The Concentration-Dependent Effects of Essential Oils on the Growth of Fusarium graminearum and Mycotoxins Biosynthesis in Wheat and Maize Grain

The presence of Fusarium fungi and their toxic metabolites in agricultural crops contributes to significant quantitative and qualitative losses of crops, causing a direct threat to human and animal health and life. Modern strategies for reducing the level of fungi and mycotoxins in the food chain tend to rely on natural methods, including plant substances. Essential oils (EOs), due to their complex chemical composition, show high biological activity, including fungistatic properties, which means that they exhibit high potential as a biological plant protection factor. The aim of this study was to determine the fungistatic activity of three EOs against F. graminearum, and the reduction of mycotoxin biosynthesis in corn and wheat grain. All tested EOs effectively suppressed the growth of F. graminearum in concentrations of 5% and 10%. Cinnamon and verbena EOs also effectively reduced the ergosterol (ERG) content in both grains at the concentration of 1%, while at the 0.1% EO concentration, the reduction in the ERG amount depended on the EO type as well as on the grain. The degree of zearalenone (ZEA) reduction was consistent with the inhibition of ERG biosynthesis, while the reduction in deoxynivalenol (DON) was not consistent with this parameter.

Antifungal Peptides and Proteins to Control Toxigenic Fungi and Mycotoxin Biosynthesis

The global challenge to prevent fungal spoilage and mycotoxin contamination on food and feed requires the development of new antifungal strategies. Antimicrobial peptides and proteins (AMPs) with antifungal activity are gaining much interest as natural antifungal compounds due to their properties such as structure diversity and function, antifungal spectrum, mechanism of action, high stability and the availability of biotechnological production methods. Given their multistep mode of action, the development of fungal resistance to AMPs is presumed to be slow or delayed compared to conventional fungicides. Interestingly, AMPs also accomplish important biological functions other than antifungal activity, including anti-mycotoxin biosynthesis activity, which opens novel aspects for their future use in agriculture and food industry to fight mycotoxin contamination. AMPs can reach intracellular targets and exert their activity by mechanisms other than membrane permeabilization. The mechanisms through which AMPs affect mycotoxin production are varied and complex, ranging from oxidative stress to specific inhibition of enzymatic components of mycotoxin biosynthetic pathways. This review presents natural and synthetic antifungal AMPs from different origins which are effective against mycotoxin-producing fungi, and aims at summarizing current knowledge concerning their additional effects on mycotoxin biosynthesis. Antifungal AMPs properties and mechanisms of action are also discussed.

Transformation of Selected Fusarium Toxins and Their Masked Forms during Malting of Various Cultivars of Wheat

This study investigated the impact of malting of six wheat cultivars inoculated with Fusarium culmorum on the dynamics of content changes of selected Fusarium toxins. The grains of all the tested cultivars showed a high content of deoxynivalenol (DON), zearalenone (ZEN), and their derivatives, whereas nivalenol (NIV) and its glucoside were found only in the Legenda cultivar. Our experiments confirmed that the malting process of wheat grain enables the secondary growth of Fusarium, and mycotoxin biosynthesis. The levels of toxins in malt were few-fold higher than those in grain; an especially high increase was noted in the case of ZEN and its sulfate as the optimal temperature and pH conditions for the biosynthesis of these toxins by the pathogen are similar to those used in the grain malting process. This is the first paper reporting that during the malting process, biosynthesis of ZEN sulfate occurs, instead of glycosylation, which is a typical modification of mycotoxins by plant detoxication enzymes.

Effects of Secondary Metabolites from Pea on Fusarium Growth and Mycotoxin Biosynthesis

Fusarium species present ubiquitously in the environment are capable of infecting a wide range of plant species. They produce several mycotoxins targeted to weaken the host plant. While infecting some resistant plants, the host can alter the expression of toxin-related genes and accumulate no/very low amounts of mycotoxins. The ability of the host plant to modulate the biosynthesis of these toxins is entirely depending on the secondary metabolites produced by the plant, often as a part of systemic acquired resistance (SAR). A major role plays in the family of metabolites called phenyl propanoids, consisting of thousands of natural products, synthesized from the phenylalanine or tyrosine amino acids through a cascade of enzymatic reactions. They are also famous for inhibiting or limiting infection through their antioxidant characteristics. The current study was aimed at identifying the differentially expressed secondary metabolites in resistant (Sokolik) and susceptible (Santana) cultivars of pea (Pisum sativum L.) and understanding their roles in the growth and mycotoxin biosynthesis of two different Fusarium species. Although metabolites such as coumarin, spermidine, p-coumaric acid, isoorientin, and quercetin reduced the growth of the pathogen, a higher level of p-coumaric acid was found to enhance the growth of F. proliferatum strain PEA1. It was also noticeable that the growth of the pathogen did not depend on their ability to produce mycotoxins, as all the metabolites were able to highly inhibit the biosynthesis of fumonisin B1 and beauvericin.

Mycosubtilin Produced by Bacillus subtilis ATCC6633 Inhibits Growth and Mycotoxin Biosynthesis of Fusarium graminearum and Fusarium verticillioides

Fusarium graminearum and Fusarium verticillioides are fungal pathogens that cause diseases in cereal crops, such as Fusarium head blight (FHB), seedling blight, and stalk rot. They also produce a variety of mycotoxins that reduce crop yields and threaten human and animal health. Several strategies for controlling these diseases have been developed. However, due to a lack of resistant cultivars and the hazards of chemical fungicides, efforts are now focused on the biocontrol of plant diseases, which is a more sustainable and environmentally friendly approach. In the present study, the lipopeptide mycosubtilin purified from Bacillus subtilis ATCC6633 significantly suppressed the growth of F. graminearum PH-1 and F. verticillioides 7600 in vitro. Mycosubtilin caused the destruction and deformation of plasma membranes and cell walls in F. graminearum hyphae. Additionally, mycosubtilin inhibited conidial spore formation and germination of both fungi in a dose-dependent manner. In planta experiments demonstrated the ability of mycosubtilin to control the adverse effects caused by F. graminearum and F. verticillioides on wheat heads and maize kernels, respectively. Mycosubtilin significantly decreased the production of deoxynivalenol (DON) and B-series fumonisins (FB1, FB2 and FB3) in infected grains, with inhibition rates of 48.92, 48.48, 52.42, and 59.44%, respectively. The qRT-PCR analysis showed that mycosubtilin significantly downregulated genes involved in mycotoxin biosynthesis. In conclusion, mycosubtilin produced by B. subtilis ATCC6633 was shown to have potential as a biological agent to control plant diseases and Fusarium toxin contamination caused by F. graminearum and F. verticillioides.

Current Approaches for Advancement in Understanding the Molecular Mechanisms of Mycotoxin Biosynthesis

Filamentous fungi are able to synthesise a remarkable range of secondary metabolites, which play various key roles in the interaction between fungi and the rest of the biosphere, determining their ecological fitness. Many of them can have a beneficial activity to be exploited, as well as negative impact on human and animal health, as in the case of mycotoxins contaminating large quantities of food, feed, and agricultural products worldwide and posing serious health and economic risks. The elucidation of the molecular aspects of mycotoxin biosynthesis has been greatly sped up over the past decade due to the advent of next-generation sequencing technologies, which greatly reduced the cost of genome sequencing and related omic analyses. Here, we briefly highlight the recent progress in the use and integration of omic approaches for the study of mycotoxins biosynthesis. Particular attention has been paid to genomics and transcriptomic approaches for the identification and characterisation of biosynthetic gene clusters of mycotoxins and the understanding of the regulatory pathways activated in response to physiological and environmental factors leading to their production. The latest innovations in genome-editing technology have also provided a more powerful tool for the complete explanation of regulatory and biosynthesis pathways. Finally, we address the crucial issue of the interpretation of the combined omics data on the biology of the mycotoxigenic fungi. They are rapidly expanding and require the development of resources for more efficient integration, as well as the completeness and the availability of intertwined data for the research community.

Effect of Naturally Occurring Compounds on Fumonisin Production and fum Gene Expression in Fusarium verticillioides

Fusarium verticillioides, one of the most common pathogens in maize, is responsible for yield losses and reduced kernel quality due to contamination by fumonisins (FBs). Two F. verticillioides isolates that differed in their ability to produce FBs were treated with a selection of eight natural phenolic compounds with the aim of identifying those that were able to decrease toxin production at concentrations that had a limited effect on fungal growth. Among the tested compounds, ellagic acid and isoeugenol, which turned out to be the most effective molecules against fungal growth, were assayed at lower concentrations, while the first retained its ability to inhibit toxin production in vitro, the latter improved both the fungal growth and FB accumulation. The effect of the most effective phenolic compounds on FB accumulation was also tested on maize kernels to highlight the importance of appropriate dosages in order to avoid conditions that are able to promote mycotoxin biosynthesis. An expression analysis of genes involved in FB production allowed more detailed insights into the mechanisms underlying the inhibition of FBs by phenolic compounds. The expression of the fum gene was generally down-regulated by the treatments; however, some treatments in the low-producing F. verticillioides strain up-regulated fum gene expression without improving FB production. This study showed that although different phenolic compounds are effective for FB reduction, they can modulate biosynthesis at the transcription level in opposite manners depending on strain. In conclusion, on the basis of in vitro and in vivo screening, two out of the eight tested phenols (ellagic acid and carvacrol) appear to be promising alternative molecules for the control of FB occurrence in maize.

Accessory Chromosome-Acquired Secondary Metabolism in Plant Pathogenic Fungi: The Evolution of Biotrophs Into Host-Specific Pathogens

Accessory chromosomes are strain- or pathotype-specific chromosomes that exist in addition to the core chromosomes of a species and are generally not considered essential to the survival of the organism. Among pathogenic fungal species, accessory chromosomes harbor pathogenicity or virulence factor genes, several of which are known to encode for secondary metabolites that are involved in plant tissue invasion. Accessory chromosomes are of particular interest due to their capacity for horizontal transfer between strains and their dynamic “crosstalk” with core chromosomes. This review focuses exclusively on secondary metabolism (including mycotoxin biosynthesis) associated with accessory chromosomes in filamentous fungi and the role accessory chromosomes play in the evolution of secondary metabolite gene clusters. Untargeted metabolomics profiling in conjunction with genome sequencing provides an effective means of linking secondary metabolite products with their respective biosynthetic gene clusters that reside on accessory chromosomes. While the majority of literature describing accessory chromosome-associated toxin biosynthesis comes from studies ofAlternariapathotypes, the recent discovery of accessory chromosome-associated biosynthetic genes inFusariumspecies offer fresh insights into the evolution of biosynthetic enzymes such as non-ribosomal peptide synthetases (NRPSs), polyketide synthases (PKSs) and regulatory mechanisms governing their expression.