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Kettle holes are able to increase the soil and air humidity around them. Therefore, they create a perfect habitat for phytopathogenic fungi of the genera Fusarium and Alternaria to develop, sporulate, and immigrate into neighboring agricultural fields. In our study, we establish transects from the edges of different kettle holes and field edges up to 50 m into the fields to analyze the abundance and diversity of pathogenic fungi in these transition zones by culture-dependent and culture-independent methods. However, in 2019 and 2020, low precipitation and higher temperatures compared to the long-time average were measured, which led to limited infections of weeds in the transition zones with Fusarium and Alternaria . Therefore, the hypothesized significantly higher infection of wheat plants next to the kettle holes by a strong spread of fungal spores was not detected. Infestation patterns of Fusarium and Alternaria fungi on weeds and wheat ears were spatially different. In total, 9 different Fusarium species were found in the transition zone. The species diversity at kettle holes differed from 0 to 6 species. The trend toward increased dryness in the northeast German agricultural landscape and its impact on the changing severity of fungal infections is discussed.

Fungi of the genus Alternaria are ubiquitous in the environment. Their mycotoxins can leach out of contaminated plants or crop debris into the soil entering the plant via the roots. We aim to evaluate the importance of this entry pathway and its contribution to the overall content of Alternaria toxins (ATs) in wheat plants to better understand the soil-plant-phytopathogen system. A hydroponic cultivation system was established and wheat plants were cultivated for up to two weeks under optimal climate conditions. One half of the plants was treated with a nutrient solution spiked with alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA), whereas the other half of the plants was cultivated without mycotoxins. Plants were harvested after 1 and 2 weeks and analyzed using a QuEChERS-based extraction and an in-house validated LC-MS/MS method for quantification of the ATs in roots, crowns, and leaves separately. ATs were taken up by the roots and transported throughout the plant up to the leaves after 1 as well as 2 weeks of cultivation with the roots showing the highest ATs levels followed by the crowns and the leaves. In addition, numerous AOH and AME conjugates like glucosides, malonyl glucosides, sulfates, and di/trihexosides were detected in different plant compartments and identified by high-resolution mass spectrometry. This is the first study demonstrating the uptake of ATs in vivo using a hydroponic system and whole wheat plants examining both the distribution of ATs within the plant compartments and the modification of ATs by the wheat plants., (© 2023. The Author(s).)

Grasses growing next to agricultural fields influence the Fusarium abundance, the species composition, and the mycotoxin accumulation of wheat plants, especially the field parts directly adjacent to grasses, are highly affected. Grasses are a more attractive and suitable habitat for Fusarium fungi compared to other arable weeds and occur at mostly every semi-natural landscape element (e.g., kettle holes, hedgerows, field-to-field-borders). In our study, we analyzed the ability of a highly Fusarium infected grass stripe (F. graminearum, F. culmorum, F. sporotrichioides) to infect an adjacent wheat field with these species. Results show that the primary inoculated Fusarium species were as well the dominant species isolated from the wheat field. Regarding transects originating from the grass stripe going into the field, the results demonstrate that wheat ears next to the infected grass stripe have a higher Fusarium abundance and furthermore show higher mycotoxin accumulation in the wheat kernels. This effect was highly promoted by irrigation. Therefore, grass stripes next to arable fields must be considered as reservoirs for fungal infections and as a source for a contamination with mycotoxins., (© 2022. The Author(s).)

To adapt to climate change, several agricultural strategies are currently being explored, including a shift in land use areas. Regional differences in microbiome composition and associated phytopathogens need to be considered. However, most empirical studies on differences in the crop microbiome focused on soil communities, with insufficient attention to the phyllosphere. In this study, we focused on wheat ears in three regions in northeastern Germany (Magdeburger Börde (MBB), Müncheberger Sander (MSA), Uckermärkisches Hügelland (UKH)) with different yield potentials, soil, and climatic conditions. To gain insight into the fungal community at different sites, we used a metabarcoding approach (ITS-NGS). Further, we examined the diversity and abundance of Fusarium and Alternaria using culture-dependent and culture-independent techniques. For each region, the prevalence of different orders rich in phytopathogenic fungi was determined: Sporidiobolales in MBB, Capnodiales and Pleosporales in MSA, and Hypocreales in UKH were identified as taxonomic biomarkers. Additionally, F. graminearum was found predominantly in UKH, whereas F. poae was more abundant in the other two regions. Environmental filters seem to be strong drivers of these differences, but we also discuss the possible effects of dispersal and interaction filters. Our results can guide shifting cultivation regions to be selected in the future concerning their phytopathogenic infection potential.

The dispersal of propagules, such as fungal spores or seeds by actively moving animals, connects and shapes communities. The dispersal of plant pathogens by arthropods might be a crucial mechanism in the spread of several crop diseases. Ground-dwelling arthropods are potential linkers between fungal communities in semi-natural and agricultural habitats by transporting propagules of Fusarium fungi. We compared the Fusarium communities on the body surface of ground-dwelling arthropods with litter in semi-natural and soil in agricultural habitats with a focus on the Fusarium community. We found three relatively distinct Fusarium communities with moderate overlap. We detected a higher richness of Fusarium species on the body surface of arthropods compared to litter and soil communities. The results suggest that the Fusarium community on the body surface of arthropods relates to the composition observed in litter and soil with limited filtering mechanisms between communities. Ground-dwelling arthropods are relevant agents for the distribution of Fusarium and therefore link fungal communities in adjacent habitats.

The spread by arthropods (zoochory) is an essential dispersal mechanism for many microorganisms, like plant pathogens. Carabid beetles are very abundant and mobile ground-dwelling insects. However, their role in the dispersal of economically relevant phytopathogens, like Fusarium and Alternaria fungi is basically unknown. We quantified the total fungal, Fusarium , and Alternaria load of carabid species collected in the transition zones between small water bodies and wheat fields by screening (i) their body surface for fungal propagules with a culture-dependent method and (ii) their entire bodies for fungal DNA with a qPCR approach. The analysis of entire bodies detects fungal DNA in all carabid beetles but Alternaria DNA in 98% of them. We found that 74% of the carabids carried fungal propagules on the body surface, of which only half (49%) carried Fusarium propagules. We identified eight Fusarium and four Alternaria species on the body surface; F.   culmorum was dominant. The fungal, Fusarium and Alternaria , load differed significantly between the carabid species and was positively affected by the body size and weight of the carabids. Carabid beetles reveal a remarkable potential to disseminate different fungi. Dispersal by ground-dwelling arthropods could affect the spatial-temporal patterns of plant disease and microorganisms in general.

Semi-natural habitats (SNHs) are becoming increasingly scarce in modern agricultural landscapes. This may reduce natural ecosystem services such as pest control with its putatively positive effect on crop production. In agreement with other studies, we recently reported wheat yield reductions at field borders which were linked to the type of SNH and the distance to the border. In this experimental landscape-wide study, we asked whether these yield losses have a biotic origin while analyzing fungal seed and fungal leaf pathogens, herbivory of cereal leaf beetles, and weed cover as hypothesized mediators between SNHs and yield. We established experimental winter wheat plots of a single variety within conventionally managed wheat fields at fixed distances either to a hedgerow or to an in-field kettle hole. For each plot, we recorded the fungal infection rate on seeds, fungal infection and herbivory rates on leaves, and weed cover. Using several generalized linear mixed-effects models as well as a structural equation model, we tested the effects of SNHs at a field scale (SNH type and distance to SNH) and at a landscape scale (percentage and diversity of SNHs within a 1000-m radius). In the dry year of 2016, we detected one putative biotic culprit: Weed cover was negatively associated with yield values at a 1-m and 5-m distance from the field border with a SNH. None of the fungal and insect pests, however, significantly affected yield, neither solely nor depending on type of or distance to a SNH. However, the pest groups themselves responded differently to SNH at the field scale and at the landscape scale. Our findings highlight that crop losses at field borders may be caused by biotic culprits; however, their negative impact seems weak and is putatively reduced by conventional farming practices., Competing Interests: The authors state no conflict of interest., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Mycotoxigenic fungal pathogens Fusarium and Alternaria are a leading cause of loss in cereal production. On wheat-ears, they are confronted by bacterial antagonists such as pseudomonads. Studies on these groups' interactions often neglect the infection process's temporal aspects and the associated priority effects. In the present study, the focus was on how the first colonizer affects the subsequent ones. In a climate chamber experiment, wheat-ears were successively inoculated with two different strains ( Alternaria tenuissima At 625, Fusarium graminearum Fg 23, or Pseudomonas simiae Ps 9). Over three weeks, microbial abundances and mycotoxin concentrations were analyzed and visualized via Self Organizing Maps with Sammon Mapping (SOM-SM). All three strains revealed different characteristics and strategies to deal with co-inoculation: Fg 23, as the first colonizer, suppressed the establishment of At 625 and Ps 9. Nevertheless, primary inoculation of At 625 reduced all of the Fusarium toxins and stopped Ps 9 from establishing. Ps 9 showed priority effects in delaying and blocking the production of the fungal mycotoxins. The SOM-SM analysis visualized the competitive strengths: Fg 23 ranked first, At 625 second, Ps 9 third. Our findings of species-specific priority effects in a natural environment and the role of the mycotoxins involved are relevant for developing biocontrol strategies.

Organismal movement is ubiquitous and facilitates important ecological mechanisms that drive community and metacommunity composition and hence biodiversity. In most existing ecological theories and models in biodiversity research, movement is represented simplistically, ignoring the behavioural basis of movement and consequently the variation in behaviour at species and individual levels. However, as human endeavours modify climate and land use, the behavioural processes of organisms in response to these changes, including movement, become critical to understanding the resulting biodiversity loss. Here, we draw together research from different subdisciplines in ecology to understand the impact of individual-level movement processes on community-level patterns in species composition and coexistence. We join the movement ecology framework with the key concepts from metacommunity theory, community assembly and modern coexistence theory using the idea of micro-macro links, where various aspects of emergent movement behaviour scale up to local and regional patterns in species mobility and mobile-link-generated patterns in abiotic and biotic environmental conditions. These in turn influence both individual movement and, at ecological timescales, mechanisms such as dispersal limitation, environmental filtering, and niche partitioning. We conclude by highlighting challenges to and promising future avenues for data generation, data analysis and complementary modelling approaches and provide a brief outlook on how a new behaviour-based view on movement becomes important in understanding the responses of communities under ongoing environmental change., (© 2020 The Authors. Biological Reviews published by John Wiley & Sons Ltd on behalf of Cambridge Philosophical Society.)

Fluorescent pseudomonads colonizing wheat ears have a high antagonistic potential against phytopathogenic fungi. To check this hypothesis, the bacterial antagonist Pseudomonas simiae 9 rif+/kan+ was spray-inoculated onto the ears of winter wheat in a locally demarcated experimental field plot. Fusarium and Alternaria fungi naturally occurring on the ears and the formation of their mycotoxins in the ripe grains were investigated. Inoculated bacteria were recovered from the plants in the inoculation cell, but not in the untreated neighboring plots or in the air above the plants. Growth of fusaria and alternaria on the ears was not influenced by the bacterial antagonist. Wheat kernels were co-inoculated in vitro with the antagonist and one mycotoxin-producing strain of Fusarium and Alternaria, respectively. Mycotoxin production was almost completely suppressed in these approaches. Concentrations of zearalenone, deoxynivalenol, alternariol, and tenuazonic acid were also significantly reduced in ripe grains in the field, but to a lesser extent than in vitro. The results of this and previous studies suggest that widespread biological control of the growth of fusaria and alternaria and their mycotoxin formation by naturally occurring pseudomonads with antagonistic activity is rather unlikely.

The effects of biochar on plant growth vary depending on the applied biochar type, study site environmental conditions, microbial species, and plant-microbial interactions. The objectives of the present study were therefore to assess 1) the response of growth parameters of lupin and root disease incidence to the application of three biochar types in a loamy sandy soil, and 2) the role of endophytic bacteria in biological control of root rot disease incidence in lupin after the amendment of soil with different biochar types. As biochar types we tested (i) hydrochar (HTC) from maize silage, (ii) pyrolysis char from maize (MBC), and (iii) pyrolysis char from wood (WBC) at three different concentrations (1%, 2%, and 3% of char as soil amendments). There were no significant effects in lupin shoot and root growth in soils amended with WBC at any of the concentrations. MBC did not affect plant growth except for root dry weight at 2% MBC. HTC char at 2% concentration, significantly increased the root dry weight of lupin by 54-75%, and shoot dry weight by 21-25%. Lupin plants grown in soil amended with 2% and 3% WBC and MBC chars showed 40-50% and 10-20% disease symptoms, respectively. Plants grown in soil without biochar and with HTC char were healthy, and no disease incidence occurred. Pseudomonas putida L2 and Stenotrophomonas pavanii L8 isolates demonstrated a disease reduction compared to un-inoculated plants under MBC and WBC amended soil that was infested with Fusarium solani .

Plant associated microbial communities have recently received a lot of attention because thought to play a fundamental role in plant health and development. Focusing on cultivated crops, optimized farming practices must consider the role of these communities when aiming at reducing the impact of pathogens and increasing yields. Typical inhabitants of plant's phyllosphere are bacteria and microscopic fungi, some of them pathogenic for the plant and dangerous for the consumers, due to the production of toxins. In order to efficiently manage the microbiome, the natural drivers regulating community assembly must be clearly understood. In our study we investigated the within field variation of the phyllosphere mycobiome of wheat ears by metabarcoding of the fungal internal transcribed sequence 1 (ITS1). We selected a field characterized by a high topographic heterogeneity, which is reflected in differences in plant productivity and fitness across it. Samples were taken from 30 sampling points laid across the field where data-loggers were placed, measuring the productivity driven under canopy microclimate. The microclimatic conditions were tested as a source of potential environmental variance. Further independent spatial structures were tested using spatial eigenvector maps (MEMs). Results show considerable differences in the phyllosphere composition across the field. The local under canopy environmental conditions at each point were strong predictors of the community composition. Independent spatial effects given by the geographical position of the sampling points showed also a weaker but significant effect. Moreover we observed different spatial responses from different fungal phyla, with results resembling those described in studies done at a regional scale. This study is the first one to investigate the spatial variation of the phyllosphere mycobiome of a commercial crop within the same field. It contributes to the study of the epidemiology and community assembly dynamics of wheat phyllosphere fungi, showing how in-field community variations are the results of different environmental and spatial processes acting simultaneously. It also shows how heterogeneous fields are a smart and useful system to investigate the ecological mechanisms regulating plant microbiome composition.

Natural landscape elements (NLEs) in agricultural landscapes contribute to biodiversity and ecosystem services, but are also regarded as an obstacle for large-scale agricultural production. However, the effects of NLEs on crop yield have rarely been measured. Here, we investigated how different bordering structures, such as agricultural roads, field-to-field borders, forests, hedgerows, and kettle holes, influence agricultural yields. We hypothesized that (a) yield values at field borders differ from mid-field yields and that (b) the extent of this change in yields depends on the bordering structure. We measured winter wheat yields along transects with log-scaled distances from the border into the agricultural field within two intensively managed agricultural landscapes in Germany (2014 near Göttingen, and 2015-2017 in the Uckermark). We observed a yield loss adjacent to every investigated bordering structure of 11%-38% in comparison with mid-field yields. However, depending on the bordering structure, this yield loss disappeared at different distances. While the proximity of kettle holes did not affect yields more than neighboring agricultural fields, woody landscape elements had strong effects on winter wheat yields. Notably, 95% of mid-field yields could already be reached at a distance of 11.3 m from a kettle hole and at a distance of 17.8 m from hedgerows as well as forest borders. Our findings suggest that yield losses are especially relevant directly adjacent to woody landscape elements, but not adjacent to in-field water bodies. This highlights the potential to simultaneously counteract yield losses close to the field border and enhance biodiversity by combining different NLEs in agricultural landscapes such as creating strips of extensive grassland vegetation between woody landscape elements and agricultural fields. In conclusion, our results can be used to quantify ecocompensations to find optimal solutions for the delivery of productive and regulative ecosystem services in heterogeneous agricultural landscapes., Competing Interests: None declared.

Aim: To observe the variation in accumulation of Fusarium and Alternaria mycotoxins across a topographically heterogeneous field and tested biotic (fungal and bacterial abundance) and abiotic (microclimate) parameters as explanatory variables., Methods and Results: We selected a wheat field characterized by a diversified topography, to be responsible for variations in productivity and in canopy-driven microclimate. Fusarium and Alternaria mycotoxins where quantified in wheat ears at three sampling dates between flowering and harvest at 40 points. Tenuazonic acid (TeA), alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), deoxynivalenol (DON), zearalenone (ZEN) and deoxynivalenol-3-Glucoside (DON.3G) were quantified. In canopy temperature, air and soil humidity were recorded for each point with data-loggers. Fusarium spp. as trichothecene producers, Alternaria spp. and fungal abundances were assessed using qPCR. Pseudomonas fluorescens bacteria were quantified with a culture based method. We only found DON, DON.3G, TeA and TEN to be ubiquitous across the whole field, while AME, AOH and ZEN were only occasionally detected. Fusarium was more abundant in spots with high soil humidity, while Alternaria in warmer and drier spots. Mycotoxins correlated differently to the observed explanatory variables: positive correlations between DON accumulation, tri 5 gene and Fusarium abundance were clearly detected. The correlations among the others observed variables, such as microclimatic conditions, varied among the sampling dates. The results of statistical model identification do not exclude that species coexistence could influence mycotoxin production., Conclusions: Fusarium and Alternaria mycotoxins accumulation varies heavily across the field and the sampling dates, providing the realism of landscape-scale studies. Mycotoxin concentrations appear to be partially explained by biotic and abiotic variables., Significance and Impact of the Study: We provide a useful experimental design and useful data for understanding the dynamics of mycotoxin biosynthesis in wheat., (© 2018 The Society for Applied Microbiology.)

Natural control of phytopathogenic microorganisms is assumed as a priority function of the commensal plant microbiota. In this study, the suitability of fluorescent pseudomonads in the phyllosphere of crop plants as natural control agents was evaluated. Under field conditions, ears of winter wheat were found to be colonized with high consistency and at a high density by pseudomonads at the late milk dough stage. Isolates of these bacteria were evaluated for their potential to protect the plants from phytopathogenic Alternaria and Fusarium fungi. More Pseudomonas isolates were antagonistically active against alternaria than against fusaria in the dual culture test. The alternaria responded species-specifically and more sensitively to bacterial antagonism than the strain-specific reacting fusaria. A total of 110 randomly selected Pseudomonas isolates were screened for genes involved in the biosynthesis of the antibiotics 2,4-diacetylphloroglucinol, phenazine-1-carboxylic acid, pyoluteorin, and pyrrolnitrin. The key gene for production of the phloroglucinol was found in none of these isolates. At least one of the genes, encoding the biosynthesis of the other antibiotics was detected in 81% of the isolates tested. However, the antagonistic effect found in the dual culture assay was not necessarily associated with the presence of these antibiotic genes. Wheat grains as natural substrate were inoculated with selected antagonistic Pseudomonas isolates and Alternaria and Fusarium strains, respectively. The fungal growth was only slightly delayed, but the mycotoxin production was significantly reduced in most of these approaches. In conclusion, the distribution of phytopathogenic fungi of the genera Alternaria and Fusarium in the field is unlikely to be inhibited by naturally occurring pseudomonads, also because the bacterial antagonists were not evenly distributed in the field. However, pseudomonads can reduce the production of Alternaria and Fusarium mycotoxins in wheat grains and thus have the potential to improve the crop quality.

Necrotrophic as well as saprophytic small-spored Alternaria (A.) species are annually responsible for major losses of agricultural products, such as cereal crops, associated with the contamination of food and feedstuff with potential health-endangering Alternaria toxins. Knowledge of the metabolic capabilities of different species-groups to form mycotoxins is of importance for a reliable risk assessment. 93 Alternaria strains belonging to the four species groups Alternaria tenuissima , A. arborescens , A. alternata , and A. infectoria were isolated from winter wheat kernels harvested from fields in Germany and Russia and incubated under equal conditions. Chemical analysis by means of an HPLC-MS/MS multi- Alternaria -toxin-method showed that 95% of all strains were able to form at least one of the targeted 17 non-host specific Alternaria toxins. Simultaneous production of up to 15 (modified) Alternaria toxins by members of the A. tenuissima , A. arborescens , A. alternata species-groups and up to seven toxins by A. infectoria strains was demonstrated. Overall tenuazonic acid was the most extensively formed mycotoxin followed by alternariol and alternariol mono methylether, whereas altertoxin I was the most frequently detected toxin. Sulfoconjugated modifications of alternariol, alternariol mono methylether, altenuisol and altenuene were frequently determined. Unknown perylene quinone derivatives were additionally detected. Strains of the species-group A. infectoria could be segregated from strains of the other three species-groups due to significantly lower toxin levels and the specific production of infectopyrone. Apart from infectopyrone, alterperylenol was also frequently produced by 95% of the A. infectoria strains. Neither by the concentration nor by the composition of the targeted Alternaria toxins a differentiation between the species-groups A. alternata , A. tenuissima and A. arborescens was possible.

Fusarium spp. and Alternaria spp., two genera of filamentous fungi, are common colonizers of the wheat phyllosphere. Both can be pathogenic and produce mycotoxins that are harmful to consumers. Their in-field infection dynamics have been a focus for the development of new control strategies. We analysed the abundance on plant ears and spore deposition patterns of Fusarium spp. and Alternaria spp. in a topographically heterogeneous field. Abundances were assessed genetically, using qPCR-based techniques, and passive spore traps were installed for quantifying the spore deposition at different plant heights. Data loggers were placed to measure the differences in microclimate across the field. Results indicate different distribution and spore deposition patterns for the two fungi. Fusarium spp. spore and genetic abundances were higher in spots with a more humid and colder under-canopy microclimate. Alternaria spp. showed the opposite trend for genetic abundance, while its spore deposition was not correlated to any of the microclimatic conditions and was more uniform across the field. Our study extends the knowledge on the dispersal and in-field infection dynamics of Fusarium spp. and Alternaria spp., important for a better understanding of the epidemiology of these wheat pathogens. It also illustrates that topographically heterogeneous fields are a suitable environment for studying the ecology of phyllosphere-colonizing fungi.

Many scientific disciplines are currently experiencing a 'reproducibility crisis' because numerous scientific findings cannot be repeated consistently. A novel but controversial hypothesis postulates that stringent levels of environmental and biotic standardization in experimental studies reduce reproducibility by amplifying the impacts of laboratory-specific environmental factors not accounted for in study designs. A corollary to this hypothesis is that a deliberate introduction of controlled systematic variability (CSV) in experimental designs may lead to increased reproducibility. To test this hypothesis, we had 14 European laboratories run a simple microcosm experiment using grass (Brachypodium distachyon L.) monocultures and grass and legume (Medicago truncatula Gaertn.) mixtures. Each laboratory introduced environmental and genotypic CSV within and among replicated microcosms established in either growth chambers (with stringent control of environmental conditions) or glasshouses (with more variable environmental conditions). The introduction of genotypic CSV led to 18% lower among-laboratory variability in growth chambers, indicating increased reproducibility, but had no significant effect in glasshouses where reproducibility was generally lower. Environmental CSV had little effect on reproducibility. Although there are multiple causes for the 'reproducibility crisis', deliberately including genetic variability may be a simple solution for increasing the reproducibility of ecological studies performed under stringently controlled environmental conditions.