Investigation on the presence of mycotoxins in seed hemp varieties.

In recent years, the cultivation of hemp (Cannabis sativa L.) in Europe has aroused interest among farmers for the potential market opportunities of its products; its cultivation has increased from 20,450 ha in 2015 to 33,020 ha in 2022. Thanks to the great versatility of this crop, there are opportunities in the food and nutraceutical fields (gluten free), cosmetics, energy and industrial sectors. As for several crops, hemp seeds may also be contaminated by fungal pathogens compromising its quality and safety. Considering the recent interest of consumers in using hemp for food purposes, in the present work, a small survey on mycotoxin contamination was carried out during 2018-2022 in hemp seed samples cultivated in Italy for food use. The results showed a limited occurrence of the most common regulated mycotoxins (aflatoxins [AFs], fumonisins [FBs], ochratoxin A [OTA], deoxynivalenol [DON] and zearalenone), but very high levels of alternariols, reaching a maximum value of 38510, 308, 226 and 288 ug/kg for tenuazonic acid [TeA], tentoxin [TEN], alternariol [AOH] and alternariol monoether, respectively. In the same period, an investigation carried out in an experimental field showed that fungal contamination and mycotoxin occurrence were influenced by different meteorological conditions and different varieties.

IoT for Monitoring Fungal Growth and Ochratoxin A Development in Grapes Solar Drying in Tunnel and in Open Air.

Optimisation of solar drying to reduce fungal growth and Ochratoxin A (OTA) contamination is a crucial concern in raisin and currant production. Stochastic and deterministic analysis has been utilized to investigate environmental indicators and drying characteristics. The analysis was performed using two seedless grape varieties (Crimson-red and Thompson-white) that were artificially inoculated with Aspergillus carbonarius during open-air and tunnel drying. Air temperature (T) and relative humidity (RH) were measured and analysed during the drying experiment, along with grape surface temperature (Ts), and water activity (aw). The grape moisture content, fungal colonization, and OTA contamination were estimated, along with the water diffusivity (Deff) and peel resistance (rpeel) to water transfer. Monitoring the surface temperature of grapes is essential in the early detection of fungal growth and OTA contamination. As surface temperature should be carried out continuously, remote sensing protocols, such as infrared sensors, provide the most efficient means to achieve this. Furthermore, data collection and analysis could be conducted through the Internet of Things (IoT), thereby enabling effortless accessibility. The average Ts of the grapes was 6.5% higher in the tunnel than in the open-air drying. The difference between the RH of air and that in the plastic crates was 16.26-17.22%. In terms of CFU/mL, comparison between white and red grapes in the 2020 and 2021 experiments showed that the red grapes exhibited significantly higher values than the white grapes. Specifically, the values for red grapes were 4.3 in 2021 to 3.4 times in 2020 higher compared to the white grapes. On the basis of the conducted analysis, it was concluded that tunnel drying provided some advantages over open-air drying, provided that hygienic and managerial requirements are met.

Metabolomics Insight into the Variety-Mediated Responses to Aspergillus carbonarius Infection in Grapevine Berries.

Limited knowledge regarding the susceptibility of grape varieties to ochratoxin A (OTA)-producing fungi is available to date. This study aimed to investigate the susceptibility of different grape varieties to Aspergillus carbonarius concerning OTA contamination and modulation at the metabolome level. Six grape varieties were selected, sampled at early veraison and ripening, artificially inoculated with A. carbonarius, and incubated at two temperature regimes. Significant differences were observed across cultivars, with Barbera showing the highest incidence of moldy berries (around 30%), while Malvasia and Ortrugo showed the lowest incidence (about 2%). OTA contamination was the lowest in Ortrugo and Malvasia, and the highest in Croatina, although it was not significantly different from Barbera, Merlot, and Sauvignon Blanc. Fungal development and mycotoxin production changed with grape variety; the sugar content in berries could also have played a role. Unsupervised multivariate statistical analysis from metabolomic fingerprints highlighted cultivar-specific responses, although a more generalized response was observed by supervised OPLS-DA modeling. An accumulation of nitrogen-containing compounds (alkaloids and glucosinolates), phenylpropanoids, and terpenoids, in addition to phytoalexins, was observed in all samples. A broader modulation of the metabolome was observed in white grapes, which were less contaminated by OTA. Jasmonates and oxylipins were identified as critical upstream modulators in metabolomic profiles. A direct correlation between the plant defense machinery and OTA was not observed, but the information was acquired and can contribute to optimizing preventive actions.

Enhancing plant defense using rhizobacteria in processing tomatoes: a bioprospecting approach to overcoming Early Blight and Alternaria toxins.

Plant growth-promoting rhizobacteria (PGPR) with antagonistic activity toward plant pathogenic fungi are valuable candidates for the development of novel plant protection products based on biocontrol activity. The very first step in the formulation of such products is to screen the potential effectiveness of the selected microorganism(s). In this study, non-pathogenic rhizobacteria were isolated from the rhizosphere of tomato plants and evaluated for their biocontrol activity against three species of mycotoxin-producing Alternaria. The assessment of their biocontrol potential involved investigating both fungal biomass and Alternaria toxin reduction. A ranking system developed allowed for the identification of the 12 best-performing strains among the initial 85 isolates. Several rhizobacteria showed a significant reduction in fungal biomass (up to 76%) and/or mycotoxin production (up to 99.7%). Moreover, the same isolates also demonstrated plant growth-promoting (PGP) traits such as siderophore or IAA production, inorganic phosphate solubilization, and nitrogen fixation, confirming the multifaceted properties of PGPRs. Bacillus species, particularly B. amyloliquefaciens and two strains of B. subtilis, showed the highest efficacy in reducing fungal biomass and were also effective in lowering mycotoxin production. Isolates such as Enterobacter ludwigii, Enterobacter asburiae, Serratia nematodiphila, Pantoea agglomerans, and Kosakonia cowanii showed moderate efficacy. Results suggest that by leveraging the diverse capabilities of different microbial strains, a consortium-based approach would provide a broader spectrum of effectiveness, thereby signaling a more encouraging resolution for sustainable agriculture and addressing the multifaceted nature of crop-related biotic challenges.

Enhancement of agri-food by-products: green extractions of bioactive molecules with fungicidal action against mycotoxigenic fungi and their mycotoxins.

Introduction: Today, alternative strategies based on the use of bioactive compounds have been proposed to reduce mycotoxin contamination and limit the use of chemical fungicides. Methods: In the present work, several by-products collected from the agri-food chain (i.e., red and white grape marc, red grapevine leaves, grape seeds and stalks, pear, apple, green beans, tomato, and spent hops) were subjected to green extraction protocols (i.e., steam distillation, Ultrasound-Assisted, and Naviglio® extraction) to obtain extracts rich in polyphenols and terpenes. Each extract was assessed in vitro for its ability to inhibit the development of the main mycotoxigenic species and related mycotoxins. Results and Discussion: Aspergillus flavus and A. carbonarius were significantly reduced by pear (from -45 to -47%) and grape marc (from -21 to -51%) extracts, while F. graminearum was shown to be highly influenced by grape stalk, pear, and grape marc extracts (-24% on average). On the contrary, F. verticillioides was inhibited only by pear (-18%) and to a very low and negligible extent by apple (-1%) and green beans (-3%). Regarding the reduction of mycotoxins, the extracts were able to inhibit OTA from 2 to 57%, AFB1 from 5 to 75%, and DON from 14 to 72%. The highest percentages of reduction were obtained against FBs (from 11 to 94%), ZEN (from 17 to 100%), and Alternaria toxins (from 7 to 96%). In conclusion, this work provided promising results for the production of bioactive extracts obtained from agri-food by-products, which could be exploited as potential biofungicides against the development of mycotoxigenic fungi and related mycotoxins.

Reduction of Mycotoxigenic Fungi Growth and Their Mycotoxin Production by Bacillus subtilis QST 713.

The use of chemical pesticides to control the occurrence of mycotoxigenic fungi in crops has led to environmental and human health issues, driving the agriculture sector to a more sustainable system. Biocontrol agents such as Bacillus strains and their antimicrobial metabolites have been proposed as alternatives to chemical pesticides. In the present work, a broth obtained from a commercial product containing Bacillus subtilis QST 713 was tested for its ability to inhibit the growth of mycotoxigenic fungi as well as reduce their mycotoxin production. Mass spectrometry analysis of Bacillus subtilis broth allowed to detect the presence of 14 different lipopeptides, belonging to the iturin, fengycin, and surfactin families, already known for their antifungal properties. Bacillus subtilis broth demonstrated to be a useful tool to inhibit the growth of some of the most important mycotoxigenic fungi such as Aspergillus flavus, Fusarium verticillioides, Fusarium graminearum, Aspergillus carbonarius, and Alternaria alternata. In addition, cell-free Bacillus subtilis broth provided the most promising results against the growth of Fusarium graminearum and Alternaria alternata, where the radial growth was reduced up to 86% with respect to the untreated test. With regard to the mycotoxin reduction, raw Bacillus subtilis broth completely inhibited the production of aflatoxin B1, deoxynivalenol, zearalenone, and tenuazonic acid. Cell-free broth provided promising inhibitory properties toward all of the target mycotoxins, even if the results were less promising than the corresponding raw broth. In conclusion, this work showed that a commercial Bacillus subtilis, characterized by the presence of different lipopeptides, was able to reduce the growth of the main mycotoxigenic fungi and inhibit the production of related mycotoxins.

Functional Study of Lipoxygenase-Mediated Resistance against Fusarium verticillioides and Aspergillus flavus Infection in Maize.

Mycotoxin contamination of maize kernels by fungal pathogens like Fusarium verticillioides and Aspergillus flavus is a chronic global challenge impacting food and feed security, health, and trade. Maize lipoxygenase genes (ZmLOXs) synthetize oxylipins that play defense roles and govern host-fungal interactions. The current study investigated the involvement of ZmLOXs in maize resistance against these two fungi. A considerable intraspecific genetic and transcript variability of the ZmLOX family was highlighted by in silico analysis comparing publicly available maize pan-genomes and pan-transcriptomes, respectively. Then, phenotyping and expression analysis of ZmLOX genes along with key genes involved in oxylipin biosynthesis were carried out in a maize mutant carrying a Mu transposon insertion in the ZmLOX4 gene (named UFMulox4) together with Tzi18, Mo17, and W22 inbred lines at 3- and 7-days post-inoculation with F. verticillioides and A. flavus. Tzi18 showed the highest resistance to the pathogens coupled with the lowest mycotoxin accumulation, while UFMulox4 was highly susceptible to both pathogens with the most elevated mycotoxin content. F. verticillioides inoculation determined a stronger induction of ZmLOXs and maize allene oxide synthase genes as compared to A. flavus. Additionally, oxylipin analysis revealed prevalent linoleic (18:2) peroxidation by 9-LOXs, the accumulation of 10-oxo-11-phytoenoic acid (10-OPEA), and triglyceride peroxidation only in F. verticillioides inoculated kernels of resistant genotypes.

Fusarium verticillioides and Aspergillus flavus Co-Occurrence Influences Plant and Fungal Transcriptional Profiles in Maize Kernels and In Vitro.

Climate change will increase the co-occurrence of Fusarium verticillioides and Aspergillus flavus, along with their mycotoxins, in European maize. In this study, the expression profiles of two pathogenesis-related (PR) genes and four mycotoxin biosynthetic genes, FUM1 and FUM13, fumonisin pathway, and aflR and aflD, aflatoxin pathway, as well as mycotoxin production, were examined in kernels and in artificial medium after a single inoculation with F. verticillioides or A. flavus or with the two fungi in combination. Different temperature regimes (20, 25 and 30 °C) over a time-course of 21 days were also considered. In maize kernels, PR genes showed the strongest induction at 25 °C in the earlier days post inoculation (dpi)with both fungi inoculated singularly. A similar behaviour was maintained with fungi co-occurrence, but with enhanced defence response at 9 dpi under 20 °C. Regarding FUM genes, in the kernels inoculated with F. verticillioides the maximal transcript levels occurred at 6 dpi at 25 °C. At this temperature regime, expression values decreased with the co-occurrence of A. flavus, where the highest gene induction was detected at 20 °C. Similar results were observed in fungi grown in vitro, whilst A. flavus presence determined lower levels of expression along the entire time-course. As concerns afl genes, considering both A. flavus alone and in combination, the most elevated transcript accumulation occurred at 30 °C during all time-course both in infected kernels and in fungi grown in vitro. Regarding mycotoxin production, no significant differences were found among temperatures for kernel contamination, whereas in vitro the highest production was registered at 25 °C for aflatoxin B1 and at 20 °C for fumonisins in the case of single inoculation. In fungal co-occurrence, both mycotoxins resulted reduced at all the temperatures considered compared to the amount produced with single inoculation.

Environmental Conditions Affecting Ochratoxin A during Solar Drying of Grapes: The Case of Tunnel and Open Air-Drying.

Drying optimization, to mitigate fungal growth and Ochratoxin A (OTA) contamination is a key topic for raisin and currant production. Specific indicators of environmental conditions and drying properties were analyzed using two seedless grape varieties (Crimson-red and Thompson-white), artificially inoculated with Aspergillus carbonarius under open air and tunnel drying. The air temperature (T), relative humidity, grape surface temperature (Ts) and water activity throughout the drying experiment, the grapes' moisture content and the fungal colonization and OTA contamination during the drying process and their interactions were recorded and critically analyzed. Drying properties such as the water diffusivity (Deff) and peel resistance to water transfer were estimated. The grapes Ts was 5-7 °C higher in tunnel vs. open air-drying; the infected grapes had higher maximum Ts vs. the control (around 4-6 °C). OTA contamination was higher in tunnel vs. open air-dried grapes, but fungal colonies showed the opposite trend. The Deff was higher in tunnel than in the open air-drying by 54%; the infected grapes had more than 70% higher Deff than the control, differences explained by factors affecting the water transport. This study highlighted CFU and OTA indicators that affect the water availability between red and white grapes during open air and tunnel drying, estimated by the Deff and peel resistance. This raises new issues for future research.

Alternaria toxins in tomato products in Northern Italy in the period 2017-2019.

The occurrence of tenuazonic acid (TeA), alternariol, alternariol monomethyl and tentoxin in tomato-based products was surveyed over the years 2017-2019. A total of 120 samples were collected from retail outlets and tomato-based food producers located in Northern Italy. After extraction and purification through prepacked columns, the mycotoxins were analysed using LC-MS/MS. A widespread contamination of TeA was found in tomato-based products, particularly in concentrated tomato paste. Other Alternaria toxins were not detected. The incidence of TeA was 78.5%, 47.4%, 55.5%, and 76.9% in concentrated tomato paste (maximum value 5955 µg kg-1), tomato sauce, tomato pulp, and ketchup, respectively. The mean level was 243 ± 725 µg kg-1 in concentrated tomato paste and below 30 µg kg-1 in the other tomato products. The contamination varied from year to year. Alternaria spp strains isolated from fresh tomatoes produced mainly TeA. This study provides further Alternaria toxins occurrence data, useful for future risk assessments.

Loss of ZmLIPOXYGENASE4 Decreases Fusarium verticillioides Resistance in Maize Seedlings.

Fusarium verticillioides is one of the most relevant fungal species in maize responsible for ear, stalk and seedling rot, as well as the fumonisin contamination of kernels. Plant lipoxygenases (LOX) synthesize oxylipins that play a crucial role in the regulation of defense mechanisms against pathogens and influence the outcome of pathogenesis. To better uncover the role of these signaling molecules in maize resistance against F. verticillioides, the functional characterization of the 9-LOX gene, ZmLOX4, was carried out in this study by employing mutants carrying Mu insertions in this gene (named as UFMulox4). In this regard, the genotyping of five UFMulox4 identified the mutant UFMu10924 as the only one having an insertion in the coding region of the gene. The impact of ZmLOX4 mutagenesis on kernel defense against F. verticillioides and fumonisin accumulation were investigated, resulting in an increased fungal susceptibility compared to the inbred lines W22 and Tzi18. Moreover, the expression of most of the genes involved in the LOX, jasmonic acid (JA) and green leaf volatiles (GLV) pathways, as well as LOX enzymatic activity, decreased or were unaffected by fungal inoculation in the mutant UFMu10924. These results confirm the strategic role of ZmLOX4 in controlling defense against F. verticillioides and its influence on the expression of several LOX, JA and GLV genes.

Monitoring Phenolic Compounds in Rice during the Growing Season in Relation to Fungal and Mycotoxin Contamination.

Total phenolic content (TPC) and several phenolic acids present in rice grains were compared with fungal infection and mycotoxin presence throughout the growing season. Samples of 4 rice varieties were collected in 2018 and 2019 at 3 different plant phenological stages. Total fungal and main mycotoxigenic fungi incidence were checked and mycotoxin content was analysed. On the same samples, TPC and the concentration of 8 main phenolic acids (chlorogenic acid, caffeic acid, syringic acid, 4-hydroxybenzoic acid (4-HBA), p-coumaric acid, ferulic acid, protocatecuic acid and gallic acid) were measured. The results showed significant differences between years for both fungal incidence and mycotoxin presence. In 2018 there was a lower fungal presence (42%) than in 2019 (57%) while, regarding mycotoxins, sterigmatocystin (STC) was found in almost all the samples and at all growing stages while deoxynivalenol (DON) was found particularly during ripening. An interesting relationship was found between fungal incidence and TPC, and some phenolic acids seemed to be more involved than others in the plant defense system. Ferulic acid and protocatecuic acid showed a different trend during the growing season depending on fungal incidence and resulted to be positively correlated with p-coumaric acid and 4-HBA that seem involved in mycotoxin containment in field.

Co-Occurrence of Moniliformin and Regulated Fusarium Toxins in Maize and Wheat Grown in Italy.

The co-occurrence of moniliformin (MON), fumonisins (FBs), and deoxynivalenol (DON) was evaluated in maize, durum, and common wheat grown in different experimental fields located in several Italian regions. MON was quantified using a LC-MS/MS method adding lanthanum ions in the mobile phase. In maize, MON contamination was widespread and considerable; the toxin was detected in almost all the samples (95.1%) and exceeded 500 and 1000 µg kg-1 in 42.0% and in 18.5% of samples, respectively. Significant positive correlation was found between MON and FB contamination levels. When there were not droughty climate conditions, a positive significant correlation was found between growing degree days (GDD) and MON values. In wheat, MON contamination was not widespread like in maize and it was lower in common wheat than in durum wheat. In durum wheat, MON was detected in 45.0% of the samples with only 6 samples (7.5%) exceeding 500 µg kg-1, while in common wheat the toxin was detected above the LOD in 18.7% of samples exceeding 100 µg kg-1 in only two samples (2.5%). No correlation was found with DON contamination. Climate conditions influenced both MON and DON occurrence.

Aspergillus flavus and Fusarium verticillioides Interaction: Modeling the Impact on Mycotoxin Production.

The influence of climate change on agricultural systems has been generally accepted as having a considerable impact on food security and safety. It is believed that the occurrence of mycotoxins will be greatly affected by future climate scenarios and this has been confirmed by recent data. Temperature (T) and CO2 increases, variation in rain intensity and distribution, as well as extreme weather events, affect the dominant fungal species in different ways, depending on their ecological needs. Therefore, the aim of this work was to study Aspergillus flavus (Af) and Fusarium verticillioides (Fv) co-occurrence in vitro in order to collect quantitative data on the effect of fungal interaction on growth and mycotoxin production and develop functions for their description. Experimental trials were organized with the cited fungi grown alone or together. They were incubated at different T regimes (10-40°C, step 5°C) for 21 days. Fungal growth was measured weekly, while AFs and FBs were quantified at the end of the incubation period. Temperature and incubation time significantly affected fungal growth both for Af and Fv (p ≤ 0.01), and a significant interaction between T and the presence of one versus both fungi influenced the amount of AFs and FBs produced. Each fungus was affected by the presence of the other fungus; in particular, Af and Fv showed a decrease in colony diameter of 10 and 44%, respectively, when they were grown together, compared to alone. The same influence was not found for mycotoxin production. In fact, the dynamics of toxin production in different temperature regimes followed a comparable trend with fungi grown alone or together, but a significant impact of inoculum × temperature interaction was highlighted. Fungal growth and toxin production in different T regimes were well described, both for AFs and FBs, by a Bete function. These results are the first attempt to model mycotoxigenic fungal co-occurrence under several T regimes; this is essential in order to improve effective prediction of growth and mycotoxin production by such fungi.

Efficacy of Azoxystrobin on Mycotoxins and Related Fungi in Italian Paddy Rice.

The efficacy of azoxystrobin was evaluated in the presence of mycotoxigenic fungi and relative mycotoxins in Italian paddy rice during the growing season in the field. Three experimental fields were considered and the applied experimental design was a strip plot with three replicates; rice samples were collected at four different growing stages. The efficacy of the fungicide treatment on rice fungal population was demonstrated with around 20% less total fungal incidence in sprayed samples compared to untreated ones; the same decrease was noted also in Fusarium spp. species but not in Aspergillus versicolor. Of the mycotoxins considered, ochratoxin A (OTA) and aflatoxins (AFBs) were never detected, deoxynivalenol (DON) was found in 46% of samples at levels always lower than 100 µg/kg, while sterigmatocystin (STC) occurred in all the paddy rice samples collected after flowering, with a maximum value of 15.5 µg/kg. Treatment with azoxystrobin was not effective in reducing DON contamination, but it had an important and significant effect on STC content, showing a decrease of 67% in the sprayed samples.

Impact of Fungi Co-occurrence on Mycotoxin Contamination in Maize During the Growing Season.

Maize is a possible host of many fungi, some of them able to produce different mycotoxins. Few studies exist on co-occurring fungi and resulting multi-mycotoxin contamination in field; for this reason, in field trials were conducted in two consecutive years to verify fungal incidence and mycotoxin production in the case of the co-occurrence of the three main mycotoxigenic fungi of maize in Italy: Aspergillus flavus, Fusarium verticillioides, and Fusarium graminearum able to produce, respectively, aflatoxin B1 (AFB1), fumonisins (FBs), and deoxynivalenol (DON). Artificial inoculation was done after silk emergence of maize and samples were collected with a 2 week schedule up to harvest time (four samplings). Fungal interaction resulted as playing a role for both fungal incidence and mycotoxins production, as did weather conditions too. Main interactions were noted between A. flavus and F. verticillioides, and between F. verticillioides and F. graminearum. In particular, as a result of fungal co-occurrence, AFB1 resulted stimulated by F. graminearum presence while no effects were noted in FBs and DON in case of F. verticillioides-F. graminearum co-occurrence. Interestingly, the co-presence of A. flavus significantly reduced both FB and DON production.

Lipids as Key Markers in Maize Response to Fumonisin Accumulation.

The present field study offers new insights into the role played by plant lipid pathways in the modulation of fumonisin accumulation in maize. Untargeted metabolomics was applied to better understand the multifactorial plant-pathogen-interaction mechanisms, including host resistance. Our results showed a significant influence from the hybrid genotype and the environmental growing conditions on fumonisin accumulation. A total of 25 significant metabolites have been identified, with glycerophospholipid and linoleic acid metabolism as the main pathways affected by the plant-pathogen interactions. This evidence highlighted the crucial role played by lipid signaling as an integrated part of the complex regulatory network in plants.

Mycotoxins and Related Fungi in Italian Paddy Rice During the Growing Season and Storage.

Mycotoxigenic fungi and relative mycotoxins contamination were monitored in Italian paddy rice samples both in field during the growing season and the first five months of storage. Three experimental fields, nine rice varieties and three sowing densities were considered; then, different lots of paddy rice were stored in warehouses at different temperature regimes. Fusarium spp. and Aspergillus spp. were found to be the fungi most likely to produce mycotoxins throughout the growing season. In particular, A. flavus and A. niger were found only rarely both in field and in post-harvest, while A. versicolor was always present although in low concentrations. Penicillium spp. strains were isolated sporadically and were found to be irrelevant in Italian rice fungal contamination. Sterigmatocystin (STC) was the main mycotoxin found in Italian rice, while aflatoxin (AFB1), deoxynivalenol (DON) and ochratoxin A (OTA) were rarely detected. Contamination generally increased from post-flowering to ripening; considering rice varieties, significant differences (p ≤ 0.01) were found in fungal contamination and STC production; no differences were observed between sowing densities. During storage, an increase in STC content was observed in higher temperature regimes, while all the other considered mycotoxins remained unchanged. These results indicated that contamination by STC, an emerging mycotoxin not legislatively regulated by the European Union, can be relevant in rice.

A Genome Wide Association Study Reveals Markers and Genes Associated with Resistance to Fusarium verticillioides Infection of Seedlings in a Maize Diversity Panel.

Fusarium verticillioides infects maize, causing ear rot, yield loss and contamination by fumonisin mycotoxins. The fungus can be transmitted via kernels and cause systemic infection in maize. Maize resistance to the fungus may occur at different developmental stages, from seedling to maturity. Resistance during kernel germination is part of the plant-pathogen interaction and so far this aspect has not been investigated. In the present study, a genome wide association study (GWAS) of resistance to Fusarium during the seedling developmental stage was conducted in a maize diversity panel using 226,446 SNP markers. Seedling germination and disease phenotypes were scored on artificially inoculated kernels using the rolled towel assay. GWAS identified 164 SNPs significantly associated with the traits examined. Four SNPs were associated with disease severity score after inoculation, 153 were associated with severity in asymptomatic kernels and 7 with the difference between the severity ratings in inoculated and non-inoculated seeds. A set of genes containing or physically near the significant SNPs were identified as candidates for Fusarium resistance at the seedling stage. Functional analysis revealed that many of these genes are directly involved in plant defense against pathogens and stress responses, including transcription factors, chitinase, cytochrome P450, and ubiquitination proteins. In addition, 25 genes were found in high linkage disequilibrium with the associated SNPs identified by GWAS and four of them directly involved in disease resistance. These findings contribute to understanding the complex system of maize-F. verticillioides and may improve genomic selection for Fusarium resistance at the seedling stage.

Fate of mycotoxins and related fungi in the anaerobic digestion process.

In this study, the possibility to manage maize contaminated with aflatoxins and fumonisins for the production of biogas was considered. This is a priority in the climate change scenario that is expected to increase the occurrence of aflatoxins in maize. The results clearly underline how the anaerobic digestion process used in biogas plants is able to reduce aflatoxin contamination, mainly when highly contaminated maize is used for feeding the biodigestors without affecting the efficiency of methane production. In particular, the higher aflatoxin contamination is, the higher is mycotoxin reduction during biodigestion, with reductions up to 95% in digestate. The co-occurring mycotoxins, fumonisins, were also reduced by around 15%. The vitality of mycotoxin producing fungi was also significantly reduced. Biogas production is therefore suggested as a good alternative use for uncompliant maize.