyoides in meadow fescue, Drechslera phlei in timothy, and Septoria nodorum in wheat were significantly reduced compared to the percentages at the start of the experiment (from 63% to 34%, from 70% to 65%, and from 15% to 1%, respectively), and that Phoma betae in beet had increased significantly (from 43% to 56%). No trends in the infection percentage were observed over the years in Drechslera spp. in barley (fluctuating between 30% and 64%) or in Alternaria brassicicola in cabbage (fluctuating between 82% and 99%), nor in pathogens with low seed infection percentages at the start of the experiment. A major part of the stored sclerotia was viable after 30 years. To avoid the spread of seed-borne diseases, it is recommended that gene banks implement routines that avoid the use of infected seeds.

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.

Seed Germination after 30 Years Storage in Permafrost.

More than 30 years ago, the Nordic Gene Bank established a long-term experiment on seeds stored under permafrost conditions in an abandoned mine corridor in Svalbard, as a tool to monitor storage life under these conditions. The study included seeds from 16 Nordic agricultural and horticultural crops, each represented by two or three cultivars (altogether 38 accessions). All seeds were ultra-dried to 3%-5% moisture before being sealed in glass tubes. Germination tests were performed in accordance with the International Seed Testing Association (ISTA) protocols. At the initiation of the experiment, the samples showed good germination with the median value at 92%. The overall picture remained stable over the first twenty to twenty-five years. However, the variation became larger over time and at 30 years, the median value had dropped to 80%. At the lower end, with a high drop in germination, we found rye, wheat, and English ryegrass. At the upper end, we found Kentucky bluegrass and cucumber. The lowest germination was found in samples with the highest initial seed moisture levels. Pre-storage conditions are likely to be of major importance for longevity.

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.

Understanding Yield Loss and Pathogen Biology to Improve Disease Management: Septoria Nodorum Blotch - A Case Study in Wheat.

The estimated potential yield losses caused by plant pathogens is up to 16% globally and most research in plant pathology aims to reduce yield loss in our crops directly or indirectly. Yield losses caused by a certain disease depend not only on disease severity, but also on the weather factors, the pathogen's aggressiveness, and the ability of the crop to compensate for reduced photosynthetic area. The yield loss-disease relationship in a certain host-pathogen system might therefore change from year to year, making predictions for yield loss very difficult at the regional or even at the farmer's level. However, estimating yield losses is essential to determine disease management thresholds at which acute control measures such as fungicide applications, or strategic measures such as crop rotation or use of resistant cultivars are economically and environmentally sensible. Legislation in many countries enforces implementation of integrated pest management (IPM), based on economic thresholds at which the costs due to a disease justify the costs for its management. Without a better understanding of the relationship between disease epidemiology and yield loss, we remain insufficiently equipped to design adequate IPM strategies that will be widely adapted in agriculture. Crop loss studies are resource demanding and difficult to interpret for one particular disease, as crops are usually not invaded by only one pest or pathogen at a time. Combining our knowledge on disease epidemiology, crop physiology, yield development, damage mechanisms involved, and the effect of management practices can help us to increase our understanding of the disease-crop loss relationship. The main aim of this paper is to review and analyze the literature on a representative host-pathogen relationship in an important staple food crop to identify knowledge gaps and research areas to better assess yield loss and design management strategies based on economic thresholds.

Population Structure of Sclerotinia subarctica and Sclerotinia sclerotiorum in England, Scotland and Norway.

Sclerotinia species are important fungal pathogens of a wide range of crops and wild host plants. While the biology and population structure of Sclerotinia sclerotiorum has been well-studied, little information is available for the related species S. subarctica. In this study, Sclerotinia isolates were collected from different crop plants and the wild host Ranuculus ficaria (meadow buttercup) in England, Scotland, and Norway to determine the incidence of Sclerotinia subarctica and examine the population structure of this pathogen for the first time. Incidence was very low in England, comprising only 4.3% of isolates while moderate and high incidence of S. subarctica was identified in Scotland and Norway, comprising 18.3 and 48.0% of isolates respectively. Characterization with eight microsatellite markers identified 75 haplotypes within a total of 157 isolates over the three countries with a few haplotypes in Scotland and Norway sampled at a higher frequency than the rest across multiple locations and host plants. In total, eight microsatellite haplotypes were shared between Scotland and Norway while none were shared with England. Bayesian and principal component analyses revealed common ancestry and clustering of Scottish and Norwegian S. subarctica isolates while English isolates were assigned to a separate population cluster and exhibited low diversity indicative of isolation. Population structure was also examined for S. sclerotiorum isolates from England, Scotland, Norway, and Australia using microsatellite data, including some from a previous study in England. In total, 484 haplotypes were identified within 800 S. sclerotiorum isolates with just 15 shared between England and Scotland and none shared between any other countries. Bayesian and principal component analyses revealed a common ancestry and clustering of the English and Scottish isolates while Norwegian and Australian isolates were assigned to separate clusters. Furthermore, sequencing part of the intergenic spacer (IGS) region of the rRNA gene resulted in 26 IGS haplotypes within 870 S. sclerotiorum isolates, nine of which had not been previously identified and two of which were also widely distributed across different countries. S. subarctica therefore has a multiclonal population structure similar to S. sclerotiorum, but has a different ancestry and distribution across England, Scotland, and Norway.

Deoxynivalenol Exposure in Norway, Risk Assessments for Different Human Age Groups.

Deoxynivalenol (DON) is the most common mycotoxin in Norwegian cereals, and DON is detected in most samples of crude cereal grain and cereal food commodities such as flour, bran, and oat flakes. The Norwegian Scientific Committee for Food Safety assessed the risk for adverse effects of deoxynivalenol (DON) in different age groups of the domestic population. This review presents the main results from the risk assessment, supplemented with some recently published data. Impairment of the immune system together with reduced feed intake and weight gain are the critical effects of DON in experimental animals on which the current tolerable daily intake was established. Based on food consumption and occurrence data, the mean exposure to DON in years with low and high levels of DON in the flour, respectively, were in the range of or up to two times the Tolerable Daily Intake (TDI) in 1-year-old infants and 2-year-old children. In years with high mean DON concentration, the high (95th-percentile) exposure exceeded the TDI by up to 3.5 times in 1-, 2- , 4-, and 9-year-old children. The assessment concluded that exceeding the TDI in infants and children is of concern. The estimated dietary DON intakes in adolescent and adult populations are in the range of the TDI or below, and are not a health concern. Acute human exposure to DON is not of concern in any age group.

Inoculum Potential of Fusarium spp. Relates to Tillage and Straw Management in Norwegian Fields of Spring Oats.

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.

Temporal Variation of Mycotoxin Producing Fungi in Norwegian Cereals.

Spring barley is grown on about half of the Norwegian cereal area. The rest of the area is equally divided between wheat and oats. Most years the domestic production provides 70%-80% of the domestic market for bread wheat. Barley and oats are mainly grown for animal feed. During the years 2008-2012, severe epidemics of Fusarium head blight have led to increased mycotoxin contamination of cereals. During that period, precipitation was above normal during anthesis and grain maturation. The most important mycotoxin producers have been F. avenaceum, F. culmorum, F. graminearum and F. langsethiae. Increased deoxynivalenol contamination of Norwegian cereals during recent years is due to severe F. graminearum epidemics.