ABSTRACTThe chestnut blight fungus Cryphonectria parasitica and its hypovirus constitute a valuable model for investigating fungal pathogenesis and cross‐kingdom virus–host interplay. To investigate how hypovirus regulates protein function at the phosphorylation level in C. parasitica, we performed a comparative phosphoproteomic analysis in the fungus with or without Cryphonectria hypovirus 1 (CHV1) infection. Comparative profiling between the wild‐type (EP155) and hypovirus‐infected (EP155/CHV1‐EP713) strains revealed 700 differentially phosphorylated sites (174 upregulated, 526 downregulated). Among these, the serine 896 and 897 sites on the endoplasmic reticulum (ER) stress‐sensing protein CpIre1 drew our particular attention, as hypovirus‐induced phosphorylation targets. Western blot analysis showed that virus‐encoded p29, p40, and p48 proteins could promote CpIre1 phosphorylation. Site‐specific mutagenesis revealed that Ser‐896 and Ser‐897 are essential for CpIre1 phosphorylation, which regulates fungal phenotypic traits, virulence, and stress tolerance in C. parasitica. Reverse‐transcription‐quantitative PCR analysis of the ER stress marker genes CpHac1 and CpBip1 confirmed that CpIre1 and its phosphorylation are essential for a functional ER stress response. Notably, hypovirus replication was significantly impaired in phospho‐deficient CpIre1 mutants, showing about 40% reduction in viral RNA accumulation, whereas phospho‐mimic mutants maintained wild‐type levels of viral RNA. This indicates that efficient hypovirus accumulation requires functional phosphorylation of CpIre1. Our findings demonstrate that hypovirus‐induced phosphorylation of CpIre1 modulates fungal ER homeostasis, pathogenicity, and viral RNA accumulation, thereby revealing a mechanism through which the virus reprogrammes its host via targeted post‐translational modification.

ABSTRACT Malassezia globosa plays a crucial role as part of the human skin’s mycobiome. However, this yeast has been detected in other niches, such as the gut. Despite being commensal, the pathogenic link in several dermatological conditions, but recently, chronic diseases such as cancer, Crohn’s disease, and Parkinson’s disease, among others, have been explored. Lipids can be involved in fungal pathogenesis, and this yeast is characterized by a significant lipid metabolic versatility, with a lack of the complex fatty acid synthase (FAS) required for the de novo synthesis of fatty acids, as it relies on lipase-releasing enzymes. Here, we assess lipid dynamics (lipids consumed vs. lipids secreted) using lipidomic analysis in the supernatant of mDixon media during two growth phases. 87 lipids within 17 classes of lipids were identified in three different lipid uptake-secretion patterns. Some lipids were characteristic, including the presence of glycochenodeoxycholic acid, glycerophospholipids (such as phosphocholine), cardiolipins, and sphingolipids (such as Cer-PI). Interestingly, sterols, bile acids, cholic acid and its derivates, some phosphocholines, fatty acyls, and cardiolipins were lipids consumed over time. The dynamic consumption of these lipids could presume an intriguing role in the metabolism of lipid processes in this yeast that could determine the interaction process and its pathogenic role.

One of the most notable aspects of Aspergillus fumigatus, and related to its dynamic adaptation, is its ability to form biofilm and produce a wide variety of secondary metabolites. The aim of this study is to advance the characterization of biofilms generated by different A. fumigatus strains across their developmental stages and analytically evaluate their structure and composition and their relationship with secondary metabolism activation. An in vitro biofilm model was standardized to investigate structural and analytical differences among strains isolated from distinct clinical settings and associated with different pathologies. We found that all tested strains could form biofilms; however, the characteristics of these structures-including total biomass, cellular viability and overall structure-varied markedly among strains under the evaluated conditions. Strains isolated from cystic fibrosis patients exhibited distinct behaviors in most conducted assays compared to other strains. These findings provide new insights into the variability of biofilm composition and may contribute to a better understanding of the role of biofilms in fungal pathogenesis, persistence and treatment resistance.

Poly(A) tail shortening by deadenylases is a central checkpoint linking mRNA fate to eukaryotic development, yet its impact on fungal pathogenesis remains unexplored. Here, we uncover that the Pan2-Pan3 deadenylase complex is a master regulator of infection in the rice blast fungus Magnaporthe oryzae. Pan2 and Pan3 form a catalytically active complex that localizes to P-bodies and globally trims poly(A) tails to enforce mRNA quality control. Deletion of either or both subunits abolishes this quality-control checkpoint, causing severe virulence loss due to arrested appressorium maturation, disrupted glycogen/lipid mobilization, and impaired autophagy. Integrating poly(A)-seq and transcriptome profiling reveals 390 mRNAs whose poly(A) tails are ≥ 5 nt longer and whose steady-state levels are elevated in the Δpan2Δpan3 mutant; among them, ATG5, GLS2, and DES1-key genes governing autophagy, ER quality control, and ROS detoxification respectively-are directly deadenylated by Pan2-Pan3. Loss of deadenylation destabilizes these mRNAs and reduces their protein output, thereby crippling infection. Our findings establish Pan2-Pan3 complex-mediated deadenylation as an essential post-transcriptional layer that orchestrates fungal virulence through stringent mRNA quality control, offering a novel target for crop protection.

Rice blast, caused by Magnaporthe oryzae, remains a primary threat to global food production, making the discovery of novel antifungal targets imperative. The Target of Rapamycin (TOR) signaling network is a prime target for therapeutic intervention, but the function of its central components in M. oryzae is not fully understood. In this study, we found that MoLst8 is indispensable for fungal growth, development, and virulence in M. oryzae. Genetic deletion of MoLST8 impairs fungal growth, conidiation, and virulence by disrupting TOR signaling, triggering excessive autophagy, and damaging plasma membrane integrity due to dysregulated lipid homeostasis. In addition, given the crucial role of MoAtg8 in M. oryzae, we identified a natural product Theaflavin-3,3'-digallate (TF3) that binds MoLst8 potently and suppresses fungal pathogenesis. Uniquely, TF3 functions as a branch-selective inhibitor, targeting TORC1 rather than TORC2. Our findings establish MoLst8 as a critical vulnerability and present TF3 as a lead compound for developing novel, target-specific fungicides.

Autophagy is an essential intracellular degradation and recycling system for macromolecules and organelles, crucial for cell survival under nutrient stress conditions. In fungi, the genes involved in vesicle assembly during autophagy have been extensively characterized. However, in the pathogen Cryptococcus neoformans, the autophagy pathway remains less understood, particularly regarding its potential connections with virulence and pathogenicity. Our previous work identified Gpp2 as a key player in the biosynthesis of the sulfur-containing amino acid methionine. Through transcriptomic analysis, we observed that through transcriptomic analysis, we observed that deletion of GPP2 in C. neoformans leads to the repression of several core autophagy genes (ATG1, ATG2, ATG4, ATG15, VPS15, and VPS30), likely as an indirect consequence of altered methionine metabolism, while upregulating PEP4 expression. Since methionine is known to repress autophagy in Saccharomyces cerevisiae, we hypothesized that this amino acid might similarly regulate autophagy in C. neoformans. Our experiments demonstrated that both endogenous and exogenous methionine inhibit the expression of autophagy-related genes not only in the wild-type H99 strain but also in gpp2Δ and gpr4Δ mutant strains. Intriguingly, we found that GPR4 deletion creates a mutant unable to sense exogenous methionine, consequently releasing the repression of autophagy genes. Furthermore, microscopic analyses revealed that methionine supplementation substantially reduces autophagosome formation compared to methionine-deprived conditions. These results lead us to conclude that methionine biosynthesis regulation in gpp2Δ strains affects autophagy similarly to S. cerevisiae; GPR4 encodes a functional methionine receptor in C. neoformans; and methionine availability directly impacts autophagic flux, where the methionine receptor Gpr4 links extracellular amino acid availability to the intracellular control of autophagy likely via the Cys3/Gpp2 regulatory axis. This work provides crucial insights into the metabolic regulation of autophagy in pathogenic fungi and opens new avenues for understanding fungal pathogenesis mechanisms.

Candida albicans causes severe mucosal and systemic infections, with hypha formation playing a key role in its virulence. Hyphal invasion via endocytosis is mediated predominantly through interactions between Als3p and the epidermal growth factor receptor (EGFR). Subsequent EGFR activation by candidalysin, a hyphal-secreted cytolytic peptide toxin encoded by the ECE1 gene, induces receptor signaling and immune responses. While EGFR ubiquitination critically regulates receptor trafficking and signaling, its involvement during C. albicans infection has remained unexplored. Here, we demonstrate that C. albicans induces EGFR ubiquitination, leading to altered trafficking and lysosomal degradation in an ECE1- and ALS3-dependent manner. This correlates with changes in EGFR ligand expression, adaptor recruitment, and protein ubiquitination in oral epithelial cells. In a mouse model of oropharyngeal candidiasis, wild-type C. albicans and ece1Δ/Δ and als3Δ/Δ mutant strains were found to differentially regulate Egfr expression, ubiquitin pathway-associated genes, and protein ubiquitination. Furthermore, conditional EGFR knockout was protective during infection. Together, our findings reveal that C. albicans infection modulates the host ubiquitin system, including direct effects on EGFR, highlighting a novel aspect of host-fungal interactions.IMPORTANCECandida albicans is a common fungal pathogen that causes both mucosal infections, such as thrush, and life-threatening systemic diseases. A key step in infection is the fungus invading epithelial tissues and activating the host epidermal growth factor receptor (EGFR). We discovered that C. albicans alters how EGFR is regulated by inducing its ubiquitination, a modification that leads to receptor degradation. This process depends on two major fungal virulence factors: the adhesin Als3p and Ece1p, the polypeptide that contains the candidalysin toxin. The fungus also broadly increases protein ubiquitination in oral epithelial cells. In a mouse model of oral infection, loss of EGFR in epithelial tissues reduced disease severity, suggesting that the receptor helps the fungus establish infection. These findings reveal a previously unrecognized strategy by which C. albicans manipulates protein ubiquitination and regulation in epithelial cells, offering new insights into fungal pathogenesis and potential therapeutic approaches that target host pathways.

The agave wilt associated with Fusarium oxysporum (Fox) is a major disease of blue agave (Agave tequilana Weber var. azul), used to produce “Tequila” in Mexico. Little is known about the A. tequilana-F. oxysporum interaction yet understanding defense mechanisms against the pathogen is necessary for control strategies. During early Fox infection, plants trigger defense mechanisms to interrupt the compatible interaction, while Fox’s pathogenesis mechanism interacts with plant response. This study evaluated plant defense mechanisms induced by Fox in A. tequilana and their interaction with fungal pathogenesis. For this, an A. tequilana pathogenic strain (FPA), and the non-A. tequilana pathogenic strains FNPA and FOL were utilized. Early defense mechanisms evaluated were hypersensitive response (HR) and cell wall strengthening in agave roots. Resistance mechanisms evaluated included pathogenesis-related proteins (PR proteins), phytoanticipins and phytoalexins. For early defense, induced HR was greater with FPA than other strains. Cell wall strengthening was found in agave roots, plants responded differentially to different strains. Initial response to FPA and FOL was similar in PR proteins, phytoalexins and phytoanticipins production. However, the response differentiated with FOL over time, indicating an incompatible interaction. The study identified effective and ineffective defense responses of A. tequilana to Fox infection, where FPA exhibited compatibility and caused unregulated ROS and PCD, early inhibition of PR activity, extensive lignification, and saponin detoxification. In contrast, this study unveiled incompatible interactions (FNPA and FOL) because of limited colonization, localized HR with suppressed ROS, early and sustained POX activation, significant callose accumulation, moderate lignification, and phenol–saponin dynamics that help in tissue containment and recovery.

Accurate identification and characterization of emerging fungal pathogens are essential for improving diagnosis and treatment. Sarocladium spinificis, though not widely recognized as a human pathogen, exhibits traits suggesting a potential role in opportunistic infections. This study provides the first detailed morphological, phylogenetic, and genomic characterization of clinical S. spinificis isolates, highlighting its ability to survive at body temperature and its antifungal resistance. Additionally, we demonstrate that these strains inhibit Coccidioides posadasii, the agent of Valley fever, suggesting ecological competition or antifungal properties. These findings contribute to understanding fungal interactions in clinical and environmental settings, with implications for fungal pathogenesis and antifungal strategies. By uncovering new aspects of S. spinificis biology, this work expands knowledge of Sarocladium species in human health and lays the foundation for future research on its ecological role, pathogenic potential, and therapeutic applications.

The growing number of patients susceptible to invasive Candida albicans infections has intensified the need for new antifungal targets in pathways essential for fungal growth and pathogenesis. Among these pathways, phosphate homeostasis has emerged as a significant determinant of virulence, yet how phosphate availability shapes cell wall structure in response to host-derived oxidative stress remains unclear. During commensal growth, C. albicans cells typically enjoy phosphate repletion and a neutral redox environment. Transitioning to invade host tissues, they simultaneously experience phosphate deprivation and intense extrinsic oxidative stress. Here, we employ solid-state NMR to render details of cell wall remodeling in response to oxidative stress, in its dependence on phosphate. Phosphate deprived cells remodel the rigid wall core and reduce hydration and polymer mobility in the absence of oxidative stress. During hydrogen peroxide exposure, highly mobile outer polysaccharides are primary interactors. In wildtype cells, some of these polymers are recruited into the rigid core, reinforcing the wall scaffold, whereas phosphate transport mutants fail to undergo this remodeling. These findings establish phosphate acquisition as a component of oxidative defense and link nutrient sensing and -availability to the mechanical resilience of the fungal cell wall, revealing an architectural vulnerability with relevance for antifungal development.

Cryptococcus neoformans and Cryptococcus gattii are encapsulated yeasts that have become increasingly common central nervous system (CNS) pathogens within an enlarging populationof immunocompromised host. CNS disease represents a major interaction between the host and the yeast. In this regard, we have examined the myriads of tools that this group of yeasts possess that allow them to become very efficient pathogens in the human CNS, causing fatal meningoencephalitis if untreated. In the age of molecular approaches to the investiagion of fungal pathogenesis, cryptococcosis has been studied extensively at the genetic level, and there is an impressive array of virulence factors that this basidiomycetous pathogen has developed to cause disease. This review details a series of virulence factors and their pathways. There is also some appreciation of the complexity of the host responses, which are so essential to the outcome of these infections.

ABSTRACTCandida albicansis a common opportunistic fungal pathogen that asymptomatically colonizes most humans. Although typically a benign commensal, dysbiosis caused by antibiotic use, immune dysfunction, or epithelial barrier disruption can trigger fungal overgrowth and infection, ranging from superficial mucosal disease to life-threatening systemic candidiasis. New preclinical infection models are needed to dissectC. albicanspathogenesisin vivoacross distinct stages of infection and with different measurable host outcomes. We previously established the planarianSchmidtea mediterraneaas an invertebrate host for studying host-pathogen interactions duringC. albicansinfection.S. mediterranearelies entirely on conserved innate immune mechanisms capable of overcoming infection with pathogenic microorganisms, including bacteria and fungi. Planarians’ remarkable regenerative capacity and accessible stem cell populations make this organism a tractable model to analyze early immune responses, tissue repair, and pathogen clearancein vivo. This model supports simultaneous analysis of fungal virulence and host transcriptional responses, providing valuable insights into infection dynamics. Here, we present an updated protocol with detailed modifications, standardized procedures, and optimized steps for infectingS. mediterraneawithC. albicans, designed to enhance reproducibility and enable systematic studies of fungal pathogenesis and host defense.SUMMARYWe present an optimized and detailed protocol for using the planarianSchmidtea mediterraneaas a model system to study host-pathogen interactions during fungal infection. This method builds on our previous procedure for infecting planarians with the human fungal pathogenCandida albicans, providing detailed guidance to enhance reproducibility and experimental consistency. The system enables comprehensive, parallel analysis of both host and pathogen responses throughout infection – from initial colonization and disease progression to diverse host outcomes, including clearance or mortality.

Sorghum anthracnose, caused by Colletotrichum sublineola, severely affects leaves, stems, and panicles. This study characterizes CsMep, a zinc-dependent metalloprotease with a conserved HEXXH motif and an N-terminal signal peptide. CsMep localizes to the cell membrane, cytoplasm, and nucleus and suppresses plant programmed cell death in an HEXXH-dependent manner. This suppression was accompanied by reduced ROS accumulation and cell necrosis along with decreased electrolyte leakage in plant tissues. Deletion of CsMep impaired fungal pathogenicity, hindering hyphal growth, sporulation, and appressorium formation, while increasing sensitivity to ZnSO4 stress. Under zinc treatment, the mutant exhibited modifications in gene expression and metabolite accumulation, including the upregulation of aromatic amino acid biosynthesis along with increased levels of L-tryptophan and protocatechuic acid. These findings indicate that CsMep is crucial for adaptation to zinc stress and for achieving full virulence, thereby highlighting its significance in fungal pathogenesis and its potential as a target for disease control.

Aspergillus fumigatus is a challenging fungal pathogen in the clinic in part due to increasing azole drug resistance. In this study, we observe that loss of the A. fumigatus gene arvA results in increased azole susceptibility and significant in vitro morphological changes highlighted by hyper-swollen conidia that yield stunted and polarity deficient hyphae. Importantly, despite these severe in vitro morphological and growth abnormalities, ΔarvA surprisingly retains full pathogenicity and virulence in two immunologically distinct murine models of invasive pulmonary aspergillosis. These results challenge our understanding of the in-host environment and how it mediates fungal morphogenesis and pathogenesis. These results, consequently, not only enhances our understanding of the role of arvA in A. fumigatus morphogenesis and drug susceptibility, but further emphasizes the importance of in vivo animal models in fully evaluating potential antifungal drug targets.

Candida albicans (C. albicans) is a conditionally pathogenic fungus in humans, with its virulence significantly modulated by alterations in the composition of commensal bacteria and the surrounding microecological environment, particularly during cohabitation with methicillin-resistant Staphylococcus aureus (MRSA). Despite this, the molecular mechanisms underlying these interactions remain inadequately elucidated. In this study, we utilized an integrative multiomics approach, including proteomics and proteomics of post-translational modifications (PTMs), to systematically examine the impact of MRSA on protein expression and PTM patterns in C. albicans. Our findings indicate that the presence of MRSA markedly influenced the expression of virulence-associated proteins and modified the phosphorylation and acetylation levels of key proteins involved in essential signaling and metabolic pathways. These modifications were predominantly associated with biological processes such as energy metabolism, metabolic reprogramming, and stress response. Functional enrichment analyses further indicated that these PTMs may play crucial roles in regulating the pathogenicity and environmental adaptability of C. albicans. Moreover, in vitro enzyme activity assays revealed that lysine acetylation induced by MRSA modulated the activities of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and homoisocitrate dehydrogenase (HIcDH). This suggests that such modifications are involved in the metabolic adaptation and functional reprogramming of C. albicans. In conclusion, this study provides novel insights into the regulation of fungal physiology mediated by MRSA through PTMs, thereby offering a new theoretical framework for understanding fungal pathogenesis and for the development of enhanced anti-infective strategies within the context of bacterial-fungal interactions.

SH3 (Src homology-3) domain-containing proteins are conserved molecular scaffolds that mediate protein-protein interaction and regulate important cellular processes in eukaryotes. However, their role in phytopathogenic fungi remain poorly understood. In this study, we systematically identified and functionally characterized SH3 domain-containing proteins in the plant fungal pathogen Fusarium graminearum. We identified 29 SH3 domain-containing proteins in F. graminearum, of which only 9 were previously characterized. We found that the remaining 20 proteins, including FgSla1, FgAip5, FgRax2, FgMcy1, FgVta, FgPin3, FgYsc84, FgSh3A, FgSh3B, FgSh3C, FgBoi1, FgRvs167, FgBzz1, FgClf, FgCyk3 and FgHof1, are required for vegetative growth, plant infection and deoxynivalenol (DON) production. Notably, the absence of FgRAX2 and FgMCY1 completely abolished DON synthesis. FgHof1 and FgRax2 serve as positive and negative regulators of conidiation, respectively, and are indispensable for sexual development. Furthermore, FgHof1 and FgCyk3 are crucial for cytokinesis and nuclear distribution, as shown by irregular septation and nuclear fragmentation in the mutant strains. Subcellular localization revealed distinct distributions of these proteins, including the cytoplasm, septa/septal pore, plasma membrane, sub-apical collar and hyphal tip, consistent with the multifaceted functions of the proteins. Remarkably, FgHof1 localizes to septal pore and its deletion causes conidial breakage along the septa. FgAip5 localizes to the hyphal tip and its absence leads to retarded growth and irregular colony edges. Interestingly, several SH3 proteins contain intrinsically disordered regions (IDRs) and form protein condensates in the cytosol. These proteins exhibited features of phase separation like condensate fusion and reemergence after photobleaching, suggesting a possible role in dynamic protein assembly. Deletion of the IDRs largely altered these features in the proteins. In summary, this study highlights the varied functions of SH3 domain-containing proteins in growth, asexual/sexual development, DON biosynthesis and pathogenicity of F. graminearum, offering new insights into the functional diversity of SH3 proteins in fungal pathogenesis and potential targets for the control of Fusarium head blight (FHB).

Fungal pathogenesis and development are regulated by transcription factors. Here, we characterized DhTmp1 in the nematode-trapping fungus Dactylellina haptotyla as a negative regulator of predation. Its deletion impaired fungal growth and stress tolerance yet enhanced pathogenicity, while overexpression yielded opposite phenotypes. Integrated transcriptomic and metabolomic analyses revealed that DhTmp1 deletion upregulated secondary metabolism biosynthetic genes, increasing the abundance of nematocidal metabolites 4-hydroxy benzaldehyde (4-HBA) and 6-chloro-1H-indole-3-carboxaldehyde (6-Cl-ICA), which exhibited potent activity against Meloidogyne incognita and Caenorhabditis elegans, respectively. The 48 h LC50 values for 4-HBA and 6-Cl-ICA against M. incognita and C. elegans values were 29.65 and 92.71 μg/mL, respectively. Furthermore, pot experiments demonstrated that ΔDhTmp1 achieved 74.33% control efficiency against M. incognita compared with the PDB-treated control, outperforming the wild-type strain by ∼20.6%. Our findings reveal that DhTmp1 negatively regulates fungal predation by modulating secondary metabolite synthesis, suggesting that its manipulation could optimize this fungus for the biocontrol of M. incognita.

Neurotropic dematiaceous fungi are primary agents of cerebral phaeohyphomycosis, a life-threatening brain infection with high mortality. However, the genomic basis underlying their virulence, stress tolerance, and antifungal resistance is poorly understood. In this study, we present high-quality hybrid genome assemblies of three major neurotropic dematiaceous fungi, Cladophialophora bantiana, Fonsecaea monophora, and Cladosporium cladosporioides, using Nanopore long-read and Illumina short-read sequencing platforms. The assembled genomes ranged from 31.5 to 39.9 Mb, with high completeness (>98.9%). Functional annotation revealed diverse coding and non-coding elements associated with stress responses, iron metabolism, and antifungal resistance. The Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses uncovered metabolic versatility, enriched xenobiotic degradation pathways, and lineage-specific functional divergence. Notably, C. bantiana and F. monophora exhibited greater genomic plasticity, higher transposable element content, and broader repertoires of virulence factors, extracellular peptidases, and secondary metabolite biosynthetic gene clusters, suggesting enhanced pathogenic potential. All three genera harbored conserved stress tolerance mechanisms, melanin biosynthesis pathways, and pathogenicity-related genes linked to immune evasion and neuroinvasion. Additionally, we identified distinct multidrug efflux transporter families linked to antifungal resistance. Orthology analysis revealed a shared core proteome alongside genus-specific adaptations likely underpinning niche specialization. While the findings highlight critical genomic features driving fungal resilience and neurotropism, functional validation through transcriptomics and phenotypic assays remains essential. Despite current limitations in experimental tractability, this work provides a foundational resource for understanding the molecular basis of fungal pathogenesis and offers valuable targets for future diagnostic and therapeutic strategies against cerebral phaeohyphomycosis and related infections.

Exserohilum rostratum is a causal agent of severe maize leaf spot, posing a threat to maize production. Carbohydrate esterase (CE) can catalyze the removal of acyl modifications from plant cell wall polysaccharides, thereby promoting polysaccharide hydrolysis. A total of 87 CE genes were identified in the E. rostratum ER1 genome. In this study, we conducted a comprehensive analysis of the E. rostratum CE (ErCE) genes, including physicochemical properties, structural features, promoter cis-acting regulatory elements, and functional analysis. Subcellular localization analysis revealed that more than half of ErCEs were located extracellular. ErCEs contain abundant conserved domains, indicating functional diversity of these proteins. The promoter region of ErCE genes contains a rich variety of cis-acting regulatory elements related to plant hormone regulation, stress response, and developmental processes. Functional enrichment analysis indicated that ErCE genes are predominantly involved in metabolic pathways. In addition, the expression pattern revealed significant changes in ErCE genes during E. rostratum infection, indicating that they play an important role in pathogen invasion and lesion expansion. Overall, this study elucidated the structural characteristics and expression patterns of the CE genes in E. rostratum, providing conditions for further exploration of their roles in fungal pathogenesis and laying the foundation for the improvement of sustainable agricultural systems using related genes.

Pore-forming toxins are potent biological weapons used across nature, from virulence factors to immune defense proteins. This study identifies K62, a little-known antifungal toxin produced by a wild yeast, as a structural and functional relative of the aerolysin family, which is well-known for forming damaging pores in cell membranes. Using structure prediction, molecular simulations, and biochemical analysis, we show that K62 assembles into large, stable pore-like complexes. Remarkably, K62 is just one member of a large and previously unrecognized family of similar toxin-like proteins found in fungi, plants, and bacteria, including pathogens that affect humans and crops. These findings uncover an unexpected evolutionary link across kingdoms, suggesting that pore-forming toxins may play a widespread role in fungal pathogenesis and microbial warfare. This work lays the foundation for understanding a new group of antifungal molecules and their potential impacts on health, agriculture, and microbial ecology.