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PubMed2025

Identification of an antifungal lipopeptide from Bacillus amyloliquefaciens HAU3 inhibiting the growth of Fusarium graminearum using preparative chromatography and 2D-NMR.

UNLABELLED: The presence of fungal contamination and its mycotoxins in animal feed is pervasive, posing a significant threat to the well-being and performance of animals, as well as the safety of animal-derived food products. In this work, we screened a strain of Bacillus amyloliquefaciens (B. amyloliquefaciens) HAU3 that exhibits efficient antifungal activity against the growth of Fusarium graminearum (F. graminearum). The antifungal activity was detected in the supernatant, with 20% sterile supernatant demonstrating an impressive antifungal rate of 98.46% against F. graminearum. The antifungal activity of the strain was evaluated through spectrum analysis and silage trials, revealing its effective antifungal activity against multiple fungal species. Furthermore, the strain is capable of degrading ZEN and its derivatives. The targeted disruption of fungal mycelial membrane was observed using scanning electron microscopy and transmission electron microscopy. Additionally, staining with the reactive oxygen species (ROS)-sensitive fluorogenic dye DCFH-DA and propidium iodide (PI) revealed that the strain induces accumulation of ROS in fungal mycelia. The active compounds underwent further separation, purification, and detection. The prominent active peak was identified through mass spectrometry and magnetic resonance spectroscopy. The molecular structure of the active compounds was predicted to be lipopeptides composed of 8 amino acids known as fengycin. The whole genome sequencing and informatics analysis unveiled a total of 13 gene clusters responsible for the synthesis of secondary metabolites. The antifungal effects of B. amyloliquefaciens HAU3 are exerted through the synthesis of fengycin, which selectively targets and compromises the integrity of fungal mycelia membranes, thereby making it a potential biocontrol agent for mitigating mycotoxin contamination in feed. IMPORTANCE: Mycotoxin contamination in animal feed, predominantly driven by Fusarium graminearum, represents a persistent threat to livestock health and food chain integrity. Here, we report the isolation of a soil-derived Bacillus amyloliquefaciens HAU3, exhibiting potent and broad-spectrum antifungal activity alongside efficient biodegradation of zearalenone and its derivatives. Mechanistic dissection reveals that fengycin, the principal bioactive metabolite, compromises fungal membrane integrity and elicits intracellular oxidative stress, culminating in hyphal collapse. Genomic profiling uncovers a diverse repertoire of biosynthetic gene clusters underpinning secondary metabolite production. These findings establish strain HAU3 as a promising microbial chassis for the development of next-generation biocontrol strategies aimed at mitigating mycotoxin burden in agroecosystems.

Bacillus amyloliquefaciens↗
PubMed2026

Membrane-associated compartmentalization of zearalenone biosynthetic enzymes and Syn2-associated zearalenone homeostasis in Fusarium graminearum.

Subcellular compartmentalization has attracted increasing attention in fungal secondary metabolism, particularly in the biosynthesis and trafficking of mycotoxins. However, the subcellular site of zearalenone (ZEA) biosynthesis and the mechanisms underlying its export in Fusarium graminearum remain poorly understood. ZEA is a polyketide mycotoxin that poses a serious threat to food safety through contamination of cereal grains and induces severe estrogenic effects in mammals. Its biosynthesis is governed by a dedicated biosynthetic gene cluster consisting of PKS4, PKS13, ZEB1, and ZEB2. In this study, we investigated the subcellular organization of the ZEA biosynthetic machinery and found that key biosynthetic enzymes accumulated in punctate structures that overlapped with small CMAC-positive vacuolar structures and were closely associated with FM4-64-labeled membranes. Furthermore, our results suggest that the syntaxin-like t-SNARE protein Syn2 contributes to extracellular ZEA accumulation and intracellular toxin homeostasis. Disruption of SYN2 abolished visible ZEA crystal formation on the hyphal surface and was associated with increased intracellular ZEA retention. This intracellular accumulation was accompanied by strong induction of the ZEA biosynthetic gene cluster and reduced cellular viability. Moreover, deletion of ZEB2 in the Δsyn2 background abolished ZEA production and restored cell viability, supporting an association between Zeb2-dependent ZEA biosynthesis and the cytotoxic phenotype of the Δsyn2 mutant. Together, our findings suggest a potential link between membrane-associated organization of ZEA biosynthetic enzymes, Syn2-associated ZEA distribution, intracellular toxin homeostasis, and fungal viability. Further studies will be required to define the precise mechanisms underlying ZEA transport and compartment function.

Fusarium graminearum↗
PubMed2026

High-variance phenome database reveals important roles of WD40 proteins in the plant pathogenic fungus Fusarium graminearum.

WD40 is a highly conserved protein domain in eukaryotes that functions as a versatile platform for protein-protein interactions and participates in diverse biological processes. We performed a genome-wide functional analysis of WD40 domain-containing proteins in Fusarium graminearum, a phytopathogenic fungus that causes severe yield losses and mycotoxin contamination in major cereal crops. Comprehensive phenotypic profiling of 119 WD40 gene deletion mutants across 22 phenotypic traits established a systematic WD40 phenome dataset, revealing the broad functional involvement of WD40 proteins and a strong correlation between sexual reproduction and virulence. Protein interaction analyses of selected WD40 proteins revealed diverse WD40-mediated interaction patterns and provided further insights into WD40-mediated protein interactions and their roles in protein complex formation. This study provides a foundation for further characterization of WD40 proteins in filamentous fungi.

Fusarium graminearum↗
PubMed2025

Microbe-induced gene silencing of fungal gene confers efficient resistance against Fusarium graminearum in maize.

UNLABELLED: Small RNAs (sRNAs), the main effectors of RNA interference (or RNA silencing, RNAi), mediate cell-autonomous and non-cell-autonomous gene silencing. The discoveries of trans-kingdom RNAi and interspecies RNAi have accelerated the development of RNAi-based crop protection technologies. Recently, based on interspecies RNAi, a practical technology termed microbe-induced gene silencing (MIGS) without the need of host genetic modification is developed for crop protection against Verticillium dahliae and Fusarium oxysporum in cotton and rice plants. In this study, we utilized MIGS technology to protect maize against Fusarium graminearum, which is responsible for maize stalk rot. An RNAi-engineered Trichoderma harzianum strain, Th-FgPmt2i, was exploited to generate double-stranded RNAs (dsRNAs) to trigger the silencing of the FgPTM2 gene. Our data verify that sRNAs generated from Th-FgPmt2i can silence the FgPMT2 gene via translational inhibition in F. graminearum. We further demonstrated that Th-FgPmt2i has a stronger capacity than does the T. harzianum chassis for protection of maize against F. graminearum. Coupled with our studies on crop protection against V. dahliae and F. oxysporum, our findings reveal that MIGS can be exploited to protect various crops against distinct fungal pathogens and has extensive applicability. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s42994-025-00212-9.

Fusarium graminearum↗
PubMed1979

[Fusarium graminearum in silages. Zearalenone production (author's transl)].

Zearalenone natural contamination of transverse sections of recently opened silages has been observed. In our in vitro experiments, there is no zearalenone production on corn by Fusarium graminearum in anaerobic jar during 3 months. However, on confined atmosphere, zearalenone production is 1.5 to 3 ppm. Contamination of silage is not possible if there are anaerobic conditions in silages.

Animal Feed↗
PubMed2026

The hidden costs of using media to mimic the hosts of Fusarium graminearum: An epigenetic perspectives.

Pathogens dynamically reprogrammed gene expression when transitioning between nonhost and host environments. Epigenetic regulation can provide a rapid and reversible mechanism for this shift. Using published data from Shao et al. (2024) and Zhao et al. (2024), we compare chromatin states in the fungus Fusarium graminearum under in vitro trichothecene mycotoxin (deoxynivalenol) inducing conditions and during wheat spike infection. This revealed striking differences in H3K4me3 and H3K27me3 landscapes with the two datasets showing limited overlap in marked genes and distinct genomic distributions. This indicates that chemically induced cultures only partially replicate the complex signals encountered in planta and emphasise the need for infection-reflective experimental designs to accurately characterise pathogenicity mechanisms.

Fusarium↗
PubMed1979

Preferential binding of radiolabeled zearalenone to a protein fraction of Fusarium roseum graminearum.

Zearalenone [6-(10-hydroxy-6-oxo-trans-1-undecenyl)beta-resorcyclic acid lactone] is a hormone produced by Fusarium spp. which regulates the sexual stage in F. roseum. 3H- and 14C-labeled zearalenone were found to bind preferentially to one of two peaks containing uncharacterized proteins obtained from the cytosol of young mycelium and resolved by gel column chromatography. The proteins were partially purified by successive resolution on Sephadex G-100, Sephadex G-200, and BioGel P-300. Free zearalenone (37%) was reisolated from the purified proteins after resolution by thin-layer chromatography or partitioning with ethyl acetate.

Fungal Proteins↗
PubMed2026

A newly established reverse genetic system for a circular RNA virus reveals new requirements for infection and its biocontrol potential.

Ambiviruses are fungal-infecting circular RNA viruses that uniquely combine viroid-like and viral features, yet the function of their conserved ORF-B protein and their effects on hosts remain unknown, hindered by the lack of a reverse genetics system. Here, we constructed the first infectious cDNA clone of an ambivirus, Fusarium graminearum ambivirus 1 (FgAV1), using a head-to-tail dimer placed downstream of a fungal promoter. FgAV1 was horizontally transmitted via hyphal anastomosis to virus-free Fusarium graminearum strains. Notably, a reverse-oriented dimer construct was also infectious and transmissible. Targeted mutagenesis revealed that both ORF-A- and ORF-B-encoded proteins and the presence of embedded ribozymes are indispensable for ambivirus replication. Our results further demonstrate that FgAV1 infection triggers a fungal RNAi response, extending the antiviral role of host sRNAs to circular RNA viruses. Furthermore, FgAV1 infection suppressed fungal growth and significantly reduced the virulence of F. graminearum on wheat. These findings provide novel insights into ambivirus replication and their potential in fungal pathogen biocontrol.

RNA Viruses↗
PubMed1977

Media for identification of Gibberella zeae and production of F-2-(Zearalenone).

Media are described for the isolaton of Fusarium graminearum in the perithecial state, Gibberella zeae, and for the production of F-2 (zearalenone) by Fusarium species. On soil extract-corn meal agar isolated medium, G. Zeae produced perithecia in 9 to 14 days under a 12-h photoperiod. Species of Fusarium were screened for F-2 production on a liquid medium. From strains that produced F-2, the yields, from stationary cultures of G. zeae and F. culmorum after 12 days of incubation, ranged from 22 to 86 mg/liter. Three strains produced no F-2. Glumatic acid, starch, yeast extract,and the proper ratio of medium volume-to-flask volume were necessary for F-2 synthesis.

Culture Media↗
PubMed2025

Integrative glycomic analysis reveals the crucial role of protein glycosylation in fungal pathogenesis.

Protein glycosylation, a co- and post-translational modification that enhances the functional diversity of the proteome, contributes to various molecular and cellular functions by transferring different polysaccharides onto proteins. During the last decade, the role of glycosylation in plant pathogenic fungi has received significant attention, and glycoproteins are expected to play essential roles in various biological processes including pathogenicity. However, the comprehensive functional genetic analyses for protein glycosylation pathways and glycan structures of phytopathogenic fungi are still largely unknown. Here, we investigated the role of protein glycosylation in Fusarium graminearum by identifying 65 putative genes involved in protein glycosylation and characterizing their functions. Through cell wall component profiling and HPLC analysis, we characterized the overall N- and O-glycan structures in F. graminearum and found that deletion of ALG3 and ALG12 led to truncated core N-glycan structures. Quantitative proteomics analysis revealed that the truncated core N-glycans, generated by the loss of two key enzymes in the initial core N-glycosylation pathway, Alg3 and Alg12, affected a wide range of glycoproteins-including transcription factors, phosphatases, kinases, peroxidases, and other proteins involved in various biological processes-ultimately impacting the virulence of F. graminearum. This study elucidates the complex roles of glycosylation, highlighting the connections among genes involved in the protein glycosylation pathway, glycans, and glycoproteins in regulating the general biology and pathogenicity of F. graminearum. It also would be the fungal glycobiology study initiative.

Glycosylation↗
PubMed1978

Variation of F-2 toxin production on different substrates.

The F-2 producing capacity of one Fusarium graminearum strain (strain No. 13) and of three hyphal tip transverse lines (a b and c) isolated from the original strain and of a mixture of these lines (a b and c) was studied in two successive years on different substrates: oats, barley, wheat, grain mixture and wheat bran. In the first year the original strain produced high amounts of F-2 but was heterogenous in toxin production. The F-2 producing capacity of one of the hyphal tip transverse lines (a) was high and that of two lines (b and c) and of the mixture of the three lines (a+b+c) was poor. In the following year the F-2 producing capacity of the fungal cultures had changed: the F-2 producing capacity of the original strain (No. 13) was greatly reduced, that of one hyphal tip transverse line (b) remained poor and that of two hyphal tip transverse lines (a and c) and of the mixture of the lines clearly increased. The F-2 production was changed in all the substrates and in about the same proportion. In general oats was the best substrate for F-2 production. The possible causes of the changed F-2 production are discussed. The estrogenic effect of F-2 produced in different substrates was studied by using as criteria the uterine weight, vaginal opening and liquid content in the uterus of immature female rats. The effect was in direct proportion to the amount of F-2 ingested by the rats. The substrate was without any influence on the physiological effect of F-2, and in this respect our results deviate from some earlier findings.

Animal Feed↗
PubMed2026

Biocontrol efficacy of Bacillus albus SSR3 for controlling postharvest fungal pathogens and mycotoxin contamination.

Sweetpotato black rot, caused by Ceratocystis fimbriata, is a major postharvest disease that leads to substantial storage losses worldwide. In this study, a salt-tolerant rhizobacterial strain, Bacillus albus SSR3, was isolated from the rhizosphere of sweetpotato grown in saline-alkali soil, with broad-spectrum antagonistic activity against postharvest fungal pathogens. LC-MS/MS analysis revealed diverse bioactive metabolites associated with its antifungal activity. Integrated transcriptomic and metabolomic analyses showed that SSR3 bioactive metabolites extensively reprogrammed fungal metabolism, particularly pathways involved in carbohydrate and amino acid metabolism, antioxidant defense, and energy production. These alterations were accompanied by disruption of cell wall and membrane integrity, excessive reactive oxygen species accumulation, and mitochondrial dysfunction, ultimately inhibiting fungal growth. Here, we also found that SSR3 bioactive metabolites effectively inhibited aflatoxin B1 production by Aspergillus flavus and deoxynivalenol accumulation in Fusarium graminearum. In vivo assays further demonstrated that SSR3 bioactive metabolites significantly reduced sweetpotato black rot severity and effectively limited fungal colonization and mycotoxin contamination in stored agricultural commodities. Collectively, our findings demonstrate that B. albus SSR3 suppresses postharvest fungal pathogens through coordinated metabolic reprogramming, oxidative stress induction, and cellular integrity disruption, highlighting its potential as a sustainable biocontrol agent for postharvest disease management.

Bacillus albus↗
PubMed1977

Incidence of zearalenone producing strains of Fusarium in barley seeds.

Barley grains at harvest in 1971, 1973 and 1974 were mycologically examined for isolates of the fungus Fusarium. These isolates were then examined for the ability to produce zearalenone. In 1974 the grain was also examined for the presence of zearalenone. The most common isolate was F. culmorum, 60 p. 100 of which in 1974, produced zearalenone. Other species isolated which produced this mycotoxin were F. moniliforme, F. graminearum, F. avenaceum, F. nivale and F. sambucinum var. coerulum, the last 3 species have not previously been recorded as producing zearalenone.

Edible Grain↗