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Maize lethal necrosis (MLN) is a maize disease caused by the maize chlorotic mottle virus (MCMV), a potyvirus which causes yield losses of 30–100%. The present study aimed to isolate nanobodies against the MCMV coat protein (CP) for the diagnosis of MLN. MCMV CP expressed in Escherichia coli was used for llama immunization. VHH (i.e. variable heavy domain of heavy chain) gene fragments were prepared from blood drawn from the immunized llama and used to generate a library in E. coli TG1 cells. MCMV specific nanobodies were selected by three rounds of phage display and panning against MCMV CP. The selected nanobodies were finally expressed in E. coli WK6 cells and purified. Eleven MCMV‐specific nanobodies were identified and shown to detect MCMV in infected maize plants. Thus, our results show that nanobodies isolated from llama immunized with MCMV CP can distinguish infected and healthy maize plants, potentially enabling development of affordable MCMV detection protocols.

Abstract Background Grasslands are the primary source of forage for ruminants. Legal restrictions on nitrogen fertilization force farmers to optimize nitrogen use efficiency (NUE) of grassland and protein quality in grass silage. Methods The present study included grassland data from 10 dairy farms in Flanders. For each grassland field, the presence of clover was determined based on the seed mixture composition. Soil and manure were sampled before the first application and analyzed to determine plant available N content. The annual NUE was calculated for each field possible (n = 28). Prewilted grass samples were taken just before ensiling (n = 65) and grass silage samples (n = 41) were collected from the same fields. Results Clover fields demonstrated improved NUE due to a lower nitrogen input from fertilizers compared to pure grassland fields (75 vs. 265 kg ha−1). The protein quality of the grass silage was positively correlated with the organic matter digestibility, sugar content, and dry matter content of the prewilted grass. Conversely, crude ash content had a negative effect. Conclusions These findings underscore the importance and potential of clover and the practical feasibility of optimizing grassland management to improve both NUE in grasslands and protein quality of the silage.

Abstract Background Triticale is making its way on dairy farms as an alternative forage crop. This requires the availability of high-yielding triticale varieties with good digestibility. Triticale forage breeding mainly focussed on biomass yield, but efforts to improve digestibility are increasing. We previously investigated the interrelationships among different quality traits in soft dough triticale: starch, acid detergent fibre and in vitro digestibility of organic matter (IVOMD) and of neutral detergent fibre (IVNDFD) of the total plant, IVNDFD and Klason lignin of the stems, and ear proportion and stem length. Here we determine the genetic control of these traits, using a genome-wide association (GWAS) approach. A total of 33,231 DArTseq SNP markers assessed in a collection of 118 winter triticale genotypes, including 101 varieties and 17 breeding lines, were used. Results The GWAS identified a total of 53 significant marker-trait associations (MTAs). The highest number of significantly associated SNP markers (n = 10) was identified for total plant IVNDFD. A SNP marker on chromosome 1A (4211801_19_C/T; 474,437,796 bp) was found to be significantly associated with ear proportion, and plant and stem IVNDFD, with the largest phenotypic variation for ear proportion (R²p = 0.23). Based on MTAs, candidate genes were identified which were of particular relevance for variation in in vitro digestibility (IVD) because they are putatively involved in plasma membrane transport, cytoskeleton organisation, carbohydrate metabolic processes, protein phosphorylation, and sterol and cell wall biogenesis. Interestingly, a xyloglucan-related candidate gene on chromosome 2R, SECCE2Rv1G0126340, was located in close proximity of a SNP significantly associated with stem IVNDFD. Furthermore, quantitative trait loci previously reported in wheat co-localized with significantly associated SNP markers in triticale. Conclusions A collection of 118 winter triticale genotypes combined with DArTseq SNP markers served as a source for identifying 53 MTAs and several candidate genes for forage IVD and related traits through a GWAS approach. Taken together, the results of this study demonstrate that the genetic diversity available in this collection can be further exploited for research and breeding purposes to improve the IVD of triticale forage.

Fusarium wilt, caused by Fusarium oxysporum f. sp. spinaciae, causes a significant challenge on vegetative spinach and seed production. Addressing this issue necessitates continuous research focused on innovative treatments and protocols through comprehensive bioassays. Recent studies have highlighted the potential of plant-based compounds in controlling fungal diseases. The present work aims to conduct a series of experiments, encompassing both in vitro and in planta assessments, to investigate the biocontrol capabilities of different essential oils (EOs) at various application rates, with the ultimate goal of reducing the incidence of Fusarium wilt in spinach. The inhibitory effect of four plant EOs (marjoram, thyme, oregano, and tea tree) was initially assessed on the spore germination of five unknown Fusarium strains. The outcomes revealed diverse sensitivities of Fusarium strains to EOs, with thyme exhibiting the broadest inhibition, followed by oregano at the highest concentration (6.66 μL/mL) in most strains. The tested compounds displayed a diverse range of median effective dose (ED50) values (0.69 to 7.53 µL/mL), with thyme and oregano consistently showing lower ED50 values. The direct and indirect inhibitory impact of these compounds on Fusarium mycelial growth ranged from ~14% to ~100%, wherein thyme and oregano consistently exhibiting the highest effectiveness. Following the results of five distinct inoculation approaches and molecular identification, the highly pathogenic strain F-17536 (F. oxysporum f.sp. spinaciae) was chosen for Fusarium wilt assessment in spinach seedlings, employing two promising EO candidates through seed and soil treatments. Our findings indicate that colonized grain (CG) proved to be a convenient and optimal inoculation method for consistent Fusarium wilt assessment under greenhouse conditions. Seed treatments with thyme and oregano EOs consistently resulted in significantly better disease reduction rates, approximately 54% and 36% respectively, compared to soil treatments (P > 0.05). Notably, thyme, applied at 6.66 µL/mL, exhibited a favorable emergence rate (ERI), exceeding seven, in both treatments, emphasizing its potential for effective disease control in spinach seedlings without inducing phytotoxic effects. This study successfully transitions from in vitro to in planta experiments, highlighting the potential incorporation of EOs into integrated disease management for Fusarium wilt in spinach production.

Intercropping is considered advantageous for many reasons, including increased yield stability, nutritional value and the provision of various regulating ecosystem services. However, intercropping also introduces diverse competition effects between the mixing partners, which can negatively impact their agronomic performance. Therefore, selecting complementary intercropping partners is the key to realizing a well-mixed crop production. Several specialized intercrop breeding concepts have been proposed to support the development of complementary varieties, but their practical implementation still needs to be improved. To lower this adoption threshold, we explore the potential of introducing minor adaptations to commonly used monocrop breeding strategies as an initial stepping stone towards implementing dedicated intercrop breeding schemes. While we acknowledge that recurrent selection for reciprocal mixing abilities is likely a more effective breeding paradigm to obtain genetic progress for intercrops, a well-considered adaptation of monoculture breeding strategies is far less intrusive concerning the design of the breeding programme and allows for balancing genetic gain for both monocrop and intercrop performance. The main idea is to develop compatible variety combinations by improving the monocrop performance in the two breeding pools in parallel and testing for intercrop performance in the later stages of selection. We show that the optimal stage for switching from monocrop to intercrop testing should be adapted to the specificity of the crop and the heritability of the traits involved. However, the genetic correlation between the monocrop and intercrop trait performance is the primary driver of the intercrop breeding scheme optimization process.

Abstract Background Growing interest in agrobiodiversity and sustainable agricultural practices has stimulated debates on diversifying cropping systems, furthering the potential for the reintroduction of underutilised crops. These crops may support multiple ecosystem services and enhance food security and agricultural value chains. This study used a systematic mapping approach to collate and summarise the state of research literature addresses the research question: What is the evidence for ecosystem service provision and economic value of underutilised crops? We focused on oats, triticale, hull-less barley, narrow-leaved lupin, buckwheat and faba beans due to their limited use in Europe, their broad gene pool, ecological benefits, and nutritional value. Method Three academic databases were used to identify research articles investigating the impacts of using the six underutilised crops of interest on outcomes including breeding, agronomic traits, nutrition and health, and economic values. In addition, current and recently completed European projects were searched to identify ongoing relevant research. After screening for relevance, data was extracted from all included articles and projects and imported into a spreadsheet for cross-tabulation and to produce descriptive statistics. Results From an initial 34,522 articles identified by the searches, 1346 relevant primary research articles containing 2229 studies were included. A total of 38 relevant European projects were identified, with 112 research results or goals relating to the six underutilised crops. Faba bean was the most common crop in both European projects and published literature. No current projects had a focus on hull-less barley. Agronomic traits were the most common primary research topic across the crops (56.39%), with oats and faba bean being well researched. Hull-less barley was the least studied crop across all topics. Within sub-topics related to specific ecosystem services, desirable traits, disease, weed and pest control all ranked highly, whilst invertebrate diversity and nitrogen fixation ranked lowest. Conclusion Primary research varies between crops and topics, with hull-less barley receiving the least interest. Key knowledge gaps were identified in all crops across all topics relating to breeding tools, breeding for desirable traits, agronomic traits of buckwheat, narrow-leaved lupin and hull-less barley, inclusion of the crops in human nutrition and health, and the socioeconomics of these crops. Evidence presented in this map could inform further research areas with these crops and aid future policy making for the inclusion of these crops in rotations and practices that could benefit all stakeholders along the food systems value chain.

To increase Europe’s self-sufficiency for protein sources, boosting plant protein production is a prerequisite. Yield variability is one of the main problems regarding the cultivation of protein crops. In this light, cereal–legume intercropping can offer a solution, as well-balanced intercropping systems are less prone to yield variations. Therefore, in this study the effects of (i) species/genotype combination, (ii) intercropping sowing densities and (iii) fertilizer regime were evaluated under Belgian (Northwestern European) conditions over three years (i.e., the 2020–2021, 2021–2022 and 2022–2023 seasons). Regarding the species combinations, winter barley x winter pea, winter wheat x winter faba bean and winter triticale x winter faba bean, it was observed that the best-performing combination varied from year to year depending on the prevailing weather conditions. A reduced sowing density (i.e., 130 seeds/m2 for the cereal partner and 20 seeds/m2 in the case of faba bean or 40 seeds/m2 in the case of pea) was sufficient to achieve competitive yields under the prevailing conditions. Inoculation with commercial Rhizobium strains did not result in an increased yield. Fertilization with one or two nitrogen fractions significantly increased the total yield thanks to a yield increase in the cereal partner; however, as a consequence, the proportion of legumes in the mixture decreased. In conclusion, it can be stated that with the investigated cereal–legume combinations, a competitive yield and qualitative protein yield can be achieved with a reduced fertilizer input.

IntroductionOver the last decade, there has been a growing interest in cereal-legume intercropping for sustainable agriculture. As a result numerous papers, including reviews, focus on this topic. Screening this large amount of papers, to identify knowledge gaps and future research opportunities, manually, would be a complex and time consuming task.Materials and methodsBibliometric analysis combined with text mining and topic modelling, to automatically find topics and to derive a representation of intercropping papers as a potential solution to reduce the workload was tested. Both common (e.g. wheat and soybean) as well as underutilized crops (e.g. buckwheat, lupin, triticale) were the focus of this study. The corpus used for the analysis was retrieved from Web of Science and Scopus on 5th September 2022 and consisted of 4,732 papers.ResultsThe number of papers on cereal-legume intercropping increased in recent years, with most studies being located in China. Literature mainly dealt with the cereals maize and wheat and the legume soybean whereas buckwheat and lupin received little attention from academic researchers. These underutilized crops are certainly interesting to be used as intercropping partners, however, additional research on optimization of management and cultivar’s choice is important. Yield and nitrogen fixation are the most commonly studied traits in cereal-legume intercropping. Last decade, there is an increasing interest in climate resilience, sustainability and biodiversity. Also the term “ecosystem services” came into play, but still with a low frequency. The regulating services and provisioning services seem to be the most studied, in contrast terms related to potential cultural services were not encountered.DiscussionIn conclusion, based on this review several research opportunities were identified. Minor crops like lupin and buckwheat need to be evaluated for their role as intercropping partners. The interaction between species based on e.g. root exudates needs to be further unraveled. Also diseases, pests and weeds in relation to intercropping deserve more attention and finally more in-depth research on the additional benefits/ecosystem services associated with intercropping systems is necessary.

Maize (Zea mays L.) is a staple food for many households in sub-Saharan Africa (SSA) and also contributes to the gross domestic product (GDP). However, the maize yields reported in most SSA countries are very low and this is mainly attributed to biotic and abiotic stresses. These stresses have been exacerbated by climate change which has led to long periods of drought or heavy flooding and the emergence of new biotic stresses. Few reports exist which compile the biotic stresses affecting maize production in SSA. Here, five major biotic stresses of maize in Kenya are presented which are attributed to high yield losses. They include Maize lethal necrosis, fall armyworm, gray leaf spot, turcicum leaf blight and desert locusts. Maize lethal necrosis and fall armyworm are new biotic stresses to the Kenyan maize farmer while gray leaf spot, and turcicum leaf blight are endemic to the region. The invasion by the desert locusts is speculated to be caused by climate change. The biotic stresses cause a reduction in maize yield of 30–100% threatening food security. Therefore, this review focuses on the cause, control measures employed to control these diseases and future prospective. There should be deliberate efforts from the government and researchers to control biotic stresses affecting maize yields as the effect of these stresses is being exacerbated by the changing climate.

Abstract Background Gaining insight into crop diversity, both at the genetic and phenotypic levels, is of prime importance for onion breeding with an enhanced yield and quality in combination with improved resistance to biotic and abiotic stresses. In the current study, 192 different onion plants, representing 16 ecotypes, were characterized using ISSR markers. Results Based on the ISSR marker profile, there was a clear grouping of the plants into 16 different ecotypes. Though the 16 populations originated from the same geographic region in Morocco, a significant genetic diversity was detected. After a genomic characterization, field trials in three different environments in Morocco were laid out. The phenotypic characterization showed that there were always significant differences between ecotypes, and for most traits, there was also a significant environmental effect and a significant interaction between environment and ecotype. The broad-sense heritability (H 2) for the phenotypic traits associated with color (L*, a*, and b*) was the largest (84.2%, 80.6%, 79.2%), demonstrating that color is conditioned primarily by genetic factors. In contrast, the H 2 for yield was the lowest (41.8%), indicating that the environment has a substantial effect on yield. In addition, there was a significant association between the presence/absence of certain bands and various phenotypic traits. Conclusion ISSR markers are a powerful tool in distinguishing onion ecotypes. In addition, significant associations between marker scores and phenotypic traits could be detected, representing particular importance for future breeding programs.

Cereal forages, such as triticale forage, progressively gain interest as alternative crop for maize. The main study objective was to investigate the variation in potential feeding value of triticale forage among maturity stage, growing season and genotype, using total plant and stem fractions. Therefore, near infrared spectroscopy (NIRS) was evaluated as fast screening tool. The prediction ability was good (ratio of prediction to deviation, RPD ≥3.0) for total plant residual moisture, starch, sugars and for stem crude ash (CAsh) and neutral detergent fibre (aNDFom); suitable for screening (2.0 ≤ RPD

In vitro root cultivation techniques based on modified root systems are often used in studies on Arbuscular Mycorrhizal Fungi (AMF). It is a simplified but powerful tool to investigate AMF root colonization and development of the extraradical mycelium. The aim of this study was to establish and characterize the in vitro culture of a Cuban strain of Rhizophagus irregularis (INCAM 11) by using transformed chicory roots. For that, superficially disinfected propagules of R. irregularis were co-culture with the hairy transformed chicory roots on Modified Strullu and Romand (MSR) medium during five months. Spore germination was observed 3-5 days after surface disinfection. The first contact between AMF hyphae and roots occurred 1 - 3 days after germination and a significant production of extensive extraradical mycelium was observed. New spore formation started within 21 - 25 days. After 5 months, 2000 spores could be observed per plate which were able to germinate, colonize, establish and reproduce again spores when associated to young transformed roots of chicory. The most frequent associated microorganism to the in vitro culture of INCAM 11 was isolated and identified as Paenibacillus sp.

Recovery of biobased fertilizers derived from manure to replace synthetic fertilizers is considered a key strategy to close the nutrients loop for a more sustainable agricultural system. This study evaluated the nitrogen (N) fertilizer value of five biobased fertilizers [i.e., raw pig manure (PM), digestate (DIG), the liquid fraction of digestate (LFD), evaporator concentrate (EVA) and ammonia water (AW)] recovered from an integrated anaerobic digestion–centrifugation–evaporation process. The shoot and root growth of maize (Zea mays L.) under biobased fertilization was compared with the application of synthetic mineral N fertilizer, i.e., calcium ammonium nitrate (CAN). The non-invasive technologies, i.e., minirhizotron and unmanned aerial vehicle (UAV) based spectrum sensing, were integrated with the classic plant and soil sampling to enhance the in-season monitoring of the crop and soil status. Results showed no significant difference in the canopy status, biomass yield or crop N uptake under biobased fertilization as compared to CAN, except a lower crop N uptake in DIG treatment. The total root length detected by minirhizotron revealed a higher early-stage N availability at the rooting zone under biobased fertilization as compared to CAN, probably due to the liquid form of N supplied by biobased fertilizers showing higher mobility in soil under dry conditions than the solid form of CAN. Given a high soil N supply (averagely 70–232 kg ha−1) in the latter growing season of this study, the higher N availability in the early growing season seemed to promote a luxury N uptake in maize plants, resulting in significantly (p < 0.05) higher N concentrations in the harvested biomass of PM, LFD and AW than that in the no-N fertilized control. Therefore, the biobased fertilizers, i.e., PM, LFD, EVA and AW have a high potential as substitutes for synthetic mineral N fertilizers, with additional value in providing easier accessible N for crops during dry seasons, especially under global warming which is supposed to cause more frequent drought all over the world.

Essential oils have been used since ancient times in traditional medicine and agri-food science to preserve food, and to combat human diseases. Essential oils (EOs) from clove and rosemary, obtained by hydro-distillation, were analyzed by GC/MS and evaluated for their antifungal activity against strains of Aspergillus tubingensis and A. luchuensis and their effect on ochratoxin A (OTA) production by these fungi. The major constituent of clove EO was eugenol (86.4%), and of rosemary EO was eucalyptol (35.9%). Mycelium growth inhibition assays showed that the EOs had dose-depend inhibition effects, which reached 36.6% for rosemary EO, and 100% for clove EO, at EO concentration of 200 μL L-1. These reductions were different for different Aspergillus strains, the essential oil type, and the strain/oil type interaction. Analyses of OTA in the culture medium extracts of Aspergillus strains was carried out using HPLC-FLD, and was confirmed by LC-MS/MS for positive controls of two OTA-producing strains of A. tubingensis. In general, OTA was reduced (from 45 to 100% reduction), except for two strains of A. tubingensis, where OTA production was stimulated.

Summary The transition to sustainable agriculture and horticulture is a societal challenge of global importance. Fertilization with a minimum impact on the environment can facilitate this. Organic fertilizers can play an important role, given their typical release pattern and production through resource recovery. Microbial fertilizers (MFs) constitute an emerging class of organic fertilizers and consist of dried microbial biomass, for instance produced on effluents from the food and beverage industry. In this study, three groups of organisms were tested as MFs: a high‐rate consortium aerobic bacteria (CAB), the microalga Arthrospira platensis (‘Spirulina’) and a purple non‐sulfur bacterium (PNSB) Rhodobacter sp. During storage as dry products, the MFs showed light hygroscopic activity, but the mineral and organic fractions remained stable over a storage period of 91 days. For biological tests, a reference organic fertilizer (ROF) was used as positive control, and a commercial organic growing medium (GM) as substrate. The mineralization patterns without and with plants were similar for all MFs and ROF, with more than 70% of the organic nitrogen mineralized in 77 days. In a first fertilization trial with parsley, all MFs showed equal performance compared to ROF, and the plant fresh weight was even higher with CAB fertilization. CAB was subsequently used in a follow‐up trial with petunia and resulted in elevated plant height, comparable chlorophyll content and a higher amount of flowers compared to ROF. Finally, a cost estimation for packed GM with supplemented fertilizer indicated that CAB and a blend of CAB/PNSB (85%/15%) were most cost competitive, with an increase of 6% and 7% in cost compared to ROF. In conclusion, as bio‐based fertilizers, MFs have the potential to contribute to sustainable plant nutrition, performing as good as a commercially available organic fertilizer, and to a circular economy.

Summary Purple non‐sulphur bacteria (PNSB) are phototrophic microorganisms, which increasingly gain attention in plant production due to their ability to produce and accumulate high‐value compounds that are beneficial for plant growth. Remarkable features of PNSB include the accumulation of polyphosphate, the production of pigments and vitamins and the production of plant growth‐promoting substances (PGPSs). Scattered case studies on the application of PNSB for plant cultivation have been reported for decades, yet a comprehensive overview is lacking. This review highlights the potential of using PNSB in plant production, with emphasis on three key performance indicators (KPIs): fertilization, resistance to stress (biotic and abiotic) and environmental benefits. PNSB have the potential to enhance plant growth performance, increase the yield and quality of edible plant biomass, boost the resistance to environmental stresses, bioremediate heavy metals and mitigate greenhouse gas emissions. Here, the mechanisms responsible for these attributes are discussed. A distinction is made between the use of living and dead PNSB cells, where critical interpretation of existing literature revealed the better performance of living cells. Finally, this review presents research gaps that remain yet to be elucidated and proposes a roadmap for future research and implementation paving the way for a more sustainable crop production.

With nematicides progressively being banned due to their environmental impact, an urgent need for novel and sustainable control strategies has arisen. Stimulation of plant immunity, a phenomenon referred to as “induced resistance” (IR), is a promising option. In this study, Cucurbitaceae COld Peeling Extracts (CCOPEs) were shown to protect rice (Oryza sativa) and tomato (Solanum lycopersicum) against the root-knot nematodes Meloidogyne graminicola and Meloidogyne incognita, respectively. Focusing on CCOPE derived from peels of melon (Cucumis melo var. cantalupensis; mCOPE), we unveiled that this extract combines an IR-triggering capacity with direct nematicidal effects. Under lab conditions, the observed resistance was comparable to the protection obtained by commercially available IR stimuli or nematicides. Via mRNA sequencing and confirmatory biochemical assays, it was proven that mCOPE-IR in rice is associated with systemic effects on ethylene accumulation, reactive oxygen species (ROS) metabolism and cell wall-related modifications. While no negative trade-offs were detected with respect to plant growth or plant susceptibility to necrotrophic pests or pathogens, additional infection experiments indicated that mCOPE may have a predominant activity toward biotrophs. In summary, the presented data illustrate a propitious potential for these extracts, which can be derived from agro-industrial waste streams.

Pre-anthesis drought is expected to greatly increase yield losses in wheat (Triticum aestivum L.), one of the most important crops worldwide. Most studies investigate the effects of pre-anthesis drought only at maturity. The physiology of the plant before anthesis and how it is affected during drought is less studied. Our study focused on physiological patterns in wheat plants during pre- and post-anthesis drought. To this end, we measured leaf xylem water potential, osmotic potential and water content in different plant parts at a high temporal frequency: every 3 days, three times a day. The experiment started just before booting until 2 weeks after flowering. Drought stress was induced by withholding irrigation with rewatering upon turgor loss, which occurred once before and once after anthesis. The goal was to investigate the patterns of osmotic adjustment, when it is used for protection against drought, and if the strategy changes during the phenological development of the plant. Our data gave no indication of daily osmotic adjustment, but did show a delicate control of the osmotic potential during drought in both leaves and stem. Under high drought stress, osmotic potential decreased to avoid further water loss. Before anthesis, rewatering restored leaf water potential and osmotic potential quickly. After anthesis, rewatering restored water potential in the flag leaves, but the osmotic potential in the stem and flag leaf remained low longer. Osmotic adjustment was thus maintained longer after anthesis, showing that the plants invest more energy in the osmotic adjustment after anthesis than before anthesis. We hypothesize that this is because the plants consider the developing ear after anthesis a more important carbohydrate sink than the stem, which is a carbohydrate sink before anthesis, to be used later as a reserve. Low osmotic potential in the stem allowed turgor maintenance, while the low osmotic potential in the flag leaf led to an increase in leaf turgor beyond the level of the control plants. This allowed leaf functioning under drought and assured that water was redirected to the flag leaf and not used to refill the stem storage.

Organic fertilizers and especially microbial biomass, also known as microbial fertilizer, can enable a paradigm shift to the conventional fertilizer-to-food chain, particularly when produced on secondary resources. Microbial fertilizers are already common practice (e.g. Bloom® and Synagro); yet microbial fertilizer blends to align the nutrient release profile to the plant's needs are, thus far, unexplored. Moreover, most research only focuses on direct fertilization effects without considering added value properties, such as disease prevention. This study has explored three promising types of microbial fertilizers, namely dried biomass from a consortium of aerobic heterotrophic bacteria, a microalga (Arthrospira platensis) and a purple non-sulfur bacterium (Rhodobacter sphaeroides). Mineralization and nitrification experiments showed that the nitrogen mineralization profile can be tuned to the plant's needs by blending microbial fertilizers, without having toxic ammonium peaks. In a pot trial with perennial ryegrass (Lolium perenne L.), the performance of microbial fertilizers was similar to the reference organic fertilizer, with cumulative dry matter yields of 5.6-6.7 g per pot. This was confirmed in a pot trial with tomato (Solanum lycopersicum L.), showing an average total plant length of 90-99 cm after a growing period of 62 days for the reference organic fertilizer and the microbial fertilizers. Moreover, tomato plants artificially infected with powdery mildew (Oidium neolycopersici), a devastating disease for the horticultural industry, showed reduced disease symptoms when A. platensis was present in the growing medium. These findings strengthen the application potential of this novel class of organic fertilizers in the bioeconomy, with a promising match between nutrient mineralization and plant requirements as well as added value in crop protection.

Rye (Secale cereale subsp. cereale L.) has long been exploited as a valuable alternative genetic resource in wheat (Triticum aestivum L.) breeding. Indeed, the introgression of rye genetic material led to significant breakthroughs in the improvement of disease and pest resistance of wheat, as well as a few agronomic traits. While such traits remain a high priority in cereal breeding, nutritional aspects of grain crops are coming under the spotlight as consumers become more conscious about their dietary choices and the food industry strives to offer food options that meet their demands. To address this new challenge, wheat breeding can once again turn to rye to look for additional genetic variation. A nutritional aspect that can potentially greatly benefit from the introgression of rye genetic material is the dietary fibre content of flour. In fact, rye is richer in dietary fibre than wheat, especially in terms of arabinoxylan content. Arabinoxylan is a major dietary fibre component in wheat and rye endosperm flours, and it is associated with a variety of health benefits, including normalisation of glycaemic levels and promotion of the gut microbiota. Thus, it is a valuable addition to the human diet, and it can represent a novel target for wheat–rye introgression breeding.

'Candidatus Liberibacter solanacearum' is an economically important pathogen in the Americas, New Zealand and Europe. The primary objective of this review is to systematically investigate the polymerase chain reaction (PCR)-based methods used for its detection in plant samples. Several databases were searched from the inception of the relevant literature up to August 2018. This review identified 53 studies that met all the inclusion criteria. The performance of the different methods was also compared, however due to data heterogeneity and insufficient evidence on the sensitivity of all assays used, a meta-analysis of the data was not possible. Nonetheless, the review indicates that the rtPCR designed to the 16S ribosomal RNA gene can be routinely employed as a fast, cost-effective, and reliable detection technique in diagnostic laboratories.

Mycotoxin contamination of cereals is a significant health risk for humans and animals, particularly in developing countries. To gain insight into food safety related to agricultural practices, we assessed levels of mycotoxin contamination in 105 samples of food grains raised and stored for consumption by rural households in the post-conflict districts of Kitgum and Lamwo in Northern Uganda. Aflatoxin, ochratoxin and deoxynivalenol (DON) contamination was assessed by quantitative enzyme-linked immunosorbent assay. Total aflatoxin in the foods analyzed varied from nd (not detected) to 68.2 μg/Kg. Ochratoxin ranged from 0.1 to 16.4 μg/Kg. DON ranged from nd to 2606 μg/Kg. The mean concentration of total aflatoxins was significantly higher (P = 0.002) in sorghum than in millet, maize and sesame seeds. Frequency of co-occurrence of two mycotoxins ranged from 8.3 to 100%, with the highest being aflatoxin and ochratoxin in sorghum. Co-occurrence of all three mycotoxins ranged from 8.3 to 35.3%, with the highest again being in sorghum. Mean levels of aflatoxins concentration in sorghum samples were 11.8 μg/Kg, exceeding the Ugandan national regulatory limits of 10 μg/Kg. Furthermore, 46.5% of the sorghum consumed in both districts exceeded this limit, and 86.1% of sorghum samples exceeded the European Union (E.U.) maximum tolerable limit of 4 μg/Kg. The Estimated Daily Intake (EDI) and Hazard Indices (HI) values were in the range of 1.2 × 10−5–91.521 and 1.3 × 10−7 to 0.0059, respectively. In conclusion, our results provide evidence of high levels of mycotoxin contamination and co-occurrence in food grains in Northern Uganda with aflatoxins and ochratoxins at high levels in all the cereal types analyzed. Consumption of cereals cultivated in this region poses no health risk of mycotoxins exposure since HI values obtained were less than 1. Keywords: Aflatoxin, Ochratoxin, Deoxynivalenol, Food grains, Food safety, Uganda

The activation and priming of plant defense upon perception of green leaf volatiles (GLVs) have often been reported. However, information as to which metabolic pathways in plants are affected by GLVs remains elusive. We report the production of reactive oxygen species in the tip of young wheat leaves followed by activation of antioxidant-related enzyme activity. In this study, we aimed to uncover metabolic signatures upon exposure to the GLV Z-3-hexenyl acetate (Z-3-HAC). By using an untargeted metabolomics approach, we observed changes in the phenylpropanoid pathways which yield metabolites that are involved in many anti-oxidative processes. Furthermore, exposure to GLV, followed by infection with Fusarium graminearum (Fg), induced significantly greater changes in the phenylpropanoid pathway compared to a sole Z-3-HAC treatment. Fragmentation of a selection of metabolites, which are significantly more upregulated in the Z-3-HAC + Fg treatment, showed D-glucose to be present as a substructure. This suggests that Z-3-HAC induces early glycosylation processes in plants. Additionally, we identified the presence of hexenyl diglycosides, which indicates that aerial Z-3-HAC is metabolized in the leaves by glycosyltransferases. Together these data indicate that GLV Z-3-HAC is taken up by leaves and incites oxidative stress. This subsequently results in the modulation of the phenylpropanoid pathway and an induction of glycosylation processes.

Acute stunting in children, liver cancer, and death often occur due to human exposure to aflatoxins in food. The severity of aflatoxin contamination depends on the type of Aspergillus fungus infecting the crops. In this study, Aspergillus species were isolated from households’ staple foods and were characterized for different aflatoxin chemotypes. The non-aflatoxigenic chemotypes were evaluated for their ability to reduce aflatoxin levels produced by aflatoxigenic A. flavus strains on maize grains. Aspergillus flavus (63%), A. tamarii (14%), and A. niger (23%) were the main species present. The A. flavus species included isolates that predominantly produced aflatoxins B1 and B2, with most isolates producing a high amount (>20 ug/µL) of aflatoxin B1 (AFB1), and a marginal proportion of them also producing G aflatoxins with a higher level of aflatoxin G1 (AFG1) than AFB1. Some non-aflatoxigenic A. tamarii demonstrated a strong ability to reduce the level of AFB1 by more than 95% when co-inoculated with aflatoxigenic A. flavus. Therefore, field evaluation of both non-aflatoxigenic A. flavus and A. tamarii would be an important step toward developing biocontrol agents for mitigating field contamination of crops with aflatoxins in Uganda.

Rhizospheric microorganisms can alter plant physiology and morphology in many different ways including through the emission of volatile organic compounds (VOCs). Here we demonstrate that VOCs from beneficial root endophytic Serendipita spp. are able to improve the performance of in vitro grown Arabidopsis seedlings, with an up to 9.3-fold increase in plant biomass. Additional changes in VOC-exposed plants comprised petiole elongation, epidermal cell and leaf area expansion, extension of the lateral root system, enhanced maximum quantum efficiency of photosystem II (Fv/Fm), and accumulation of high levels of anthocyanin. Notwithstanding that the magnitude of the effects was highly dependent on the test system and cultivation medium, the volatile blends of each of the examined strains, including the references S. indica and S. williamsii, exhibited comparable plant growth-promoting activities. By combining different approaches, we provide strong evidence that not only fungal respiratory CO2 accumulating in the headspace, but also other volatile compounds contribute to the observed plant responses. Volatile profiling identified methyl benzoate as the most abundant fungal VOC, released especially by Serendipita cultures that elicit plant growth promotion. However, under our experimental conditions, application of methyl benzoate as a sole volatile did not affect plant performance, suggesting that other compounds are involved or that the mixture of VOCs, rather than single molecules, accounts for the strong plant responses. Using Arabidopsis mutant and reporter lines in some of the major plant hormone signal transduction pathways further revealed the involvement of auxin and cytokinin signaling in Serendipita VOC-induced plant growth modulation. Although we are still far from translating the current knowledge into the implementation of Serendipita VOCs as biofertilizers and phytostimulants, volatile production is a novel mechanism by which sebacinoid fungi can trigger and control biological processes in plants, which might offer opportunities to address agricultural and environmental problems in the future.

Late blight caused by the oomycete Phytophthora infestans is considered the biggest threat to potato farming worldwide. For susceptible cultivars, the disease is often managed by frequent applications of fungicides to reduce yield loss. The use of bio-based compounds that interfere with biologically active systems is an innovative strategy for improving disease management. In the present work, the control of P. infestans infection on potatoes by potassium phosphite (KPhi) combined with recommended and reduced doses of active ingredients (Ais) from different fungicides was evaluated. The protective effects of different combinations were initially assessed in vivo and subsequently compared with a greenhouse screening. The active ingredients cyazofamid (CFD) and mancozeb (MCB), used at recommended and reduced doses, were less effective at reducing P. infestans infections than when combined with KPhi. In greenhouse trials, CFD, mandipropamid (MPD) and MCB at recommended doses were the most effective treatments when combined with KPhi; meanwhile, the combination of KPhi with azoxystrobin (AZ), benthiavalicarb-isopropyl/mancozeb (ISO/MCB), and CFD at reduced doses exhibited strong protective activity compared to other similar combinations. This decreased the severity of infection by P. infestans up to ~89%. Greenhouse experiments also demonstrated that a combination of KPhi and CFD at both doses caused the highest reduction in disease severity (up to ~90%) within 35 days of infection. In microplot experiments, KPhi delayed the progression of late blight in susceptible potato varieties; therefore, in the combined treatments AUDPC values were significantly lower than those obtained after applications with CFD doses, providing sufficient protection against late blight. Our data suggest that optimizing the formulation with addition of KPhi could result in a lower recommended dose. This would result in a reduction of the active compounds of the fungicides in potato farming. Furthermore, the impact of KPhi on late blight development makes it a potential component for incorporation into an integrated pest management system.

Abstract Objective Nodding syndrome (NS) is a type of epilepsy characterized by repeated head-nodding seizures that appear in previously healthy children between 3 and 18 years of age. In 2012, during a WHO International Meeting on NS in Kampala, Uganda, it was recommended that fungal contamination of foods should be investigated as a possible cause of the disease. We therefore aimed to assess whether consumption of fungal mycotoxins contributes to NS development. Results We detected similar high levels of total aflatoxin and ochratoxin in mostly millet, sorghum, maize and groundnuts in both households with and without children with NS. Furthermore, there was no significant association between concentrations of total aflatoxin, ochratoxin and doxynivalenol and the presence of children with NS in households. In conclusion, our results show no supporting evidence for the association of NS with consumption of mycotoxins in contaminated foods.

Abstract Background The mycotoxin deoxynivalenol (DON) is highly prevalent in cereals in moderate climates and therefore pigs are often exposed to a DON-contaminated diet. Pigs are highly susceptible to DON and intake of DON-contaminated feed may lead to an altered immune response and may influence the pathogenesis of specific bacterial diseases. Therefore, the maximum guidance level in feed is lowest in this species and has been set at 900 μg/kg feed by the European Commission. This study aimed to determine the effect of in-feed administration of a moderately high DON concentration (1514 μg/kg) on the severity of an experimental Mycoplasma hyopneumoniae (M. hyopneumoniae) infection in weaned piglets. Fifty M. hyopneumoniae-free piglets were assigned at 30 days of age [study day (D)0] to four different groups: 1) negative control group (NCG; n = 5), 2) DON-contaminated group (DON; n = 15), 3) DON-contaminated and M. hyopneumoniae-inoculated group (DONMHYO; n = 15), 4) M. hyopneumoniae-inoculated group (MHYO; n = 15). The piglets were fed the experimental diets ad libitum for five weeks and were monitored during this period and euthanized at day 35 [27 days post infection (DPI)] or 36 (28 DPI). The main parameters under investigation were macroscopic lung lesions (MLL) at euthanasia, respiratory disease score (RDS) from day 8 until day 35, histopathologic lesions and log copies of M. hyopneumoniae DNA detected by qPCR, determined at the day of euthanasia. Results No significant difference was obtained for MLL at euthanasia, RDS (8–35), histopathologic lung lesions and log copies of M. hyopneumoniae DNA in the DONMHYO and MHYO group and consequently, no enhancement of the severity of the M. hyopneumoniae infection could be detected in the DONMHYO compared to the MHYO group. Conclusions Under present conditions, the findings imply that feed contaminated with DON (1514 μg/kg) provided to weaned pigs for five weeks did not increase the severity of an experimental M. hyopneumoniae infection. Further research is needed to investigate the impact of DON on M. hyopneumoniae infections in a multi-mycotoxin and multi-pathogen environment.

Yield losses of crops due to plant pathogens are a major threat in all agricultural systems. In view of environmental issues and legislative limitations for chemical crop protection products, the need to design new environmentally friendly disease management strategies has gained interest. Despite the unique capability of green leaf volatiles (GLVs) to suppress a broad spectrum of plant pathogens, their capacity to control the potato late-blight-causing agent Phytophthora infestans has not been well studied. This study addresses the potential role of the GLV Z-3-hexenyl acetate (Z-3-HAC) in decreasing the severity of late blight and the underlying gene-based evidence leading to this effect. Nine-week-old potato plants (Solanum tuberosum L.) were exposed to Z-3-HAC before they were inoculated with P. infestans genotypes at different time points. These pre-exposed potato plants exhibited slower disease development after infection with the highly pathogenic genotype of P. infestans (EU-13-A2) over time. Qualitative assessment showed that the exposed, infected plants possessed significantly lower sporulation intensity and disease severity compared to the control plants. Hypersensitive response (HR)-like symptoms were observed on the treated leaves when inoculated with different pathogen genotypes. No HR-like lesions were detected on the untreated leaves after infection. It was shown that the transcript levels of several defense-related genes, especially those that are involved in reactive oxygen species (ROS) production pathways were significantly expressed in plants at 48 and 72 h postexposure to the Z-3-HAC. The current work provides evidence on the role of Z-3-HAC in the increased protection of potato plants against late blight through plant immunity and offers new opportunities for the sustainable control of potato diseases.

Maize silage, which in Europe is the main feed for dairy cattle in winter, can be contaminated by mycotoxins. Mycotoxigenic Fusarium spp. originating from field infections may survive in badly sealed silages or re-infect at the cutting edge during feed-out. In this way, mycotoxins produced in the field may persist during the silage process. In addition, typical silage fungi such as Penicillium spp. and Aspergillus spp. survive in silage conditions and produce mycotoxins. In this research, 56 maize silages in Flanders were sampled over the course of three years (2016–2018). The concentration of 22 different mycotoxins was investigated using a multi-mycotoxin liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, and the presence of DNA of three Fusarium spp. (F. graminearum, F. culmorum and F. verticillioides) was analyzed in a selection of these samples using quantitative polymerase chain reaction (qPCR). Every maize silage contained at least two different mycotoxins. Nivalenol (NIV) and deoxynivalenol (DON) were the most prevalent (both in 97.7% of maize silages), followed by ENN B (88.7%). Concentrations often exceeded the EU recommendations for DON and zearalenone (ZEN), especially in 2017 (21.3% and 27.7% of the maize silages, respectively). No correlations were found between fungal DNA and mycotoxin concentrations. Furthermore, by ensiling maize with a known mycotoxin load in a net bag, the mycotoxin contamination could be monitored from seed to feed. Analysis of these net bag samples revealed that the average concentration of all detected mycotoxins decreased after fermentation. We hypothesize that mycotoxins are eluted, degraded, or adsorbed during fermentation, but certain badly preserved silages are prone to additional mycotoxin production during the stable phase due to oxygen ingression, leading to extremely high toxin levels.

Worldwide fungal contamination leads to both quantitative and qualitative grain losses during crop growth and/or storage. A greater proportion of grains contamination with toxins often occurs in sub-Saharan Africa, where control measures are limited. We determined fungal diversity and their toxin production ability in household grains meant for human consumption to highlight the risk of mycotoxin exposure among people from northern Uganda. The study underlines the high diversity of fungi that group into 15 genera; many of which are plant pathogens with toxigenic potential. Fusarium verticillioides was the most common fungal species isolated from household grains. The study also indicates that northern Uganda is favored by a high proportion of toxigenic isolates of F. verticillioides, F. andiyazi, and F. proliferatum, which are characterized by a high fumonisins production capability. The fumonisins production ability was not dependent on the species, grain types, and haplotype group to which the isolates belong. The contamination of most household grains with fungi capable of producing a high amount of toxin shows that most people are exposed to an elevated amount of mycotoxins, which shows the frequent problems with mycotoxins that have been reported in most parts of sub-Saharan Africa.

Ruminal microbiota of cattle are not able to detoxify all mycotoxins. In addition, detoxification can be hampered by adverse ruminal conditions (e.g., low ruminal pH). Hence, in the cattle husbandry, mycotoxin binders and modifiers could be used to prevent animal exposure to mycotoxins. In this study, an in vitro rumen model, including feed matrix, was established as first screening tool to test the efficacy of five products claiming to detoxify mycotoxins. The detoxifiers had different modes of action: (a) binding (three products); (b) enzymatic detoxification of zearalenone (ZEN; one product, ZenA); and (c) bacterial transformation of trichothecenes (one product, BBSH 797). For the mycotoxin binders, the binding to the mycotoxins enniatin B (ENN B), roquefortine C (ROQ-C), mycophenolic acid (MPA), deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEN) were tested at a dose recommended by the manufacturers. The in vitro model demonstrated that all binders adsorbed ENN B to a certain extent, while only one of the binders also partially adsorbed ROQ-C. The binders did not change the concentrations of the other mycotoxins in the ruminal fluid. The enzyme ZenA detoxified ZEN very quickly and prevented the formation of the more toxic metabolite α-zearalenol (α-ZEL), both at normal (6.8) and low ruminal pH (5.8). The addition of BBSH 797 enhanced detoxification of DON and NIV, both at normal and low ruminal pH. The in vitro rumen model demonstrated that the addition of ZenA seems to be a very promising strategy to prevent estrogenic effects of ZEN contaminated feed, and BBSH 797 is efficient in the detoxification of trichothecenes.

Ruminants are generally considered to be less susceptible to the effects of mycotoxins than monogastric animals as the rumen microbiota are capable of detoxifying some of these toxins. Despite this potential degradation, mycotoxin-associated subclinical health problems are seen in dairy cows. In this research, the disappearance of several mycotoxins was determined in an in vitro rumen model and the effect of realistic concentrations of those mycotoxins on fermentation was assessed by volatile fatty acid production. In addition, two hypotheses were tested: (1) a lower rumen pH leads to a decreased degradation of mycotoxins and (2) rumen fluid of lactating cows degrade mycotoxins better than rumen fluid of non-lactating cows. Maize silage was spiked with a mixture of deoxynivalenol (DON), nivalenol (NIV), enniatin B (ENN B), mycophenolic acid (MPA), roquefortine C (ROQ-C) and zearalenone (ZEN). Fresh rumen fluid of two lactating cows (L) and two non-lactating cows (N) was added to a buffer of normal pH (6.8) and low pH (5.8), leading to four combinations (L6.8, L5.8, N6.8, N5.8), which were added to the spiked maize substrate. In this study, mycotoxins had no effect on volatile fatty acid production. However, not all mycotoxins fully disappeared during incubation. ENN B and ROQ-C disappeared only partially, whereas MPA showed almost no disappearance. The disappearance of DON, NIV, and ENN B was hampered when pH was low, especially when the inoculum of non-lactating cows was used. For ZEN, a limited transformation of ZEN to α-ZEL and β-ZEL was observed, but only at pH 6.8. In conclusion, based on the type of mycotoxin and the ruminal conditions, mycotoxins can stay intact in the rumen.

Mycotoxins are well-known contaminants of several food- and feedstuffs, including silage maize for dairy cattle. Climate change and year-to-year variations in climatic conditions may cause a shift in the fungal populations infecting maize, and therefore alter the mycotoxin load. In this research, 257 maize samples were taken from fields across Flanders, Belgium, over the course of three years (2016−2018) and analyzed for 22 different mycotoxins using a multi-mycotoxin liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. DNA of Fusarium graminearum, F. culmorum and F. verticillioides was quantified using the quantitative polymerase chain reaction (qPCR). Multi-mycotoxin contamination occurred frequently, with 47% of samples containing five or more mycotoxins. Nivalenol (NIV) was the most prevalent mycotoxin, being present in 99% of the samples, followed by deoxynivalenol (DON) in 86% and zearalenone (ZEN) in 50% of the samples. Fumonisins (FUMs) were found in only 2% of the samples in the wet, cold year of 2016, but in 61% in the extremely hot and dry year of 2018. Positive correlations were found between DON and NIV and between F. graminearum and F. culmorum, among others. FUM concentrations were not correlated with any other mycotoxin, nor with any Fusarium sp., except F. verticillioides. These results show that changing weather conditions can influence fungal populations and the corresponding mycotoxin contamination of maize significantly, and that multi-mycotoxin contamination increases the risk of mycotoxicosis in dairy cattle.

Ruminants are less susceptible to the effects of mycotoxins than monogastric animals as their rumen microbiota are claimed to degrade and/or deactivate at least some of these toxic compounds. However, the mycotoxin degradation is not well-known yet. For this, a sensitive, specific, and accurate analytical method is needed to determine mycotoxins in the rumen fluid. This study aims to develop and thoroughly validate an ultra-performance liquid chromatography tandem mass spectrometry method for the quantitative determination in the rumen fluid of some of the most relevant mycotoxins found in maize silage in Western Europe: deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEN), mycophenolic acid (MPA), roquefortine C (ROQ-C) and enniatin B (ENN B), as well as their metabolites deepoxy-deoxynivalenol (DOM-1), α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalanone (ZAN), α-zearalanol (α-ZAL) and β-zearalanol (β-ZAL). As feed is often present in the rumen fluid samples, the potential interaction of feed particles with the mycotoxin extraction and analysis was investigated. Extraction recovery and matrix effects were determined in the rumen fluid with and without maize silage. Differences in those parameters between rumen fluid alone and rumen fluid with maize silage highlight the importance of using matrix-matched calibration curves for the quantification of mycotoxins in rumen fluid samples. A cross-validation of the method with rumen fluid and maize silage demonstrates that this analytical method can be applied in research on rumen fluid samples to investigate the degradation of the reported mycotoxins by rumen microbiota if matrix-matched calibration is performed.

The mycotoxin deoxynivalenol (DON), produced by several Fusarium spp., acts as a virulence factor and is essential for symptom development after initial wheat infection. Accumulating evidence shows that the production of this secondary metabolite can be triggered by diverse environmental and cellular signals, implying that it might have additional roles during the life cycle of the fungus. Here, we review data that position DON in the saprophytic fitness of Fusarium, in defense and in the primary C and N metabolism of the plant and the fungus. We combine the available information in speculative models on the role of DON throughout the interaction with the host, providing working hypotheses that await experimental validation. We also highlight the possible impact of control measures in the field on DON production and summarize the influence of abiotic factors during processing and storage of food and feed matrices. Altogether, we can conclude that DON is a very important compound for Fusarium to cope with a changing environment and to assure its growth, survival, and production of toxic metabolites in diverse situations.

It is recognized that mycotoxins can cause a variety of adverse health effects in animals, including altered gastrointestinal barrier function. It is the aim of the present study to determine whether mycotoxin-contaminated diets can alter the oral bioavailability of the antibiotics doxycycline and paromomycin in pigs, and whether a mycotoxin adsorbing agent included into diets interacts with those antibiotics. Experiments were conducted with pigs utilizing diets that contained blank feed, mycotoxin-contaminated feed (T-2 toxin or deoxynivalenol), mycotoxin-contaminated feed supplemented with a glucomannan mycotoxin binder, or blank feed supplemented with mycotoxin binder. Diets with T-2 toxin and binder or deoxynivalenol and binder induced increased plasma concentrations of doxycycline administered as single bolus in pigs compared to diets containing blank feed. These results suggest that complex interactions may occur between mycotoxins, mycotoxin binders, and antibiotics which could alter antibiotic bioavailability. This could have consequences for animal toxicity, withdrawal time for oral antibiotics, or public health.

Fusarium graminearum can cause Giberella Ear Rot (GER) and seedling blight in maize, resulting in major yield losses. Besides GER, the infected grains are consequently contaminated with multiple mycotoxins of F. graminearum. Zearalenone and trichothecenes, such as deoxynivalenol and its acetylated forms, are among the major mycotoxins associated with F. graminearum infection in maize. In the current work, we explored the effect of the endophytic fungal genera of Epicoccum and Sordaria, to control F. graminearum infection in comparative trials with Piriformospora spp., an elusive endophytic genus. Furthermore, we investigated the effect of these endophytes on zearalenone, deoxynivalenol, and 15-acetyldeoxynivalenol levels using in vitro and in planta assays. As plants are endowed with several detoxification mechanisms comprising e.g., glucosylation of trichothecenes, the effect of the isolated fungal endophytes on the deoxynivalenol-3-glucoside level was also assessed. In general, results showed a considerable variability in the antifungal activity, both among species and among isolates within one species. Additionally, the effect on mycotoxin levels was variable, and not necessarily related to the antifungal activity except for zearalenone levels which were consistently reduced by the endophytes. These results highlight the great potential of certain endophytic fungal strains as new biocontrol agents in agricultural science.

Fusarium verticillioides is the most common fungal pathogen associated with maize ear rot in Tanzania. In a two-year trial, we investigated the efficacy of crop protection (insecticide and/or fungicide) and fertilizer (nitrogen and/or phosphorus) treatments in reducing the occurrence of F. verticillioides and its mycotoxins in maize grown in Tanzania. Seasonal differences were seen to have a substantial influence on the incidence and severity of insect infestation, Fusarium ear and kernel rot, biomass of F. verticillioides and contamination with fumonisins. With regard to the application of fertilizers, it was concluded that the impact on maize stalk borer injury, Fusarium symptoms and fumonisin levels was not significant, whereas crop protection significantly reduced maize damage. The application of an insecticide was most effective in reducing insect injury and as a result of the reduced insect injury the insecticide treatment also resulted in a significant decrease in Fusarium symptoms. In 2014, fumonisin levels were also significantly lower in maize treated with an insecticide. Additionally, significant positive correlations between insect damage and Fusarium symptoms were observed. In conclusion, this study clearly shows that application of an insecticide alone or in combination with a fungicide at anthesis significantly reduces insect damage and consequently reduces F. verticillioides infection and associated fumonisin contamination.

Fusarium head blight is a disease caused by a complex of Fusarium species. F. poae is omnipresent throughout Europe in spite of its low virulence. In this study, we assessed a geographically diverse collection of F. poae isolates for its genetic diversity using AFLP (Amplified Fragment Length Polymorphism). Furthermore, studying the mating type locus and chromosomal insertions, we identified hallmarks of both sexual recombination and clonal spread of successful genotypes in the population. Despite the large genetic variation found, all F. poae isolates possess the nivalenol chemotype based on Tri7 sequence analysis. Nevertheless, Tri gene clusters showed two layers of genetic variability. Firstly, the Tri1 locus was highly variable with mostly synonymous mutations and mutations in introns pointing to a strong purifying selection pressure. Secondly, in a subset of isolates, the main trichothecene gene cluster was invaded by a transposable element between Tri5 and Tri6. To investigate the impact of these variations on the phenotypic chemotype, mycotoxin production was assessed on artificial medium. Complex blends of type A and type B trichothecenes were produced but neither genetic variability in the Tri genes nor variability in the genome or geography accounted for the divergence in trichothecene production. In view of its complex chemotype, it will be of utmost interest to uncover the role of trichothecenes in virulence, spread and survival of F. poae.

Aerobic spoilage of silages occurs frequently and is undesirable because it reduces both its nutritive and hygienic quality. Silage inoculants containing heterofermentative lactic acid bacteria, like Lactobacillus buchneri, have already been proven to improve aerobic stability by augmented production of acetic acid, which inhibits yeasts. In this study, the effect of L. buchneri on fermentation characteristics and aerobic stability of alfalfa-ryegrass silage, red clover silage and maize silage was assessed using microsilos. Two dosages, 1×105 and 3×105 cfu g-1 of fresh matter, were compared to untreated control silage. Inoculation with L. buchneri clearly altered the fermentation characteristics of alfalfa-ryegrass and red clover silage, resulting in a significantly higher aerobic stability at both dosages. The effects of L. buchneri inoculation on maize silage were less clear, but nevertheless the aerobic stability of maize silage inoculated with 1×105 cfu g-1 of fresh matter was significantly higher compared to the untreated silage.

The omnipresent group of pesticide adjuvants are often referred to as "inert" ingredients, a rather misleading term since consumers associate this term with "safe". The upcoming new EU regulation concerning the introduction of plant protection products on the market (EC1107/2009) includes for the first time the demand for information on the possible negative effects of not only the active ingredients but also the used adjuvants. This new regulation requires basic toxicological information that allows decisions on the use/ban or preference of use of available adjuvants. In this study we obtained toxicological relevant information through a multiple endpoint reporter assay for a broad selection of commonly used adjuvants including several solvents (e.g. isophorone) and non-ionic surfactants (e.g. ethoxylated alcohols). The used assay allows the toxicity screening in a mechanistic way, with direct measurement of specific toxicological responses (e.g. oxidative stress, DNA damage, membrane damage and general cell lesions). The results show that the selected solvents are less toxic than the surfactants, suggesting that solvents may have a preference of use, but further research on more compounds is needed to confirm this observation. The gene expression profiles of the selected surfactants reveal that a phenol (ethoxylated tristyrylphenol) and an organosilicone surfactant (ethoxylated trisiloxane) show little or no inductions at EC(20) concentrations, making them preferred surfactants for use in different applications. The organosilicone surfactant shows little or no toxicity and good adjuvant properties. However, this study also illustrates possible genotoxicity (induction of the bacterial SOS response) for several surfactants (POEA, AE, tri-EO, EO FA and EO NP) and one solvent (gamma-butyrolactone). Although the number of compounds that were evaluated is rather limited (13), the results show that the used reporter assay is a promising tool to rank commonly used agricultural adjuvants based on toxicity and toxic mode of action data.